A synergistic lipid-lowering composition and a lipid-lowering health drink

Abstract

The present invention provides a synergistic lipid-lowering composition and a lipid-lowering health drink, belonging to the field of food technology. The present invention uses blue indigo fruit, black fungus and hawthorn as main raw materials, and can achieve the best lipid-lowering effect by providing a synergistic lipid-lowering composition with a specific ratio, which is better than the individual components. And through rat animal experiments, a new type of lipid-lowering nutritional functional drink was finally developed. The present invention can overcome the problem of the lack of synergistic lipid-lowering nutritional functional preparations using blue indigo fruit as raw materials in the prior art. The sensory score of the lipid-lowering liquid preparation prepared by the present invention is 92.3, with excellent quality, and is very suitable for industrial promotion. The active ingredients of the nutritional functional drink of the present invention will not be lost due to the necessary product sterilization process, and can be put into large-scale production. The present invention can enhance the commercial value of blue indigo fruit and provide new ideas for the development of blue indigo fruit products.

Description

Technical Field

[0001] The invention belongs to the technical field of food, and in particular relates to a synergistic lipid-lowering composition and a lipid-lowering health drink. Background Art

[0002] With the progress of industrial construction and the vigorous development of social economy, environmental pollution is becoming more and more serious, the pace of life is accelerating, and people's physical and mental health problems are becoming more and more serious under high-intensity work pressure. Metabolic disorders such as high cholesterol, obesity, and hyperlipidemia have become the norm in society. The incidence of diseases such as coronary heart disease, atherosclerosis, and myocardial infarction is also gradually increasing, which is a social problem that cannot be ignored. At present, drugs, surgery, and diet control are mainly used in the clinic to treat hyperlipidemia, but most of these therapies have the hazards of reducing the body's immunity, large toxic side effects, and easy recurrence. Among them, the most widely used statins can also cause adverse reactions such as liver damage and gastrointestinal damage. Many studies have emphasized the lipid-lowering benefits of phytochemicals, which are multi-component, multi-target, and have relatively low toxic effects. Many in vitro and animal studies have shown that the consumption of these bioactive phytochemicals can significantly improve hypertension, low-density lipoprotein oxidation, lipid peroxidation, total plasma antioxidant capacity, and dyslipidemia.

[0003] The fresh blue indigo fruit tastes sour and astringent, and is not accepted by consumers, but it is rich in active substances such as anthocyanins, anthocyanins, flavonols, polyphenolic acids, etc. The anthocyanin content is much higher than that of plant products such as blueberry, sea buckthorn, and bilberry, and can prevent liver damage.

[0004] So far, there is no synergistic lipid-lowering preparation using blue indigo fruit as raw material. Summary of the invention

[0005] In view of this, the object of the present invention is to provide a synergistic lipid-lowering composition and a lipid-lowering health drink. The present invention extracts blue indigo fruit to solve the problem that blue indigo fruit has a sour taste and is not accepted by consumers in the prior art, and achieves a synergistic lipid-lowering effect by combining black fungus extract and hawthorn extract to solve the problem that there is a lack of synergistic lipid-lowering preparations using blue indigo fruit as raw materials in the prior art.

[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0007] The invention provides a synergistic blood lipid lowering composition, comprising the following components in a volume ratio: caerulea edulis extract: black fungus extract: hawthorn extract of 3-5:15-17:4-6.

[0008] Preferably, the concentration of the blue loquat fruit extract is 1-2 g / mL, the concentration of the black fungus extract is 0.05-0.20 g / mL, and the concentration of the hawthorn extract is 0.1-0.4 g / mL.

[0009] Preferably, the blue honeysuckle fruit extract, black fungus extract or hawthorn extract is obtained by ultrasonic water extraction using blue honeysuckle fruit, black fungus or hawthorn as raw materials.

[0010] Preferably, the water extraction process comprises the following steps:

[0011] The black fungus is crushed and sieved, mixed with water for a first ultrasonic extraction, and then centrifuged for a first time to obtain a first black fungus supernatant and a first black fungus residue;

[0012] The first black fungus filter residue is mixed with water to perform a second ultrasonic extraction, and after the extraction, a second centrifugation is performed to obtain a second black fungus supernatant and a second black fungus filter residue;

[0013] The first black fungus supernatant and the second black fungus supernatant are combined and concentrated to obtain a black fungus extract;

[0014] The blue indigo fruit is subjected to a wall breaking treatment, mixed with water for a first ultrasonic extraction, and then subjected to a first centrifugation to obtain a first blue indigo fruit supernatant and a first blue indigo fruit residue;

[0015] The first filter residue of blue Indigo Fruit is mixed with water for a second ultrasonic extraction, and then centrifuged for a second time to obtain a second supernatant of blue Indigo Fruit and a second filter residue of blue Indigo Fruit;

[0016] The first supernatant of blue Indigo Carpa and the second supernatant of blue Indigo Carpa are combined and concentrated to obtain a blue Indigo Carpa extract;

[0017] The hawthorn is de-cored, the wall is broken, mixed with water for a first ultrasonic extraction, and after the extraction, a first centrifugation is performed to obtain a first hawthorn supernatant and a first hawthorn residue;

[0018] The first hawthorn residue is mixed with water for a second ultrasonic extraction, and then centrifuged for a second time to obtain a second hawthorn supernatant and a second hawthorn residue;

[0019] The first hawthorn supernatant and the second hawthorn supernatant are combined and concentrated to obtain an extract.

[0020] Preferably, the black fungus is sieved with a mesh number of 30 to 50, and the total weight ratio of the black fungus, blue indigo fruit, hawthorn and water is 1:3 to 50; the temperature of each ultrasonic extraction is 35 to 60° C., and the time of each ultrasonic extraction is 40 to 70 minutes; the concentration is concentrated by rotary evaporation, the concentration temperature is 40 to 55° C., and the volume after concentration is 1 / 4 to 1 / 6 of the original volume.

[0021] The present invention provides the use of the synergistic lipid-lowering composition in the preparation of medicines or nutritional functional foods for treating hyperlipidemia.

[0022] The present invention also provides a beverage of the synergistic lipid-lowering composition, which further comprises auxiliary materials, including erythritol, potassium sorbate, pectinase, soybean polysaccharide, and CMC.

[0023] Preferably, the percentage of the auxiliary materials added to the beverage is as follows: erythritol is added in an amount of 12-15%, pectinase is added in an amount of 0.3-0.7%, soybean polysaccharide is added in an amount of 0.3-0.7%, CMC is added in an amount of 0.2-0.5%, and potassium sorbate is added in an amount of 0.01-0.05%.

[0024] Compared with the prior art, the present invention has the following beneficial effects: the present invention uses blue indigo fruit, black fungus and hawthorn as main raw materials, and successfully develops a new type of lipid-lowering nutritional functional beverage. The synergistic lipid-lowering composition with a specific ratio provided by the present invention can achieve the best lipid-lowering effect, which is better than the individual components. The blue indigo fruit extract, black fungus extract and hawthorn extract have a synergistic effect in lowering blood lipids. Experiments have shown that the sensory score of the lipid-lowering beverage prepared by the present invention is 92.3, with excellent quality, and is very suitable for industrial promotion. After pasteurization, the active ingredients of the lipid-lowering nutritional functional beverage of the present invention will not be lost due to the necessary product sterilization process, and can be put into large-scale production. The present invention can promote the deep processing of blue indigo fruit, enhance the commercial value of blue indigo fruit, provide new ideas for the development of blue indigo fruit products, and lay the foundation for local pillar industries.

[0025] The present invention has been further tested and proved that the lipid-lowering nutritional functional drink provided by the present invention is not toxic to rats, and alleviates the weight gain of hyperlipidemia rats to a certain extent. After 28 days of oral administration of LAC to hyperlipidemia rats, the symptoms of high body weight and high blood routine indicators of hyperlipidemia in each dose administration group were effectively alleviated, among which the hLAC group had the best effect. The organ indexes of each administration group were alleviated compared with the model group, and the lesions could be alleviated to a certain extent. The LAC provided by the present invention has a significant lipid-lowering effect, and helps to restore the lipid metabolism function of liver tissue by improving the redox state of liver tissue in rats with hyperlipidemia induced by a high-fat diet. The results of liver HE and Oil Red O staining and the determination of liver function indicators can prove that LAC can protect liver tissue, thereby improving liver function. LAC can also significantly upregulate the levels of serum adiponectin and lecithin cholesterol acyltransferase in lipid metabolism, significantly downregulate the expression of leptin and fatty acid synthase and the level of free fatty acids in rats, improve the blood lipid level in rats, and thus alleviate the symptoms of hyperlipidemia at multiple targets. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 is the standard curve of sodium glycocholate;

[0027] Figure 2 is the standard curve of sodium taurocholate;

[0028] Figure 3 The results are the binding rates of the blue loquat extract with sodium glycocholate and sodium taurocholate;

[0029] Figure 4 The results are the binding rates of black fungus extract with sodium glycocholate and sodium taurocholate;

[0030] Figure 5 The results are the binding rates of hawthorn extract with sodium glycocholate and sodium taurocholate;

[0031] Figure 6 It is the combination rate of black fungus and hawthorn compound with sodium glycocholate and sodium taurocholate;

[0032] Figure 7 The results are the binding rates of different ratios of the lipid-lowering composition and sodium glycocholate;

[0033] Figure 8 The results are the binding rates of different ratios of the lipid-lowering composition and sodium taurocholate;

[0034] Fig. 9 is the standard curve of total polysaccharide content;

[0035] Fig.10 is the standard curve of total flavonoids content;

[0036] Fig.11The particle size distribution of the lipid-lowering beverage before sterilization;

[0037] Fig.12 The particle size distribution of the lipid-lowering beverage after sterilization;

[0038] Fig.13 To analyze the static rheological properties of blood lipid-lowering beverages;

[0039] Fig.14 To analyze the dynamic rheological properties of lipid-lowering beverages;

[0040] Fig.15 This is the EPR spectrum of the lipid-lowering drink's scavenging effect on hydroxyl free radicals before sterilization;

[0041] Fig.16 This is the EPR spectrum of the sterilized lipid-lowering beverage's scavenging effect on hydroxyl free radicals;

[0042] Fig.17 The concentration of LAC crude drug and hydroxyl radical scavenging rate at different dilution multiples;

[0043] Fig.18 The concentration of LAC crude drug and the scavenging rate of ABTS free radicals at different dilution multiples;

[0044] Fig.19 To determine the concentration and reducing power of LAC crude drug at different dilution multiples;

[0045] Fig. 20 Serum TG analysis for modeling mixed hyperlipidemia in SD rats;

[0046] Fig.21 Serum TC analysis was performed to model mixed hyperlipidemia in SD rats;

[0047] Fig. 22 Serum LDL-C analysis for modeling mixed hyperlipidemia in SD rats;

[0048] Fig.23 Serum HDL-C analysis for modeling mixed hyperlipidemia in SD rats;

[0049] Fig.24 The weight changes of rats in each group during oral gavage;

[0050] Fig.25 The changes of food intake of rats in each group during oral gavage;

[0051] Fig.26 This is the change of water intake of rats in each group during oral gavage;

[0052] Fig. 27 The effect of LAC on serum TG levels in hyperlipidemic rats;

[0053] Fig.28 The effect of LAC on serum TC level in hyperlipidemic rats;

[0054] Fig.29 The effect of LAC on serum LDL-C level in hyperlipidemic rats;

[0055] Fig.30 The effect of LAC on serum HDL-C level in hyperlipidemic rats;

[0056] Fig.31 The effect of LAC on the level of TG in the liver of hyperlipidemic rats;

[0057] Fig.32 The effect of LAC on TC level in the liver of hyperlipidemic rats;

[0058] Fig.33 The effect of LAC on the level of LDL-C in the liver of hyperlipidemic rats;

[0059] Fig.34 The effect of LAC on the HDL-C level in the liver of hyperlipidemic rats;

[0060] Fig.35 The effect of LAC on serum aspartate aminotransferase level in hyperlipidemic rats;

[0061] Fig.36 The effect of LAC on the level of aspartate aminotransferase in the liver of hyperlipidemic rats;

[0062] Fig.37 The effect of LAC on serum alanine aminotransferase level in hyperlipidemic rats;

[0063] Fig.38 The effect of LAC on the level of alanine aminotransferase in the liver of hyperlipidemic rats;

[0064] Fig.39 The effect of LAC on the morphology of liver tissue in hyperlipidemic rats by HE staining;

[0065] Fig.40 The effect of LAC on Oil Red O staining of liver tissue morphology in hyperlipidemic rats;

[0066] Fig.41 The effect of LAC on the morphology of brown adipose tissue in hyperlipidemic rats by HE staining;

[0067] Fig.42 The effect of LAC on serum superoxide dismutase level in hyperlipidemic rats;

[0068] Fig.43 The effect of LAC on the level of superoxide dismutase in the liver of hyperlipidemic rats;

[0069] Fig.44 The effect of LAC on the level of serum peroxidation product malondialdehyde in hyperlipidemic rats;

[0070] Fig.45 The effect of LAC on the level of malondialdehyde, a peroxidation product, in the liver of hyperlipidemic rats;

[0071] Fig.46 The effect of LAC on the total antioxidant capacity of serum in hyperlipidemic rats;

[0072] Fig.47 The effect of LAC on the total antioxidant capacity of the liver in hyperlipidemic rats;

[0073] Fig.48 The effect of LAC on serum leptin level in hyperlipidemic rats;

[0074] Fig.49 The effect of LAC on serum adiponectin level in hyperlipidemic rats;

[0075] Fig.50 The effect of LAC on the level of serum lecithin cholesterol acyltransferase in hyperlipidemic rats;

[0076] Fig.51 The effect of LAC on serum fatty acid synthase levels in hyperlipidemic rats;

[0077] Fig.52 The effect of LAC on the level of fatty acid synthase in the liver of hyperlipidemic rats;

[0078] Fig.53 The effect of LAC on serum free fatty acid levels in hyperlipidemic rats;

[0079] Fig.54 Effect of LAC on free fatty acid levels in the liver of hyperlipidemic rats. DETAILED DESCRIPTION

[0080] The present invention provides a synergistic lipid-lowering composition, comprising the following components in a volume ratio: blue loquat fruit extract: black fungus extract: hawthorn extract of 3-5:15-17:4-6, preferably 3.5-4.5:15.5-16.5:4.5-5.5, and more preferably 4:16:5.

[0081] In the present invention, the concentration of the blue honeysuckle extract is 1-2 g / mL, preferably 1.25 g / mL; the concentration of the black fungus extract is 0.05-0.20 g / mL, preferably 0.125 g / mL; the concentration of the hawthorn extract is 0.1-0.4 g / mL, preferably 0.25 g / mL; and the concentrations are all crude drug concentrations.

[0082] In the present invention, the black fungus extract is prepared by the following steps: crushing and sieving the black fungus, mixing it with water for the first ultrasonic extraction, and performing the first centrifugation after the extraction to obtain a first black fungus supernatant and a first black fungus residue; mixing the first black fungus residue with water for the second ultrasonic extraction, and performing the second centrifugation after the extraction to obtain a second black fungus supernatant and a second black fungus residue; combining the first black fungus supernatant and the second black fungus supernatant, and concentrating them to obtain the black fungus extract. In the present invention, the black fungus is sieved with a mesh number of 30 to 50, preferably 35 to 45, and further preferably 40 meshes, the total weight ratio of the black fungus to water is 1:3 to 50, preferably 1:20 to 45; the weight ratio of the black fungus raw material and water mixed in the first ultrasonic extraction is 1:10 to 30, preferably 1:20; the temperature of each ultrasonic extraction is 35 to 60° C., preferably 45 to 55° C., and the time of each ultrasonic extraction is 40 to 70 min, preferably 55 to 65 min; the centrifugal speed is 500 to 2000 rpm, preferably 1000 to 1500 rpm; the centrifugal time is 5 to 20 min, preferably 10 to 15 min; the concentration is carried out by rotary evaporation, the concentration temperature is 40 to 55° C., preferably 47 to 52° C., and the volume after concentration is 1 / 4 to 1 / 6 of the original volume, preferably 1 / 5.

[0083] In the present invention, the blue indigo fruit extract is prepared by the following steps: performing a wall-breaking treatment on blue indigo fruit, mixing with water for a first ultrasonic extraction, and performing a first centrifugation after the extraction to obtain a first blue indigo fruit supernatant and a first blue indigo fruit residue; mixing the first blue indigo fruit residue with water for a second ultrasonic extraction, and performing a second centrifugation after the extraction to obtain a second blue indigo fruit supernatant and a second blue indigo fruit residue; combining the first supernatant and the second supernatant, and concentrating to obtain the blue indigo fruit extract. In the present invention, the total weight ratio of the blue indigo fruit to water is 1:3-50, preferably 1:4-20; the weight ratio of the blue indigo fruit raw material and water mixed in the first ultrasonic extraction is 1:1-5, preferably 1:2; the temperature of each ultrasonic extraction is 35-60°C, preferably 37-45°C, and the time of each ultrasonic extraction is 40-70min, preferably 42-58min; the wall breaking is processed by a wall breaking machine, and the stirring power is 300-700W, preferably 400-600W; the centrifugal speed is 500-2000rpm, preferably 1000-1500rpm; the centrifugal time is 5-20min, preferably 10-15min; the concentration is preferably concentrated by rotary evaporation, the rotary evaporation concentration temperature is 40-55°C, preferably 45-50°C, and the volume after concentration is 1 / 4-1 / 6 of the original volume, preferably 1 / 5.

[0084] In the present invention, the hawthorn extract is prepared by the following steps: removing the core of the hawthorn, performing a wall-breaking treatment, mixing with water for a first ultrasonic extraction, and performing a first centrifugation after the extraction to obtain a first hawthorn supernatant and a first hawthorn residue; mixing the first hawthorn residue with water for a second ultrasonic extraction, and performing a second centrifugation after the extraction to obtain a second hawthorn supernatant and a second hawthorn residue; combining the first supernatant and the second supernatant, and concentrating to obtain an extract; the total weight ratio of the hawthorn to water is 1:3-50, preferably 1:10-30; the weight ratio of the hawthorn raw material to water in the first ultrasonic extraction is 1:5-15, preferably 1:10; each ultrasonic extraction is performed The temperature of the ultrasonic extraction is 35-60°C, preferably 45-55°C, and the time of each ultrasonic extraction is 40-70min, preferably 55-65min; the wall breaking is carried out by a wall breaking machine, and the stirring power is 300-700W, preferably 400-600W; the centrifugal speed is 500-2000rpm, preferably 1000-1500rpm; the centrifugal time is 5-20min, preferably 10-15min; the concentration is preferably carried out by rotary evaporation concentration, and the rotary evaporation concentration temperature is 40-55°C, preferably 47-52°C, and the volume after concentration is 1 / 4-1 / 6 of the original volume, preferably 1 / 5.

[0085] The present invention provides the use of the synergistic lipid-lowering composition in the preparation of a medicine or nutritional functional food for treating hyperlipidemia, wherein the medicine or nutritional functional food preferably uses the synergistic lipid-lowering composition of the present invention as the sole active ingredient; the dosage form of the medicine of the present invention is preferably powder, tablet, granule, capsule, solution, emulsion, suspension or injection, etc., and the dosage form of the nutritional functional food is preferably honey paste, dew, soft capsule, powder, fresh juice, hard capsule, tablet, tea, oral liquid, wine or granule, etc.; the medicine or nutritional functional food of the present invention preferably also includes auxiliary materials, and the types of the auxiliary materials can be fillers, sweeteners or flavoring agents, disintegrants, stabilizers, lubricants, adhesives, coatings, colorants, preservatives, etc.

[0086] The present invention also provides a beverage of the synergistic lipid-lowering composition, which further comprises auxiliary materials, including erythritol, potassium sorbate, pectinase, soybean polysaccharide, and CMC.

[0087] In the present invention, the percentage of the added auxiliary materials to the beverage is as follows: the added amount of erythritol is 12-15%, preferably 13-14%, the added amount of pectinase is 0.3-0.7%, preferably 0.4-0.6%; the added amount of soybean polysaccharide is 0.3-0.7%, preferably 0.4-0.6%; the added amount of CMC is 0.2-0.5%, preferably 0.3-0.4%; the added amount of potassium sorbate is 0.01-0.05%, preferably 0.02-0.04%; the rest is a synergistic lipid-lowering composition.

[0088] In the present invention, the blood lipid lowering nutritional functional beverage is sterilized by a pasteurization process, the temperature of the pasteurization process is 75 to 85° C., and the time is 25 to 35 minutes.

[0089] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0090] Example 1

[0091] After washing and breaking the wall of the blue indigo fruit, accurately weigh 10.00 g, add water at a material-water ratio of 1:4, perform ultrasonic-assisted extraction at 40°C for 45 min, extract twice, centrifuge to obtain the supernatant, combine the secondary supernatant filtrates, concentrate by rotary evaporation at 50°C, and store in a 4°C refrigerator for later use.

[0092] After washing and crushing the black fungus, pass it through a 40-mesh sieve, accurately weigh 10.00 g, add water at a material-water ratio of 1:40, perform ultrasonic-assisted extraction at 50°C for 60 min, extract twice, centrifuge to obtain the supernatant, combine the two supernatant filtrates, concentrate by rotary evaporation at 50°C, and store in a 4°C refrigerator for later use.

[0093] After washing, removing the core and breaking the wall of the hawthorn, accurately weigh 10.00 g, add water at a material-water ratio of 1:20, perform ultrasonic-assisted extraction at 50°C for 60 min, extract twice, centrifuge to obtain the supernatant, combine the two supernatant filtrates, concentrate by rotary evaporation at 50°C, and store in a 4°C refrigerator for later use.

[0094] The blue honeysuckle fruit extract, the black fungus extract and the hawthorn extract were mixed in a volume ratio of 4:16:5 to obtain a synergistic lipid-lowering composition.

[0095] Example 2

[0096] After washing and breaking the wall of the blue indigo fruit, accurately weigh 10.00 g, add water at a material-water ratio of 1:3, perform ultrasonic-assisted extraction at 40°C for 40 min, extract twice, centrifuge to obtain the supernatant, combine the secondary supernatant filtrates, concentrate by rotary evaporation at 50°C, and store in a 4°C refrigerator for later use.

[0097] After washing and crushing the black fungus, pass it through a 50-mesh sieve, accurately weigh 10.00 g, add water at a material-water ratio of 1:45, perform ultrasonic-assisted extraction at 45°C for 70 min, extract twice, centrifuge to obtain the supernatant, combine the two supernatant filtrates, concentrate by rotary evaporation at 50°C, and store in a 4°C refrigerator for later use.

[0098] Wash and crush the hawthorn, pass it through a 50-mesh sieve, accurately weigh 10.00 g, add water at a material-water ratio of 1:25, perform ultrasonic-assisted extraction at 45°C for 70 min, extract twice, centrifuge to obtain the supernatant, combine the two supernatant filtrates, concentrate by rotary evaporation at 50°C, and store in a 4°C refrigerator for later use.

[0099] The blue honeysuckle fruit extract, the black fungus extract and the hawthorn extract were mixed in a volume ratio of 3:16:5 to obtain a synergistic lipid-lowering composition.

[0100] Example 3

[0101] After washing and breaking the wall of the blue indigo fruit, accurately weigh 10.00 g, add water at a material-water ratio of 1:5, perform ultrasonic-assisted extraction at 40°C for 50 min, extract twice, centrifuge to obtain the supernatant, combine the secondary supernatant filtrates, concentrate by rotary evaporation at 50°C, and store in a 4°C refrigerator for later use.

[0102] After washing and crushing the black fungus, pass it through a 30-mesh sieve, accurately weigh 10.00 g, add water at a material-water ratio of 1:35, perform ultrasonic-assisted extraction at 55°C for 65 min, extract twice, centrifuge to obtain the supernatant, combine the two supernatant filtrates, concentrate by rotary evaporation at 50°C, and store in a 4°C refrigerator for later use.

[0103] After washing, removing the core and breaking the wall of the hawthorn, accurately weigh 10.00 g, add water at a material-water ratio of 1:30, perform ultrasonic-assisted extraction at 55°C for 65 min, extract twice, centrifuge to obtain the supernatant, combine the secondary supernatant filtrates, concentrate by rotary evaporation at 50°C, and store in a 4°C refrigerator for later use.

[0104] The blue honeysuckle fruit extract, the black fungus extract and the hawthorn extract were mixed in a volume ratio of 5:16:5 to obtain a synergistic lipid-lowering composition.

[0105] Comparative Example 1

[0106] The only difference from Example 1 is that the blue loquat fruit extract, the black fungus extract and the hawthorn extract are mixed in a volume ratio of 2:16:5 to obtain a lipid-lowering composition.

[0107] Comparative Example 2

[0108] The only difference from Example 1 is that the blue loquat fruit extract, the black fungus extract and the black fungus extract are mixed in a volume ratio of 6:16:5 to obtain a lipid-lowering composition.

[0109] Example 4

[0110] The synergistic lipid-lowering composition of Example 1 was taken, 13% erythritol, 0.4% pectinase, 0.4% soybean polysaccharide, 0.3% CMC, and 0.02% potassium sorbate were added, and the mixture was sterilized by a pasteurization process at 75° C. for 25 min to obtain a lipid-lowering beverage.

[0111] Example 5

[0112] Take the synergistic lipid-lowering composition of Example 2, add 14% erythritol, 0.5% pectinase, 0.5% soybean polysaccharide, 0.4% CMC, and 0.03% potassium sorbate, and sterilize it using a pasteurization process at 80°C for 30 minutes to obtain a lipid-lowering beverage.

[0113] Example 6

[0114] Take the synergistic lipid-lowering composition of Example 3, add 15% erythritol, 0.6% pectinase, 0.6% soybean polysaccharide, 0.5% CMC, and 0.04% potassium sorbate, and sterilize it using a pasteurization process at 85°C for 35 minutes to obtain a lipid-lowering beverage.

[0115] Experimental Example 1: Verification of synergistic effect

[0116] Drawing of standard curves of sodium glycocholate and sodium taurocholate: Weigh appropriate amounts of sodium glycocholate (M=487.6) and sodium taurocholate (M=537.6) respectively and place them in a beaker, add PBS (0.1M) buffer at pH 6.3 to fully dissolve and use a 100mL volumetric flask to make up the volume, shake well, and prepare a reference solution; then dilute with PBS solution to obtain standard solutions with different mass concentration gradients. Take 2.5mL of the above standard solutions with different gradients in 10mL stoppered test tubes respectively, add 7.5mL of 60% H2SO4, place in a water bath at 70℃ for 25min, take out and place in an ice bath for 5min, use an ultraviolet spectrophotometer to measure the absorbance of the solution in each test tube at 387nm, use the bile salt concentration as the horizontal coordinate and the measured absorbance as the vertical coordinate, and draw the standard curves of sodium glycocholate and sodium taurocholate, as shown below: Figure 1 , 2 The above solutions were prepared and used immediately.

[0117] Determination of the binding rate of sodium glycocholate and sodium taurocholate: Take 2 mL of the extract in a beaker, add 3 mL of a 10 mg / mL pepsin solution (prepared with 0.1 M PBS buffer at pH = 6.3) and 1 mL of a 0.01 mol / L HCl solution to each beaker, and digest in a 37°C constant temperature shaking incubator for 1 h (simulating the gastric digestion environment); adjust the solution to pH 6.3 with 0.1 mol / L NaOH, add 4 mL of a 10 mg / mL trypsin solution, and digest in a 37°C constant temperature shaking incubator for 1 h (simulating the gastrointestinal environment). After the in vitro digestion is completed, add 4 mL of sodium glycocholate and sodium taurocholate reference solution to the beaker, shake in a 37°C constant temperature shaking incubator for 1 h (90 rpm), then centrifuge at 6000 r / min for 20 min in a centrifuge, take 2.5 mL of the supernatant into a 10 mL stoppered test tube, add 7.5 mL of 60% H2SO4, bathe in 70°C water for 25 min, take out and bathe in ice for 5 min, and use a UV spectrophotometer to measure the absorbance of the solution in each test tube at 387 nm. Each sample is repeated three times, and the binding rate is calculated according to the following formula.

[0118]

[0119] In formula I: c1 is the initial bile salt concentration in the reaction solution (mg / mL), and c2 is the remaining bile salt concentration in the solution after the reaction is completed (mg / mL).

[0120] Figure 3 The binding capacity curves of blue honeysuckle extract with different concentrations of crude drug and two bile salts are shown in Figure 2. The IC values ​​of blue honeysuckle for sodium glycocholate and sodium taurocholate are shown in Figure 2. 50 The values ​​were 6.27 mg / mL and 6.49 mg / mL; Figure 4 The IC values ​​of black fungus for sodium glycocholate and sodium taurocholate were determined by the determination of the binding rates of black fungus extracts with different concentrations of raw drugs and two bile salts. 50 The values ​​were 9.99 mg / mL and 10.39 mg / mL; Figure 5 The binding capacity curves of hawthorn extract with different raw drug concentrations and two bile salts are shown in Figure 2. The IC values ​​of hawthorn for sodium glycocholate and sodium taurocholate are shown in Figure 2. 50 The values ​​were 19.28 mg / mL and 14.41 mg / mL; Figure 6 The purpose of this study was to determine the binding rate of the extract of black fungus and hawthorn complex with different amounts of raw herbs and sodium glycocholate and sodium taurocholate. The IC values ​​of black fungus and hawthorn complex on sodium glycocholate and sodium taurocholate were 50 The values ​​were 9.34 mg / mL and 9.39 mg / mL respectively. Figure 7The results are the binding rates of different ratios of the lipid-lowering composition and sodium glycocholate. Among them, Example 1 has the highest binding rate with sodium glycocholate. Its IC 50 The value is 5.41 mg / mL. Figure 8 The results are the binding rates of different ratios of the lipid-lowering composition and sodium taurocholate. Among them, Example 1 has the highest binding rate with sodium taurocholate. Its IC 50 The value is 5.39 mg / mL.

[0121] Among them, the IC values ​​of hawthorn extract on sodium glycocholate and sodium taurocholate were 50 The IC values ​​of Lonicera edulis extract on sodium glycocholate and sodium taurocholate were the largest. 50 The value is the smallest, indicating that among the three, hawthorn has the worst binding ability with bile salt, and blue indigo fruit has the best binding ability with bile salt. However, after mixing equal volumes of black fungus and hawthorn extract, its IC 50 The value is less than the IC of any single component 50 The CI combined indexes of the binding rate of the black fungus and hawthorn compound with sodium glycocholate and sodium taurocholate calculated by the Chou-Talalay model method were 0.709 and 0.778, respectively, both satisfying CI < 1, and the result is less than 1, which further illustrates the synergistic effect of the black fungus and hawthorn compound. The CI combined indexes of the binding rate of the lipid-lowering composition of Example 1 with sodium glycocholate and sodium taurocholate calculated by the Chou-Talalay model method were 0.562 and 0.574, respectively, both satisfying CI < 1, and the result is less than 1, which further illustrates the synergistic effect between the three raw materials in the lipid-lowering composition, and the synergistic effect is better than that of the black fungus and hawthorn compound.

[0122] Experimental Example 2 Sensory Evaluation Experiment

[0123] The blood lipid lowering drinks (referred to as LAC) obtained in Examples 4, 5, and 6 were evaluated.

[0124] 10 students were selected to conduct sensory evaluation from four aspects: color, flavor, texture, and taste. The total score was 100 points. The evaluation criteria are shown in Table 1. All participants in the sensory evaluation were randomly selected and were required to be healthy, not anorexic, and have good sensory evaluation ability. The evaluation was conducted three times under the same conditions to reduce experimental errors.

[0125] Table 1 Sensory scoring standards for blood lipid-lowering beverages

[0126]

[0127]

[0128] As shown in Table 2, the sensory score of the lipid-lowering beverage in Example 4 is 85.4, the sensory score of the lipid-lowering beverage in Example 6 is 87.9, and the sensory score of the lipid-lowering beverage in Example 5 is 92.3, which are of excellent quality and very suitable for industrial promotion.

[0129] Experimental Example 3 Determination of active substances

[0130] 1. Determination of anthocyanin content

[0131] Compared with purified anthocyanins, crude extracts are more thermally stable due to the interaction with certain nutrients (proteins, polysaccharides) in crude extracts. Anthocyanins are more stable at low pH, and the acidic conditions of LAC also play a certain protective role in the stability of anthocyanins. In order to control microorganisms, the pasteurization process was used to determine and analyze the anthocyanin content of LAC before and after sterilization. The hydrochloric acid methanol method was used to determine the anthocyanin content and evaluate the rationality of the sterilization process. The hydrochloric acid methanol method was used to determine the anthocyanin content. Accurately draw 200μL of the sample, place it in a 25mL volumetric flask, add about 20mL of 2% hydrochloric acid methanol solution, treat it in a 200W ultrasonic water bath at 20℃ for 30min, place it at room temperature for 10min, and then use 2% hydrochloric acid methanol solution to make up the volume, centrifuge it at 3500r for 15min, take the supernatant to determine the absorbance value, and substitute it into the following formula to calculate the content.

[0132]

[0133] In formula II: A is absorbance, DF is sample dilution factor, M is relative molecular mass of cy-3-glu (449.2), V1 is fixed volume of test solution, ε is extinction coefficient of cy-3-glu (26900), m is sample mass (mg), and L is light path length of cuvette.

[0134] The results showed that the anthocyanin content in LAC before sterilization was 74.4±3.14mg / 100mL, and the anthocyanin content in LAC after sterilization was 71.3±2.31mg / 100mL. Comparative analysis found that the anthocyanin content in LAC decreased slightly after pasteurization, but it was not significant (P>0.05). It is inferred that since anthocyanin is unstable and easily decomposed at high temperature, but this phenomenon is not significant, it can be seen that the pasteurization process of the present invention can retain the anthocyanin content in LAC to the maximum extent.

[0135] 2. Determination of polysaccharide content

[0136] With reference to SN / T 4260-2015, the polysaccharide content was determined by the phenol-sulfuric acid method. 1.0 g of glucose standard dried to constant weight at 105°C was accurately weighed, dissolved and diluted, and then fixed to volume in a 100 mL volumetric flask with pure water to prepare a 10 mg / mL standard stock solution. 1.0 mL of the stock solution was taken and fixed to 100 mL to prepare a 0.1 mg / mL glucose standard solution. 0 / 0.2 / 0.4 / 0.6 / 0.8 / 1.0 mL of 0.1 mg / mL glucose standard solution was taken and placed in a 20 mL stoppered test tube and filled to 1.0 mL with distilled water. 1.0 mL of 5% phenol solution was added to the test solution, and then 5.0 mL of concentrated sulfuric acid was quickly added and allowed to stand for 10 min. The reaction solution was fully mixed using a vortex oscillator, and then the test tube was placed in a 100°C water bath for 20 min. After the mixture was cooled to room temperature, the supernatant was aspirated and the absorbance was measured at 490 nm to draw a standard curve. Fig. 9 Then, 1.0 mL of the sample solution was drawn into a 20 mL stoppered test tube, and the polysaccharide content in the LAC before and after sterilization was compared and analyzed by drawing a standard curve.

[0137] The results showed that the standard curve y = a + b * x was obtained by linear fitting of the test results, where a = 9.22697 ± 0.43458, b = 0.04602 ± 0.02302, R 2 =0.99972, indicating that the standard curve has a good fitting degree. The polysaccharide content in LAC before sterilization is 6.03±0.94Glucose g / 100mL, and the polysaccharide content in LAC after sterilization is 6.12±0.88Glucose g / 100mL. Comparative analysis found that the polysaccharide content in LAC increased slightly after pasteurization, but it was not significant (P>0.05). Since polysaccharides have good thermal stability, it is speculated that sterilization treatment makes polysaccharide extraction more sufficient. It can be seen that the pasteurization process in the present invention will not affect the polysaccharide content in LAC.

[0138] 3. Determination of flavonoid content

[0139] The total flavonoid content of LAC was determined by the rutin control method. 0.0, 1.0, 2.0, 3.0, 4.0, 5.0, and 6.0 mL of rutin reference solution were accurately drawn and placed in 25 mL stoppered graduated tubes, respectively. Pure water was added to 6 mL (V3). 1.0 mL of 5% sodium nitrite solution was added, shaken, and allowed to stand for 6 min. 1.0 mL of 10% aluminum nitrate solution was added, shaken, and allowed to stand for 6 min. 10.0 mL of sodium hydroxide solution was added, shaken, and then distilled water was added to the scale. Shake, and allowed to stand for 15 min. The solution prepared with 0.0 mL of reference solution was used as the blank. The absorbance values ​​were measured at a wavelength of 510 nm. With absorbance as the ordinate (A) and reference concentration as the abscissa (mg / mL), a standard curve was drawn (see Fig.10 ). Then, draw 2.0 mL (M) of the test sample into a 25 mL (V1) volumetric flask, add 60% ethanol to dissolve and dilute to the scale, shake well, and determine the total flavonoid concentration in the LAC sample according to the method of determining the standard curve, and substitute it into the following formula to calculate the total flavonoid content.

[0140]

[0141] In formula III: X is the percentage of total flavonoids in the sample, rutin (C 27 H 30 O 16 ), C is the concentration of total flavonoids in the test solution read on the standard curve, V1 is the sample volume, V2 is the volume of the test solution absorbed, V3 is the color development volume, and M is the sample sampling amount.

[0142] The results showed that the standard curve y = a + b * x was obtained by linear fitting of the measurement results, where a = -0.00369 ± 0.0044, b = 0.00284 ± 3.79931E -5 , R 2 =0.99911, indicating that the standard curve has a good fitting degree. The flavonoid content in LAC before sterilization is 123.81±4.32Rutin mg / 100mL, and the flavonoid content in LAC after sterilization is 112.85±8.19Rutin mg / 100mL. Comparative analysis found that the flavonoid content in LAC decreased after pasteurization. It is speculated that the sterilization treatment destroyed a small amount of flavonoid activity, but it was not significant (P>0.05). It can be seen that the pasteurization process of the present invention will not affect the flavonoid content in LAC.

[0143] Experimental Example 4: Stability Analysis

[0144] 1. Particle size analysis

[0145] Particle size analysis is a necessary means to detect the stability of liquid preparations. The larger the particle size, the easier it is to aggregate, affecting the stability of the liquid preparation system. The smaller the particle size, the better the product stability. The particle size distribution of LAC before and after pasteurization can be seen in Fig.11 and Fig.12 .

[0146] The results showed that the average particle size of the LAC liquid preparation before pasteurization was 913.9nm and the PDI was 0.389. The average particle size of the LAC liquid preparation after pasteurization was 927.7nm and the PDI was 0.315. The PDI before and after sterilization was less than 1. Therefore, it was shown that the distribution of LAC was relatively uniform before and after pasteurization, the system was stable and not easy to aggregate, and it had good stability. Although the particle size increased after heat treatment, the difference was not significant (P>0.05). By comparison, it can be seen that the PDI value decreased from 0.389 to 0.315, indicating that the particle size distribution of LAC was more uniform after pasteurization, the particle polymerization degree distribution was narrower, the molecular weight difference became smaller, and the stability was improved. In summary, LAC still has a good particle size and centrifugal stability coefficient after pasteurization, which indicates that its stability is better.

[0147] 2. Other physical properties

[0148] The soluble solids (TSS), pH, turbidity and centrifugal stability coefficient of LAC before and after pasteurization were determined. TSS was determined according to GB / T12143-2008. The pH value of LAC samples was measured at ambient temperature (25±1°C) using a digital pH meter (FZ-600T; Chengdu Century Ark Technology Co., Ltd., Chengdu, China); the centrifugal stability coefficient was determined using a spectrophotometer (WFJ7200; Shanghai Lianke Instrument Co., Ltd.), with the detection wavelength set to 720nm, and the 100-fold diluted LAC and the solution centrifuged at 4000r / min for 10min were measured to obtain the absorbance X1 and X2. The stability coefficient W is the ratio of X2 to X1. If W≥90%, it indicates good stability. The changes in the stability of LAC before and after pasteurization were evaluated, and the results are shown in Table 3 below:

[0149] Table 3 Comparison of basic physical properties before and after pasteurization

[0150]

[0151] Note: Different superscript letters in the same column indicate significant differences between the two (p<0.05).

[0152] The results show that the centrifugal stability coefficient of LAC increased from 90.41% to 95.45% after pasteurization, and the solid components in each component were suspended in LAC, indicating that its stability increased. The pH value of LAC changed from 3.45±0.06 to 3.41±0.01 after pasteurization, and the difference was not significant (P>0.05). The TSS content of LAC changed from 15.37±0.31% to 15.02±0.08% after pasteurization, and the difference was not significant (P>0.05). The absorbance of LAC when measuring turbidity after pasteurization decreased from 0.145±0.003 to 0.124±0.004, and the difference was significant (P<0.05). The reduction in turbidity has a positive effect on the stability of liquid preparations. Combined with the test results of the above physical properties, it is shown that pasteurization can improve the stability of LAC.

[0153] 3. Rheological properties analysis

[0154] Static rheology (i.e. shear viscosity) mainly studies the types of food fluids; shear viscosity is the viscosity generally referred to in practical liquid foods. The shear viscosity value has a certain correlation with sample stability and shelf life, and is also closely related to pipeline delivery rate and energy consumption during the production process.

[0155] like Fig.13 As shown, as the shear rate increases at 0.1s -1 ~100s -1 As the shear rate increases, the viscosity of LAC decreases, which indicates that LAC is a pseudoplastic fluid with shear-thinning behavior. As can be seen from the figure, the viscosity value changes significantly at first. As the shear rate increases, the weak three-dimensional network structure between LAC particles is disrupted, the intermolecular force decreases, and the viscosity decreases. -1 After that, the viscosity value remained basically stable with little change, indicating that the three-dimensional structure had been completely destroyed.

[0156] Dynamic rheology mainly studies the changes in the response values ​​of the energy storage model and the loss model under different frequencies and other conditions. The main indicators involved are the storage modulus (G'), loss modulus (G") and loss tangent. Among them, G' mainly reflects the elastic properties of the sample; G" mainly reflects the viscous properties of the sample; the loss tangent is the ratio of G" and G', which mainly reflects the viscosity-elasticity ratio of the sample.

[0157] like Fig.14As shown, with the increase of oscillation frequency, the storage modulus (G') and loss modulus (G") of LAC gradually increase, indicating that its viscoelasticity is enhanced within this range. In the range of 0.1Hz to 100Hz, G'>G", and the loss tangent value of the sample is less than 1, indicating that the elasticity of the LAC liquid preparation dominates the viscoelasticity, showing the characteristics of a suspension emulsion, similar to most foods and beverages.

[0158] Experimental Example 5 Analysis of Antioxidant Properties in Vitro

[0159] 1. Hydroxyl radical scavenging rate

[0160] 50 μL of LAC samples with different concentrations (8, 4, 2, 1, 0.5 mg / mL) were measured, and 40 μL of 0.2 mmol / LFeSO4 aqueous solution, 40 μL of double distilled water and 40 μL of 0.1 mol / LDMPO aqueous solution were added. After mixing, 40 μL of 0.1% H2O2 was added. After mixing, the sample was taken immediately, and the supernatant was injected into the Teflon capillary with a micro syringe. The capillary was folded in half and inserted into the bottom of the paramagnetic quartz sample tube. The sample tube was inserted into the center of the resonant cavity for EPR detection. The whole process was completed within 3 minutes. The EPR experimental parameters were set as follows: the central magnetic field was 3510G, the sweep width was 100G, the modulation frequency was 100KHz, the modulation amplitude was 1.0G, the magnification was 2.00e+004, the number of scans was 1, the microwave power was 2.017mW, the time constant was 163.84ms, and the sweep time was 41.943s.

[0161] Hydrogen peroxide (H2O2) can enter the body through human breathing, eyes or skin contact, and quickly decompose to produce hydroxyl radicals (·OH). ·OH is the most harmful reactive oxygen in nature, which can induce lipid peroxidation in the human body and damage DNA.

[0162] Electron paramagnetic resonance spectroscopy (EPR) can identify and characterize ·OH by electron spin trapping. This method has high sensitivity (<10 -9 M). 5,5-pyrroline-N-oxide (DMPO) is usually added as a free radical scavenger to evaluate the ·OH level in the reaction system. In this study, the ·OH scavenging effect of LAC was detected using EPR technology. The results are shown in Fig.15 , 16 shown.

[0163] Depend on Fig.15 , 16It can be seen that the spectrum has a quartet with a peak height ratio of 1:2:2:1 and a hyperfine coupling constant of aN=aH=14.9G, which determines that the peak is the ·OH capture peak. It can be seen that there is no significant difference between the non-sterilized and sterilized LAC samples at each dilution factor. The peak height in the spectra of each sample group to be tested has changed significantly, but the overall peak shape is similar. By comparison, it is found that the blank group has a significantly higher hydroxyl radical peak intensity than other sample groups due to the lack of LAC, which indicates that LAC has a certain scavenging effect on ·OH.

[0164] like Fig.17 As shown in the figure, at different dilution multiples, different concentrations of raw drug LAC showed good hydroxyl radical scavenging ability. The hydroxyl radical scavenging rate IC corresponding to unsterilized LAC 50 The value is 44.69 mg / mL; the hydroxyl radical scavenging rate IC of sterilized LAC 50 The value was 50.55 mg / mL. And as the dilution multiple increased and the concentration of LAC crude drug decreased, the hydroxyl radical scavenging rate decreased, showing a certain dose relationship.

[0165] 2. ABTS free radical scavenging rate

[0166] The ABTS method is often used to detect the antioxidant capacity of food. Antioxidant ingredients can reduce ABTS free radicals. Take 6 numbered test tubes, add 0.1mL of polysaccharide solution of different concentrations and 1.9mL of ABTS free radical working solution, vortex mix and react at room temperature for 6 minutes, and measure the absorbance value A1 at a wavelength of 734nm; replace the polysaccharide solution with 0.1mL of double distilled water, and measure its absorbance value as A0 under other conditions; replace the ABTS free radical working solution with 1.9mL of double distilled water, and measure the absorbance value A2 as background under other conditions. Calculate the scavenging rate of polysaccharide components for ABTS free radicals according to the following formula.

[0167]

[0168] like Fig.18As shown in the figure, as the dilution factor increases, the scavenging ability of LAC for ABTS free radicals continues to decrease. When the dilution factor is 80 times, the scavenging rate of unsterilized LAC for ABTS free radicals is 58.14%, and the scavenging rate of sterilized LAC for ABTS free radicals is 56.40%, and there is no significant difference between the two (P>0.05). When the dilution factor is 40 times, the scavenging rate of unsterilized LAC for ABTS free radicals is 91.75%, and the scavenging rate of sterilized LAC for ABTS free radicals is 89.92%. It can be concluded that after pasteurization, the scavenging ability of LAC for ABTS free radicals has a certain decrease, but it is not obvious (P>0.05). When the dilution factor of LAC is reduced from 40 times to 1 times, the scavenging ability of LAC for ABTS free radicals is also gradually increasing, and the scavenging rate is almost higher than 90%. At the same time, it was measured that the scavenging rate of LAC stock solution on ABTS free radicals before and after pasteurization was 98.14%, indicating that LAC has a strong scavenging ability on ABTS free radicals. It also verifies that the pasteurization process in the present invention will not affect the scavenging ability of LAC on ABTS free radicals.

[0169] 3. Restoration

[0170] The strength of reducing power can indirectly reflect the strength of the sample's antioxidant activity. This method is often used to quickly evaluate the total antioxidant capacity of various foods and liquid beverages. Add 1.0 mL of sample solution, 2.5 mL of 0.2 mol / L pH 6.6 phosphate buffer solution, and 2.5 mL of 1.0% potassium ferrocyanide solution to a test tube. Mix well and place in a water bath at 50°C for 20 minutes. Add 2.5 mL of 10.0% trichloroacetic acid solution, 2.5 mL of distilled water, and 0.5 mL of 0.1% ferric chloride solution, mix well, and react for 10 minutes. Measure the absorbance A at 700 nm, using distilled water as a blank.

[0171] like Fig.19 As shown, as the dilution factor increases, the reducing power of LAC continues to decrease. When the dilution factor is 5 times, the reducing power absorbance value of unsterilized LAC is 1.827, and the reducing power absorbance value of sterilized LAC is 1.809. It can be concluded that the reducing power of LAC decreases to a certain extent after pasteurization, but it is not obvious (P>0.05). It can be concluded that LAC has good reducing power, and it also verifies that the pasteurization process in the present invention does not affect the reducing power of LAC.

[0172] Experimental Example 6: Improvement of Physiological and Biochemical Indexes in Rats with Hyperlipidemia

[0173] Establishment and grouping of experimental animal model: Healthy male clean-grade SD rats were purchased from Liaoning Changsheng Biotechnology Co., Ltd., license number: SCXK (Liao) 2020-0001, 3-4 weeks old, adaptive feeding for 1 week, environmental temperature 22 ± 2 ° C, humidity 60% ± 5%, 12h light / dark cycle. After 4 weeks of feeding with ordinary feed, 25 rats were randomly selected for high-fat feeding for 4 weeks; another 5 rats were taken as blank control group (Normal) and fed with ordinary feed. After 4 weeks, blood was collected from the tail vein, left at room temperature for 2 hours to wait for serum precipitation, centrifuged at 3500r / min for 20 minutes, and the supernatant was taken to measure the four blood routine levels of serum TG, TC, LDL-C, and HDL-C in each group of rats. After processing with SPSS data analysis software, when the blood routine levels of rats fed with high-fat feed were significantly different from those of rats fed with conventional maintenance feed, the hyperlipidemia rat model was confirmed to be successful, and the remaining blood routine values ​​did not meet the diagnostic criteria for hyperlipidemia, and the experiment was eliminated.

[0174] 30 rats were divided into 6 groups: blank group (Noraml group), model group (Model group), simvastatin positive control group (Simvas group), sample low-dose intervention group (lLAC group), medium-dose intervention group (mLAC group) and high-dose intervention group (hLAC group). The dose groups were given 5, 10 and 15 times concentrated LAC samples (samples obtained in Example 1) by gavage according to the dose of 5mL / kg in the low, medium and high dose groups, respectively, the positive control group was given simvastatin (0.9mg / kg·bw), and the other groups were gavaged with equal volume of distilled water. The gavage lasted for 28 days, and the rats' food intake and water intake were recorded every day, and the rats' body weight was measured every 7 days.

[0175] Blood sample collection: Rats in each group were fasted overnight, and injected with about 800 μL of 10% chloroform in the abdomen for anesthesia the next morning. The rats were randomly dissected and whole blood was collected from the abdominal vein, about 10-20 mL. The rats were left standing at room temperature for 2 hours to wait for serum to precipitate, centrifuged at 3500 r / min for 20 minutes, and the supernatant was stored at -20°C.

[0176] Collection of tissue specimens: Rats were intravenously injected with 1.5 mL of 10% chloral hydrate solution. After the rats were paralyzed, their eyes were distracted, and their mood was depressed, the liver, kidney, heart, spleen, pancreas, thymus, brown fat, colon, and epididymis were dissected and rinsed in pre-cooled saline. The kidney, right lobe of the liver, brown fat, and colon (5 in each group) were temporarily placed in a centrifuge tube filled with 4% paraformaldehyde fixative, marked for HE staining, and the pathological morphological changes of the tissues were observed. The remaining tissues were quickly frozen in liquid nitrogen and quickly transferred to a -80°C refrigerator for subsequent determination, and then the rats were killed by dislocating the neck.

[0177] Effect on rat organ tissue morphology (HE staining): The liver and brown adipose tissue were fixed with 4% paraformaldehyde fixative, taken out and placed in an embedding box, rinsed with running water for 12 hours, dehydrated with gradient alcohol, and embedded in paraffin after replacing the alcohol with xylene to make it transparent. After sectioning, the sections were ironed with hot water to avoid wrinkles, dried, dewaxed with xylene, and rinsed with gradient concentrations of alcohol. Then, they were soaked in distilled water, stained with hematoxylin and eosin, and sealed and observed and photographed under a Philips EM400T transmission electron microscope.

[0178] Effect on rat organ tissue morphology (Oil Red O staining): The frozen liver sections (thickness 4-8 μm) were dried at room temperature for 20 min, then incubated in 100% isopropanol solution for 5 min, taken out and incubated in 0.5% Oil Red O staining solution in a 60°C oven for 8 min, then immersed in 85% isopropanol solution for 3 min, rinsed with double dehydration, and then stained in hematoxylin solution for 1 min. After rinsing with double dehydration, the sections were sealed with sealing media, and observed and photographed under a microscope at 200 times magnification.

[0179] Determination of lipid metabolism indexes: The experiment used serum and liver tissue homogenate supernatant to determine the lipid metabolism indexes of each group of rats. The detected indexes were: total cholesterol (TC), triglycerides (TG), low-density lipoprotein (LDL-C), and high-density lipoprotein (HDL-C). Follow the steps in the kit instructions.

[0180] ELISA determination of key lipid metabolism enzymes and substances: The experiment used serum and liver tissue homogenate supernatant to determine the conventional lipid metabolism indicators of rats in each group. The indicators tested were: leptin (LEP), adiponectin (ADPN), lecithin cholesterol acyltransferase (LCAT), fatty acid synthase (FAS), free fatty acids (FFAs). Follow the steps in the kit instructions.

[0181] Determination of antioxidant indexes: The experiment used serum and liver tissue homogenate supernatant to determine the antioxidant indexes of each group of rats. The detected indexes were: superoxide dismutase (SOD), malondialdehyde (MDA), and total antioxidant capacity (T-AOC). Follow the steps in the kit instructions.

[0182] Determination of liver function indexes: The experiment used serum and liver tissue homogenate supernatant to determine the liver function indexes of rats in each group. The detected indexes were aspartate aminotransferase (AST) and alanine aminotransferase (ALT). The steps in the kit instructions were followed.

[0183] 1. Modeling and analysis of mixed hyperlipidemia in SD rats: Hyperlipidemia is caused by abnormal lipid metabolism, which is usually manifested as increased serum total triglycerides (TG), total cholesterol (TC), low-density lipoprotein (LDL-C) or decreased serum high-density lipoprotein (HDL-C). Fig. 20 , 21 , 22, and 23 are the four routine serum biochemical indices measured at the end of the modeling period of SD rats. It can be seen that after 4 weeks of high-fat feeding, the serum TG, TC, and LDL-C concentrations of SD rats in each group increased significantly (P < 0.01), and the serum HDL-C concentration decreased significantly (P < 0.01). Compared with the serum indicators of SD rats in the Normal group, the serum TG, TC, and LDL-C concentrations of SD rats in each group of high-fat modeling increased by 161.25%, 130.38%, and 351.72%, respectively, and the HDL-C concentration decreased by 44.66%, indicating that the high-fat model was well modeled and LAC and simvastatin positive administration could be performed.

[0184] 2. Effect on organ index: Organ weight can reflect whether the animal's organs have inflammatory reactions or lesions and the degree of development. The liver, kidney, heart, spleen, pancreas, thymus, and testis indexes were used to evaluate the repair effect of different doses of LAC on the damage of important organs of lipid metabolism in SD rats. The results are shown in Table 4:

[0185] Table 4 Organ indexes of rats in each group

[0186]

[0187]

[0188] Note: Normal: blank control group; Model: hyperlipidemia model group; lLAC: low-dose sample group; mLAC: medium-dose sample group; hLAC: high-dose sample group; Simvas: simvastatin positive control group. Different superscript letters in the same row represent significant differences between the two groups (p<0.05).

[0189] From the statistical data in the above table, it can be seen that the heart organ index of SD rats in the Model group was significantly increased compared with the Nomal group (P<0.05), and the liver, kidney, spleen, pancreas, and testis organ indexes were extremely significantly increased (P<0.01). It may be due to the obesity of SD rats after high-fat feeding, which caused pathological enlargement of organs, and inflammatory response in rats, resulting in an increase in organ indexes, among which the liver had the largest organ index, indicating that the liver is the largest organ in SD rats. The organ indexes of each drug-administered group were alleviated compared with the model group, which may have alleviated the lesions to a certain extent. Among them, the change trend of the liver organ index was the most obvious. Compared with the Model group, the liver organ indexes of each LAC dose group and the Simvas group decreased by 17.53%, 25.58%, 43.83%, and 42.40%, respectively, indicating that the LAC dose group showed a certain dose dependence.

[0190] 3. Effect on rat weight: Before modeling, SD rats had shiny fur and good mental state. After modeling with high-fat diet, the fur of the rats became dark yellow, the spirit was depressed, the mental state was relatively inactive, the food intake increased, the water intake increased, and the body weight increased significantly. SD rats were gavaged with LAC for 4 consecutive weeks, and the body weight was measured at the end of the 1st, 2nd, 3rd, and 4th weeks of the gavage period. The results are as follows: Fig.24 As shown. Except for the Normal group, there was no significant difference in the body weight of SD rats fed with high-fat diet at the 0th week of gavage period. There were significant differences between each group and the Normal group (P<0.01), indicating that the high-fat model was well established. With the increase of gavage days, the body weight difference between the groups gradually increased. The average body weight of SD rats in the Model group increased by 19.95% (P<0.01) compared with the average body weight of SD rats in the Normal group. The average body weight of SD rats in the low, medium and high dose groups of LAC and the Simvas group decreased by 5.33% (P<0.01), 7.09% (P<0.01), 12.19% (P<0.01), and 11.10% (P<0.01) respectively compared with the Model group. Each dose group had different degrees of improvement.

[0191] 4. Effect on rats' food intake and water intake: Hyperlipidemia rats have symptoms of polyphagia and polydipsia. The food intake and water intake of the Model group were significantly increased (P<0.05). The changes in the food intake of rats in each group during intragastric administration were shown in Fig.25 , changes in water intake Fig.26 .from Fig.25 It can be found that the daily food intake of the Normal group increased steadily with the growth of rats, and the food intake began to stabilize in the second week of gavage; however, the food intake of the Model group was nearly 40% higher than that of the Normal group. Compared with the Model group, the food intake of each drug-treated group was improved, among which the food intake of the hLAC group decreased the most (P<0.05). The daily food intake of each rat in the Normal group, Model group, and hLAC group in the fourth week of gavage was 20.91±1.24g, 27.86±1.47g, and 22.23±1.06g, respectively. Compared with the Model group, the food intake of the hLAC group decreased by an average of about 20.21%.

[0192] like Fig.26 As shown in the results, the water intake of the Normal group was normal, with an average of 28.02±2.21 mL per rat per day, while the water intake of the hyperlipidemia rats increased significantly (P<0.05), with an average of 41.52±2.09 mL in the Model group. The high-dose group of the LAC group had the greatest symptom relief, with an average of 33.31±2.25 mL, which was about 19.77% lower than that of the Model group.

[0193] From the above results, it can be concluded that after 28 days of oral administration of LAC, the symptoms of polyphagia and polydipsia in hyperlipidemic rats were effectively alleviated in all dosage groups, among which the hLAC group had the best effect.

[0194] 5. Effect of LAC on serum TG, TC, LDL-C, and HDL-C lipid metabolism index concentrations: After the gavage period, the routine serum lipid metabolism indexes of SD rats in each group were measured. Fig. 27 , 28 As shown in Figures 29 and 30, the concentrations of four biochemical indexes, namely TG, TC, LDL-C and HDL-C in serum of each group of SD rats were measured after the gavage period.

[0195] Compared with the Model group, the serum TG concentrations of SD rats in the LAC dose groups and Simvas group decreased by 21.46%, 49.87%, 71.21%, and 72.98%, respectively. The differences between each group and the Model group were extremely significant (P < 0.01). As the LAC dose concentration increased, the TG decrease level was more obvious, showing a dose-dependent manner. Compared with the Normal group, there was no significant difference in the TG levels of the hLAC dose group and the Simvas group, indicating that the TG level was close to the normal level.

[0196] Compared with the Model group, the serum TC concentrations of SD rats in the LAC dose groups and Simvas group decreased by 18.40%, 36.56%, 53.64% and 54.96%, respectively, with extremely significant differences (P < 0.01). With the increase of LAC dose concentration, the serum TC level was reduced more significantly, showing a dose-dependency.

[0197] Compared with the Model group, the serum LDL-C concentrations of SD rats in the LAC dose groups and Simvas group decreased by 28.77%, 58.38%, 69.66% and 71.28%, respectively. The differences between each group and the Model group were extremely significant (P < 0.01), indicating that each group had different degrees of improvement in serum LDL-C concentration indicators.

[0198] Compared with the Model group, serum HDL-C concentrations in each group increased significantly (P < 0.01). Compared with the Normal group, serum HDL-C concentrations in the Model group decreased by 75.00%, LAC dose groups decreased by 56.82%, 32.39%, and 19.32%, respectively, and Simvas group decreased by 2.27%, which was not significantly different from the Normal group.

[0199] 6. Effect of LAC on the concentrations of lipid metabolism indicators in the liver, including TG, TC, LDL-C, and HDL-C: The liver also contains a small amount of TG, TC, LDL-C, and HDL-C. After the gavage period, the supernatant of liver tissue homogenate was taken to measure the concentrations of four biochemical indicators, including serum TG, TC, LDL-C, and HDL-C, in each group.

[0200] TG is the core of lipoprotein, a fat molecule composed of glycerol and fatty acids. It is present in high levels in chylomicrons (CM) and very low-density lipoproteins (VLDL). TG levels can usually be reduced by promoting TG decomposition or inhibiting TG formation. Fig.31 As shown, compared with the Model group, the TG concentration in the liver of SD rats in each LAC dose group and Simvas group decreased by 18.46%, 34.88%, 49.16%, and 62.22%, respectively. The mLAC, hLAC, and Simvas groups were significantly different from the Model group (P < 0.01), and the lLAC was significantly different from the Model group (P < 0.05). As the LAC dose concentration increased, the TG decrease level was more obvious, showing a dose-dependent manner. Among them, the hLAC dose group returned to normal levels, with no significant difference from the Normal group.

[0201] Endogenous TC is the main source of TC in the human body. TC can be synthesized by cells of various organs in the body, and the liver is the most important place. Fig.32 As shown, compared with the Model group, the TC concentrations in the liver of SD rats in each LAC dose group and Simvas group decreased by 22.72%, 34.27%, 53.52% and 69.21%, respectively. The differences between each group and the Model group were extremely significant (P < 0.01). With the increase of LAC dose concentration, the serum TC level was reduced more significantly, showing a dose-dependency.

[0202] like Fig.33 As shown, the liver LDL-C concentrations of SD rats in each LAC dose group and Simvas group decreased by 19.07%, 33.62%, 56.12% and 64.03% respectively compared with the Model group. The differences between each group and the Model group were extremely significant (P < 0.01), indicating that each group had different degrees of improvement in serum LDL-C concentration indicators.

[0203] like Fig.34As shown in the figure, compared with the Model group, the serum HDL-C concentration in the mLAC, hLAC, and Simvas groups increased significantly (P < 0.01), and there was no significant difference in the lLAC dose group. Compared with the Normal group, the serum HDL-C concentration in the Model group decreased by 76.95%, the LAC dose groups decreased by 78.20%, 54.00%, and 23.53%, respectively, and the Simvas group decreased by 20.62%. Except for the lLAC dose group, all groups showed different degrees of improvement.

[0204] 7. Effect of LAC on biochemical indices of liver function in SD rats: Clinically, the detection indices of liver function include aspartate aminotransferase (AST), alanine aminotransferase (ALT), transaminase ratio (DRR) and alkaline phosphatase (ALP). Therefore, the effect of LAC on ALT, AST and transaminase ratio DRR (De Ritis ratio) of SD rats in each group was determined, where DRR = AST / ALT.

[0205] AST is mainly distributed in myocardium, liver, kidney and other tissues. When tissue cells are damaged, AST in the cytoplasm will be released into the serum. At the same time, due to the destruction of liver tissue cells, the AST concentration in the liver also rises sharply. Fig.35 As shown in the figure, the serum AST concentration in the Model group increased by 76.82% compared with the Normal group, which was a very significant increase (P < 0.01). Compared with the Model group, the lLAC group showed a significant downward trend (P < 0.05), and the other groups showed a very significant downward trend (P < 0.01). The serum AST concentrations in the lLAC, mLAC, hLAC dose groups and Simvas group decreased by 6.78%, 19.48%, 31.36% and 33.86% respectively compared with the Model group; Fig.36 As shown in the results, the liver AST concentration in the Model group increased by 92.09% significantly (P < 0.01) compared with the Normal group, which, together with the change in serum AST concentration, showed that the liver of SD rats in the Model group was damaged and the AST concentration increased significantly. The liver AST concentration in each dose group decreased by 17.72%, 37.15%, 43.10%, and 43.00% significantly (P < 0.01) compared with the Model group, but there were still significant differences compared with the Normal group, indicating that the blood and liver AST concentration indicators of SD rats were improved to a certain extent after LAC gavage.

[0206] ALT is mainly distributed in liver cells and is the most sensitive test indicator for liver damage. Under normal circumstances, the serum ALT concentration is low. When liver cells are damaged, the ALT in the liver cells is released into the serum, causing the concentration to increase. At the same time, due to the destruction of liver tissue cells, the ALT concentration in the liver also rises sharply. Fig.37As shown in the figure, the serum ALT concentration in the Model group increased significantly (P < 0.01) by 66.17% compared with the Normal group. Compared with the Model group, the serum ALT concentrations in the lLAC, mLAC, hLAC dose groups and Simvas group decreased significantly (P < 0.01), decreasing by 12.65%, 20.44%, 26.09% and 26.36%, respectively. Fig.38 As shown in the data, the liver ALT concentration in the Model group increased by 42.30% significantly (P < 0.01) compared with that in the Normal group, which synergistically with the change in serum ALT concentration indicated that the liver of SD rats in the Model group was damaged and the ALT concentration increased significantly. The liver ALT concentrations in the lLAC, mLAC, hLAC dose groups and Simvas group showed a very significant downward trend compared with that in the Model group (P < 0.01), decreasing by 9.75%, 15.78%, 18.75% and 20.16%, respectively, showing a good dose-dependency. However, there were still significant differences compared with the Normal group, indicating that the blood and liver ALT concentration indicators of SD rats were improved to a certain extent after LAC gavage.

[0207] 8. Effect of LAC on the morphology of liver tissue in SD rats by HE staining: Fig.39 As shown in the figure, the liver lobules of SD rats in the Normal group were regular, the cell size and shape were basically the same, the cell boundaries were clearly visible and densely arranged, the structure was normal, and no inflammatory cell infiltration was found. In the Model group, the liver lobules were disordered, the cells were disordered, and there was point and sheet necrosis, loose cytoplasm, blurred cell boundaries, large intercellular spaces, cell necrosis, a large number of atrophy or dissolution of the nucleus, cell vacuolation, etc. There were uneven numbers and sizes of round fat vacuoles distributed in the cytoplasm of hepatocytes. Compared with the Model group, the other groups were improved, among which the hLAC dose group had the best recovery effect. The number of hepatocytes was radially distributed with the central vein as the center, the outline of the liver lobules was clear, the hepatocytes were tightly arranged, and the complete liver tissue structure was still maintained, indicating that LAC administration has a good repair effect on the liver state of SD rats under a high-fat diet.

[0208] 9. Effect of LAC on Oil Red O staining of liver tissue morphology in SD rats: Fig.40As shown in the figure, the Normal group showed almost no staining with Oil Red O dye, and the liver morphology of SD rats was normal, while in the Model group, darker orange-red lipid droplets were observed, and flakes of dark orange-red were observed in the necrotic area, indicating that there was structural disorder and fat droplet deposition in the liver, showing diffuse fatty degeneration of hepatocytes. Compared with the Model group, each LAC group showed different degrees of improvement. Among them, the liver morphology of SD rats in the hLAC group was normal, and only a small number of hepatocytes contained fine dot-shaped powder-stained lipid droplets, which returned to normal levels, indicating that LAC has a good ability to clear excess lipids in the liver of SD rats.

[0209] 10. Effect of LAC on the morphology of brown adipose tissue in SD rats by HE staining: Fig.41 As shown, the cells in the brown adipose tissue of the Normal group were small and uniform, with multiple smaller lipid droplets in the cytoplasm; the cells in the brown adipose tissue of the Model group were of different sizes and had a disordered structure. Larger single-vesicle adipocytes were visible in the Model group, with a larger lipid droplet in the center of the cell, thin cytoplasm surrounding the lipid droplet, a flat nucleus located at the edge, and the cell was vacuolar.

[0210] The brown adipocytes in each LAC administration group were arranged more neatly than those in the Model group, and the cell sizes were more uniform. Smaller single-vesicle adipocytes could be observed, and the number of adipocytes per unit area was larger. The number of cell nuclei in the field of view gradually increased with the increase of LAC crude drug concentration, and the volume of lipid droplets decreased.

[0211] 11. Effect of LAC on the antioxidant function of SD rats: The free radical oxidation and antioxidant function of normal organisms are often in a dynamic balance. SOD is an important antioxidant enzyme in the body, which can effectively prevent the body's peroxidation reaction. MDA is the end product of the oxidation reaction and has certain cytotoxicity. Its content can reflect the body's oxidation degree. Therefore, the effect of LAC on the antioxidant enzyme system index superoxide dismutase SOD, peroxidation product MDA and total antioxidant capacity (T-AOC) of SD rats was determined, and the effect of LAC on the lipid-lowering function of SD rats was analyzed from the perspective of antioxidant function.

[0212] like Fig.42 , 43As shown in the results, the serum SOD level in the Model group was significantly reduced by 32.88% (P < 0.01) compared with the Normal group. Compared with the Model group, the serum SOD level in the lLAC group was significantly reduced by 6.43% (P < 0.01), and the mLAC group, hLAC group and Simvas group were significantly reduced by 16.54%, 30.84% ​​and 34.56%, respectively. The liver SOD level in the Model group was significantly reduced by 57.37% (P < 0.01) compared with the Normal group. Compared with the Model group, the liver SOD levels in each group were significantly increased (P < 0.01), among which the serum SOD levels in the hLAC group and Simvas group were only reduced by 14.93% and 15.11% compared with the Normal group. Among them, there was no significant difference in the SOD levels of the hLAC group and Simvas group in serum and liver, indicating that LAC has a better effect under high-dose gavage conditions and has a certain dose dependence.

[0213] like Fig.44 , 45 As shown, compared with the Normal group, the serum MDA level in the Model group increased significantly by 46.98% (P < 0.01), indicating that the serum MDA substance level in the Model group increased significantly. Compared with the Model group, the MDA levels in the LAC dose group and Simvas group after oral gavage showed an extremely significant downward trend (P < 0.01), decreasing by 7.54%, 13.85%, 21.26, and 27.57%, respectively; compared with the Normal group, the serum MDA level in the Model group increased extremely significantly by 128.22% (P < 0.01), indicating that the liver MDA level in the Model group increased significantly. Compared with the Model group, the liver MDA levels in the LAC low, medium, and high dose groups and Simvas group showed an extremely significant downward trend (P < 0.01), decreasing by 13.70%, 24.18%, 43.87%, and 53.18%, respectively, showing a certain dose-related effect. The recovery effect of serum and liver MDA levels was best under high-dose LAC oral gavage, which was close to normal levels.

[0214] like Fig.46 , 47As shown in the data, compared with the Normal group, the serum T-AOC level in the Model group decreased significantly by 59.00% (P < 0.01), and the serum T-AOC levels in the LAC administration groups and Simvas group decreased by 37.47%, 22.14%, 10.11%, and 2.43% respectively compared with the Normal group. The serum T-AOC levels in each group improved to varying degrees, and the serum T-AOC level in the hLAC group recovered most significantly, indicating that high-dose LAC gavage is helpful to improve the total antioxidant capacity of the serum of SD rats. Compared with the Normal group, the liver T-AOC level in the Model group decreased significantly by 65.49% (P < 0.01). The liver T-AOC levels in the LAC groups and Simvas group decreased by 36.75%, 330.05%, 5.46% and 22.66% respectively compared with the Normal group. The liver T-AOC levels in each group improved to varying degrees, and had a certain dose-dependency. Among them, the liver T-AOC level in the hLAC group returned to normal levels (P > 0.05), indicating that high-dose LAC gavage can restore the total antioxidant capacity of the liver of SD rats.

[0215] 12. Effect of LAC on key metabolic enzymes in SD rat serum: Leptin can suppress appetite, promote energy consumption, and reduce fat synthesis, and is crucial to fat synthesis and metabolism in SD rats; serum adiponectin (ADPN) promotes fat utilization and consumes energy by regulating the level of inflammatory factors. It also plays a key role in regulating the body's lipid metabolism function, maintaining blood lipid balance in the body, and improving the development of hyperlipidemia; lecithin cholesterol acyltransferase (LCAT) is synthesized by the liver and released into the blood. It is an enzyme that catalyzes in plasma, transferring the unsaturated fatty acids at the C2 position of HDL-C lecithin to free cholesterol to generate lysophosphatidylcholine and cholesterol esters. LCAT often binds to HDL-C, has high activity on the surface of HDL-C particles and plays a catalytic role. Its level in plasma can reflect the synthesis capacity of HDL-C.

[0216] Fig.48The effect of LAC on the serum leptin level of SD rats. After high-fat diet feeding, the average serum leptin content of SD rats in the Model group was 1038.6pg / mL, which was significantly higher than 588.8pg / mL in the Normal group (P<0.01), indicating that hyperlipidemia can cause a significant increase in leptin levels (P<0.01). After LAC intervention, the serum leptin content of SD rats was significantly lower than that of the Model group, showing a certain dose relationship. Compared with the Model group, the leptin level in the lLAC group was 962.0 pg / mL, significantly decreased by 7.37%. The leptin levels in the mLAC group, hLAC group, and Simvas group were 856.7 pg / mL, 619.6 pg / mL, and 711.5 pg / mL, respectively, extremely significantly decreased by 17.51%, 40.34%, and 31.49%, respectively (P<0.01). There was no significant difference in the leptin level between the hLAC group and the Normal group (P>0.05), which proved that LAC can alleviate the leptin resistance of SD rats with hyperlipidemia and can return to normal levels at a certain concentration of raw medicine.

[0217] Fig.49 The effect of LAC on the level of adiponectin in the serum of SD rats. After high-fat diet feeding, the average content of adiponectin in the serum of SD rats in the Model group was 638.4 ng / mL, which was significantly lower than 1634.1 ng / mL in the Normal group (P<0.01), indicating that hyperlipidemia can cause a significant decrease in adiponectin levels (P<0.01). After intervention with different concentrations of LAC, the serum adiponectin content of SD rats was significantly restored compared with the Model group. Compared with the Model group, there was a very significant increase (P<0.01). The adiponectin levels of the lLAC group, mLAC group, hLAC group, and Simvas group were 844.5 ng / mL, 988.9 ng / mL, 1413.1 ng / mL, and 1585.4 ng / mL, respectively. Among them, the adiponectin level in the hLAC group was the highest, proving that LAC can repair the adiponectin level of SD rats with hyperlipidemia at a certain concentration of raw drugs.

[0218] Fig.50The effect of LAC on the level of lecithin cholesterol acyltransferase in the serum of SD rats. The effect of different experimental groups on the content of lecithin cholesterol acyltransferase in the serum of SD rats is shown in the figure. After high-fat diet feeding, the average LCAT content in the serum of SD rats in the Model group was 10.33μmol / L, which was significantly lower than 18.94μmol / L in the Normal group (P<0.01), indicating that hyperlipidemia can cause a significant decrease in LCAT levels (P<0.01). After intervention with different concentrations of LAC, the serum LCAT content of SD rats was significantly restored compared with the Model group. Compared with the Model group, the LCAT levels of the lLAC group, mLAC group, hLAC group, and Simvas group were 13.19 μmol / L, 16.16 μmol / L, 17.57 μmol / L, and 17.60 μmol / L, respectively, which were significantly increased by 27.65%, 56.43%, 70.10%, and 70.35%, respectively. Among them, the hLAC group had the highest LCAT level, which was not significantly different from that of the Simvas group (P>0.05), proving that LAC can repair the LCAT level of hyperlipidemic SD rats at a certain concentration of crude drug.

[0219] 13. Effect of LAC on FAS and FFAs in SD rats: Fatty acid synthase (FAS) is a key enzyme involved in cellular fat metabolism, and hyperlipidemia can lead to increased FAS activity in the body. Free fatty acids (FFAs) are substances broken down from neutral fat. When the energy required for body activities is exhausted, adipose tissue will break down neutral fat into free fatty acids to be used as energy. Increased free fatty acids can lead to abnormal liver fat metabolism and cause the risk of fatty liver. The liver is the core organ of lipid metabolism in higher animals. Hepatocytes mainly take in free fatty acids from the blood, and also take in free fatty acids converted from sugar metabolism in the blood, and then synthesize triglycerides, phospholipids and cholesterol esters.

[0220] Fig.51 , 52 The effect of LAC on the level of fatty acid synthase in serum and liver of SD rats. Fig.51As shown in the results, after high-fat diet feeding, the level of fatty acid synthase in the serum of SD rats in the Model group was 124.83 nmol / L, which was significantly higher than that of rats in the Normal group (34.23 nmol / L (P<0.01), indicating that hyperlipidemia can cause a significant increase in serum fatty acid synthase levels (P<0.01). After intervention with different concentrations of LAC, the content of serum fatty acid synthase in SD rats was significantly lower than that in the Model group. Compared with the Model group, the fatty acid synthase levels of the lLAC group, mLAC group, hLAC group, and Simvas group were 114.66 nmol / L, 73.18 nmol / L, 54.92 nmol / L, and 61.57 nmol / L, respectively, which were extremely significantly reduced by 8.15%, 41.38%, 56.01%, and 50.68%, respectively. Among them, the fatty acid synthase level of the hLAC group was the lowest, and there was no significant difference between it and the Simvas group (P>0.05). The gap between the lLAC group and the mLAC group was large, which may be because when the concentration of LAC crude drug reached the level of the mLAC group, LAC had a significant effect on liver repair, thereby repairing liver cells and reducing the release of fatty acid synthase. This proves that at a certain crude drug concentration, LAC can repair the serum fatty acid synthase level of hyperlipidemia SD rats.

[0221] Effects of different experimental groups on the level of fatty acid synthase in the liver of SD rats Fig.52 As shown in the results, the level of fatty acid synthase in the liver of SD rats in the Model group was 1.174 nmol / L, which was significantly higher than 0.611 nmol / L in rats in the Normal group (P<0.01), indicating that hyperlipidemia can cause a significant increase in the level of fatty acid synthase in the liver (P<0.01). After intervention with different concentrations of LAC, the content of fatty acid synthase in the liver of SD rats was significantly reduced compared with the Model group. Compared with the Model group, the fatty acid synthase levels of the lLAC group, mLAC group, hLAC group, and Simvas group were 0.922nmol / L, 0.837nmol / L, 0.699nmol / L, and 0.706nmol / L, respectively, which were significantly reduced by 21.52%, 28.77%, 40.41%, and 39.91%, respectively. Among them, the fatty acid synthase level in the hLAC group was the lowest, and there was no significant difference between it and the Normal group (P>0.05), which proved that LAC at a certain concentration of crude drug can repair the serum fatty acid synthase of hyperlipidemic SD rats to normal levels.

[0222] Fig.53 , 54 The effect of LAC on the free fatty acid levels in the serum and liver of SD rats. Fig.53As shown in the figure, after high-fat diet feeding, the level of free fatty acids in the serum of SD rats in the Model group was 4.021 nmol / mL, which was significantly higher than 0.781 nmol / mL in the Normal group (P<0.01), indicating that hyperlipidemia can cause a significant increase in serum free fatty acid levels (P<0.01). After intervention with different concentrations of LAC, the content of serum free fatty acids in SD rats was significantly reduced compared with the Model group. Compared with the Model group, the free fatty acid levels of the lLAC group, mLAC group, hLAC group, and Simvas group were 2.347 nmol / mL, 1.834 nmol / mL, 1.044 nmol / mL, and 1.429 nmol / mL, respectively, which were significantly reduced by 41.61%, 54.38%, 74.04%, and 64.44%, respectively. Among them, the free fatty acid level of the hLAC group was the lowest, and there was no significant difference between it and the Normal group (P>0.05). The level of the lLAC group decreased more significantly compared with the Model group, which may be because when the concentration of LAC crude drug reached the level of the lLAC group, LAC had a significant repair effect on liver lipid metabolism, proving that LAC can repair the serum free fatty acid level of hyperlipidemic SD rats at a certain crude drug concentration.

[0223] Effects of different experimental groups on free fatty acid levels in the liver of SD rats Fig.54 As shown in the results, the free fatty acid level in the liver of SD rats in the Model group was 2.145 nmol / mL, which was significantly higher than 0.784 nmol / mL in the Normal group (P<0.01), indicating that hyperlipidemia can cause a significant increase in the free fatty acid level in the liver (P<0.01). After intervention with different concentrations of LAC, the free fatty acid content in the liver of SD rats was significantly reduced compared with the Model group. Compared with the Model group, the free fatty acid levels of the lLAC group, mLAC group, hLAC group, and Simvas group were 1.797 nmol / mL, 1.599 nmol / mL, 1.005 nmol / mL, and 1.145 nmol / mL, respectively, which were significantly reduced by 16.23%, 25.44%, 53.16%, and 46.62%, respectively. Among them, the free fatty acid level in the hLAC group was the lowest, proving that LAC can repair the serum free fatty acid level of SD rats with hyperlipidemia at a certain concentration of LAC.

[0224] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A synergistic lipid-lowering composition, characterized in that: The composition comprises the following volume ratios: blue indigo fruit extract: black fungus extract: hawthorn extract of 3-5:15-17:4-6; The concentration of the blue honeysuckle fruit extract is 1-2 g / mL, the concentration of the black fungus extract is 0.05-0.20 g / mL, and the concentration of the hawthorn extract is 0.1-0.4 g / mL.

2. The synergistic lipid-lowering composition according to claim 1, characterized in that: The blue indigo fruit extract, black fungus extract or hawthorn extract is obtained by using blue indigo fruit, black fungus or hawthorn as raw materials through ultrasonic water extraction.

3. The synergistic lipid-lowering composition according to claim 2, characterized in that: The ultrasonic water extraction process comprises the following steps: The black fungus is crushed and sieved, mixed with water for a first ultrasonic extraction, and then centrifuged for a first time to obtain a first black fungus supernatant and a first black fungus residue; The first black fungus filter residue is mixed with water to perform a second ultrasonic extraction, and after the extraction, a second centrifugation is performed to obtain a second black fungus supernatant and a second black fungus filter residue; The first black fungus supernatant and the second black fungus supernatant are combined and concentrated to obtain a black fungus extract; The blue indigo fruit is subjected to a wall breaking treatment, mixed with water for a first ultrasonic extraction, and then subjected to a first centrifugation to obtain a first blue indigo fruit supernatant and a first blue indigo fruit residue; The first filter residue of blue Indigo Fruit is mixed with water for a second ultrasonic extraction, and then centrifuged for a second time to obtain a second supernatant of blue Indigo Fruit and a second filter residue of blue Indigo Fruit; The first supernatant of blue Indigo Carpa and the second supernatant of blue Indigo Carpa are combined and concentrated to obtain a blue Indigo Carpa extract; The hawthorn is de-cored, the wall is broken, mixed with water for a first ultrasonic extraction, and after the extraction, a first centrifugation is performed to obtain a first hawthorn supernatant and a first hawthorn residue; The first hawthorn residue is mixed with water for a second ultrasonic extraction, and then centrifuged for a second time to obtain a second hawthorn supernatant and a second hawthorn residue; The first hawthorn supernatant and the second hawthorn supernatant are combined and concentrated to obtain an extract.

4. The synergistic lipid-lowering composition according to claim 3, characterized in that: The black fungus is sieved with a mesh number of 30-50, and the total weight ratio of the black fungus, blue indigo fruit, hawthorn and water is 1:3-50.

5. The synergistic lipid-lowering composition according to claim 3, characterized in that: During the ultrasonic extraction of black fungus, blue indigo fruit or hawthorn, the temperature of each ultrasonic extraction is 35-60°C, and the time of each ultrasonic extraction is 40-70 minutes.

6. The synergistic lipid-lowering composition according to claim 3, characterized in that: The concentration is carried out by rotary evaporation, the concentration temperature is 40-55° C., and the volume after concentration is 1 / 4-1 / 6 of the original volume.

7. Use of the synergistic lipid-lowering composition according to any one of claims 1 to 6 in the preparation of a drug for treating hyperlipidemia or a nutritional functional food.

8. A beverage containing the synergistic lipid-lowering composition according to any one of claims 1 to 6, characterized in that: The beverage further comprises auxiliary materials, which include erythritol, potassium sorbate, pectinase, soybean polysaccharide and CMC.

9. The beverage according to claim 8, characterized in that The percentage of the auxiliary materials added to the beverage is as follows: the amount of erythritol added is 12-15%, the amount of pectinase added is 0.3-0.7%, the amount of soybean polysaccharide added is 0.3-0.7%, the amount of CMC added is 0.2-0.5%, and the amount of potassium sorbate added is 0.01-0.05%.

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