Lactobacillus paracasei compound beverage, preparation method and application of lactobacillus paracasei

Abstract

The invention discloses a lactobacillus paracasei compound beverage, a preparation method and application of lactobacillus paracasei. The beverage comprises 20-50 v% of a lactobacillus paracasei bacterial suspension and 50-80 v% of a plant extract, the preservation number of the lactobacillus paracasei SWFU-D16 is CCTCC (China Center for Type Culture Collection) NO: M2023709, and the lactobacillus paracasei SWFU-D16 is preserved in the China Center for Type Culture Collection on May 9, 2023. According to the present invention, the Lacticaseibacillus paracasei SWFU-D16 separated from Yunnan characteristic traditional food (dairy cake), the hawthorn, the pawpaw, the roxburgh rose, the black tea and the like are prepared into the composite beverage, and the beverage has characteristics of high nutrition value, spleen and stomach regulating, blood fat level regulating, intestinal flora regulating and other functions, and meets the requirements of people on taste, nutrition, health, health preserving and other aspects.

Description

Technical Field

[0001] This invention belongs to the field of food processing technology, specifically relating to a Lactobacillus paracasei compound beverage, its preparation method, and the application of Lactobacillus paracasei. Background Technology

[0002] Lactic acid bacteria play a vital role in human health and life. Besides improving the flavor and texture of food, they also regulate the balance of the gut microbiota, enhance immunity, and delay aging. Studies have shown that lactic acid bacteria are crucial for maintaining the balance of the human gut microbiota and lipid metabolism.

[0003] Some traditional fermented foods in my country contain abundant lactic acid bacteria resources, providing favorable conditions for screening lactic acid bacteria strains with excellent lipid-lowering functions. Milk cake, a traditional Yunnan specialty food made from goat milk through natural fermentation, has not only preserved a rich and diverse range of lactic acid bacteria strains during its long-term natural domestication and process optimization, but has also fostered many lactic acid bacteria strains with unique biological characteristics.

[0004] With improved living standards and a faster pace of life, people are consuming more energy than they are exercising, which can easily lead to elevated blood lipid levels and intestinal dysfunction. Statistics show that the prevalence of dyslipidemia among Chinese adults is 40.40%. Hyperlipidemia cannot be cured with medication, and most medications have side effects, impacting people's quality of life and health. Summary of the Invention

[0005] To reduce the harm of lipid-lowering drugs to the human body and develop lipid-lowering food products with no toxic side effects, this invention isolates lactic acid bacteria with lipid-lowering effects from traditionally fermented goat milk cake and combines them with medicinal herbs that have spleen-strengthening, lipid-lowering, and immunity-enhancing effects to develop a nutritious, delicious, and health-promoting sugar-free lactic acid bacteria beverage. The specific technical solution provided by this invention is as follows:

[0006] In a first aspect, the present invention provides a compound beverage of Lactobacillus paracasei and plant extract with lipid-lowering effect.

[0007] Preferably, the Lactobacillus paracasei SWFU-D16 has the accession number CCTCC NO: M2023709 and was deposited at the China Center for Type Culture Collection on May 9, 2023.

[0008] Preferably, the compound beverage comprises 20-50 v% of the Lactobacillus paracasei suspension and 50-80 v% of the plant extract.

[0009] Preferably, the proportion of Lactobacillus paracasei suspension in the compound beverage is 20v%, 25v%, 30v%, 35v%, 40v%, 45v%, and 50v%, and the corresponding proportion of the plant extract is 80v%, 75v%, 70v%, 65v%, 60v%, 55v%, and 50v%.

[0010] Furthermore, the compound beverage comprises 50 v% of Lactobacillus paracasei suspension and 50 v% of plant extract.

[0011] Preferably, the bacterial count of the *Lactobacillus paracasei* suspension is 1 × 10⁻⁶. 8 ~1×10 9 CFU / mL.

[0012] Furthermore, the plant extracts include hawthorn extract, black tea extract, prickly pear extract, and papaya extract.

[0013] Preferably, the volume fractions of the hawthorn extract are 10-30 parts, the volume fractions of the black tea extract are 30-50 parts, the volume fractions of the prickly pear extract are 10-30 parts, and the volume fractions of the papaya extract are 5-20 parts.

[0014] Preferably, the plant extract further includes citric acid, erythritol, sucralose, pectin, and carrageenan.

[0015] Preferably, in the plant extract, the concentration of citric acid is 2-5 g / L, the concentration of erythritol is 20-100 g / L, the concentration of sucralose is 0.1-0.3 g / L, the concentration of pectin is 0.1-1 g / L, and the concentration of carrageenan is 0.1-1 g / L.

[0016] In a second aspect, the present invention provides a method for preparing the compound beverage described in the first aspect, characterized in that: the preparation method includes the following steps:

[0017] Using Lactobacillus paracasei SWFU-D16 with accession number CCTCC NO: M2023709 as the strain, a Lactobacillus paracasei bacterial suspension was prepared.

[0018] Add dried hawthorn to hot water, extract and filter to obtain hawthorn extract;

[0019] Prickly pear was added to drinking water, pulped, and filtered to obtain prickly pear extract;

[0020] Papaya slices were added to the second hot water, and the extract was obtained by soaking and filtering.

[0021] Black tea is brewed with hot water (the third type of hot water) and then filtered to obtain a black tea extract.

[0022] The hawthorn extract, the prickly pear extract, the papaya extract and the black tea extract are mixed according to the first preset ratio. Then, citric acid, erythritol, sucralose, pectin and carrageenan are added. After mixing, the mixture is sterilized at high temperature to obtain the plant extract.

[0023] The compound beverage is prepared by mixing and homogenizing the Lactobacillus paracasei suspension with the plant extract according to the second preset ratio.

[0024] Preferably, the ratio of dried hawthorn to the first hot water is 1:20-30 (w:v), for example, 1:20, 1:22, 1:25, 1:28, or 1:30.

[0025] Preferably, the temperature of the first hot water is 70-80℃, for example, 70℃, 73℃, 75℃, 78℃, or 80℃.

[0026] Preferably, the first hot water extraction time is 5-10 minutes, for example, 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes, or 10 minutes.

[0027] Preferably, the ratio of prickly pear to drinking water is 1:20-30 (w:v), for example, 1:20, 1:22, 1:25, 1:28, or 1:30.

[0028] Preferably, the drinking water is at room temperature.

[0029] Preferably, the ratio of papaya slices to the second hot water is 1:20-30 (w:v), for example, 1:20, 1:22, 1:25, 1:28, or 1:30.

[0030] Preferably, the temperature of the second hot water is 90-95℃, for example 90℃, 91℃, 92℃, 93℃, 94℃, or 95℃.

[0031] Preferably, the second hot water extraction time is 60-80 minutes, for example, 60 minutes, 65 minutes, 70 minutes, 75 minutes, or 80 minutes.

[0032] Preferably, the ratio of the black tea to the third hot water is 1:60-80 (w:v), for example, 1:60, 1:65, 1:70, 1:75, or 1:80.

[0033] Preferably, the temperature of the third hot water is 90-95℃, for example, 90℃, 91℃, 92℃, 93℃, 94℃, or 95℃.

[0034] Preferably, the brewing time is 1-3 minutes, for example, 1 minute, 1.5 minutes, 2 minutes, 2.5 minutes, or 3 minutes.

[0035] Preferably, the first preset ratio is (v%) of hawthorn extract: (v%) of black tea extract: (v%) of prickly pear extract: (v%) of papaya extract.

[0036] Furthermore, the first preset ratio is 10-30:30-50:10-30:5-20.

[0037] Preferably, in the plant extract, the concentration of citric acid is 2-5 g / L, the concentration of erythritol is 20-100 g / L, the concentration of sucralose is 0.1-0.3 g / L, the concentration of pectin is 0.1-1 g / L, and the concentration of carrageenan is 0.1-1 g / L.

[0038] Preferably, the second preset ratio is Lactobacillus paracasei suspension (v%): plant extract (v%).

[0039] Furthermore, the second preset ratio is 20-50:50-80.

[0040] Thirdly, the present invention provides the application of the compound beverage described in the first aspect in lowering blood lipids.

[0041] Preferably, the application includes at least one of the following applications:

[0042] (1) Reduce TC, TG and LDL-C, and increase HDL-C;

[0043] (2) Degradation of triglycerides;

[0044] (3) Regulate the expression of lipid metabolism-related genes in the liver, wherein the genes are preferably LPL, PPAR-α, CYP7A1 and / or CPT1, and further, the regulation is upregulation;

[0045] (4) Enhance the diversity of gut microbiota;

[0046] (5) Increase the thickness of the digestive tract mucosa.

[0047] Fourthly, this invention provides the use of Lactobacillus paracasei SWFU-D16, with accession number CCTCC NO: M2023709, in the preparation of foods, health products, and / or pharmaceuticals that lower blood lipids.

[0048] Preferably, the application is the same as the application content of the third aspect.

[0049] Preferably, the food also includes permitted additives.

[0050] Preferably, the health product also includes excipients that are permitted to be added to the health product.

[0051] Preferably, the feed also includes additives that are permitted to be added to the feed.

[0052] Preferably, the drug further includes excipients that are permitted to be added to the drug.

[0053] Fifthly, the present invention provides the use of Lacticaseibacillus paracasei SWFU-D16 bacterial suspension in the preparation of foods, health products, feeds and / or drugs that lower blood lipids.

[0054] Preferably, the application is the same as the application content of the third aspect.

[0055] Preferably, the food also includes permitted additives.

[0056] Preferably, the health product also includes excipients that are permitted to be added to the health product.

[0057] Preferably, the feed also includes additives that are permitted to be added to the feed.

[0058] Preferably, the drug further includes excipients that are permitted to be added to the drug.

[0059] The beneficial effects of this invention are:

[0060] The Lactobacillus paracasei SWFU-D16 strain in this invention was isolated from a traditional Yunnan specialty food (milk cake). Compared with existing lactic acid bacteria strains, SWFU-D16 has a stronger effect on lowering blood lipids and restoring intestinal microbial species.

[0061] The beverage, made from SWFU-D16 and blended with hawthorn, papaya, prickly pear, black tea, and other ingredients, is highly nutritious. This blended beverage also has functions such as regulating the spleen and stomach, adjusting blood lipid levels and gut microbiota, thus meeting people's needs in terms of taste, nutrition, health, and wellness. Attached Figure Description

[0062] Figure 1 The effects of SWFU-D16 on body weight and Lee's index in high-fat mice;

[0063] Figure 2 Fat vacuoles in hepatocytes of high-fat mice;

[0064] Figure 3This refers to fatty degeneration within liver tissue cells of high-fat mice.

[0065] Figure 4 The effect of SWFU-D16 on the mRNA expression levels of lipid metabolism-related genes LPL, PPAR-α, CYP7A1, CPT1 and PPAR-γ in the liver tissue of hyperlipidemic mice;

[0066] Figure 5 The effect of SWFU-D16 on the Alpha diversity index of gut microbiota in high-fat mice;

[0067] Figure 6 The effect of SWFU-D16 on the species taxonomy system in the intestines of high-fat mice;

[0068] Figure 7 The effect of SWFU-D16 on species taxonomic branching in the intestines of high-fat mice;

[0069] Figure 8 The effect of SWFU-D16 on LDA values ​​of intestinal species in high-fat mice;

[0070] Figure 9 The effect of compound beverages on the expression of adipogenesis genes in 3T3-L1 preadipocytes.

[0071] The strain of this invention was deposited on May 9, 2023, with accession number CCTCC NO: M2023709. It is classified and named *Lactaseibacillus paracasei* SWFU-D16, and the depository is the China Center for Type Microbiology, located at Wuhan University, China, 430072, China. Detailed Implementation

[0072] The technical solution of the present invention will be further described below with reference to embodiments and accompanying drawings. The advantages and features of the present invention will become clearer as the description unfolds. However, it should be understood that the embodiments are merely exemplary and do not constitute a limitation on the scope of the present invention.

[0073] It should be noted that, unless otherwise specified, the experimental methods used in the following embodiments are conventional methods in the art.

[0074] Example 1: Effects of SWFU-D16 on blood lipids in mice

[0075] 1.1 Culture of SWFU-D16

[0076] After activating the preserved SWFU-D16 strain, it was inoculated into MRS liquid medium at a 1% inoculum and cultured at 37°C, yielding a bacterial count of 2.0 × 10⁻⁶. 11 CFU / mL bacterial suspension.

[0077] 1.2 Mouse feeding experiment

[0078] Seventy-two mice weighing 20g were randomly divided into six groups: control group, model group, low-dose group, medium-dose group, high-dose group, and drug group. The control group was fed a low-fat control diet and saline; the model group was fed a 60% high-fat model diet and saline; and the low-dose group was fed a 60% high-fat model diet and a drug containing 1×10⁻⁶ mg / L. 8 The medium-dose group was fed a 60% high-fat model diet and a diet containing 1×10 CFU / mL SWFU-D16 in physiological saline; the medium-dose group was fed a 60% high-fat model diet and a diet containing 1×10 CFU / mL SWFU-D16 in physiological saline. 9 CFU / mL SWFU-D16 in physiological saline; the high-dose group was fed a 60% high-fat model diet and a diet containing 1×10 10 The control group was fed physiological saline with CFU / mL SWFU-D16; the drug group was fed a 60% high-fat model diet and cholestyramine powder (1g / kg).

[0079] Table 1 Animal Feed Formulation

[0080]

[0081] Table 2 Grouping of Animal Experiments

[0082]

[0083] Each group had free access to water. Mice were administered 0.4 mL of fluid per mouse via gavage daily in the morning as described above. Daily food intake and weekly body weight were recorded. After 10 weeks of feeding, mice in each group were anesthetized with sodium pentobarbital via intraperitoneal injection, and blood was collected from the heart, feces, and liver. The following analyses were performed:

[0084] The liver, heart, kidneys, spleen, perirenal fat, and epididymal fat of mice were collected and weighed, and the organ indices of the mice were calculated. The experimental results are shown below. Figure 1 .

[0085] Depend on Figure 1 As can be seen, the body weight of mice in the high-fat model group, low-dose SWFU-D16 group, medium-dose SWFU-D16 group, high-dose SWFU-D16 group, and drug group was higher than that of the control group, indicating that the mouse high-fat model was successfully established. Compared with the high-fat model group, the body weight of mice in the low-dose, medium-dose, and high-dose SWFU-D16 groups decreased with increasing SWFU-D16 dosage, and the difference reached a significant level. There was no significant difference in body weight between the medium-dose and high-dose SWFU-D16 groups and the drug group. This indicates that SWFU-D16 can effectively reduce mouse body weight and achieve the same effect as the drug.

[0086] Furthermore, the Lee's index of mice in the model group, low-dose SWFU-D16 group, medium-dose SWFU-D16 group, drug group, and high-dose SWFU-D16 group decreased sequentially. The differences between the medium-dose SWFU-D16 group, high-dose SWFU-D16 group, and drug group and the model group reached statistical significance, and the difference between the high-dose SWFU-D16 group and the drug group was close to statistical significance. This indicates that oral administration of SWFU-D16 can reduce body fat gain in mice under a high-fat diet.

[0087] Morphological observations of the same areas of mouse liver and epididymal fat were performed using an optical microscope. The results are shown in [Figure number missing]. Figure 2 and Figure 3 In the high-fat diet model group, the liver tissue of mice exhibited microvesicular steatosis and high fat content, with numerous fat vacuoles observed around blood vessels, leading to cell swelling and disruption of cell membrane integrity. After intervention with SWFU-D16, steatosis in the mouse liver tissue was reduced, fat vacuoles within hepatocytes decreased, and cell morphology was similar to that of the control group. In the high-fat diet model group, epididymal adipocytes were large and had thin cell membranes. In contrast, the epididymal adidymal tissue of mice treated with SWFU-D16 was denser than that of the model group, and the average size of adipocytes was similar to that of the control group. This indicates that SWFU-D16 can alleviate adipocyte degeneration and hypertrophy induced by a high-fat diet.

[0088] The levels of triglycerides (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) in mouse serum were measured, and the results are shown in Table 3.

[0089] Table 3. Serum biochemical indicators of mice

[0090]

[0091] Table 3 shows that the TC, TG, and LDL-C levels in the high-fat model group, low-dose SWFU-D16 group, medium-dose SWFU-D16 group, and high-dose SWFU-D16 group were all higher than those in the control group, while the HDL-C levels were all lower than those in the control group, indicating that the mouse high-fat model was successfully established. Compared with the high-fat model group, the TC, TG, and LDL-C levels in the low-dose, medium-dose, and high-dose SWFU-D16 groups decreased with increasing SWFU-D16 dosage, while the HDL-C level increased with increasing SWFU-D16 dosage. The TC levels in the low-dose, medium-dose, and high-dose SWFU-D16 groups and the drug group were significantly different from those in the model group. There were no significant differences between the low-dose and medium-dose SWFU-D16 groups and the drug group, but the difference between the high-dose SWFU-D16 group and the drug group was significant. The TG levels in the SWFU-D16 medium-dose group, SWFU-D16 high-dose group, and drug group were significantly different from those in the model group, while the difference between the SWFU-D16 high-dose group and the drug group was not significant. The LDL-C levels in the SWFU-D16 low-dose group, SWFU-D16 medium-dose group, SWFU-D16 high-dose group, and drug group were significantly different from those in the model group, while the differences between the SWFU-D16 medium-dose group, SWFU-D16 high-dose group, and drug group were not significant. The HDL-C levels in the SWFU-D16 medium-dose group, SWFU-D16 high-dose group, and drug group were significantly different from those in the model group, while the difference between the SWFU-D16 high-dose group and the drug group was significant. This indicates that SWFU-D16 can effectively reduce serum lipid levels in mice and thus play a role in reducing obesity.

[0092] Analysis of gene expression related to liver lipid metabolism in mice; results are shown below. Figure 4 .

[0093] Depend on Figure 4It can be seen that, compared with the high-fat model group, the mRNA expression levels of LPL, PPAR-α, CYP7A1, and CPT1 in the livers of mice in the low-dose, medium-dose, and high-dose SWFU-D16 groups increased with increasing SWFU-D16 dosage, while the expression level of PPAR-γ decreased with increasing SWFU-D16 dosage. PPARs are ligand-activated receptors in the nuclear hormone receptor family and are involved in the physiological processes of various diseases, including obesity. Among them, PPAR-α plays an important role in maintaining lipid metabolism in organisms. SWFU-D16 intervention upregulated the expression level of PPAR-α in mice in the high-fat model group, indicating that the lactic acid bacteria SWFU-D16 can normalize the transport and oxidation of free fatty acids and reduce lipid accumulation; PPAR-γ regulates the proliferation and differentiation of adipocytes. SWFU-D16 intervention downregulated the expression level of PPAR-γ, indicating that the lactic acid bacteria SWFU-D16 can reduce the proliferation and differentiation of adipocytes, making them similar to those of normal mice. CPT1 is a crucial rate-limiting enzyme in fatty acid oxidation, and its level changes are closely related to the development of obesity. SWFU-D16 intervention upregulated CPT1 expression, indicating that the lactic acid bacteria SWFU-D16 can promote fatty acid oxidation metabolism in mice. LPL is a key enzyme in lipid metabolism; increased LPL expression can increase serum high-density lipoprotein cholesterol levels and decrease cholesterol content. SWFU-D16 intervention upregulated LPL expression in high-fat model mice, indicating that SWFU-D16 regulates and inhibits the increase in serum cholesterol in mice by affecting LPL expression. CYP7A1 is the rate-limiting enzyme for bile acid synthesis, catalyzing the breakdown of cholesterol into bile acids in the liver and preventing obesity. SWFU-D16 intervention upregulated CYP7A1 gene expression in high-fat model mice, indicating that SWFU-D16 can promote cholesterol breakdown and reduce mouse weight. Therefore, the mRNA regulation levels in the SWFU-D16 group were close to or exceeded those in the drug group, indicating that SWFU-D16 has a significant lipid-lowering effect.

[0094] Analysis of the gut microbiota of mice was performed, and the results are shown in the table below. Figures 5-8 .

[0095] Depend on Figure 5 It can be seen that the Chao, observed species and Shannon indices of mice fed a high-fat diet were low, while the higher the Shannon index, the higher the community diversity. This indicates that a high-fat diet will reduce the abundance of microorganisms in the mouse gut. However, the abundance of lactic acid bacteria SWFU-D16 after gavage showed a slight upward trend, indicating that the intake of lactic acid bacteria SWFU-D16 improved the diversity of the gut microbiota.

[0096] Depend on Figure 6 It can be seen that the gut microbiota at the genus level in the six groups of mice were mainly composed of *Ruminiclostridium 9*, *Helicobacter*, *Akkermansia spp*, *Bacteroides*, and *Desulfovibrio*. Among them, the relative abundance of *Ruminiclostridium* was significantly increased after intervention with lactic acid bacteria SWFU-D16. This indicates that lactic acid bacteria SWFU-D16 prevents harmful substances from passing through the digestive tract by increasing the thickness of the mouse digestive tract mucosa, thereby playing a role in preventing hyperlipidemia.

[0097] like Figure 7 and Figure 8 As shown, the biomarkers with an LDA Score greater than 5 are Firmicutes, Clostridia, Clostridiales, Bacteroidetes, Bacteroidia, and Bacteroidales.

[0098] Example 2: Preparation of Compound Beverage Sample 1

[0099] 2.1 Culture of SWFU-D16

[0100] The specific cultivation process is the same as in 1.1 of Example 1.

[0101] 2.2 Preparation of plant extract

[0102] 2.2.1 Grind dried hawthorn into powder, and extract it with hot water at 75℃ for 7 minutes at a ratio of 1:25 (w:v). Filter the solution to obtain hawthorn extract.

[0103] 2.2.2 Fresh prickly pear was mixed with drinking water at room temperature at a ratio of 1:25 (w:v), pulped and filtered to obtain prickly pear extract.

[0104] 2.2.3 The papaya slices were soaked in a 95℃ hot water bath for 70 minutes at a ratio of 1:25 (w:v) and then filtered after cooling to obtain papaya extract.

[0105] 2.2.4 Take Dianhong tea leaves, brew them with 95℃ hot water at a ratio of 1:70 (w:v) and cover for 2 minutes, then filter to obtain black tea extract.

[0106] 2.2.5 The SWFU-D16 bacterial suspension prepared in Experiment 2.1 was serially diluted 10-fold to obtain a viable count of 1×10⁻⁶. 9 SWFU-D16 bacterial suspension at CFU / mL.

[0107] 2.2.6 First, mix 20 parts hawthorn extract, 40 parts black tea extract, 20 parts prickly pear extract, and 10 parts papaya extract by volume. Then, add citric acid, erythritol, sucralose, pectin, and carrageenan to the mixture at concentrations of 3 g / L, 50 g / L, 0.15 g / L, 0.75 g / L, and 0.4 g / L, respectively. After mixing thoroughly, sterilize at 90°C for 30 seconds, then cool to room temperature for later use.

[0108] 2.2.7 Take the above-mentioned sterilized plant extract according to the proportions in Table 4, and add 1×10 9 A lactic acid bacteria compound beverage sample was prepared by mixing CFU / mL SWFU-D16 bacterial suspension evenly.

[0109] Table 4. Combination of Compound Beverages

[0110]

[0111] 2.2.8 The lipid-lowering ability of the compound beverages No. 1-6 prepared in 2.2.7 was determined using a triglyceride kit. A 2.0% polyvinyl alcohol solution and vegetable oil were ultrasonically treated and added to the sample solution as triglyceride sources. After thorough mixing, the working solution was added, and the mixture was incubated at 37℃ for 10 min. The absorbance was measured at 500 nm using a blank tube for zeroing, and the degradation rate of triglycerides was calculated. Simultaneously, the color, flavor, and texture of the compound beverages No. 1-6 were sensory evaluated. The results are shown in Table 5.

[0112] Table 5. Lipid-lowering rate and sensory scores of compound beverages

[0113]

[0114] As shown in Table 5, the compound beverage sample 3 has a significantly stronger lipid-lowering ability, a light brown color, and unique aromas and flavors of hawthorn, prickly pear and black tea. It has a pleasantly sweet and sour taste and its overall weighted score is significantly better than other formulas.

[0115] Example 3: Preparation of comparative samples of compound beverages

[0116] The specific experimental procedure is the same as that of the compound beverage sample 3 in Example 2, except that SWFU-D16 is replaced with Lactobacillus plantarum (ATCC 8014).

[0117] Example 4: Effects of compound beverage on adipogenesis genes in 3T3-L1 preadipocytes

[0118] After resuscitation, medium change, and passage, 3T3-L1 preadipocytes were cultured at a rate of 2×10⁻⁶. 4Cells were seeded at a density of 10 cells / well in 24-well plates and cultured until the cell density reached over 90%. Preadipocytes were induced in MDI medium for 2 days. 10 μL of water, compound beverage sample 3, compound beverage sample 4, SWFU-D16 bacterial suspension, extract of medicinal and edible plants, and sample from Example 3 were added to each well, respectively. Cells were then maintained in basal medium supplemented with 10 μg / mL insulin, with the medium being replaced every 2 days. Induction continued until day 8, when significant lipid droplet formation was observed. RNA was extracted and cDNA was synthesized. Quantitative real-time PCR was used to analyze changes in adipogenesis and differentiation genes in 3T3-L1 cells. The results are shown in [Figure number missing]. Figure 9 .

[0119] The differentiation of preadipocytes into adipocytes is influenced by adipogenesis-related factors C / EBPα and C / EBPβ, and is also related to lipid metabolism genes acetyl-CoA carboxylase (ACC) and fatty acid synthase (FAS). Lipid-lowering drugs or foods often inhibit the differentiation of 3T3-L1 preadipocytes by suppressing adipogenesis and adipocyte formation. Figure 9 As shown, all five experimental groups reduced the expression levels of adipogenesis genes C / EBPα and C / EBPβ, and adipogenesis genes FAS and ACC. Compared with compound beverage sample 4, SWFU-D16 bacterial suspension, medicinal and edible plant extract, and the compound beverage sample of Example 3, compound beverage sample 3 showed significantly better reduction in the expression levels of the four genes. This indicates that the Lactobacillus paracasei SWFU-D16-medicinal and edible plant compound beverage prepared in this invention, derived from dairy cake, has a strong lipid-lowering effect.

[0120] The above description is merely an embodiment of the present invention and does not limit the patent scope of the present invention. Any equivalent structural transformations made using the content of the present invention specification, or direct or indirect applications in other related technical fields, are similarly included within the patent protection scope of the present invention.

Claims

1. A compound beverage of Lactobacillus paracasei and plant extract with lipid-lowering effect, characterized in that: The compound beverage comprises 20-50 v% of Lactobacillus paracasei suspension and 50-80 v% of plant extract. The Lactobacillus paracasei SWFU-D16, with accession number CCTCCNO: M2023709, was deposited at the China Center for Type Culture Collection on May 9, 2023.

2. The compound beverage according to claim 1, characterized in that: The compound beverage comprises 20-50 v% of the Lactobacillus paracasei suspension and 50-80 v% of the plant extract.

3. The compound beverage according to claim 3, characterized in that: The compound beverage comprises 50 v% of the Lactobacillus paracasei suspension and 50 v% of the plant extract.

4. The compound beverage according to any one of claims 1-3, characterized in that: The bacterial count of the Lactobacillus paracasei suspension was 1×10⁻⁶. 8 ~1×10 9 CFU / mL.

5. The compound beverage according to any one of claims 1-3, characterized in that: The plant extracts include, by volume, 10-30 parts hawthorn extract, 30-50 parts black tea extract, 10-30 parts prickly pear extract, and 5-20 parts papaya extract.

6. The compound beverage according to claim 5, characterized in that: The plant extract also includes citric acid, erythritol, sucralose, pectin, and carrageenan; The concentration of citric acid is 2-5 g / L, the concentration of erythritol is 20-100 g / L, the concentration of sucralose is 0.1-0.3 g / L, the concentration of pectin is 0.1-1 g / L, and the concentration of carrageenan is 0.1-1 g / L.

7. A method for preparing a compound beverage of Lactobacillus paracasei and plant extract with lipid-lowering effect, characterized in that: The preparation method includes the following steps: Using Lactobacillus paracaseiSWFU-D16 with accession number CCTCC NO: M2023709 as the strain, a Lactobacillus paracasei bacterial suspension was prepared. Add dried hawthorn to hot water, extract and filter to obtain hawthorn extract; Prickly pear was added to drinking water, pulped, and filtered to obtain prickly pear extract; Papaya slices were added to the second hot water, and the extract was obtained by soaking and filtering. Black tea is brewed with hot water (the third type of hot water) and then filtered to obtain a black tea extract. The hawthorn extract, the prickly pear extract, the papaya extract and the black tea extract are mixed according to the first preset ratio. Then, citric acid, erythritol, sucralose, pectin and carrageenan are added. After mixing, the mixture is sterilized at high temperature to obtain the plant extract. The compound beverage is prepared by mixing and homogenizing the Lactobacillus paracasei suspension with the plant extract according to the second preset ratio.

8. The preparation method according to claim 7, characterized in that: The ratio of dried hawthorn to the first hot water is 1:20-30 (w:v), the temperature of the first hot water is 70-80℃, and the extraction time of the first hot water is 5-10 minutes. The ratio of prickly pear to drinking water is 1:20-30 (w:v), and the drinking water temperature is room temperature; The ratio of papaya slices to the second hot water is 1:20-30 (w:v), the temperature of the second hot water is 90-95℃, and the extraction time of the second hot water is 60-80 min; The ratio of black tea to the third hot water is 1:60-80 (w:v), the temperature of the third hot water is 90-95℃, and the brewing time is 1-3 minutes.

9. The use of the compound beverage according to any one of claims 1-6 in lowering blood lipids.

10. The application of Lactobacillus paracasei SWFU-D16 with accession number CCTCC NO: M2023709 in the preparation of foods, health products, feeds and / or drugs that lower blood lipids.

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