Preparation method of broccoli seed aqueous extract compound with liver protection effect

Abstract

The invention discloses a preparation method of a broccoli seed aqueous extract compound with a liver protection effect, and belongs to the field of traditional Chinese medicine. The compound is prepared from broccoli seed aqueous extract, radix puerariae extract, emblic leafflower fruit extract and resistant dextrin. According to the preparation method disclosed by the invention, the raw materials are all common medicinal and edible health food raw materials, and a large number of experiments prove that the compounded compound disclosed by the invention has the effects of well improving alcoholic liver injury, acetaminophen-induced liver injury and fatty liver, is safe and effective, and does not have other toxic or side effects.

Description

Technical Field

[0001] The invention relates to the field of traditional Chinese medicine, and in particular to a method for preparing a broccoli seed water extract complex with liver-protecting effect. Background Art

[0002] The liver is the largest detoxification organ in animals and the main site of drug metabolism. Acetaminophen is currently the most widely used antipyretic and analgesic, and its use at the recommended therapeutic concentration is considered safe, but as a drug ingredient, acetaminophen exists in many different drugs, and excessive use can cause acute liver damage or even acute liver failure.

[0003] According to known data, acetaminophen has the risk of damaging the liver and inducing blood cancer, and taking too much can also cause serious damage to the body. The incidence of acetaminophen-induced liver damage is high and the consequences are serious, and there are currently few drugs to treat it. Therefore, it is very important to explore more effective and safer drugs to prevent and treat liver damage caused by acetaminophen.

[0004] Alcoholic liver disease is a liver-damaging disease caused by long-term, large-scale drinking of various alcoholic beverages. It is a public health issue that has attracted worldwide attention. If patients with alcoholic fatty liver disease do not actively prevent and treat it, the disease will directly worsen and develop into alcoholic hepatitis and liver fibrosis, and even cirrhosis. At present, Chinese patients with alcoholic liver disease have severe liver damage, of which 28.8% have alcoholic hepatitis and 37.4% have alcoholic cirrhosis.

[0005] Broccoli is known as the crown of vegetables. It is rich in a variety of nutrients and active ingredients needed by the human body. In particular, the water extract of broccoli seeds is rich in glucoraphanin, which has multiple biological activities such as antioxidant, antibacterial, anticancer and immunomodulatory, and is an ideal raw material for natural functional products. As a common alcohol-relieving health product, Pueraria root can effectively antagonize the lipid peroxidation damage of liver tissue caused by alcohol. It can decompose the acetaldehyde toxicity generated by the metabolism of ethanol into the liver, prevent the weakening of the inhibitory function of alcohol on the brain, inhibit the absorption of alcohol by the stomach and intestines, and promote the metabolism and excretion of alcohol in the blood. Emblica emblica is rich in nutrients and has been designated by the World Health Organization as one of the three health plants promoted for cultivation in the world. It has high medicinal value. Modern medical research shows that the fruit of Emblica emblica is rich in antioxidants such as vitamin C, vitamin E, SOD, etc., which has a significant effect on protecting the liver, can effectively reduce total cholesterol and weight gain index in the blood, and has a certain preventive effect on atherosclerosis. It can remove superoxide free radicals, reduce their damage to the body, and delay the aging process. To this end, the present invention provides a method for preparing a broccoli seed water extract complex having a liver-protecting effect, so as to solve the defects and shortcomings of the prior art. Summary of the invention

[0006] In view of the above problems, the present invention provides a method for preparing a broccoli seed water extract complex having a liver-protecting effect.

[0007] In order to achieve the above-mentioned purpose, the present invention adopts the following technical solutions:

[0008] A method for preparing a broccoli seed water extract composite having a liver-protecting effect, wherein the composite is prepared from the following raw materials in parts by weight:

[0009] 10-40 parts of broccoli seed water extract, 5-30 parts of kudzu root extract, 5-30 parts of emblica fruit extract, and 5-10 parts of resistant dextrin;

[0010] The method for preparing the broccoli seed water extract comprises the following steps:

[0011] (1) firstly crush the broccoli seeds, add water and mix, treat under ultrasound for 5 minutes, then add pH adjusting solution and adjust the pH to 6.15-6.5;

[0012] (2) adding enzymatic hydrolysis solution, controlling the enzymatic hydrolysis temperature at 25-35° C., and filtering after enzymatic hydrolysis to obtain a filtrate and a filter residue;

[0013] (3) concentrating the filtrate under reduced pressure to a powdery solid to obtain a broccoli seed aqueous extract;

[0014] The preparation method of the kudzu root extract comprises the following steps:

[0015] (1) Crush the kudzu root into powder, add water, boil over high heat, then simmer over low heat for 2-3 hours, filter to obtain a decoction and a residue, add water to the residue and continue to decoct, filter and combine the decoctions obtained from the two decoctions;

[0016] (2) concentrating the decoction under reduced pressure at 2.0-2.5 MPa to 1 / 5 of the original volume, then adding 60-65% ethanol to precipitate the decoction, filtering impurities and then precipitating again, and continuing to filter to obtain a filtrate;

[0017] (3) adding water to the filtrate and then evaporating and concentrating it to a powder solid to obtain a kudzu root extract;

[0018] Wherein, the preparation method of the emblica extract comprises the following steps:

[0019] The emblica fruit is crushed by a grinder, and then water is added to soak and mix, and then a pH adjusting solution is added to adjust the pH to 7.65-7.7, and then a composite enzyme is added to mix evenly, and then enzymolysis is performed at a temperature of 45-55° C. for 1-1.5 hours, and an enzymolysis solution is obtained by filtration, and then the obtained enzymolysis solution is evaporated and concentrated into a powder solid to obtain an emblica fruit extract;

[0020] The complex enzyme is a mixture of cellulase and pectinase of equal quality.

[0021] Preferably, the composite is made of the following raw materials in parts by weight:

[0022] 15-35 parts of broccoli seed water extract, 8-25 parts of kudzu root extract, 8-25 parts of emblica fruit extract, and 6-9 parts of resistant dextrin.

[0023] Preferably, the composite is made of the following raw materials in parts by weight:

[0024] 20-30 parts of broccoli seed water extract, 10-20 parts of kudzu root extract, 10-20 parts of emblica fruit extract, and 7-8 parts of resistant dextrin.

[0025] Preferably, the composite is made of the following raw materials in parts by weight:

[0026] 25 parts of broccoli seed water extract, 15 parts of kudzu root extract, 15 parts of emblica fruit extract, and 7 parts of resistant dextrin.

[0027] Preferably, the preparation method is:

[0028] The broccoli seed water extract, kudzu root extract, emblica fruit extract and resistant dextrin are weighed in parts by weight, mixed evenly with a traditional Chinese medicine powder grinder, and dried to obtain the powder.

[0029] Preferably, the invention is used in the prevention and treatment of alcoholic liver damage and acetaminophen-induced liver damage.

[0030] Due to the adoption of the above technical solution, the beneficial effects of the present invention are:

[0031] The formula of the invention is scientific and reasonable, and adopts common medicinal and edible raw materials and health food raw materials. After a large number of experimental verifications, the compound of the invention can well improve alcoholic liver damage, acetaminophen-induced liver damage, and improve fatty liver, and is safe and effective without other toxic and side effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 :Data graph showing the effects of each group of complexes on the activity of serum alanine aminotransferase (ALT) in mice;

[0033] Figure 2 :Data graph showing the effects of each group of complexes on serum aspartate aminotransferase (AST) activity in mice;

[0034] Figure 3 :Data graph showing the effects of each group of complexes on low-density lipoprotein (LDL) in mouse serum;

[0035] Figure 4: Data graph showing the effects of each group of complexes on high-density lipoprotein (HDL) in mouse serum. DETAILED DESCRIPTION

[0036] To make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention will be clearly and completely described below in combination with the embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0037] Embodiment 1:

[0038] A method for preparing a broccoli seed water extract complex with liver protection effect, wherein the weight proportions of raw materials are: 25 parts of broccoli seed water extract, 15 parts of kudzu root extract, 15 parts of emblica fruit extract, and 7 parts of resistant dextrin;

[0039] The steps for preparing the above-mentioned complex are:

[0040] (1) Weigh the above-mentioned parts by weight of broccoli seed water extract, kudzu root extract, and emblica fruit extract, mix them evenly, and sieve through 150 mesh;

[0041] (2) Weigh the resistant dextrin, mix it evenly with the raw materials in step (1), mix it with a traditional Chinese medicine powder grinder for 5 minutes, and cool it to obtain a composite for use.

[0042] Embodiment 2:

[0043] A method for preparing a broccoli seed water extract complex having a liver-protecting effect, wherein the weight proportions of the raw materials are:

[0044] 1. 20 parts of broccoli seed water extract, 25 parts of kudzu root extract, 25 parts of emblica fruit extract, and 9 parts of resistant dextrin;

[0045] 2. Preparation steps:

[0046] (1) Weigh the above-mentioned parts by weight of broccoli seed water extract, kudzu root extract, and emblica fruit extract, mix them evenly, and sieve through 150 mesh;

[0047] (2) Weigh the resistant dextrin, mix it evenly with the raw materials in step (1), mix it with a traditional Chinese medicine powder grinder for 5 minutes, and cool it to obtain a composite for use.

[0048] 1. Experiment on the protective effect of the complex on alcoholic liver damage in mice

[0049] Experimental method: 48 mice were selected and after 1 week of adaptive feeding, they were randomly divided into 4 groups, with 12 mice in each group, namely blank control group, model group, experimental group 1, and experimental group 2. Except for the blank group, the mice in other experimental groups were gavaged with 50% ethanol solution (the ethanol doses in the first 3 weeks were 2.0g / kg.bw, 2.4g / kg.bw, and 2.8g / kg.bw, respectively, and the ethanol gavage dose after the 4th week was 3.2g / kg.bw), while the mice in the blank group were gavaged with distilled water of the same volume as ethanol; 1 hour later, the blank group and the model group were gavaged with distilled water at 6mL / kg.bw, and the other experimental groups were gavaged with 6mL / kg.bw of complex CMC-Na solution (complex content was 10%), and this was done once in the morning and evening for a total of 6 weeks.

[0050] Among them, experimental group 1 and experimental group 2 respectively use the composites prepared in Examples 1 and 2;

[0051] 1. The effect on the activity of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in mouse serum was recorded and statistically analyzed. Figure 1 and Figure 2 ;

[0052] Among them, * compared with the model group, the difference was significant (P<0.05); ** compared with the model group, the difference was extremely significant (P<0.01).

[0053] Based on the above results, it can be concluded that compared with the model group, experimental groups 1 and 2 had significant inhibitory effects on AST and ALT activities, and the effect of experimental group 1 was significantly better than that of experimental group 2, showing a significant difference (P < 0.05), indicating that the complex has a good liver protection effect, and the broccoli seed water extract has a significant synergistic effect on the liver protection effect (P < 0.05).

[0054] 2. Effects on lipid metabolism in mice

[0055] Alcoholic liver injury mice are generally accompanied by pathological features of hyperlipidemia. The pathological condition of alcoholic fatty liver can be reflected to a certain extent by measuring the levels of lipids such as triglycerides (TG), cholesterol (TC), low-density lipoprotein (LDL), and high-density lipoprotein (HDL) in mouse serum. The data on the effects of the complex on mouse TG, TC, LDL, and HDL were recorded and statistically analyzed. Figure 3 and Figure 4 .

[0056] Among them, * compared with the model group, the difference was significant (P<0.05); ** compared with the model group, the difference was extremely significant (P<0.01).

[0057] Based on the above results, it can be concluded that compared with the model group, experimental group 1 and experimental group 2 can significantly reduce the TG, TC and LDL levels in mouse serum and increase the HDL content, indicating that the complex can significantly reduce the TG, TC and LDL levels caused by alcoholic liver damage (P < 0.05), while increasing the HDL level, and the broccoli seed water extract has a significant synergistic effect on this effect (P < 0.05).

[0058] 2. Experimental study on the protective effect of the compound on acetaminophen-induced liver injury in mice

[0059] Experimental method: 48 mice were selected and after 1 week of adaptive feeding, they were randomly divided into 4 groups, with 12 mice in each group, namely blank control group, model group, experimental group 1 and experimental group 2. The mice in other experimental groups except the blank group and the model group were gavaged with the complex CMC-Na solution (the complex content was 10%). The blank group and the model group were gavaged with an equal amount of solvent (CMC-Na solution) once in the morning and evening for 15 consecutive days. 1 hour after the last administration, the mice in other groups except the blank control group were intraperitoneally injected with 400 mg / kg of acetaminophen once to replicate the acetaminophen mouse acute drug-induced liver injury model.

[0060] The experimental groups 1 and 2 used the composites prepared in Examples 1 and 2 respectively.

[0061] 1. Effects on the activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in mouse serum

[0062] Table 1 Effects of the complex on the activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in mouse serum (u / L);

[0063] Group AST ALT Blank control group 42.38±6.51* 21.38±6.20* Model Group 7029.01±253.72 6755.33±326.56 Experimental Group 1 3752.37±142.52* 2940.91±204.88* Experimental Group 2 4947.21±156.33*# 4507.02±409.19*#

[0064] Among them, # was significantly different compared with the two experimental groups (P<0.05); * was significantly different compared with the model group (P<0.05).

[0065] Based on the above results, it can be concluded that compared with the model group, experimental groups 1 and 2 had significant inhibitory effects on AST and ALT activities (P < 0.05), and the effect of experimental group 1 was significantly better than that of experimental group 2, showing a significant difference (P < 0.05), indicating that the complex has a significant protective effect on acetaminophen-induced liver damage in mice, and broccoli seed water extract has a significant synergistic effect on this effect (P < 0.05).

[0066] 2. Effects on MDA and SOD content in mouse liver

[0067] Table 2 Effects of the complex on the activity of MDA and SOD in mouse serum

[0068]

[0069]

[0070] Among them, # was significantly different compared with the two experimental groups (P<0.05); * was significantly different compared with the model group (P<0.05).

[0071] Polyunsaturated fatty acids in the body will produce lipid peroxidation products such as MDA, hydroxyl, carbonyl and new oxygen free radicals under the attack of oxygen free radicals, causing cell damage. The measurement of MDA content can reflect the degree of lipid peroxidation in liver tissue, thereby indirectly reflecting the degree of liver cell damage. Superoxide dismutase (SOD) is an important component of the antioxidant system in tissues, and also plays an important role in scavenging free radicals in the body, preventing free radical damage and anti-oxidation.

[0072] Based on the above results, it can be seen from Table 2 that the complex can significantly reduce the increase in MDA content caused by acetaminophen liver damage in mice (P < 0.05), and can significantly increase the activity of SOD enzyme in the body (P < 0.05), and broccoli seed water extract has a significant synergistic effect on this effect (P < 0.05).

[0073] 3. Effects on the levels of TNF-α and IL-6 in mouse serum

[0074] Studies have shown that IL-1 plays an important role in liver damage. High concentrations of IL-1 can cause the liver to synthesize a large amount of acute phase proteins, further aggravating liver cell damage. Tumor necrosis factor (TNF-α) is an important pro-inflammatory factor mainly produced by Kupffer cells in the liver. It can directly cause cytotoxicity and lead to liver cell death.

[0075] Table 3 Effects of the complex on the levels of TNF-α and IL-6 in mouse serum

[0076]

[0077]

[0078] Based on the above results, it can be seen from Table 3 that the complex can significantly reduce the increase of TNF-α caused by acetaminophen liver damage in mice (P < 0.05), and can also significantly increase the level of IL-6 in the body (P < 0.05), and the broccoli seed water extract has a significant synergistic effect on this effect (P < 0.05).

[0079] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that the technical solutions described in the aforementioned embodiments may still be modified, or some of the technical features may be replaced by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

[0080] It should be noted that, in this article, the terms "comprises", "includes" or any other variations thereof are intended to cover non-exclusive inclusion, so that a process, method, article or apparatus that includes a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or apparatus.

[0081] This article uses specific examples to illustrate the principles and implementation methods of the present invention. The above examples are only used to help understand the method and core ideas of the present invention. The above is only a preferred implementation of the present invention. It should be pointed out that due to the limitations of textual expression and the objective existence of infinite specific structures, ordinary technicians in this technical field can make several improvements, modifications or changes without departing from the principles of the present invention, and can also combine the above technical features in an appropriate manner; these improvements, modifications, changes or combinations, or the direct application of the inventive concept and technical solution to other occasions without improvement, should be regarded as the protection scope of the present invention.

Claims

1. A method for preparing a broccoli seed water extract complex having a liver-protecting effect, characterized in that: The composite is made from the following raw materials in parts by weight: 10-40 parts of broccoli seed water extract, 5-30 parts of kudzu root extract, 5-30 parts of emblica fruit extract, and 5-10 parts of resistant dextrin; The method for preparing the broccoli seed water extract comprises the following steps: (1) firstly crush the broccoli seeds, add water and mix, treat under ultrasound for 5 minutes, then add pH adjusting solution to adjust the pH to 6.15-6.5; (2) adding enzymatic hydrolysis solution, controlling the enzymatic hydrolysis temperature at 25-35° C., and filtering after enzymatic hydrolysis to obtain a filtrate and a filter residue; (3) concentrating the filtrate under reduced pressure to a powdery solid to obtain a broccoli seed aqueous extract; The preparation method of the kudzu root extract comprises the following steps: (1) Crush the kudzu root into powder, add water, boil over high heat, then simmer over low heat for 2-3 hours, filter to obtain a decoction and a residue, add water to the residue and continue to decoct, filter and combine the decoctions obtained from the two decoctions; (2) concentrating the decoction under reduced pressure at 2.0-2.5 MPa to 1 / 5 of the original volume, then adding 60-65% ethanol to precipitate the decoction, filtering impurities and then precipitating again, and continuing to filter to obtain a filtrate; (3) adding water to the filtrate and then evaporating and concentrating it to a powder solid to obtain a kudzu root extract; Wherein, the preparation method of the emblica extract comprises the following steps: The emblica fruit is crushed by a grinder, and then water is added to soak and mix, and then a pH adjusting solution is added to adjust the pH to 7.65-7.7, and then a composite enzyme is added to mix evenly, and then enzymolysis is performed at a temperature of 45-55° C. for 1-1.5 hours, and an enzymolysis solution is obtained by filtration, and then the obtained enzymolysis solution is evaporated and concentrated into a powder solid to obtain an emblica fruit extract; The complex enzyme is a mixture of cellulase and pectinase of equal quality.

2. The method for preparing the broccoli seed water extract complex with liver protection effect according to claim 1, characterized in that: The composite is made from the following raw materials in parts by weight: 15-35 parts of broccoli seed water extract, 8-25 parts of kudzu root extract, 8-25 parts of emblica fruit extract, and 6-9 parts of resistant dextrin.

3. The method for preparing the broccoli seed water extract complex with liver protection effect according to claim 1, characterized in that: The composite is made from the following raw materials in parts by weight: 20-30 parts of broccoli seed water extract, 10-20 parts of kudzu root extract, 10-20 parts of emblica fruit extract, and 7-8 parts of resistant dextrin.

4. The method for preparing the broccoli seed water extract complex with liver protection effect according to claim 1, characterized in that: The composite is made from the following raw materials in parts by weight: 25 parts of broccoli seed water extract, 15 parts of kudzu root extract, 15 parts of emblica fruit extract, and 7 parts of resistant dextrin.

5. The method for preparing the broccoli seed water extract complex with liver protection effect according to claims 1-4, characterized in that: The preparation method is: The broccoli seed water extract, kudzu root extract, emblica fruit extract and resistant dextrin are weighed in parts by weight, mixed evenly with a traditional Chinese medicine powder grinder, and dried to obtain the powder.

6. The method for preparing the broccoli seed water extract complex having liver protection effect according to claim 5, characterized in that: Used in the prevention and treatment of alcoholic liver damage and acetaminophen-induced liver damage.

Images