Huangqin fermented water wine and preparation method and application thereof

Abstract

The present application relates to the technical field of microbial fermentation, in particular to a kind of Huangjing fermented water wine and its preparation method and application.The Huangjing fermented water wine is a kind of brewing wine using new soil koji to ferment Huangjing and waxy rice mixture, without distillation, Huangjing bioactivity is retained, Huangjing fermented water wine has the effect of regulating intestinal flora, reducing lipid and antioxidant, and can also reduce the harmful effects of alcohol on the brain.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological medicine, and particularly relates to a fermented water wine of rhizoma polygonati, a preparation method and application thereof. BACKGROUND

[0002] Rhizoma polygonati is first recorded in Shang Bie Lu: it is used for tonifying the middle energizer, removing wind and damp, and soothing five zang organs. It can also be used for lightening the body, prolonging life, and preventing hunger. Since ancient times, rhizoma polygonati has been considered as a medicine for nourishing the body, a tonic for the spleen and stomach, and a correct product for tonifying the spleen yin. In the present application, the fermentation technology of water wine is applied to the research and development of a new product of rhizoma polygonati. The new product is prepared by using rhizoma polygonati and glutinous rice as raw materials and through newhua fermentation. The new product can improve the taste of rhizoma polygonati products, make better use of rhizoma polygonati, and retain the characteristics and nutritional components of natural substances in rhizoma polygonati. More forward-looking research is needed to support whether the fermented water wine of rhizoma polygonati can be used as a functional food in daily life. SUMMARY

[0003] To solve the above technical problems, the present application aims to provide a fermented water wine of rhizoma polygonati with rhizoma polygonati efficacy, inexpensive raw materials, and stable quality, a production process and application thereof. The fermented water wine of rhizoma polygonati retains its original components and efficacy without distillation. After the action of microorganisms and enzymes, the taste is improved, the product color is beautiful, and the efficacy is further improved. To achieve the above technical effects, the present application adopts the following technical solutions:

[0004] A preparation method of a fermented water wine of rhizoma polygonati: newhua is taken and mixed with embryo material, then after mixing, sugarization and fermentation are carried out for 5-7 days, then cold boiled water is added for secondary fermentation for 3-7 days, so that the sugar content is 100-180 g / L, then pressure filtration, sterilization and clarification are carried out to obtain the fermented water wine of rhizoma polygonati.

[0005] The raw materials of the newhua are rice powder: chaffy knotweed herb: Salix matsudana Koidz: licorice = 0.8-1.5: 0.2-0.4: 0.08-0.12: 0.08-0.12. First, the licorice is decocted with water, then the rice, chaffy knotweed and Salix matsudana Koidz are crushed to pass through a 100-mesh sieve and mixed uniformly, then the licorice decoction is added to the crushed mixture of the rice, chaffy knotweed herb and Salix matsudana Koidz, cold boiled water is added, 2-4% of mother powder is added, the mother powder contains 3000-3500 rhizopus spores / g, 2000-2400 yeast bacteria / g and 2500-3000 lactic acid bacteria / g, and then it is mixed and made into pills for cultivation.

[0006] Further, the newhua is made into pills with a diameter of 3-5 cm, and incubated at 25 DEG C until the water content of the pills is 20%, the saccharifying enzyme activity of the newhua is 3000-3500 U / g, the rhizopus spores are 2000-3000 / g, the yeast bacteria are 1000-1400 / g, and the lactic acid bacteria are 1500-2000 / g.

[0007] Preferably, the ratio of rice powder, sparganium, salix and licorice is 1:0.3:0.1:0.1.

[0008] The preparation method,

[0009] 97.80wt%-98.98wt% glutinous rice is soaked for 8-12 hours, 1.02wt%-2.20wt% of rhizoma polygonati is added, mixed, cooked until cooked, and then spread to 25-30℃ to serve as the embryo material; 0.20wt%-0.30wt% of new soil koji is taken and mixed with 99.7wt%-99.8wt% of the embryo material for fermentation.

[0010] Preferably, 98.04wt% glutinous rice is soaked for 8-12 hours, 1.96wt% of rhizoma polygonati is added, mixed, cooked until cooked, and then spread to 25-30℃ to serve as the embryo material; 0.25wt% of new soil koji is taken and mixed with 99.75wt% of the embryo material for fermentation.

[0011] The preparation method, when fermenting, the mixed material is poured into a ceramic jar and moved to a room with a temperature of 25-28℃, saccharification and fermentation are carried out for 5-7 days, then the fermented material is moved to a ceramic jar, 0.50-1.0L of cold boiled water is added to the fermented material for secondary fermentation, and the fermented material is fermented for 3-7 days at 25-28℃ to make the sugar content reach 100-180g / L, then the fermented material is pressed, sterilized, and clarified to obtain the fermented water wine.

[0012] The application also provides a fermented water wine of rhizoma polygonati prepared by the above method.

[0013] The application also provides an application of the fermented water wine of rhizoma polygonati in preparing food for regulating the intestinal flora function of human or animals, improving blood lipid and oxidative stress of human or animals.

[0014] The application also provides an application of the fermented water wine of rhizoma polygonati in preparing food for reducing the harmful effects of alcohol on the brain.

[0015] Through sensory evaluation, component analysis and animal experiments, it is found that the fresh rhizoma polygonati fermented water wine is superior to ordinary water wine, and the nine-steaming-nine-preparing rhizoma polygonati fermented water wine is superior to the fresh rhizoma polygonati fermented water wine.

[0016] In order to ensure the fermentation quality of the fermented water wine of rhizoma polygonati, the rhizoma polygonati is mixed with glutinous rice for cooking, so that the microorganisms in the raw materials are inactivated, and the action conditions of the microorganisms in the new soil koji are met.

[0017] By adding sparganium, Homonoia riparia Lour and licorice to rice powder, the balanced growth of microorganisms in the process of preparing new soil koji is regulated, the fermentation requirements of water wine are met, and the lactic acid bacteria in the water wine are important bacteria for forming the aroma of the water wine in the ripening process of the water wine.

[0018] In the production of the new Huazhu koji, the rhynchocoris, salix, and licorice are added to make the saccharification bacteria and the yeast bacteria grow coordinately during the production of the koji, and to ensure the growth of the lactic acid bacteria, so that the reliable quality fermented koji of the application is obtained.

[0019] The new Huazhu koji is used to meet the production requirements of the fermented water wine of the rhizoma polygonati, and the fresh rhizoma polygonati or the nine-steaming and nine-preparing rhizoma polygonati is mixed with the glutinous rice for fermentation, so that the utilization of the rhizoma polygonati and the water wine is improved through the action of the microorganisms and the enzymes, the fermented water wine of the rhizoma polygonati is produced, the fermented water wine of the fresh rhizoma polygonati does not have the peculiar smell of the fresh rhizoma polygonati, and the color of the fermented water wine of the nine-steaming and nine-preparing rhizoma polygonati is better.

[0020] The fermented water wine of the rhizoma polygonati has the application of regulating the intestinal flora function of a human or an animal, improving the blood fat and the oxidation stress of the human or the animal, and can also reduce the harmful effects of alcohol on the brain. BRIEF DESCRIPTION OF DRAWINGS

[0021] Figure 1 Effects of the fermented water wine of the rhizoma polygonati provided for the embodiment 3 of the application on four indexes of blood fat of mice are shown in the following table.

[0022] In the table, (1A) is the TG content in the serum of the mice in each group, (1B) is the TCHO content in the serum of the mice in each group, (1C) is the LDL-C content in the serum of the mice in each group, and (1D) is the HDL-C content in the serum of the mice in each group.

[0023] Figure 2 Effects of the fermented water wine of the rhizoma polygonati provided for the embodiment 3 of the application on the oxidation stress factors MDA, SOD, and GSH-Px in the liver of mice are shown in the following table.

[0024] In the table, (2A) is the MDA content in the liver of the mice in each group, (2B) is the SOD content in the liver of the mice in each group, and (2C) is the GSH-Px content in the liver of the mice in each group.

[0025] Figure 3 Effects of the fermented water wine of the rhizoma polygonati provided for the embodiment 3 of the application on BDNF and GABA in the brain of mice are shown in the following table.

[0026] (3A) is the BDNF content in the brain of the mice in each group, and (3B) is the GABA content in the brain of the mice in each group.

[0027] *: P<0.05; **: P<0.01.

[0028] Figure 4 Effects of the fermented water wine of the rhizoma polygonati provided for the embodiment 3 of the application on the number of ASV of mice are shown in the following table.

[0029] Figure 5Effect of the fermented water wine of Rhizoma polygonati provided for the embodiment 3 of the present application on the Beta diversity of mice.

[0030] Wherein, figure (5A) is PCoA analysis; figure (5B) is NMDS analysis.

[0031] Figure 6 Effect of the fermented water wine of Rhizoma polygonati provided for the embodiment 3 of the present application on the characteristic bacteria of mice. DETAILED DESCRIPTION

[0032] The embodiments of the technical solutions of the present application will be described in detail below with reference to the drawings. The following embodiments are only used to more clearly illustrate the technical solutions of the present application, and therefore only serve as examples, and cannot limit the protection scope of the present application. The test methods used in the following embodiments are conventional methods unless otherwise specified; and the materials, reagents, etc. used are commercially available reagents and materials unless otherwise specified.

[0033] The microorganisms used in the embodiments of the present application are conventional strains, which are purchased from the China Industrial Microbial Culture Collection Center (CICC):

[0034] Rhizopus oryzae is used for Rhizopus, and the strain number is CICC 41435.

[0035] Saccharomyces cerevisiae is used for Saccharomyces, and the strain number is CICC 1308.

[0036] Lactobacillus plantarum is used for Lactobacillus, and the strain number is CICC 22696.

[0037] The method of the present application includes but is not limited to the above specific strains.

[0038] Embodiment 1:

[0039] In order to obtain the fermented water wine of Rhizoma polygonati, new soil molds are used for the production of fermented water wine of Rhizoma polygonati:

[0040] (1)98.04wt% glutinous rice, 1.96wt% Huang- jing (fresh or nine steaming and nine processing Huang- jing), mix well, cook until done, then spread and cool to 28°C, as the embryo material. Take 0.25wt% Xinhua daqu, mix with 99.75wt% embryo material, then pour into ceramic jar and move to room temperature 28°C room, saccharify and ferment for 6 days, then move to ceramic jar, add cold boiled water to the raw material and cold boiled water mass volume ratio of 1kg:0.8L for secondary fermentation, ferment for 6 days at 28°C, sugar content 100-180g / L, filter, sterilize, clarify, store.

[0041] Xinhua daqu raw material is rice powder: rhubarb: Homonoia riparia Lour: licorice = 1:0.3:0.1:0.1, first licorice with water decoction, then the rice, rhubarb, Homonoia riparia Lour leaf stem are crushed and mixed, then the licorice water decoction is placed in the crushed mixture of rice, rhubarb, Homonoia riparia Lour leaf stem, add cold boiled water to 60-70% moisture content, add 3% daqu starter (mother powder) (composed of 3000-3500 rhizopus spores / g, 2000-2400 yeast / g, 2500-3000 lactic acid bacteria / g), mix well, make into 3-5cm diameter pills, constant temperature culture at 25°C until the water content of the pills is 20%, Xinhua daqu has saccharifying enzyme activity of 3000-3500U / g, 2000-3000 rhizopus spores / g, 1000-1400 yeast / g, 1500-2000 lactic acid bacteria / g.

[0042] (2)98.04wt% glutinous rice, 1.96wt% Huang- jing (fresh or nine steaming and nine processing Huang- jing), mix well, cook until done, then spread and cool to 28°C, as the embryo material. Take 0.25wt% Xinhua daqu, mix with 99.75wt% embryo material, then pour into ceramic jar and move to room temperature 28°C room, saccharify and ferment for 6 days, then move to ceramic jar, add cold boiled water to the raw material and cold boiled water mass volume ratio of 1kg:0.8L for secondary fermentation, ferment for 6 days at 28°C, sugar content 100-180g / L, filter, sterilize, clarify, store.

[0043] The raw materials of Rhizopus yeast koji are rice powder: chonglou grass: Homonoia riparia Lour: licorice = 1:0.3:0.1:0.1. First, the licorice is decocted with water, and then the rice, chonglou grass and Homonoia riparia Lour are crushed and mixed uniformly. The licorice decoction is added to the crushed mixture of rice, chonglou grass and Homonoia riparia Lour, and cold water is added to make the water content 60-70%. Rhizopus and yeast mother powder (Rhizopus spores 3000-3500 / g, yeast 2000-2400 / g) are added and mixed uniformly to make 3-5 cm diameter pills. The pills are incubated at 25°C until the water content is 20%. The saccharifying enzyme activity in the Rhizopus koji is 3200-3400 U / g, the Rhizopus spores are 2000-3000 / g, and the yeast is 1000-1400 / g. In this embodiment, the preparation process of each water wine is evaluated by single factor experiment. The sensory evaluation test in the following test process is based on the sensory requirements specified in GB / T 13662-2018 "Rice Wine". The specific contents are shown in Table 1:

[0044] The optimal component ratio and preparation process are used for preparation, and the physicochemical indexes and nutritional component indexes are determined. The experimental methods and test results are as follows:

[0045] 2.8.1 Total sugar determination

[0046] Determined according to the method specified in GB / T 13662-2018 "Rice Wine".

[0047] 2.8.2 Non-sugar solid determination

[0048] Determined according to the method specified in GB / T 13662-2018 "Rice Wine".

[0049] 2.8.3 Alcohol content determination

[0050] Determined according to the method specified in GB 5009.225-2016 "National Food Safety Standard Determination of Ethanol Concentration in Wine".

[0051] 2.8.4 Amino acid nitrogen determination

[0052] Determined according to the method specified in GB / T 13662-2018 "Rice Wine".

[0053] 2.8.5 Total polyphenol determination

[0054] Determined according to the method specified in GB / T 4928-2008 "Beer Analysis Method".

[0055] 2.8.6 Total ester determination

[0056] Determination was carried out according to the method specified in GB / T 10345-2022 “Analysis Methods for Baijiu”.

[0057] 2.8.7 Total acid determination

[0058] Determination was carried out according to the method specified in GB / T 13662-2018 “Yellow Rice Wine”.

[0059] 2.8.8 Total flavonoids determination

[0060] Determination was carried out according to the method specified in Appendix G of GB / T 12143-2008 “General Analysis Methods for Beverages”.

[0061] 2.8.9 Polysaccharide determination

[0062] Determination was carried out according to the method specified in Appendix A of GB / T 18672-2014 “Medicinal Wine”.

[0063] 2.8.10 Total saponins determination

[0064] Determination was carried out according to the method specified in the second part of “Technical Guidelines for the Test and Evaluation of Physical and Chemical and Hygienic Indicators of Health Foods” (2020 edition) “Test Methods for Efficacy Components / Marker Components (Second Method)”.

[0065] Table 1 Sensory quality, physicochemical indicators and nutritional components of each water wine

[0066]

[0067] After sensory and component analysis, the water wine produced by Xinhua soil starter had a richer aroma than the water wine produced by Rhizopus yeast starter. The taste of the Huangjing fermented water wine produced by Xinhua soil starter was mellow, without astringency and irritation. Fresh Huangjing and nine-steaming and nine-processing Huangjing were both suitable for producing Huangjing fermented water wine, but the nine-steaming and nine-processing Huangjing fermented water wine was superior to the fresh Huangjing fermented water wine. Table 1.

[0068] Example 2:

[0069] The raw materials of the new Huazitou starter are rice powder: chaff: salix: liquorice = 1:0.3:0.1:0.1. The liquorice is first decocted with water, and then the rice, chaff and salix are mixed and crushed. The decoction of liquorice is added to the mixture of rice, chaff and salix, and cold water is added to reach a water content of 60-70%. 3% of the new Huazitou starter (mother powder) (the mother powder is composed of 3000-3500 rhizopus spores / g, 2000-2400 yeast bacteria / g and 2500-3000 lactic acid bacteria / g) is added and mixed, and then the mixture is made into pills with a diameter of 3-5 cm. The pills are incubated at 25°C until the water content is 20%. The glucoamylase activity of the new Huazitou starter is 3000-3500 U / g, the rhizopus spore content is 2000-3000 / g, the yeast bacteria content is 1000-1400 / g, and the lactic acid bacteria content is 1500-2000 / g.

[0070] By adding chaff, salix (Homonoia riparia Lour) and liquorice to the rice powder, the balanced growth of microorganisms during the preparation of the new Huazitou starter is adjusted, and the fermentation requirements of the water wine are met. The lactic acid bacteria in the new Huazitou starter are important bacteria for the formation of the aroma of the water wine during the aging process of the water wine.

[0071] When the raw materials are rice powder: bran: liquorice = 1:0.3:0.1, the liquorice is first decocted with water, and then the rice powder and bran are mixed. The decoction of liquorice is added to the mixture of rice powder and bran, and cold water is added to reach a water content of 60-70%. 3% of the new Huazitou starter (mother powder) (the mother powder is composed of 3000-3500 rhizopus spores / g, 2000-2400 yeast bacteria / g and 2500-3000 lactic acid bacteria / g) is added and mixed, and then the mixture is made into pills with a diameter of 3-5 cm. The pills are incubated at 25°C until the water content is 20%. The detection results show that the glucoamylase activity of the starter is 2500-3000 U / g, the rhizopus spore content is 1800-2500 / g, the yeast bacteria content is 800-1200 / g, the lactic acid bacteria content is 400-900 / g, and the bacillus content is 1900-2100 / g. When the starter is used for water wine production, the water wine has an excessive sour taste and an odor due to the influence of the bacillus in the starter.

[0072] It can be seen that, during the preparation of the new Huazitou starter, the addition of chaff, salix and liquorice ensures the coordinated growth of saccharifying bacteria and yeast bacteria during the preparation of the starter, and guarantees the growth of lactic acid bacteria, so that the fermentation starter with reliable quality is obtained.

[0073] Example 3

[0074] In order to verify the efficacy of the product of the present application, male Kunming mice were used as experimental materials, and the above-mentioned optimal component ratio and preparation process were used to prepare the Huangjing fermented water wine. The pharmacodynamics of the prepared Huangjing fermented water wine was verified, including:

[0075] 2.1 Experimental animals and feeding

[0076] SPF level male Kunming mice 48, 6 weeks old, purchased from Hunan Slike Jingda Experimental Animal Co., Ltd., using 60 Co irradiation of experimental mice growth and reproduction feed feeding (Hunan University of Chinese Medicine provided by the experimental animal center).

[0077] 2.2 Experimental method

[0078] After 48 male Kunming mice were adaptively fed in a suitable environment (room temperature 23-25℃, relative humidity 50%-70%, clean and quiet) for 4d, they were randomly divided into blank control group (CCN, 12), ordinary water wine group (except that no Huangjing was added, which was consistent with the Huangjing fermented water wine of the application) (CCA, 12), fresh Huangjing fermented water wine group (CRP, 12), nine steaming nine processing Huangjing fermented water wine group (CPP, 12), numbered and caged, 6 / cage.

[0079] The blank control group was given 0.35mL of sterile water at room temperature, and the ordinary water wine group, fresh Huangjing fermented water wine group and nine steaming nine processing Huangjing fermented water wine group were given 0.35mL of ordinary water wine, fresh Huangjing fermented water wine and nine steaming nine processing Huangjing fermented water wine at room temperature, respectively, for 42d.

[0080] 2.3 Experimental results

[0081] 2.3.1 Effect of Huangjing fermented water wine on four items of blood lipids in mice

[0082] Four items of blood lipids are one of the commonly used means for checking blood lipid abnormalities in clinics, which specifically include triglyceride (TG), total cholesterol (TCHO), high-density lipoprotein (HDL-C) and low-density lipoprotein (LDL-C). Determining the content of blood lipids in serum can be used to evaluate the level of fat metabolism in mice.

[0083] In this embodiment, the content of TCHO, TG, LDL-C and HDL-C was determined by microplate method, and the specific operation was carried out according to the instruction manual of the kit.

[0084] The experimental results are shown in Figure 1 The experimental results show that:

[0085] As shown in Figure 1 A, compared with the CCN group, the TG content of each Huangjing fermented water wine group was reduced, while the TG content of the CCA group was increased, but the difference was not significant (P>0.05), and by comparing the TG content in the serum of mice in different groups, it was found that: CPP group < CRP group < CCN group < CCA group.

[0086] As shown in Figure 1As shown in B, compared with the CCN group, the TCHO content in each group of Polygonatum fermented water wine was decreased, while the TCHO content in the CCA group was increased, but the difference was not significant (P>0.05). By comparing the TCHO content in the serum of mice in different groups, the results showed that: CPP group < CRP group < CCN group < CCA group.

[0087] like Figure 1 As shown in Figure C, compared with the CCN group, the LDL-C content of each Polygonatum fermented water wine group decreased, while the LDL-C content of the CCA group increased, with extremely significant differences (CCN group vs CPP group, P < 0.01; CCA group vs CPP group, P < 0.01). By comparing the LDL-C content in the serum of mice in different groups, the results showed that: CPP group < CRP group < CCN group < CCA group.

[0088] like Figure 1 As shown in Figure D, compared with the CCN group, the HDL-C content in each water-wine group was significantly increased (CCN group vs CPP group, P < 0.05; CCA group vs CPP group, P < 0.05; CRP group vs CPP group, P < 0.05). By comparing the HDL-C content in the serum of mice in different groups, the results were: CCN group < CCA group < CRP group < CPP group.

[0089] In summary, it can be inferred that the nine-times-steamed and nine-times-processed Polygonatum fermented wine has the best effect on lowering blood lipids in mice, followed by fresh Polygonatum fermented wine, while ordinary wine has the worst effect.

[0090] 2.3.2 Effects of Polygonatum sibiricum fermented rice wine on oxidative stress factors MDA, SOD, and GSH-Px in mouse liver

[0091] Superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) are important components of the body's antioxidant system. SOD catalyzes the dismutation of superoxide anion free radicals to produce oxygen and hydrogen peroxide, playing a crucial role in the body's oxidation-antioxidant balance, protecting the body from oxidative damage. Its levels, to a certain extent, reflect the body's ability to remove oxygen free radicals and are closely related to the occurrence and development of many diseases. GSH-Px specifically catalyzes the reaction of reduced glutathione with reactive oxygen species to produce oxidized glutathione, thereby protecting biological membranes from reactive oxygen species damage, maintaining normal cell function, and possessing abilities such as protecting the liver, enhancing immunity, antagonizing the damage of harmful metal ions, and increasing the body's resistance to radiation. Free radicals act on lipids, causing peroxidation reactions, with the final oxidation product being malondialdehyde (MDA). MDA can cause cross-linking and polymerization of biomolecules such as proteins and nucleic acids, and has cytotoxic effects. When lipid peroxidation in the body is enhanced and excessive oxygen free radicals are generated, the body's antioxidant enzymes such as SOD and GSH-Px will increase, thereby clearing the excess oxygen free radicals and maintaining a dynamic balance between oxidation and antioxidation in the body.

[0092] This embodiment uses enzyme-linked immunosorbent assay (ELISA) to determine the levels of MDA, SOD, and GSH-Px in the liver. The specific operation is performed according to the kit instructions.

[0093] Experimental results are as follows Figure 2 As shown, the experimental results indicate that:

[0094] like Figure 2 As shown in Figure A, compared with the CCN group, the MDA content in each water-based alcohol group was significantly or extremely significantly reduced (CCN group vs. CPP group, P < 0.01; CCA group vs. CPP group, P < 0.05; CRP group vs. CPP group, P < 0.05). By comparing the MDA content in the liver of mice in different groups, the results showed that: CPP group < CRP group < CCA group < CCN group.

[0095] like Figure 2 As shown in B, compared with the CCN group, the SOD content in each water-wine group was significantly or extremely significantly increased (CCN group vs CRP group, P < 0.05; CCN group vs CPP group, P < 0.01; CCA group vs CPP group, P < 0.05). By comparing the SOD content in the liver of mice in different groups, the results showed that: CCN group < CCA group < CRP group < CPP group.

[0096] like Figure 2As shown in Table C, compared with the CCN group, the GSH-Px content of each water wine group was significantly or extremely significantly increased (CCN group vs CRP group, P<0.05; CCN group vs CPP group, P<0.01; CCA group vs CPP group, P<0.05). By comparing the GSH-Px content in the liver of mice in different groups, it was found that CCN group < CCA group < CRP group < CPP group.

[0097] In summary, it can be inferred that the antioxidant capacity of mice after intragastric administration of nine steaming and nine processing fermented water wine is the highest, followed by fresh fermented water wine, and ordinary water wine is the worst.

[0098] 2.3.3 Effect of fermented water wine of Polygonatum on BDNF and GABA in the brain of mice

[0099] Brain-derived neurotrophic factor (BDNF) plays a key role in neuronal survival, differentiation, and synaptic plasticity, and can affect learning and memory ability, and has potential therapeutic effects on cognitive changes related to neurodegenerative diseases. Gamma-aminobutyric acid (GABA) is an inhibitory neurotransmitter in the central nervous system and is one of the most important neurotransmitters in brain tissue. Its role is to reduce neuronal activity, prevent nerve cells from overheating, calm nerves, and reduce anxiety. GABA and BDNF not only maintain neuronal function, but also improve hippocampal neurogenesis. Moreover, there is a mutual regulation between GABA and BDNF. GABA can promote the transport and expression level of BDNF, and BDNF can promote the transcriptional activity of GABA.

[0100] In this example, the content of BDNF and GABA in the brain was determined by enzyme-linked immunosorbent assay, and the specific operation was performed according to the kit instructions.

[0101] The experimental results are shown in Table C. Figure 3 The experimental results show that:

[0102] As shown in Table C, compared with the CCN group, the GSH-Px content of each water wine group was significantly or extremely significantly increased (CCN group vs CRP group, P<0.05; CCN group vs CPP group, P<0.01; CCA group vs CPP group, P<0.05). By comparing the GSH-Px content in the liver of mice in different groups, it was found that CCN group < CCA group < CRP group < CPP group. Figure 3Compared with the CCN group, the BDNF and GABA contents of each group of Huangqi fermented water wine were increased, while the BDNF and GABA contents of the CCA group were significantly or extremely significantly reduced (BDNF: CCN group vs CRP group, P<0.01; CCN group vs CPP group, P<0.01; CCA group vs CRP group, P<0.01; CCA group vs CPP group, P<0.01. GABA: CCN group vs CRP group, P<0.05; CCN group vs CPP group, P<0.01; CCA group vs CRP group, P<0.05; CCA group vs CPP group, P<0.01). By comparing the BDNF and GABA contents in the brains of mice in different groups, it was found that CCA group<CCN group<CRP group<CPP group.

[0103] Based on the above, it can be inferred that the protective effect of intragastric administration of nine steaming and nine processing Huangqi fermented water wine on the hippocampus of mice is the best, followed by fresh Huangqi fermented water wine, and ordinary water wine is the worst. Huangqi fermented water wine may reduce the harmful effects of alcohol on the brain.

[0104] 2.3.4 Observation of the effect of Huangqi fermented water wine on intestinal flora of mice

[0105] After the mice were killed by cervical dislocation, they were immediately placed on a clean bench, and the small intestine was taken as a detection sample. The chyme content was squeezed out with sterilized forceps and separately numbered into 1.5 mL sterilized EP tubes, which were then quickly frozen with liquid nitrogen and subsequently stored in a-80℃ refrigerator for 16S rRNA gene high-throughput sequencing.

[0106] The total DNA of intestinal content samples of each group of mice was extracted according to the DNA extraction kit (MNNucleoSpin 96Soi). The concentration of the extracted DNA was determined using a Nanodrop, and the purity of the extracted DNA was detected by 1.8% agarose gel electrophoresis. The V3+V4 high variable region of the 16S rRNA gene was selected as the PCR amplification region, and the qualified DNA was amplified using 16S rRNA universal primers (338F / 806R): upstream primer 338F (5'-ACTCCTACGGGAGGCAGCA-3') and downstream primer 806R (5'-GGACTACHVGGGTWTCTAAT-3'). The amplification of 10 μL system includes: VnF (10 μmol / L) 0.3 μL, VnR (10 μmol / L) 0.3 μL, KOD FX Neo Buffer 5 μL, dNTP (2 mmol / L each) 2 μL, KOD FX Neo 0.2 μL, and ddH2O to 10 μL. The amplification reaction conditions are as follows: 95℃ pre-denaturation for 5 min, then 25 cycles of 95℃ denaturation for 30 s, 50℃ annealing for 30 s, 72℃ extension for 40 s, finally 72℃ extension for 7 min, and 4℃ termination reaction.

[0107] The PCR products were mixed according to the mass ratio of 1:1 according to the electrophoresis quantification (ImageJ software) results. After mixing, the OMEGA DNA purification kit was used for column purification. After 1.8% agarose gel electrophoresis at 120V for 40 min, the target fragment was cut and recovered using the Monarch DNA gel recovery kit.

[0108] SPSS 25.0 software was used for statistical analysis. All measurement data were expressed as mean ± standard deviation (mean ± SD). When the data were normally distributed, independent sample t test was used for comparison between two groups; one-way analysis of variance was used for comparison among multiple groups, and LSD method was used for comparison between groups. Otherwise, Mann-Whitney U test was used for comparison between two groups, and Kruskal-Wallis H test was used for comparison among multiple groups. The test level was set as α = 0.05. When P < 0.05, the result was statistically significantly different; when P < 0.01, the result was extremely statistically significantly different.

[0109] 2.3.4.1 Effect of Huangjing fermented water wine on ASV number of mice

[0110] The data after quality control was denoised using the DADA2 method in QIIME2 (version 2020.6), and 0.005% of the total number of sequences was used as the threshold to filter the amplicon sequence variants (ASVs) by default. As shown in Figure 4 , the CCN group had a total of 951 ASVs, of which 620 were unique ASVs; the CCA group had a total of 647 ASVs, of which 356 were unique ASVs; the CRP group had a total of 1514 ASVs, of which 1151 were unique ASVs; and the CPP group had a total of 957 ASVs, of which 616 were unique ASVs. The four groups had a total of 137 ASVs Figure 4 . The results show that after gavage with ordinary water wine, the total number of ASVs in mice decreases and the number of unique ASVs decreases accordingly, while after gavage with fresh Huangjing fermented water wine, the total number of ASVs in mice increases and the number of unique ASVs increases accordingly, and after gavage with nine steaming and nine processing Huangjing fermented water wine, the total number of ASVs in mice has no obvious change, indicating that the intake of different water wines significantly changes the types and diversity of intestinal microorganisms in mice, thereby affecting the effect of intestinal flora on the body.

[0111] 2.3.4.2 Effect of Huangjing Fermented Water Wine on Beta Diversity of Mice

[0112] The experimental results are shown in Figure 5 , based on the Bray-Curtis distance algorithm to evaluate beta diversity, the microbial community composition of different groups is significantly different (P = 0.019), which is also confirmed by anosim. The closer the distance between the two groups on the coordinate axis, the more similar the community composition of the two groups in the corresponding dimension. The PCoA analysis result shows Figure 5 A), the contribution rate of the horizontal coordinate PCoA1 is 35.28%, and the contribution rate of the vertical coordinate PCoA2 is 22.62%. The samples of the CCN group partially overlap with the samples of the CRP group and the samples of the CPP group, but there is no overlapping part with the samples of the CCA group. Obviously, compared with the CCN group, the community structure of the intestinal content flora of the CRP group and the CPP group has changed significantly but also has certain similarities, while the community structure of the intestinal content flora of the CCA group is obviously different from that of the CCN group. The NMDS analysis Figure 5 B) is consistent with the PCoA analysis result. It shows that water wine affects the function of the body by changing the intestinal flora.

[0113] 2.3.4.3 Effect of Huangjing Fermented Water Wine on Characteristic Bacterial Genera of Mice

[0114] To further verify the regulatory effects of different alcoholic beverages on the intestinal flora of mice, LEfSe analysis was used with a linear judgment analysis (LDA) score >2 as the screening criterion to compare multiple groups and identify characteristic bacterial genera with significant differences between groups.

[0115] like Figure 6 As shown, the CCN, CCA, CRP, and CPP groups contain 5, 1, 2, and 10 characteristic bacterial genera, respectively. Among them, Staphylococcus, Christensenellaceae R 7 group, and unclassified Enterobacteriaceae are characteristic genera enriched in the CCN group; unclassified Bacilli is a characteristic genera enriched in the CCA group; Hafnia obesumbacterium and Lachnospiraceae NK4A136 group are characteristic genera enriched in the CRP group; and unclassified Desulfovibrionaceae, Aeromonas, and unclassified Oscillospiraceae are characteristic genera enriched in the CPP group.

[0116] The gut microbiota influences various aspects of health, including disease, nutrition, drug efficacy, and immunity. In summary, fermented Polygonatum sibiricum wine exerts its lipid-lowering and antioxidant effects by regulating the gut microbiota and influencing related functional factors in the body, and may also reduce the harmful effects of alcohol on the brain.

Claims

1. A preparation method of fermented water wine of Rhizoma Polygonati, characterized by comprising the following steps: The soaked glutinous rice is mixed with the rhizoma polygonati, and then cooked until done, and then spread to cool down to serve as the embryo material; the new Xinhua koji is taken and mixed with the embryo material, and then the mixture is used for saccharification and fermentation for 5-7 days, and then cold boiled water is added for secondary fermentation for 3-7 days, so that the sugar content is 100-180 g / L, and then the mixture is filtered, sterilized, and clarified to obtain the product; ​ The raw materials of the new Xinhua koji are rice powder: polygonum hydropiper: salix matsudana: licorice = 0.8-1.5: 0.2-0.4: 0.08-0.12: 0.08-0.12, the licorice is first boiled in water, and then the rice, polygonum hydropiper, and salix matsudana are crushed to pass through a 100-mesh sieve and mixed evenly, the licorice water decoction is then added to the crushed mixture of the rice, polygonum hydropiper, and salix matsudana, cold boiled water is added, and 2-4% of the mother powder is added, the mother powder contains 3000-3500 rhizopus spores / g, 2000-2400 yeast bacteria / g, and 2500-3000 lactic acid bacteria / g, the mixture is mixed evenly, and then 3-5 cm diameter pellets are made, and the pellets are incubated at 25°C until the water content is 20%, the sugar content of the new Xinhua koji is 3000-3500 U / g, the rhizopus spore content is 2000-3000 / g, the yeast bacteria content is 1000-1400 / g, and the lactic acid bacteria content is 1500-2000 / g, and the new Xinhua koji is obtained.

2. The preparation method according to claim 1, characterized in that, The ratio of rice powder: polygonum hydropiper: salix matsudana: licorice is 1:0.3:0.1:0.

1.

3. The preparation method according to claim 1, characterized in that, 97.80wt%-98.98wt% of the glutinous rice is soaked for 8-12 hours, 1.02wt%-2.20wt% of the rhizoma polygonati is added, and the mixture is mixed evenly and cooked until done, and then spread to cool down to 25-30°C to serve as the embryo material; 0.20wt%-0.30wt% of the new Xinhua koji is mixed with 99.7wt%-99.8wt% of the embryo material for fermentation.

4. The preparation method according to claim 3, characterized in that, 98.04wt% of the glutinous rice is soaked for 8-12 hours, 1.96wt% of the rhizoma polygonati is added, and the mixture is mixed evenly and cooked until done, and then spread to cool down to 25-30°C to serve as the embryo material; 0.25wt% of the new Xinhua koji is mixed with 99.75wt% of the embryo material for fermentation.

5. The preparation method according to claim 1, characterized in that, During fermentation, the mixture is poured into a ceramic jar and moved to a room at a room temperature of 25-28°C, and then the mixture is used for saccharification and fermentation for 5-7 days, and then moved to a ceramic jar, cold boiled water is added at a material to water ratio of 1:0.50-1.0, and then the mixture is used for secondary fermentation at 25-28°C for 3-7 days, so that the sugar content is 100-180 g / L, and then the mixture is filtered, sterilized, and clarified to obtain the product.

6. A fermented water wine of Polygonatum, characterized in that, The product is prepared by the method of any one of claims 1-5.

7. Use of the rhizoma polygonati fermented water wine of claim 6 in the preparation of a food for regulating the intestinal flora function of a human or an animal or improving the blood lipid of a human or an animal.

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