Microecological preparation capable of regulating lipid metabolism disorder and oxidative stress and preparation method thereof

Abstract

The invention discloses a microecological preparation capable of regulating lipid metabolism disorder and oxidative stress and a preparation method thereof, and belongs to the technical field of biological preparations. The invention aims to provide a microecological preparation for regulating lipid metabolism disorder and oxidative stress, which is prepared by taking grape seed procyanidine as a raw material and matching with compound yeast in an auxiliary manner, is used for regulating metabolic disorder diseases and has the effects of regulating lipid metabolism disorder and oxidative stress, and a preparation method thereof. The preparation comprises grape seed procyanidine, a compound bacteria solution and a fermentation culture medium. The microecological preparation is simple in preparation step and low in cost, the average polymerization degree of proanthocyanidins can be effectively reduced through mixed fermentation, the proanthocyanidins are stably dissolved in a compound bacteria solution and are easy to absorb, and therefore lipid metabolism disorder of an organism is effectively adjusted, and oxidative stress of the organism is improved. The procyanidine has an inhibiting effect on common pathogenic bacteria in gastrointestinal tracts of organisms, can help the probiotics to colonize in the gastrointestinal tracts, regulate the steady state of microorganisms in the gastrointestinal tracts, promote the growth of effective microbial communities in the gastrointestinal tracts and enhance the immunity of the organisms.

Description

Technical Field

[0001] The invention belongs to the technical field of biological preparations. Background Art

[0002] As a major grape producer, my country produces a large number of grapes every year. The other parts of grapes except the pulp are generally discarded as waste, but these discarded parts contain a large number of polyphenols, such as proanthocyanidins, phenolic acids and flavonoids. Every year, grapes used for processing will produce a large amount of discarded grape seed byproducts, which are generally used as fertilizers or animal feeds. These grape seeds also bring great pressure to environmental protection. Grape seeds account for 3%-6% of the weight of the fruit and contain rich chemical components. Among them, the content of polyphenols is relatively high, accounting for 4%. Grape seed proanthocyanidin extract has become the main source of industrial proanthocyanidin due to its high proanthocyanidin content. Therefore, extracting proanthocyanidins from grape seeds and developing their new value is in line with the national ecological civilization development strategy, and has broad market prospects and important economic value.

[0003] Proanthocyanidins are a type of polyphenol rich in phenolic hydroxyl groups, which have significant effects on regulating lipid metabolism and anti-oxidation. Proanthocyanidins also have the potential to improve the quality of livestock and poultry meat. However, the degree of polymerization of crude proanthocyanidins from plants is usually high, the bioavailability of proanthocyanidin polymers is very low, and large amounts of accumulation can lead to frequent intestinal inflammation and endanger the health of the body. Therefore, obtaining proanthocyanidins with a low degree of polymerization is beneficial to the body's absorption and improves the utilization rate of the product.

[0004] Saccharomyces cerevisiae is a probiotic that promotes animal growth and enhances immunity. It can produce essential amino acids and multiple vitamins required by animals. In addition, Saccharomyces cerevisiae itself has excellent antioxidant capacity and can promote the dissolution and depolymerization of proanthocyanidins. Candida utilis is certified by the U.S. Food and Drug Administration (FDA) as a beneficial bacteria that can be used in food, and is also widely used in industrial food production in my country. It has a fast proliferation rate and is rich in protein and vitamin B. It can increase the protein content of fermentation products and improve their smell and palatability.

[0005] The occurrence and development of metabolic disorders such as fatty liver, obesity, diabetes and hyperlipidemia are closely related to oxidative stress and lipid metabolism disorders. Obesity and related metabolic disorders are characterized by excessive lipid accumulation in peripheral tissues and lipid peroxidation. Relieving the body's oxidative stress and lipid metabolism disorders can effectively alleviate the occurrence and development of diseases such as obesity. Therefore, the development of microecological preparations with high-efficiency antioxidant and lipid metabolism improvement capabilities is crucial in production practice. Summary of the invention

[0006] The purpose of the present invention is to prepare a microecological preparation for regulating lipid metabolism disorder and oxidative stress by using grape seed proanthocyanidins as raw materials and compound yeast as auxiliary for regulating metabolic disorder diseases and a preparation method thereof.

[0007] The preparation of the present invention comprises grape seed proanthocyanidins, composite bacterial liquid and fermentation medium; The composite bacterial liquid is fermented by mixing Saccharomyces cerevisiae and Candida utilis, with the addition ratio of 3:2; The fermentation medium is composed of 28-30 parts by weight of glucose, 9-11 parts by weight of peptone, 4-6 parts by weight of yeast extract powder, 1-2 parts by weight of dimethyl hydrogen phosphate, 1-3 parts by weight of ammonium citrate, 0.15-0.3 parts by weight of magnesium sulfate, and 950-1000 parts by weight of water; Grape seed proanthocyanidin powder, composite bacterial liquid and fermentation medium are mixed and added into a fermentation tank for fermentation, wherein 45-65 weight parts of grape seed proanthocyanidin and 20-40 weight parts of composite bacterial liquid are added. 2-3 weight parts of fermentation medium need to be added during mixed fermentation in the tank. The fermentation temperature is 25-32°C, the stirring speed is 120-140 r / min, the pH is 5.8-6.2, the fermentation time is 18-24 h, and the product after the fermentation is a microecological preparation.

[0008] The purity of the grape seed proanthocyanidins of the invention is 80-90%, and the average degree of polymerization is 6.9-9.2.

[0009] The microecological preparation of the present invention has simple preparation steps and low cost. The average degree of polymerization of proanthocyanidins can be effectively reduced through mixed fermentation, so that the proanthocyanidins can be stably dissolved in the composite bacterial liquid and easily absorbed, thereby effectively regulating the body's lipid metabolism disorder and improving the body's oxidative stress. Proanthocyanidins have an inhibitory effect on common pathogenic bacteria in the gastrointestinal tract of the body, can help probiotics colonize in the gastrointestinal tract, regulate the homeostasis of gastrointestinal microorganisms, promote the growth of beneficial gastrointestinal flora, and enhance the body's immunity. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] Figure 1 HE staining of the liver of mice in each group of the prevention experiment, where the experimental groups were control group, probiotic preparation group, high-fat group and high-fat probiotic preparation group; Figure 2 HE staining of epididymal fat of mice in each group of the prevention experiment, where the experimental groups were control group, probiotic preparation group, high-fat group and high-fat probiotic preparation group; Figure 3 HE staining of the liver of mice in each treatment experiment group, where the experimental groups are control group, probiotic preparation group, high-fat group and high-fat probiotic preparation group; Figure 4 HE staining of epididymal fat of mice in each experimental group, including the control group, proecological preparation group, high-fat group and high-fat proecological preparation group. DETAILED DESCRIPTION

[0011] The present invention discloses the preparation and application of a microecological preparation capable of regulating lipid metabolism disorder and oxidative stress. The microecological preparation is prepared by fermenting grape seed proanthocyanidin powder, fermentation medium and composite bacterial liquid, wherein the composite bacterial liquid is mixed and fermented by saccharomyces cerevisiae and Candida utilis to obtain a composite bacterial liquid containing beneficial bacteria and their metabolites. The addition ratio of saccharomyces cerevisiae to Candida utilis is 3:2. The microecological preparation provided by the present invention reduces the average degree of polymerization of proanthocyanidins in commercial crude grape seed proanthocyanidin powder by mixed fermentation, and can effectively regulate the body's lipid metabolism disorder and improve antioxidant activity. The microecological preparation has low cost, good stability, and is easy to use directly in production practice, thereby enhancing the immune ability of the animal body and improving economic benefits.

[0012] The present invention provides the preparation and application of a microecological preparation capable of regulating lipid metabolism disorder and oxidative stress, which has the advantages of regulating lipid metabolism disorder and improving antioxidant activity, and can effectively reduce the incidence of diseases such as obesity caused by lipid metabolism disorder and oxidative stress damage.

[0013] The invention is prepared by fermenting grape seed proanthocyanidin powder, fermentation medium and composite bacterial liquid.

[0014] The composite bacterial liquid is fermented by mixing Saccharomyces cerevisiae and Candida utilis, with the addition ratio being 3:2.

[0015] The Candida utilis and Saccharomyces cerevisiae preserved in the laboratory were resuscitated and subcultured. After their activities were restored, the bacterial solutions were adjusted to the turbidity of No. 1 McFarland turbidimetric tube using physiological saline. Then, the Saccharomyces cerevisiae culture solution and the Candida utilis culture solution were mixed in a ratio of 3:2 to obtain a composite bacterial solution.

[0016] The fermentation medium components include: 28-30 parts by weight of glucose, 9-11 parts by weight of peptone, 4-6 parts by weight of yeast extract powder, 1-2 parts by weight of dimethyl hydrogen phosphate, 1-3 parts by weight of ammonium citrate, 0.15-0.3 parts by weight of magnesium sulfate and 950-1000 parts by weight of water.

[0017] Grape seed proanthocyanidin powder, composite bacterial liquid and fermentation medium are mixed and added into a fermentation tank for fermentation, wherein 45-65 weight parts of grape seed proanthocyanidin and 20-40 weight parts of composite bacterial liquid are added. 2-3 weight parts of fermentation medium need to be added during mixed fermentation in the tank. The fermentation temperature is 25-32°C, the stirring speed is 120-140 r / min, the pH is 5.8-6.2, the fermentation time is 18-24 h, and the product after the fermentation is a microecological preparation.

[0018] According to the characteristics of saccharomyces cerevisiae and Candida utilis, the components such as carbon source, nitrogen source, growth factor and inorganic salt in the fermentation medium are screened, and the influence of these components on the average polymerization degree of proanthocyanidins is analyzed. The components that significantly affect the average polymerization degree of proanthocyanidins in the fermentation medium are screened by Plackett-Burman experiment and a regression equation is established. When the lowest value of the average polymerization degree of proanthocyanidins is determined by Box-behnken response surface analysis, the components of the fermentation medium are optimized to obtain the fermentation medium. The fermentation medium components are composed of 28-30 weight parts of glucose, 9-11 weight parts of peptone, 4-6 weight parts of yeast extract powder, 1-2 weight parts of dimethyl hydrogen phosphate, 1-3 weight parts of ammonium citrate, 0.15-0.3 weight parts of magnesium sulfate, and 950-1000 weight parts of water. According to the above optimized fermentation medium composition and concentration, the fermentation medium containing grape seed proanthocyanidins was prepared, and Saccharomyces cerevisiae and Candida utilis were inoculated according to the above inoculation ratio, and the fermentation time, fermentation temperature, stirring speed and fermentation pH value at the lowest average polymerization degree of proanthocyanidins were screened in turn. Then, the fermentation culture conditions were optimized by Box-behnken response surface analysis: fermentation temperature was 25-32 ℃, stirring speed was 120-140 r / min, initial pH was 5.8-6.2, and fermentation time was 18-24 h.

[0019] Embodiment 1: The weight proportions of grape seed proanthocyanidin powder, composite bacterial solution and fermentation medium are: 45 parts, 20 parts and 2 parts respectively; wherein the composite bacterial solution is mixed and fermented by saccharomyces cerevisiae and Candida utilis, and the addition ratio is 3:2; the weight proportions of the components constituting the fermentation medium are: 28 parts of glucose, 9 parts of peptone, 4 parts of yeast extract powder, 1 part of dimethyl hydrogen phosphate, 1 part of ammonium citrate, 0.15 parts of magnesium sulfate and 950 parts of water. The mixing and preparation method is the same as above.

[0020] Embodiment 2: The weight proportions of grape seed proanthocyanidin powder, composite bacterial solution and fermentation medium are: 50 parts, 30 parts and 2.5 parts; wherein the composite bacterial solution is mixed and fermented by saccharomyces cerevisiae and Candida utilis, and the addition ratio is 3:2; the weight proportions of the components constituting the fermentation medium are: 29 parts of glucose, 10 parts of peptone, 5 parts of yeast extract powder, 1.5 parts of dimethyl phosphate, 2 parts of ammonium citrate, 0.2 parts of magnesium sulfate and 1000 parts of water. The mixing and preparation method is the same as above.

[0021] Embodiment 3: The weight proportions of grape seed proanthocyanidin powder, composite bacterial solution and fermentation medium are: 65 parts, 40 parts and 3 parts respectively; the composite bacterial solution is mixed and fermented by saccharomyces cerevisiae and Candida utilis, and the addition ratio is 3:2; the weight proportions of the components constituting the fermentation medium are: 30 parts of glucose, 11 parts of peptone, 6 parts of yeast extract powder, 2 parts of dimethyl hydrogen phosphate, 3 parts of ammonium citrate, 0.3 parts of magnesium sulfate and 1000 parts of water. The mixing and preparation method is the same as above.

[0022] The purity of the grape seed proanthocyanidins of the present invention is 80-90%, and the average degree of polymerization is 6.9-9.2.

[0023] The experimental methods and experimental materials in the following examples are conventional methods and conventional materials unless otherwise specified. The quantitative experiments were all repeated three times.

[0024] The invention provides a microecological preparation for regulating lipid metabolism disorder and oxidative stress, comprising 45-65 parts by weight of grape seed proanthocyanidins, 20-40 parts by weight of composite bacterial liquid and 2-3 parts by weight of fermentation medium.

[0025] Experimental example 1. Recovery and subculture of bacterial strains The Saccharomyces cerevisiae and Candida utilis preserved in the laboratory were revived and subcultured to restore the activity of the strains.

[0026] Optimization of fermentation medium composition The components of the fermentation medium after optimization are as follows: 28-30 parts by weight of glucose, 9-11 parts by weight of peptone, 4-6 parts by weight of yeast extract powder, 1-2 parts by weight of dimethyl hydrogen phosphate, 1-3 parts by weight of ammonium citrate, 0.15-0.3 parts by weight of magnesium sulfate, and 950-1000 parts by weight of water.

[0027] 3. Mixed fermentation 45-65 parts by weight of grape seed proanthocyanidins and 20-40 parts by weight of composite bacterial liquid are added when entering the tank for mixed fermentation. The fermentation temperature is 25-32°C, the stirring speed is 120-140 r / min, the pH is 5.8-6.2, the fermentation time is 18-24 h, and the product after the fermentation is the microecological preparation.

[0028] 4. Application experiment of microecological preparations The SPF-grade C57BL / 6 mice and ordinary mouse maintenance feed used in this experiment were obtained from Liaoning Changsheng Biotechnology Co., Ltd., and a high-fat mouse feed containing 60% fat (D12492, Research Diets) was used to induce the obese mouse model.

[0029] Data 1: Preventive effects of proanthocyanidins on lipid metabolism disorders and oxidative stress in high-fat diet-induced obese mice.

[0030] In this experiment, 20 male C57BL / 6 mice of 6-8 weeks of age and similar weight were randomly divided into 4 groups, namely ① control group (normal food + normal saline), ② probiotic preparation group (normal food + probiotic preparation), ③ high-fat group (high-fat diet + normal saline) and ④ high-fat probiotic preparation group (high-fat diet + probiotic preparation). The volume of normal saline and probiotic preparation for gavage was 20 µL per gram of body weight of mice. For 8 consecutive weeks, groups ① and ② were provided with free normal diet and free drinking water, and groups ③ and ④ were provided with free high-fat diet and free drinking water. During these 8 weeks, groups ① and ③ were gavaged with 20 µL of normal saline per gram of body weight of each mouse every other day, and groups ② and ④ were gavaged with 20 µL of probiotic preparation per gram of body weight of each mouse. The weight of the mice was recorded every week during the experiment, and the growth status of the mice was observed. At the end of the experiment, the mice were fasted for 12 hours, and then the eyeballs were removed for blood collection and autopsy sampling. The liver, subcutaneous fat and epididymal fat of mice were weighed, and their blood biochemical indicators were tested: triglyceride (TG), total cholesterol (TC), low-density lipoprotein (LDL-C), high-density lipoprotein (HDL-C) and blood glucose (Glu); oxidative stress related indicators: superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA) and glutathione peroxidase (GSH); and liver injury indicators: aspartate aminotransferase (AST) and alanine aminotransferase (ALT). Another part of the liver tissue and adipose tissue was fixed with 4% paraformaldehyde and made into HE-stained sections for pathological morphological observation.

[0031] Table 1 Detection of related indicators in the experiment of proanthocyanidin microecological preparations preventing lipid metabolism disorders, oxidative stress and liver damage in mice Control group Microecological preparation group High fat group High-fat microecological preparation group Weight (g) <![CDATA[28.52 ± 0.68 a ]]> <![CDATA[28.99 ± 0.45 a ]]> <![CDATA[42.24 ± 1.24 b ]]> <![CDATA[31.91 ± 0.63 a ]]> Liver (g) <![CDATA[1.39 ± 0.05 a ]]> <![CDATA[1.54 ± 0.02 a ]]> <![CDATA[2.01 ± 0.11 b ]]> <![CDATA[1.57 ± 0.04 a ]]> Subcutaneous fat (g) <![CDATA[0.37 ± 0.02 a ]]> <![CDATA[0.33 ± 0.01 a ]]> <![CDATA[1.34 ± 0.07 b ]]> <![CDATA[0.90 ± 0.08 c ]]> Epididymal fat (g) <![CDATA[0.38 ± 0.03 a ]]> <![CDATA[0.33 ± 0.02 a ]]> <![CDATA[1.66 ± 0.05 b ]]> <![CDATA[1.07 ± 0.05 c ]]> TG (mmol / L) <![CDATA[0.75 ± 0.05 a ]]> <![CDATA[0.65 ± 0.24 a ]]> <![CDATA[1.40 ± 0.10 b ]]> <![CDATA[0.96 ± 0.06 a ]]> TC (mmol / L) <![CDATA[3.88 ± 0.22 a ]]> <![CDATA[3.70 ± 0.15 a ]]> <![CDATA[6.48 ± 0.43 b ]]> <![CDATA[5.35 ± 0.28 b ]]> LDL-C (mmol / L) <![CDATA[2.91 ± 0.13 a ]]> <![CDATA[3.00 ± 0.22 a ]]> <![CDATA[5.32 ± 0.13 b ]]> <![CDATA[4.24 ± 0.15 c ]]> HDL-C (mmol / L) <![CDATA[1.44 ± 0.05 a ]]> <![CDATA[1.40 ± 0.04 a ]]> <![CDATA[2.03 ± 0.12 b ]]> <![CDATA[1.47 ± 0.08 a ]]> SOD (U / mg) <![CDATA[86.69 ± 2.41 a ]]> <![CDATA[91.31 ± 2.62 a ]]> <![CDATA[55.48 ± 3.47 b ]]> <![CDATA[76.03 ± 6.60 a ]]> CAT (U / mg) <![CDATA[6.85 ± 0.52 a ]]> <![CDATA[7.35 ± 0.53 a ]]> <![CDATA[3.21 ± 0.53 b ]]> <![CDATA[5.47 ± 0.43 a ]]> MDA (nmol / mg) <![CDATA[5.58 ± 0.47 a ]]> <![CDATA[5.07 ± 0.26 a ]]> <![CDATA[9.28 ± 0.49 b ]]> <![CDATA[6.94 ± 0.55 c ]]> GSH (µmol / g) <![CDATA[27.23 ± 2.67 a ]]> <![CDATA[28.40 ± 3.35 a ]]> <![CDATA[11.92 ± 1.24 b ]]> <![CDATA[24.11 ± 1.79 a ]]> Glu (mmol / L) <![CDATA[5.70 ± 0.26 a ]]> <![CDATA[5.74 ± 0.24 a ]]> <![CDATA[8.90 ± 0.42 b ]]> <![CDATA[6.52 ± 0.24 a ]]> AST (U / L) <![CDATA[29.56 ± 2.32 a ]]> <![CDATA[28.43 ± 2.02 a ]]> <![CDATA[68.29 ± 3.35 b ]]> <![CDATA[40.31 ± 1.04 a <!-- 4 -->]]> ALT (U / L) <![CDATA[24.82 ± 2.02 a ]]> <![CDATA[24.67 ± 2.30 a ]]> <![CDATA[48.76 ± 3.40 b ]]> <![CDATA[33.20 ± 2.53 a ]]> .

[0032] The results are expressed as mean ± standard error, the sample size is 5, and different lowercase letters in the same row indicate significant differences ( P < 0.05). Figure 1 HE staining of the livers of mice in each group of the prevention experiment, from left to right: control group, proecological preparation group, high-fat group, and high-fat proecological preparation group. Figure 2 HE staining of epididymal fat of mice in each group of the prevention experiment, from left to right: control group, probiotic preparation group, high-fat group, and high-fat probiotic preparation group.

[0033] In the experiment of the preventive effect of proanthocyanidin microecological preparation on lipid metabolism disorder and oxidative stress in obese mice induced by high-fat diet, the mice grew well and did not die or have abnormal conditions. The experimental results showed that high-fat diet can successfully induce obesity in mice. The body weight, liver weight, subcutaneous and epididymal fat weight of high-fat mice were significantly higher than those of control mice ( P < 0.05). There were no significant differences in body weight, liver weight, subcutaneous fat weight, epididymal fat weight, blood biochemical indexes, oxidative stress indexes, and liver injury indexes between the mice in the probiotic preparation group and the control group ( P > 0.05). The blood biochemical indexes TG, LDL-C, HDL-C and Glu in the high-fat microecological preparation group were significantly lower than those in the high-fat group ( P < 0.05); the antioxidant related indicators SOD, CAT and GSH were significantly higher than those in the high fat group ( P < 0.05), and the oxidative stress index MDA was significantly lower than that in the high-fat group ( P < 0.05); and liver injury indicators AST and ALT were significantly lower than those in the high-fat group ( P < 0.05). Figure 1 Liver and Figure 2 HE staining of epididymal fat showed that the addition of probiotics can significantly improve fatty degeneration of liver tissue and reduce the size of adipocytes in adipose tissue, proving that probiotics can effectively alleviate the state of lipid metabolism disorder. This result confirms that proanthocyanidin probiotics can effectively prevent lipid metabolism disorder and oxidative stress in mice.

[0034] Data 2: Therapeutic effects of proanthocyanidin microecological preparation on lipid metabolism disorders and oxidative stress in high-fat diet-induced obese mice.

[0035] In this experiment, 20 male C57BL / 6 mice of 6-8 weeks of age and similar weight were randomly divided into 4 groups, namely ① control group (normal food + normal saline), ② probiotic preparation group (normal food + probiotic preparation), ③ high-fat group (high-fat diet + normal saline) and ④ high-fat probiotic preparation group (high-fat diet + probiotic preparation). The volume of normal saline and probiotic preparation for gavage was 20 µL per gram of body weight of mice. Groups ① and ② were provided with free normal diet and free drinking water for 10 consecutive weeks, and groups ③ and ④ were provided with free high-fat diet and free drinking water. Starting from the 9th week, daily gavage was performed for 14 consecutive days. Each mouse in groups ① and ③ was gavaged with 20 µL of normal saline per gram of body weight, and each mouse in groups ② and ④ was gavaged with 20 µL of probiotic preparation per gram of body weight. The weight of the mice was recorded every week during the experiment, and the growth status of the mice was observed. At the end of the experiment, the mice were fasted for 12 hours, and then the eyeballs were removed for blood collection and autopsy sampling. The liver weight, subcutaneous fat and epididymal fat of mice were weighed, and their blood biochemical indicators were tested: TG, TC, LDL-C, HDL-C and Glu; oxidative stress related indicators: SOD, CAT, MDA and GSH; and liver damage indicators: AST and ALT. Another part of the liver tissue and adipose tissue was fixed with 4% paraformaldehyde and made into HE-stained sections for pathological morphological observation.

[0036] Table 2 Detection of related indicators in the experiment of proanthocyanidin microecological preparations treating lipid metabolism disorders, oxidative stress and liver damage in mice Control group Microecological preparation group High fat group High-fat microecological preparation group Weight (g) <![CDATA[29.14 ± 0.77 a ]]> <![CDATA[30.06 ± 0.43 a ]]> <![CDATA[47.60 ± 1.36 b ]]> <![CDATA[34.29 ± 0.44 c ]]> Liver (g) <![CDATA[1.38 ± 0.05 a ]]> <![CDATA[1.47 ± 0.03 ab ]]> <![CDATA[1.90 ± 0.08 c ]]> <![CDATA[1.63 ± 0.04 b ]]> Subcutaneous fat (g) <![CDATA[0.36 ± 0.02 a ]]> <![CDATA[0.34 ± 0.02 a ]]> <![CDATA[1.38 ± 0.08 b ]]> <![CDATA[0.89 ± 0.07 c ]]> Epididymal fat (g) <![CDATA[0.36 ± 0.03 a ]]> <![CDATA[0.31 ± 0.01 a ]]> <![CDATA[1.63 ± 0.07 b ]]> <![CDATA[1.08 ± 0.05 c ]]> TG (mmol / L) <![CDATA[0.77 ± 0.04 a ]]> <![CDATA[0.69 ± 0.03 a ]]> <![CDATA[1.41 ± 0.09 b ]]> <![CDATA[0.93 ± 0.04 a ]]> TC (mmol / L) <![CDATA[3.81 ± 0.15 a ]]> <![CDATA[3.82 ± 0.14 a ]]> <![CDATA[6.20 ± 0.26 b ]]> <![CDATA[5.37 ± 0.33 b <!-- 5 -->]]> LDL-C (mmol / L) <![CDATA[2.78 ± 0.13 a ]]> <![CDATA[2.76 ± 0.13 a ]]> <![CDATA[5.12 ± 0.09 b ]]> <![CDATA[4.12 ± 0.08 c ]]> HDL-C (mmol / L) <![CDATA[1.40 ± 0.05 a ]]> <![CDATA[1.34 ± 0.04 a ]]> <![CDATA[2.16 ± 0.16 b ]]> <![CDATA[1.49 ± 0.04 a ]]> SOD (U / mg) <![CDATA[84.87 ± 3.03 a ]]> <![CDATA[88.25 ± 1.77 a ]]> <![CDATA[57.62 ±2 .95 b ]]> <![CDATA[73.52 ± 4.71 a ]]> CAT (U / mg) <![CDATA[7.02 ± 0.73 a ]]> <![CDATA[7.42 ± 0.68 a ]]> <![CDATA[2.67 ± 0.47 b ]]> <![CDATA[5.15 ± 0.45 a ]]> MDA (nmol / mg) <![CDATA[5.43 ± 0.40 a ]]> <![CDATA[5.11 ± 0.24 a ]]> <![CDATA[9.24 ± 0.59 b ]]> <![CDATA[7.04 ± 0.49 a ]]> GSH (µmol / g) <![CDATA[26.18 ± 2.72 a ]]> <![CDATA[30.08 ± 3.30 a ]]> <![CDATA[12.04 ± 1.37 b ]]> <![CDATA[24.23 ± 1.64 a ]]> Glu (mmol / L) <![CDATA[5.4 ± 0.32 a ]]> <![CDATA[5.7 ± 0.28 a ]]> <![CDATA[8.14 ± 0.45 b ]]> <![CDATA[6.56 ± 0.26 a ]]> AST (U / L) <![CDATA[31.89 ± 2.85 a ]]> <![CDATA[31.30 ± 2.27 a ]]> <![CDATA[71.87 ± 4.32 b ]]> <![CDATA[43.68 ± 2.39 a ]]> ALT (U / L) <![CDATA[25.75 ± 2.85 a ]]> <![CDATA[23.90 ± 1.87 a ]]> <![CDATA[47.46 ± 3.38 b ]]> <![CDATA[33.29 ± 2.10 a ]]> .

[0037] The results are expressed as mean ± standard error, the sample size is 5, and different lowercase letters in the same row indicate significant differences ( P < 0.05). Figure 3 HE staining of the livers of mice in each group of the treatment experiment, from left to right: control group, proecological preparation group, high-fat group, and high-fat proecological preparation group. Figure 4 HE staining of epididymal fat of mice in each group of the treatment experiment, from left to right: control group, proecological preparation group, high-fat group and high-fat proecological preparation group.

[0038] In the experiment of the therapeutic effect of proanthocyanidin microecological preparation on lipid metabolism disorder and oxidative stress in obese mice induced by high-fat diet, the mice grew well and did not die or have abnormal conditions. The experimental results showed that high-fat diet can successfully induce obesity in mice. The body weight, liver weight and fat weight of mice in the high-fat group were significantly higher than those of mice in the control group ( P < 0.05). There were no significant differences in body weight, liver weight, subcutaneous fat weight, epididymal fat weight, blood biochemical indexes, oxidative stress indexes, and liver injury indexes between the mice in the probiotic preparation group and the control group ( P> 0.05). The blood biochemical indexes TG, LDL-C, HDL-C and Glu in the high-fat microecological preparation group were significantly lower than those in the high-fat group ( P < 0.05); the antioxidant related indicators SOD, CAT and GSH were significantly higher than those in the high fat group ( P < 0.05), and the oxidative stress index MDA was significantly lower than that in the high-fat group ( P < 0.05) and liver damage indicators AST and ALT were significantly lower than those in the high-fat group ( P < 0.05). Figure 3 Liver and Figure 4 HE staining results of epididymal fat showed that the addition of probiotics can significantly improve fatty degeneration of liver tissue and reduce the size of adipocytes in adipose tissue, proving that probiotics can treat lipid metabolism disorders in mice. This result confirms that proanthocyanidin probiotics can significantly improve lipid metabolism disorders and oxidative stress in mice.

[0039] In summary, the microecological preparation of the present invention has good safety, and the microecological preparation obtained by mixed fermentation of proanthocyanidins and composite bacterial liquid can prevent and treat lipid metabolism disorders and oxidative stress in mice.

Claims

1. A microecological preparation for regulating lipid metabolism disorders and oxidative stress and a preparation method thereof, characterized in that: It includes grape seed proanthocyanidins, compound bacterial liquid and fermentation medium; The composite bacterial liquid is mixed and fermented by Saccharomyces cerevisiae and Candida utilis, and the addition ratio is 3:2; The fermentation medium is composed of 28-30 parts by weight of glucose, 9-11 parts by weight of peptone, 4-6 parts by weight of yeast extract powder, 1-2 parts by weight of dimethyl hydrogen phosphate, 1-3 parts by weight of ammonium citrate, 0.15-0.3 parts by weight of magnesium sulfate, and 950-1000 parts by weight of water; Grape seed proanthocyanidin powder, composite bacterial liquid and fermentation medium are mixed and added into a fermentation tank for fermentation, wherein 45-65 weight parts of grape seed proanthocyanidin and 20-40 weight parts of composite bacterial liquid are added. 2-3 weight parts of fermentation medium need to be added during mixed fermentation in the tank. The fermentation temperature is 25-32°C, the stirring speed is 120-140 r / min, the pH is 5.8-6.2, the fermentation time is 18-24h, and the product after the fermentation is completed is the microecological preparation.

2. The microecological preparation for regulating lipid metabolism disorders and oxidative stress and the preparation method according to claim 1, characterized in that: The purity of grape seed proanthocyanidins is 80-90%, and the average degree of polymerization is 6.9-9.2.

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