Application of bacteroides fragilis 839 in preparation of health food, food composition or medicine for improving hepatotoxicity caused by antituberculosis medicine

Abstract

The invention provides application of bacteroides fragilis 839 in preparation of health food, food composition or medicine for improving hepatotoxicity caused by antituberculosis medicine. The bacteroides fragilis BF839 is adopted for the first time in the world to improve the mouse hepatotoxicity caused by the antituberculous drug, and it is found that the bacteroides fragilis BF839 can reduce or relieve the mouse hepatotoxicity caused by the antituberculous drug and has the advantages of being efficient and safe.

Description

Technical Field

[0001] The present invention relates to the use of Bacteroides fragilis 839 in the preparation of health-care foods, food compositions or medicines for improving hepatotoxicity caused by anti-tuberculosis drugs. Background Art

[0002] Tuberculosis is one of the most important infectious diseases in the world. Under the standard anti-tuberculosis drug treatment regimen (isoniazid + rifampicin + pyrazinamide + ethambutol), approximately 85% of patients will have sputum bacteria converted to negative within 2 months of treatment. However, various types of adverse drug reactions may occur during anti-tuberculosis treatment. Among them, drug-induced liver injury caused by anti-tuberculosis drugs is the most common, and its incidence has shown a clear upward trend worldwide, from 5.07% in 1999 to 29.40% in 2020 (Nannan W, Xinyu C, Zhuolu H, et al. Incidence and Temporal Trend of Antituberculosis Drug-Induced Liver Injury: A Systematic Review and Meta-Analysis. J Trop Med, 2022: 266878). The incidence rate among hospitalized patients can be as high as 40% (Gu Jin, Lin Minggui, Tang Shenjie. Interpretation of Updated Key Points in the "Guidelines for the Diagnosis and Treatment of Drug-Induced Liver Injury Caused by Anti-Tuberculosis Drugs (2024 Edition)" [J / OL]. Journal of Tuberculosis and Lung Diseases, 1-8). Commonly used hepatoprotective drugs in clinical practice include bicyclol, glycyrrhizic acid derivatives, glutathione, silymarin, polyene phosphatidylcholine, and ursodeoxycholic acid, but their overall efficacy is poor. Some patients are forced to discontinue anti-tuberculosis treatment, thus compromising the overall effectiveness of tuberculosis treatment. More effective strategies to improve liver function are needed.

[0003] The liver is the core of material metabolism and has an important impact on human body functions. It is also an important organ for drug or poison metabolism and detoxification. In recent years, due to the close physiological and pathological connection between the intestine and the liver, the concept of the "gut-liver axis" has been widely accepted. Maintaining the balance of the intestinal ecology may support the liver's anti-inflammatory and antioxidant responses and prevent acute hepatotoxicity. Bacteroides fragilis 839 (Totem Prebiotic Liquid) is a Chinese patented strain (patent number of the same family: CN1029857C) cultured from neonatal feces by Chinese microbiologist Zhang Jijie in September 1983. Preliminary studies have found that it has the effect of improving the function of immune T cells, preventing and treating intestinal and respiratory diseases, and is a non-toxic and harmless Bacteroides fragilis. However, research on improving the hepatotoxicity of mice caused by anti-tuberculosis drugs has not been reported. The present invention found that it can significantly improve the hepatotoxicity caused by anti-tuberculosis drugs. Summary of the Invention

[0004] The object of the present invention is to provide the use of Bacteroides fragilis 839 in the preparation of health foods, food compositions or medicines for improving hepatotoxicity caused by anti-tuberculosis drugs.

[0005] To achieve the above-mentioned purpose, the technical solution adopted is: use of Bacteroides fragilis 839 in the preparation of health foods, food compositions or medicines for improving hepatotoxicity caused by anti-tuberculosis drugs.

[0006] Preferably, the health food, food composition or medicine is in the form of tablets, capsules, oral liquid or lyophilized powder.

[0007] Preferably, each 1 ml or 10 g of health food, food composition or medicine contains 10 8 CFU.

[0008] Preferably, the health food, food composition or medicine comprises Bacteroides fragilis 839 and an acceptable carrier thereof.

[0009] Preferably, the acceptable carrier comprises at least one of skim milk, lactose, glucose, sucrose, inulin, oligofructose, sorbitol, mannose, trehalose, starch, gum arabic, calcium phosphate, alginate, gelatin, calcium silicate, fine crystalline cellulose, polyvinyl pyrrolidone, water, syrup, methylcellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, magnesium stearate and mineral oil.

[0010] Preferably, the anti-tuberculosis drug includes at least one of rifampicin, isoniazid, pyrazinamide and ethambutol.

[0011] Preferably, the improving hepatotoxicity caused by anti-tuberculosis drugs refers to reducing or alleviating the hepatotoxicity caused by anti-tuberculosis drugs.

[0012] Beneficial effects:

[0013] This invention is the first in the world to use Bacteroides fragilis BF839 to improve the liver toxicity of mice caused by anti-tuberculosis drugs. It was found that Bacteroides fragilis BF839 can reduce the liver toxicity of mice caused by anti-tuberculosis drugs, and is highly effective and safe. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 : Effects of BF839 on liver index in mice. Note: **: P < 0.01 compared with the drug model group and the normal control group. Note: A: Normal control group; B: Drug model group; C: Drug + BF839 group; D: BF839 group.

[0015] Figure 2Detection of antioxidant markers in mouse liver tissue; Note: A: Normal control group; B: Drug model group; C: Drug + BF839 group; D: BF839 group. (A) Changes in SOD activity in mouse liver tissue, *: Compared with group A, P < 0.05, #: Compared with group B, P < 0.05; (B) MDA content in mouse liver tissue, **: Compared with group A, P < 0.01; #: Compared with group B, P < 0.05.

[0016] Figure 3 Detection of inflammatory markers in mouse liver tissue; Note: A: Normal control group; B: Drug model group; C: Drug + BF839 group; D: BF839 group. (A) TNF-α content in mouse liver, **: Compared with group A, P < 0.01; #: Compared with group B, P < 0.05. (B) IL-6 content in mouse liver, **: Compared with group A, P < 0.01; ##: Compared with group B, P < 0.01. (C) IL-1β content in mouse liver, *: Compared with group A, P < 0.05; #: Compared with group B, P < 0.05.

[0017] Figure 4 :Effects of BF839 on the pathological morphology of liver tissue in mice; Note: A: normal control group; B: drug model group; C: BF839+drug model group; D: BF839 group. DETAILED DESCRIPTION

[0018] In order to better illustrate the purpose, technical solutions and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments.

[0019] The Bacteroides fragilis 839 used in this example is Bacteroides fragilis 839BF839, which was deposited with the China General Microbiology Center (CGMCC) on May 3, 1990, with the deposit number CGMCC No. 0157. The deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The Bacteroides fragilis 839 bacterial solution (Totem Probiotic Solution) was provided by Dalian Totem Bioengineering Co., Ltd.

[0020] Example 1 Experiment on the Effect of Bacteroides fragilis BF839 on Anti-TB Drug-Induced Hepatotoxicity in Mice 1. Animal Model Preparation and Grouping

[0021] Modeling groups: 40 healthy adult male Kunming mice were fed an adaptive diet for one week, and their health was observed. Five animals were placed in each cage, and the cages were lined with sterilized cork shavings. The animal laboratory was maintained at a temperature of 23-25°C, a relative humidity of 50-60%, approximately 12 air changes per hour, and a 12-hour light / 12-hour dark cycle. Animals were fed dry, stick-shaped chow and had free access to water. The mice were randomly divided into a normal control group, a drug model group, a BF839+drug model group, and a BF839 group, with 10 mice in each group.

[0022] 2. Intervention methods

[0023] The drug model group was gavaged with anti-tuberculosis drugs (rifampicin 0.2 mg / kg bw + isoniazid 0.1 mg / kg bw + pyrazinamide 0.42 mg / kg bw + ethambutol 0.2 mg / kg bw), 10 ml / kg bw; the bacterial solution protection group was given drugs and supplemented with BF839 bacterial solution (10 8 cfu / ml), 0.2 ml / d was administered orally to each mouse; the BF839 group was given only BF839 solution, while the control group was given an equal volume of distilled water orally for 30 consecutive days.

[0024] The day before the end of the experiment, mouse feces were collected in a sterile environment and stored at -80°C for 16s rDNA sequencing analysis. After the last dose, the animals were fasted but not watered for 16 hours, and blood was collected from the eyeball under anesthesia. The serum was separated and stored at -80°C for the detection of liver biochemical indicators. After the animals were sacrificed, the liver and spleen were removed and weighed. The right lobe of the liver and jejunum were placed in formalin solution, fixed, and sliced ​​for HE staining to observe the pathological changes in liver tissue. The remaining liver was packaged and stored at -80°C for the detection of the antioxidant indicators SOD, MDA and inflammatory response factors TNF-α, IL-6, and IL-1β.

[0025] 3. Observation indicators and detection methods

[0026] 3.1 Calculation of mouse liver and spleen index

[0027] After removing the mouse liver and spleen, rinse the surface of the organ with saline, drain the excess water, and weigh them using an electronic balance to calculate the organ index. The organ index calculation formula is: Organ coefficient = organ mass / total body weight.

[0028] 3.2 Liver biochemical index detection

[0029] Serum ALT, AST, ALP, DBiL, TBiL, and TBA levels were measured using a Hitachi 7600-110 biochemical automatic analyzer. Liver MDA, GSH, and SOD levels were measured according to the kit instructions, and TNF-α, IL-6, and IL-1β levels were measured using Elisa.

[0030] 3.3 Observation of pathological changes in mouse liver tissue

[0031] The tissues were routinely paraffin-embedded, sectioned, and stained with HE. The pathological changes of liver tissue were observed under a light microscope.

[0032] 4. Statistical processing

[0033] SPSS 13.0 software was used, and all experimental data were expressed as x ± s. One-way analysis of variance was performed, and p < 0.05 was considered statistically significant.

[0034] 5. Results

[0035] 5.1 Effects of BF839 on Liver and Spleen Indexes in Mice

[0036] Compared with the normal group, the liver organ index of the model group increased significantly, and the difference was statistically significant (P<0.01), indicating that the liver was damaged and swollen and the weight increased, and the anti-tuberculosis drug-induced liver injury model was established; compared with the model group, the liver organ index of the BF839 bacterial solution group showed a downward trend, but the difference was not statistically significant. Figure 1 Compared with the control group, the spleen organ coefficients of mice in each group did not change significantly (P>0.05) 5.2 Effects of BF839 on the levels of ALT, AST, ALP, DBiL, TBiL, and TBA in mouse serum

[0037] Compared with the normal control group, the serum AST and ALT activities of mice in the drug model group were significantly increased (P<0.05, P<0.01). Compared with the drug model group, the serum ALT and AST activities of mice in the BF839+drug model group were significantly decreased (P<0.05, P<0.01), indicating that BF839 has a certain hepatoprotective effect. The results are shown in Table 1.

[0038] Table 1. Effects of BF839 on serum transaminase activity in mice (n=10, x±s)

[0039]

[0040] Note: *: compared with the normal control group, P < 0.05; **: compared with the normal control group, P < 0.01; #: compared with the drug model group, P < 0.05, ##: compared with the drug model group, P < 0.01

[0041] Compared with the normal control group, the ALP, DBiL, TBiL, and TBA levels in the drug model group were significantly increased (P < 0.01). Compared with the drug model group, the BF839+drug model group, with the exception of ALP, showed a significant decrease in all other indicators (P < 0.05), suggesting that BF839 has a protective effect against liver damage in mice. The results are shown in Table 2.

[0042] Table 2. Effects of BF839 on direct bilirubin, total bilirubin, total bile acid, and alkaline phosphatase in mouse serum (n=10, x±s)

[0043]

[0044] Note: Compared with the normal group, *: compared with the normal control group, P < 0.05; **: compared with the normal control group, P < 0.01; #: compared with the drug model group, P < 0.05, ##: compared with the drug model group, P < 0.01

[0045] Effects of 5.3BF839 on SOD activity and MDA levels in mouse liver tissue

[0046] Compared with the normal control group, the SOD activity unit of the drug model group was significantly decreased (P < 0.05), and the MDA level was significantly increased (P < 0.01); compared with the drug model group, the SOD of the drug + BF839 bacterial solution group was significantly increased (P < 0.05), and the MDA level was significantly decreased (P < 0.05), and the differences were statistically significant (P < 0.05); compared with the normal control group, the SOD of the BF839 group was slightly increased, and the MDA level was slightly decreased, but there was no statistical significance (P > 0.05); the results are shown in Figure 2 .

[0047] Effects of 5.4BF839 on the levels of inflammatory factors TNF-α, IL-6, and IL-1β in mouse liver tissue

[0048] Compared with the normal control group, the levels of inflammatory factors TNF-α, IL-6, and IL-1β in the liver tissue of mice in the drug model group were significantly increased. Compared with the drug model group, the levels of TNF-α, IL-6, and IL-1β in the drug + BF839 group were significantly decreased (P < 0.01 or (P < 0.05); the results are shown in Figure 3 .

[0049] Effects of 5.5BF839 on pathological changes of liver tissue in mice

[0050] The normal control group ( Figure 4 A) The liver lobule structure is normal, most of the liver cells have normal morphology, and a few have mild edema. Figure 4B) Hepatocytes showed varying degrees of edema (light staining of the hepatocyte cytoplasm) and fatty degeneration (fatty vacuoles were seen in the hepatocyte cytoplasm). Figure 4 C) Some hepatocytes showed mild edema and fatty degeneration. Figure 4 D) The liver lobule structure is normal, and the liver cells show moderate to severe edema. Compared with the normal control group, the model group showed changes in liver damage, and the model was established; the degree of liver damage in the BF839+ model group mice was alleviated; see Figure 4 .

[0051] This invention is the first in the world to use Bacteroides fragilis BF839 to improve the liver toxicity of mice caused by anti-tuberculosis drugs. It was found that Bacteroides fragilis BF839 can reduce or alleviate the liver toxicity of mice caused by anti-tuberculosis drugs, and is highly effective and safe.

[0052] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. Use of Bacteroides fragilis 839 in the preparation of health foods, food compositions or medicines for improving hepatotoxicity caused by anti-tuberculosis drugs.

2. The use according to claim 1, characterized in that The health food, food composition or medicine is in the form of tablets, capsules, oral liquids or freeze-dried powders.

3. The use according to claim 1, characterized in that Each 1ml or 10g of health food, food composition or medicine contains 10 live bacteria of Bacteroides fragilis 839 8 CFU.

4. The use according to claim 1, characterized in that The health food, food composition or medicine includes Bacteroides fragilis 839 and an acceptable carrier thereof.

5. The use according to claim 4, characterized in that The acceptable carrier includes at least one of skim milk, lactose, glucose, sucrose, inulin, oligofructose, sorbitol, mannose, trehalose, starch, gum arabic, calcium phosphate, alginate, gelatin, calcium silicate, fine crystalline cellulose, polyvinyl pyrrolidone, water, syrup, methylcellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, magnesium stearate and mineral oil.

6. The use according to claim 1, characterized in that The anti-tuberculosis drug includes at least one of rifampicin, isoniazid, pyrazinamide and ethambutol.

7. The use according to claim 1, characterized in that The improvement of hepatotoxicity caused by anti-tuberculosis drugs refers to reducing or alleviating the hepatotoxicity caused by anti-tuberculosis drugs.

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