Traditional Chinese medicine composition for controlling fat in body, dispelling effects of alcohol and protecting liver and preparation method of traditional Chinese medicine composition

Abstract

The invention belongs to the technical field of traditional Chinese medicines, mainly relates to a preparation method of a traditional Chinese medicine composition, and particularly relates to a traditional Chinese medicine composition for controlling body fat, dispelling effects of alcohol and protecting liver and a preparation method thereof. The invention aims to provide a traditional Chinese medicine composition which has the effects of controlling body fat, dispelling the effects of alcohol and protecting liver and has an auxiliary protection effect on chemical liver injury. Meanwhile, research finds that the composition also has the effect of improving anoxia tolerance. The composition comprises the following medicinal materials in parts by weight: 6-20 parts of orange peel, 9-30 parts of phaseolus calcaratus, 5-20 parts of semen cassiae and 10-30 parts of dandelion root.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicines, and mainly relates to a preparation method of a traditional Chinese medicine composition, and in particular to a traditional Chinese medicine composition for controlling body fat, sobering up, and protecting the liver, and a preparation method thereof. Background Art

[0002] With the rapid development of modern society and the rapid changes in lifestyle, obesity and liver health issues have become major challenges in the field of global public health. Obesity is not only related to external image, but also deeply related to the risk of chronic diseases such as cardiovascular health and diabetes, and its impact cannot be ignored. At the same time, chemical liver damage, as a common liver disease, has complex causes, covering food contaminants, alcohol consumption, adverse drug reactions and other aspects, posing a serious threat to human health.

[0003] Although there are a wide variety of products in the current weight loss market, most of them have been criticized for their significant side effects and poor long-lasting effects. In contrast, Chinese medicine for weight loss has gradually won market favor with its mild nature. According to Chinese medicine theory, the root cause of obesity lies in spleen deficiency, dampness accumulation, and phlegm stagnation. By harmonizing the spleen and stomach functions and promoting the discharge of dampness and phlegm, the goal of healthy weight loss can be achieved. However, the Chinese medicine weight loss prescriptions currently on the market mostly focus on weight loss effects, and pay insufficient attention to the protection of key organs such as the liver.

[0004] Although existing treatments for chemical liver damage include drug intervention and nutritional support, their efficacy is often limited and may be accompanied by adverse reactions, making it difficult to meet the comprehensive needs of patients. In view of this situation, it is particularly important and urgent to develop a Chinese medicine compound that combines liver protection and weight loss assistance. This type of innovative drug aims to scientifically combine a variety of Chinese medicinal materials to not only effectively promote weight management, but also enhance the liver's detoxification and repair capabilities, providing patients with obesity and chemical liver damage with a safer and more comprehensive treatment option. Summary of the invention

[0005] In view of the above problems, the present invention aims to provide a Chinese medicine composition that has the function of controlling body fat and assisting in protecting chemical liver damage. Meanwhile, it is found through research that the composition also has the function of improving hypoxia tolerance. The components of the composition include: 6-20 parts of orange peel, 9-30 parts of red beans, 5-20 parts of cassia seeds, and 10-30 parts of dandelion roots.

[0006] In a first aspect, the present invention provides a method for preparing a Chinese medicine composition.

[0007] S1, weighing 6-20g of orange peel, 9-30g of red bean, 5-20g of cassia seed, and 10-30g of dandelion root, and then washing, drying and crushing them for subsequent use;

[0008] S2. Bifidobacterium breve CICC-6204 was inoculated on glucose blood liver agar medium and cultured at 37°C for 12-18 hours to activate the strain. The activated Bifidobacterium breve was transferred to a 500-1000 mL glucose blood liver liquid medium shake flask and fermented for another 8 hours to obtain 1.2-1.5×10 8 cfu / mL of bacterial solution;

[0009] S3, Lactobacillus crispatus ATCC-33820 was inoculated on MRS agar medium and cultured at 37℃ for 12-18h to activate the strain. The activated Lactobacillus crispatus was transferred to a 500-1000mL MRS liquid medium shake flask and fermented for another 8h to finally obtain 1.8-2.2×10 8 cfu / mL of bacterial solution;

[0010] S4, after mixing the raw materials prepared in step S1, add 15-20 mL of the bacterial solution of Bifidobacterium breve CICC-6204 prepared in step S2 and 15-20 mL of the bacterial solution of Lactobacillus crispatus ATCC-33820 prepared in step S3, and ferment anaerobically at 37°C for 20-24 h. After the fermentation is completed, use 3% H 2 O 2 The solution inactivates Bifidobacterium breve and Lactobacillus crispatus in the fermentation product, thereby obtaining the desired Chinese medicine fermentation product;

[0011] S5. Add 8-10 times of 60-75% ethanol to the fermented Chinese medicine prepared in step S4, and distill in a constant temperature water bath for 3 hours, repeat twice, combine the filtrate and concentrate until there is no alcohol taste, so as to obtain the Chinese medicine composition of the present invention.

[0012] Preferably: in step S1 of the present invention, orange peel, red bean, cassia seed and dandelion root are crushed into 80-120 mesh particles respectively.

[0013] Preferably, in step S1 of the present invention, 10-15 g of orange peel, 15-20 g of red bean, 8-12 g of cassia seed, and 15-18 g of dandelion root are used.

[0014] Preferably, the glucose blood-liver agar medium used in step S2 of the present invention comprises: 3 g beef extract powder, 10 g prion peptone, 5 g tryptone, 3 g plant peptone, 5 g yeast extract powder, 150 mL liver extract, 10 g glucose, 0.5 g soluble starch, 10 mL solution A (K 2 HPO 4 10g, KH 2 PO 4 10 g, distilled water 100 mL), solution B 5 mL (MgSO 4 7H 2O 4g, NaCl 0.2g, FeSO 4 7H 2 O 0.2g, MnSO 4 ·2H 2 O 0.2g, distilled water 100mL), 5% L-cysteine ​​hydrochloride solution 10mL, Tween-80 1g, agar 15g, defibrinated sheep blood 50mL, distilled water 825mL.

[0015] Preferably: the components of the MRS agar medium used in step S3 of the present invention are: 10g peptone, 10g beef extract powder, 5g yeast extract powder, 20g glucose, 1g Tween, 5g sodium acetate, 2g triammonium citrate, 2g dipotassium hydrogen phosphate, 0.2g magnesium sulfate heptahydrate, 0.05g sulfuric acid tetrahydrate, 15g agar, 1000mL distilled water, and the pH is adjusted to 6.2-6.5; the components of the glucose blood liver agar medium are: 2.5g peptone, 1.5g beef extract, 10g glucose blood liver, 18g agar, and 1000mL distilled water.

[0016] Preferably, in step S4 of the present invention, the volumes of the Bifidobacterium breve bacterial solution and the Lactobacillus crispatus bacterial solution are both 18 mL.

[0017] Preferably, the time of anaerobic fermentation in step S4 of the present invention is 22 hours.

[0018] Preferably, the temperature of the constant temperature water bath distillation in step S5 of the present invention is 80°C.

[0019] In the second aspect, the Chinese medicine composition prepared by the above method has an auxiliary protective effect on chemical liver damage, helps to control body fat and improves hypoxia tolerance.

[0020] In a third aspect, the Chinese medicine composition of the present invention can be used in the preparation of health products or medicines.

[0021] The present invention has the following beneficial effects:

[0022] 1. The synergistic fermentation of orange peel and cassia seed in the present invention has an auxiliary protective effect on chemical liver damage.

[0023] 2. The synergistic effect of the adzuki bean fermented liquid and the dandelion root fermented liquid in the present invention helps to control body fat.

[0024] 3. The synergistic effect of the orange peel fermentation liquid, the adzuki bean fermentation liquid and the dandelion root fermentation liquid in the present invention has the effect of improving the tolerance to hypoxia.

[0025] 4. The orange peel of the present invention has the effects of regulating qi and strengthening the spleen, and can be used to relieve symptoms such as indigestion caused by drinking, and has the effect of sobering up.

[0026] 5. The Chinese medicine composition of the present invention is all derived from natural plants and has multiple biological activities. Compared with Western medicine, the Chinese medicine composition has more effects, higher safety and fewer side effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0028] Figure 1 This is a comparison chart of the fat coefficients of mice in each group after 30 days of administration in Example 2.

[0029] Figure 2 This is a comparison chart of serum TC concentrations of mice in each group after 30 days of administration in Example 2.

[0030] Figure 3 This is a comparison chart of serum TG concentrations of mice in each group after 30 days of administration in Example 2.

[0031] Figure 4 This is a comparison chart of the hypoxia-tolerant survival time of each group of mice in Example 3.

[0032] The following describes the embodiments of the present invention by specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention. It should be noted that the following embodiments and features in the embodiments can be combined with each other without conflict.

[0033] Tangerine peel, adzuki bean, cassia seed, and dandelion root were purchased from Bozhou Fushuotang Biotechnology Co., Ltd.; Bifidobacterium breve CICC-6204 was purchased from China Industrial Microbiological Culture Collection Center; Lactobacillus crispatus ATCC-33820 was purchased from Wuhan Huizao Biotechnology Co., Ltd.

[0034] Example 1

[0035] This example demonstrates that under fermentation conditions, the synergistic fermentation of orange peel and cassia seed has an auxiliary protective effect on chemical liver damage.

[0036] This embodiment provides a Chinese medicine composition, and the specific steps are as follows:

[0037] S1. Weigh 10 g of orange peel, 15 g of red bean, 8 g of cassia seed, and 15 g of dandelion root, then wash, dry, and crush into 80-mesh particles for subsequent use;

[0038] S2, Bifidobacterium breve CICC-6204 was inoculated on glucose blood liver agar medium and cultured at 37°C for 12 hours to activate the strain. The activated Bifidobacterium breve was transferred to a 500 mL glucose blood liver liquid medium shake flask and fermented for another 8 hours to finally obtain 1.2×10 8 cfu / mL of bacterial solution;

[0039] S3, Lactobacillus crispatus ATCC-33820 was inoculated on MRS agar medium and cultured at 37°C for 12 hours to activate the strain. The activated Lactobacillus crispatus was transferred to a 500 mL MRS liquid medium shake flask and fermented for another 8 hours to obtain 1.8×10 8 cfu / mL of bacterial solution;

[0040] S4, after mixing the raw materials prepared in step S1, 18 mL of the bacterial solution of Bifidobacterium breve CICC-6204 prepared in step S2 and 18 mL of the bacterial solution of Lactobacillus crispatus ATCC-33820 prepared in step S3 were added, and the mixture was fermented anaerobically at 37°C for 22 h. After the fermentation was completed, 3% H 2 O 2 The solution inactivates Bifidobacterium breve and Lactobacillus crispatus in the fermentation product, thereby obtaining the desired Chinese medicine fermentation product;

[0041] S5. Add 8 times of 75% ethanol to the fermented Chinese medicine prepared in step S4, and distill in a constant temperature water bath at 80° C. for 3 h. Repeat twice, combine the filtrates and concentrate until there is no alcohol taste, thereby obtaining a Chinese medicine composition.

[0042] Comparative Example 1: In step S4, orange peel is not added, and the remaining steps are consistent with Example 1.

[0043] Comparative Example 2: In step S4, Cassia seed was not added, and the remaining steps were kept consistent with Example 1.

[0044] Comparative Example 3: In step S2, Bifidobacterium breve CICC-6204 was not added, and the remaining steps were kept consistent with Example 1.

[0045] Comparative Example 4: In step S3, Lactobacillus crispatus ATCC-33820 was not added, and the remaining steps were kept consistent with Example 1.

[0046] Comparative Example 5: In step S2, Bifidobacterium breve CICC-6204 was replaced with Bifidobacterium breve CICC-6091 (purchased from China Industrial Microbiological Culture Collection Center), and the remaining steps were kept consistent with Example 1.

[0047] Comparative Example 6: In step S3, Lactobacillus crispatus CICC-25191 was replaced with Lactobacillus crispatus CICC-24879 (purchased from China Industrial Microbiological Culture Collection Center), and the remaining steps were kept consistent with Example 1.

[0048] Study on the effect of Chinese medicine combination on CCl 4 Protective effect in acute liver injury

[0049] 100 SPF-grade ICR mice, weighing 18-22 g, were purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd. and were stably raised in a constant temperature and oxygen supply environment at 25±2°C for one week before the experiment officially began.

[0050] The mice were randomly divided into 10 groups, namely, a blank group, a model group, a positive control group, an example 1 group, and comparative examples 1-6 groups. The positive control group was intragastrically administered with bifendate at a dose of 100 mg / (kg·d); the example 1 group and the comparative examples 1-6 groups were intragastrically administered with the corresponding Chinese medicine composition, which was diluted with purified water to a concentration of 5% v / v and then intragastrically administered at a dose of 10 mL / (kg·d); the blank group and the model group were intragastrically administered with an equal amount of purified water for 3 consecutive weeks. One hour after the last administration, except for the blank group, which was intraperitoneally injected with 10 mL / kg of peanut oil, the other groups were intraperitoneally injected with 2 g / L CCl 4 Peanut oil 10mL / kg, then fasting but not water for 16h. Kill the mice and remove the liver. Place the liver in a petri dish filled with 4℃ saline and gently shake to wash away the blood on the surface. Use filter paper to absorb the surface water, weigh it, and calculate the liver coefficient.

[0051] Liver coefficient = liver mass / mouse mass × 100%

[0052] After the mice were killed, the eyeballs were removed and blood was collected. The blood samples were centrifuged at 4°C and 3500 r / min for 10 min, and serum was collected. The ALT, AST, and TBIL concentrations were determined according to the operating specifications of the kit (ALT, AST, and TBIL kits were purchased from Shanghai Jiachu Bioengineering Co., Ltd.).

[0053] Table 1

[0054] Group Number of mice Liver coefficient (%) Blank group 10 4.41±0.10 Model Group 10 5.01±0.13 Positive control group 10 4.44±0.15 Example 1 Group 10 4.42±0.12 Comparative Example 1 10 4.71±0.17 Comparative Example 2 10 4.68±0.16 Comparative Example 3 10 4.67±0.17 Comparative Example 4 10 4.64±0.15 Comparative Example 5 10 4.62±0.14 Comparative Example 6 10 4.61±0.12

[0055] Table 2

[0056] Group ALT activity / (U / L) AST activity / (U / L) TBIL concentration / (umol / mg) Blank group 41.56±2.66 115.02±7.78 3.12±0.04 Model Group 945.81±89.72 859.22±75.90 4.26±0.09 Positive control group 493.77±60.75 624.59±58.29 3.52±0.05 Example 1 Group 486.56±51.34 606.27±63.65 3.27±0.08 Comparative Example 1 785.34±69.89 748.45±74.78 4.19±0.10 Comparative Example 2 773.27±67.57 738.78±73.79 4.04±0.09 Comparative Example 3 768.39±67.37 729.35±71.55 3.95±0.08 Comparative Example 4 759.73±66.40 717.38±70.27 3.87±0.08 Comparative Example 5 751.48±66.29 704.57±71.59 3.79±0.07 Comparative Example 6 743.50±64.58 894.28±70.68 3.62±0.05

[0057] Table 1 is a comparison of the liver coefficients of mice in each group after 3 weeks of administration. The liver coefficients of mice in the model group were significantly different from those in the blank group, indicating that CCl 4 It can induce hepatocyte edema, thereby promoting the increase of liver coefficient; compared with the model group, the liver coefficient of mice in the comparative examples 1-6 groups was relatively reduced, while the liver coefficient of mice in the embodiment 1 group was the smallest, indicating that the comparative examples 1-6 groups and the embodiment 1 group were sensitive to CCl 4 It has a certain alleviating effect on liver enlargement caused by liver poisoning, and the effect of Example 1 group is the best.

[0058] Table 2 is a comparison of the ALT, AST activity and TBI concentration in the serum of each group of mice after 3 weeks of administration. 4 After the treatment, the ALT, AST activities and TBI concentration in the serum increased significantly, indicating that the liver cells suffered serious damage; compared with the model group, the serum ALT, AST activities and TBI concentrations of the mice in the comparative examples 1-6 groups were relatively reduced, while the serum ALT, AST activities and TBI concentrations of the mice in the embodiment 1 group were the smallest, indicating that the comparative examples 1-6 groups and the embodiment 1 group can inhibit the increase of ALT, AST activities and TBI concentrations to a certain extent, reduce the damage degree of CCL to the mouse liver, and the effect of the embodiment 1 group is the best.

[0059] In summary, Example 1 group CCl 4 The protective effect on acute liver damage caused by orange peel is the best, which confirms that the synergistic fermentation of orange peel and cassia seed has a good auxiliary protective effect on chemical liver damage.

[0060] Example 2

[0061] This example demonstrates that the synergistic effect of adzuki bean fermented liquid and dandelion root fermented liquid helps control body fat.

[0062] This embodiment provides a Chinese medicine composition, and the specific steps are as follows:

[0063] S1, weigh 12g of orange peel, 17g of red bean, 10g of cassia seed, and 16g of dandelion root, then wash, dry, and crush into 100-mesh particles for subsequent use;

[0064] S2, Bifidobacterium breve CICC-6204 was inoculated on glucose blood liver agar medium and cultured at 37°C for 15 hours to activate the strain. The activated Bifidobacterium breve was transferred to a 700 mL glucose blood liver liquid medium shake flask and fermented for another 8 hours to obtain 1.3×10 8 cfu / mL of bacterial solution;

[0065] S3, Lactobacillus crispatus ATCC-33820 was inoculated on MRS agar medium and cultured at 37°C for 15 h to activate the strain. The activated Lactobacillus crispatus was transferred to a 700 mL MRS liquid medium shake flask and fermented for another 8 h to obtain 2.0×10 8 cfu / mL of bacterial solution;

[0066] S4, after mixing the raw materials prepared in step S1, 18 mL of the bacterial solution of Bifidobacterium breve CICC-6204 prepared in step S2 and 18 mL of the bacterial solution of Lactobacillus crispatus ATCC-33820 prepared in step S3 were added, and the mixture was fermented anaerobically at 37°C for 22 h. After the fermentation was completed, 3% H 2 O 2 The solution inactivates Bifidobacterium breve and Lactobacillus crispatus in the fermentation product, thereby obtaining the desired Chinese medicine fermentation product;

[0067] S5. Add 9 times of 70% ethanol to the fermented Chinese medicine prepared in step S4, and distill in a constant temperature water bath at 80° C. for 3 h. Repeat twice, combine the filtrates and concentrate until there is no alcohol taste, thereby obtaining a Chinese medicine composition.

[0068] Comparative Example 7: In step S4, no adzuki bean was added, and the remaining steps were consistent with Example 2.

[0069] Comparative Example 8: In step S4, dandelion root was not added, and the remaining steps were kept consistent with Example 2.

[0070] Study on the effect of Chinese medicine combination on controlling body fat in mice

[0071] 50 SPF-grade ICR mice, weighing 18-22 g, were purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd. and were stably raised in a constant temperature and oxygen supply environment at 25±2°C for one week before the experiment officially began.

[0072] Mice were randomly divided into 5 groups, namely, a blank group, a model group, an embodiment 2 group, and a comparative example 7-8 group. The blank group was supplied with a basic feed (provided by Hebei Demu Pet Food Co., Ltd.), and the other groups were supplied with a high-fat feed (each 100g of high-fat feed was mixed with 79g of basic feed, 10g of lard, 10g of egg yolk powder, and 1g of cholesterol). At the same time, the embodiment 2 group and the comparative example 7-8 group were gavaged with the corresponding Chinese medicine composition every day, and the Chinese medicine composition was diluted with pure water to a concentration of 5% v / v and then gavaged, and the gavage dose was 10mL / kg; the blank control group and the model group were replaced with an equal amount of pure water, and after continuous gavage for 30d, they were fasted for 24h, and the eyeballs were removed for blood collection. After standing in a constant temperature water bath at 37°C for 30min, they were centrifuged at 3000r / min for 15min, and the supernatant was taken to obtain serum, and the concentrations of TC and TG were determined according to the operating specifications of the kit (TC and TG kits were purchased from Shanghai Jiachu Bioengineering Co., Ltd.).

[0073] After removing the eyeballs, the fat around the genitals was removed and weighed to calculate the fat coefficient of each group of mice.

[0074] Fat coefficient = fat mass around mouse genitals / mouse mass × 100%

[0075] Figure 1 This is a comparison chart of the fat coefficients of mice in each group after 30 days of administration. As can be seen from the figure, compared with the blank group, the fat coefficient of mice in the model group increased significantly; compared with the model group, the fat coefficient of mice in the control group 7-8 was slightly lower, and the fat coefficient of mice in the Example 2 group was the lowest, indicating that the Example 2 group had a significant inhibitory effect on fat accumulation and obesity induced by a high-fat diet.

[0076] Figure 2 and Figure 3 The figures are the comparison of serum TC and TG concentrations of mice in each group after 30 days of administration. As can be seen from the figure, compared with the blank group, the serum TC and TG concentrations of mice in the model group were significantly increased; compared with the model group, the serum TC and TG concentrations of mice in the comparative example 7-8 groups were slightly lower, and the serum TC and TG concentrations of mice in the example 2 group were the lowest, indicating that the example 2 had a significant inhibitory effect on the increase of TC and TG caused by a high-fat diet.

[0077] In summary, the Example 2 group has the effect of inhibiting fat accumulation and controlling body fat. It is confirmed that the synergistic effect of the fermented red bean liquid and the fermented dandelion root liquid is helpful to control body fat.

[0078] Example 3

[0079] This example demonstrates that the synergistic effect of orange peel fermentation liquid, adzuki bean fermentation liquid, and dandelion root fermentation liquid has an oxygen-deficiency tolerance effect.

[0080] This embodiment provides a Chinese medicine composition, and the specific steps are as follows:

[0081] S1. Weigh 15 g of orange peel, 20 g of red bean, 12 g of cassia seed, and 18 g of dandelion root, then wash, dry, and crush into 120-mesh particles for subsequent use;

[0082] S2. Bifidobacterium breve CICC-6204 was inoculated on glucose blood liver agar medium and cultured at 37°C for 18 hours to activate the strain. The activated Bifidobacterium breve was transferred to a 1000 mL glucose blood liver liquid medium shake flask and fermented for another 8 hours to obtain 1.5×10 8 cfu / mL of bacterial solution.

[0083] S3, Lactobacillus crispatus ATCC-33820 was inoculated on MRS agar medium and cultured at 37°C for 18 hours to activate the strain. The activated Lactobacillus crispatus was transferred to a 1000 mL MRS liquid medium shake flask and fermented for another 8 hours to obtain 2.2×10 8 cfu / mL of bacterial solution.

[0084] S4, after mixing the raw materials prepared in step S1, 18 mL of the bacterial solution of Bifidobacterium breve CICC-6204 prepared in step S2 and 18 mL of the bacterial solution of Lactobacillus crispatus ATCC-33820 prepared in step S3 were added, and the mixture was fermented anaerobically at 37°C for 22 h. After the fermentation was completed, 3% H 2 O 2 The solution inactivates Bifidobacterium breve and Lactobacillus crispatus in the fermentation product, thereby obtaining the desired Chinese medicine fermentation product;

[0085] S5. Add 10 times of 75% ethanol to the fermented Chinese medicine prepared in step S4, and distill in a constant temperature water bath at 80° C. for 3 h. Repeat twice, combine the filtrates and concentrate until there is no alcohol taste, thereby obtaining a Chinese medicine composition.

[0086] Comparative Example 9: In step S4, it is chosen not to add orange peel, and the remaining steps are kept consistent with Example 3.

[0087] Comparative Example 10: In step S4, no adzuki bean was added, and the remaining steps were consistent with Example 3.

[0088] Comparative Example 11: In step S4, dandelion root is not added, and the remaining steps are consistent with Example 3.

[0089] Study on the effect of Chinese medicine combination on hypoxia tolerance in mice

[0090] 50 SPF-grade ICR mice, weighing 18-22 g, were purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd. and were stably raised in a constant temperature and oxygen supply environment at 25±2°C for one week before the experiment officially began.

[0091] Mice were randomly divided into 5 groups, namely, negative control group, Example 3 group, and Comparative Examples 9-11 groups. Example 3 group and Comparative Examples 9-11 groups were gavaged with the corresponding Chinese medicine composition, which was diluted with pure water to a concentration of 5% v / v and then gavaged, with a gavage dose of 10 mL / (kg·d). The negative control group was gavaged with an equal amount of distilled water. The gavage was continued for 30 days. One hour after the last gavage, each group of mice was placed in a 250 mL ground-mouth bottle containing 5 g of soda lime (one mouse per bottle), and the bottle mouth was sealed with vaseline. A stopwatch was immediately used to time the mice, and respiratory arrest was used as an indicator to calculate the time when the mice died due to lack of oxygen.

[0092] Figure 4 It is a comparison chart of the hypoxia tolerance survival time of each group of mice at normal pressure. It can be seen intuitively from the figure that compared with the model group mice, there is no significant difference in the hypoxia tolerance survival time of the mice in the comparative examples 9-11 groups at normal pressure, and the hypoxia tolerance survival time of the mice in the example 3 group at normal pressure is significantly increased, indicating that the example 3 group can significantly prolong the hypoxia tolerance survival time of the mice at normal pressure, and has a good hypoxia tolerance effect, which confirms that the synergistic effect of the tangerine peel fermentation liquid, the red bean fermentation liquid, and the dandelion root fermentation liquid has a hypoxia tolerance effect.

[0093] The above-mentioned embodiments only express the specific implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the protection scope of the present invention. It should be pointed out that, for those skilled in the art, without departing from the concept of the present invention, any other changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principle of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. A Chinese medicine composition, characterized in that: The Chinese medicine composition is obtained by fermentation and extraction of Chinese medicinal materials; the Chinese medicinal materials include 6-20 parts of orange peel, 9-30 parts of red beans, 5-20 parts of cassia seeds, and 10-30 parts of dandelion roots by weight; the composite strain includes Bifidobacterium breve CICC-6204 and Lactobacillus crispatus ATCC-33820.

2. The Chinese medicine composition according to claim 1, which has an auxiliary protective effect on chemical liver damage, helps control body fat and improves hypoxia tolerance.

3. A method for preparing a Chinese medicine composition for controlling body fat and protecting the liver according to claims 1-2, wherein the specific preparation method is as follows: S1, weighing 6-20g of orange peel, 9-30g of red bean, 5-20g of cassia seed, and 10-30g of dandelion root, and then washing, drying and crushing them for subsequent use; S2. Bifidobacterium breve CICC-6204 was inoculated on glucose blood liver agar medium and cultured at 37°C for 12-18 hours to activate the strain; the activated Bifidobacterium breve was transferred to a 500-1000 mL glucose blood liver liquid medium shake flask and fermented for another 8 hours to finally obtain 1.2-1.5×10 8 cfu / mL of Bifidobacterium breve CICC-6204 bacterial solution; S3, Lactobacillus crispatus ATCC-33820 was inoculated on MRS agar medium and cultured at 37°C for 12-18h to activate the strain; the activated Lactobacillus crispatus was transferred to a 500-1000mL MRS liquid medium shake flask and fermented for another 8h to finally obtain 1.8-2.2×10 8 cfu / mL of Lactobacillus crispatus ATCC-33820 bacterial solution; S4, after mixing the raw materials prepared in step S1, adding 15-20 mL of the Bifidobacterium breve CICC-6204 bacterial solution prepared in step S2 and 15-20 mL of the Lactobacillus crispatus ATCC-33820 bacterial solution prepared in step S3, and fermenting anaerobically at 37° C. for 20-24 h. After the fermentation is completed, the Bifidobacterium breve and Lactobacillus crispatus in the fermentation product are inactivated with 3% H2O2 solution to obtain the desired Chinese medicine fermentation product; S5. Add 8-10 times of 60-75% ethanol to the fermented Chinese medicine prepared in step S4, distill in a constant temperature water bath for 3 hours, repeat twice, combine the filtrate and concentrate until there is no alcohol taste, so as to obtain a Chinese medicine composition.

4. The method for preparing a Chinese medicine composition for controlling body fat and protecting the liver according to claim 3, characterized in that: In the step S1, 10-15 g of orange peel, 15-20 g of red bean, 8-12 g of cassia seed, and 15-18 g of dandelion root are used.

5. The method for preparing a Chinese medicine composition for controlling body fat and protecting the liver according to claim 3, characterized in that: The components of the glucose blood-liver agar medium used in step S2 are: 3g beef extract powder, 10g prion peptone, 5g tryptone, 3g plant peptone, 5g yeast extract powder, 150mL liver extract, 10g glucose, 0.5g soluble starch, 10mL solution A (10g K2HPO4, 10g KH2PO4, 100mL distilled water), 5mL solution B (4g MgSO4·7H2O, 0.2g NaCI, 0.2g FeSO4·7H2O, 0.2g MnSO4·2H2O, 100mL distilled water), 10mL 5% L-cysteine ​​hydrochloride solution, 1g Tween-80, 15g agar, 50mL defibrinated sheep blood, and 825mL distilled water.

6. The method for preparing a Chinese medicine composition for controlling body fat and protecting the liver according to claim 3, characterized in that: The components of the MRS agar medium used in step S3 are: 10 g of peptone, 10 g of beef extract powder, 5 g of yeast extract powder, 20 g of glucose, 1 g of Tween, 5 g of sodium acetate, 2 g of triammonium citrate, 2 g of dipotassium hydrogen phosphate, 0.2 g of magnesium sulfate heptahydrate, 0.05 g of manganese sulfate monohydrate, 15 g of agar, and 1000 mL of distilled water.

7. The method for preparing a Chinese medicine composition for controlling body fat and protecting the liver according to claim 3, characterized in that: In step S4, the volume of the Bifidobacterium breve bacterial solution and the Lactobacillus crispatus bacterial solution are both 18 mL.

8. The method for preparing a Chinese medicine composition for controlling body fat and protecting the liver according to claim 3, characterized in that: The time of anaerobic fermentation in step S4 is 22 hours.

9. The method for preparing a Chinese medicine composition for controlling body fat and protecting the liver according to claim 3, characterized in that: The temperature of the constant temperature water bath distillation in step S5 is 80°C.

10. The method for preparing a Chinese medicine composition for controlling body fat and protecting the liver according to claim 3, characterized in that: In step S1, orange peel, adzuki bean, cassia seed and dandelion root are crushed into 80-120 mesh particles respectively.

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