Application of Bifidobacterium animalis

Abstract

The present invention relates to the field of Bifidobacterium animalis, and discloses the use of Bifidobacterium animalis or foods and drugs containing Bifidobacterium animalis for preventing and / or treating atrial fibrillation. The present invention fills the gap in the prior art for preventing and treating atrial fibrillation in offspring caused by cold exposure during pregnancy, and innovatively proposes the specific use of Bifidobacterium animalis for preventing and treating atrial fibrillation in offspring caused by cold exposure during pregnancy.

Description

Technical Field

[0001] The present invention relates to the field of Bifidobacterium, and more particularly to the application of Bifidobacterium animalis. Background Art

[0002] Numerous studies have confirmed that Bifidobacterium has potential effects in various diseases such as Parkinson's disease, colitis, diabetes, and tumors. Bifidobacterium can improve intestinal integrity, reduce endotoxin and trimethylamine levels, restore the content of Th17 and Treg cells in colonic lymph nodes and aorta, and significantly reduce the blood pressure level of salt-sensitive hypertensive rats. Bifidobacterium breve can alleviate cognitive impairment by regulating the gut microbiota and microbial metabolites in Alzheimer's disease mice. Bifidobacterium longum also improves ovariectomy-induced bone loss by enhancing the anti-osteoclast and immunomodulatory potential of regulatory B cells. Recent studies have shown that Bifidobacterium breve M-16V significantly inhibits Th2 and Th17 lymphocyte subsets in mice exposed to nano-polystyrene. At the same time, Bifidobacterium breve M-16V can activate MyD88 expression and promote the production of Th1-related cytokine IL-12. Bifidobacterium animalis MSMC83 improves oxidative stress and gut microbiota in D-galactose-induced rats.

[0003] Bifidobacterium has been increasingly widely used in various diseases. Research shows that compared with healthy infants of the same age, the abundance of Bifidobacterium in the intestines of infants with severe acute malnutrition is significantly reduced. The results of placebo-controlled randomized trials show that administering commercial infant Bifidobacterium strains (EVC001) and human milk oligosaccharides can significantly increase the abundance of Bifidobacterium in infants' intestines, reduce intestinal inflammation markers, promote infant weight gain, and improve the state of infant malnutrition. Meta-analysis shows that Bifidobacterium quadruple viable bacteria combined with mesalamine has satisfactory effects in the treatment of ulcerative colitis in China and has relatively high safety. A randomized double-blind experiment on the elderly with chronic constipation found that Bifidobacterium longum BB536 significantly improved the constipation symptoms of the elderly, suggesting that Bifidobacterium longum is a potential treatment strain for constipation. Bifidobacterium also plays an important role in the field of tumor treatment. The latest research report shows that developing a new drug delivery system with Bifidobacterium as a carrier can achieve targeted delivery of anti-tumor drugs and effectively inhibit tumor growth. Supplementing Bifidobacterium animalis can enhance the anti-tumor effect of irinotecan by regulating the intestinal flora and inflammatory response, suggesting that Bifidobacterium animalis can play an important role as a nutritional supplement in anti-tumor treatment. In addition, a single-center randomized double-blind trial proved that the intervention of Bifidobacterium animalis subsp. lactis MH-02 significantly relieved the symptoms of defecation difficulty in patients undergoing endoscopic mucosal resection of colonic polyps and shortened the recovery time of intestinal function. Bifidobacterium animalis has been approved as a medical probiotic and has various functions, including promoting the balance of the intestinal flora in infants and young children, improving resistance, preventing allergic diseases such as infantile eczema, reducing the infection rate of respiratory diseases, promoting the maturation of the immune system in infants, promoting growth and development, preventing diarrhea, preventing rotavirus infection, reducing the occurrence of constipation and necrotizing enterocolitis, etc.

[0004] In recent years, people have paid more attention to the impact of life exposures during pregnancy on the health of offspring. A large number of studies have proved that life exposures during pregnancy can change the composition and structure of the offspring's intestinal flora, thus increasing the risk of various diseases in the offspring. The present invention creatively discovers that cold exposure during pregnancy has a significant impact on the offspring's intestinal flora and increases the susceptibility of the offspring to atrial fibrillation. Summary of the Invention

[0005] To solve the above technical problems in the prior art, the present invention provides the application of Bifidobacterium animalis.

[0006] The specific solution adopted by the present invention is: the application of Bifidobacterium animalis or foods and drugs containing Bifidobacterium animalis for the prevention and / or treatment of atrial fibrillation.

[0007] The prevention and / or treatment of atrial fibrillation in offspring is atrial fibrillation in offspring caused by cold exposure during pregnancy.

[0008] The food is any one of dietary supplements, prebiotics, solid beverages, and dairy products.

[0009] The prebiotic is fructooligosaccharide or xylooligosaccharide or spirulina.

[0010] The solid beverage is fruit juice.

[0011] The dairy product is any one or several of yogurt, fermented milk with flavor or cheese.

[0012] The food is in the form of pills, capsules, tablets, powders, or liquids.

[0013] The dose of Bifidobacterium animalis contained in the drug is the viable count ≥ 4 billion per bag.

[0014] The present invention has the following beneficial effects compared with the prior art:

[0015] The present invention discloses that Bifidobacterium animalis can prevent the occurrence of atrial fibrillation in offspring caused by cold exposure during pregnancy by regulating immune homeostasis, reducing circulatory and atrial inflammatory responses, and Bifidobacterium animalis can be used as a target for the prevention and treatment of atrial fibrillation. Bifidobacterium animalis is applicable to the occurrence of atrial fibrillation in offspring caused by cold exposure during pregnancy, regulates the body's immune homeostasis, and reduces the body's inflammatory level. The present invention fills the gap in the prior art for preventing and treating atrial fibrillation in offspring caused by cold exposure during pregnancy, and innovatively proposes the specific application of Bifidobacterium animalis for preventing and treating atrial fibrillation in offspring caused by cold exposure during pregnancy. Description of the Drawings

[0016] Figure 1 Results of electrophysiological detection of the atria of rats in the room-temperature offspring group and the cold offspring group - representative electrocardiogram of atrial pacing in rats;

[0017] Figure 2 Results of electrophysiological detection of the atria of rats in the room-temperature offspring group and the cold offspring group - atrial fibrillation induction rate;

[0018] Figure 3 Results of electrophysiological detection of the atria of rats in the room-temperature offspring group and the cold offspring group - atrial fibrillation duration;

[0019] Figure 4 Results of electrophysiological detection of the atria of rats in the room-temperature offspring group and the cold offspring group - effective refractory period of the atria (n = 10);

[0020] Figure 5 Results of atrial fibrosis and apoptosis in the room-temperature offspring group and the cold offspring group of rats - representative diagram of collagen deposition observed by Masson staining in the atrial tissues of rats in the room-temperature offspring group and the cold offspring group, magnification = 200 times, scale bar = 50 μm;

[0021] Figure 6 Results of atrial fibrosis and apoptosis in the room-temperature offspring group and the cold offspring group of rats - statistical chart of collagen deposition;

[0022] Figure 7For the atrial fibrosis and apoptosis of room temperature offspring and cold offspring rats - Representative diagrams of apoptotic cells in the atria of rats in the room temperature offspring group and the cold offspring group shown by Tunel staining;

[0023] Figure 8 For the atrial fibrosis and apoptosis of room temperature offspring and cold offspring rats - Diagram of the percentage of apoptotic cells (n = 4);

[0024] Figure 9 For the expression of proteins related to atrial fibrosis and apoptosis in room temperature offspring and cold offspring rats - Representative bands of TGF-β1 and α-SMA protein expression in rat atrial tissue;

[0025] Figure 10 For the expression of proteins related to atrial fibrosis and apoptosis in room temperature offspring and cold offspring rats - Quantitative statistical diagram of TGF-β1 and α-SMA protein expression;

[0026] Figure 11 For the expression of proteins related to atrial fibrosis and apoptosis in room temperature offspring and cold offspring rats - Representative bands of Bcl2 and BAX protein expression in rat atria;

[0027] Figure 12 For the expression of proteins related to atrial fibrosis and apoptosis in room temperature offspring and cold offspring rats - Quantitative statistical diagram of Bcl2 and BAX protein expression (n = 6);

[0028] Figure 13 For reducing the susceptibility of cold offspring rats to atrial fibrillation by supplementing Bifidobacterium animalis - Representative electrocardiogram of atrial pacing in rats;

[0029] Figure 14 For reducing the susceptibility of cold offspring rats to atrial fibrillation by supplementing Bifidobacterium animalis - Diagram of the induction rate of atrial fibrillation in rats;

[0030] Figure 15 For reducing the susceptibility of cold offspring rats to atrial fibrillation by supplementing Bifidobacterium animalis - Duration of atrial fibrillation in rats;

[0031] Figure 16 For the atrial fibrosis and apoptosis of rats in the solvent control group and the Bifidobacterium animalis supplementation group - Representative diagram of Masson staining of rat atrial tissue

[0032] Figure 17 For the atrial fibrosis and apoptosis of rats in the solvent control group and the Bifidobacterium animalis supplementation group - Statistical diagram of collagen deposition

[0033] Figure 18 For the atrial fibrosis and apoptosis of rats in the solvent control group and the Bifidobacterium animalis supplementation group - Representative diagram of Tunel staining of rat atrial tissue

[0034] Figure 19For the percentage of atrial fibrosis and apoptosis - apoptotic cells in rats of the solvent control group and the group supplemented with Bifidobacterium animalis (n = 4);

[0035] Figure 20 For the expression of proteins related to atrial fibrosis and apoptosis in cold offspring rats of the solvent control group and the group supplemented with Bifidobacterium animalis - representative protein bands of TGF-β1 and α-SMA in rat atria;

[0036] Figure 21 For the expression of proteins related to atrial fibrosis and apoptosis in cold offspring rats of the solvent control group and the group supplemented with Bifidobacterium animalis - quantitative statistical charts of TGF-β1 and α-SMA protein expression;

[0037] Figure 22 For the expression of proteins related to atrial fibrosis and apoptosis in cold offspring rats of the solvent control group and the group supplemented with Bifidobacterium animalis - representative protein bands of Bcl2 and BAX in rat atria;

[0038] Figure 23 For the expression of proteins related to atrial fibrosis and apoptosis in cold offspring rats of the solvent control group and the group supplemented with Bifidobacterium animalis - quantitative statistical charts of Bcl2 and BAX protein expression (n = 6);

[0039] Figure 24 For the percentage of immune T cell subsets in rats of the solvent control group and the group supplemented with Bifidobacterium animalis - percentages of CD4 T cells and CD8 T cells in rat spleens;

[0040] Figure 25 For the percentage of immune T cell subsets in rats of the solvent control group and the group supplemented with Bifidobacterium animalis - statistical chart of the CD4 / CD8 T cell ratio in rat spleens;

[0041] Figure 26 For the percentage of immune T cell subsets in rats of the solvent control group and the group supplemented with Bifidobacterium animalis - percentages of CD4 T cells and CD8 T cells in rat blood;

[0042] Figure 27 For the percentage of immune T cell subsets in rats of the solvent control group and the group supplemented with Bifidobacterium animalis - statistical chart of the CD4 / CD8 T cell ratio in rat blood (n = 4);

[0043] Figure 28 For the levels of inflammatory factors in rat plasma and atria - levels of IFN-γ, TNF-α, IL-5 and IL-6 in rat plasma (n = 8)

[0044] Figure 29 For the levels of inflammatory factors in rat plasma and atria - levels of IFN-γ, TNF-α, IL-5 and IL-6 in rat atria (n = 8). Detailed implementation methods

[0045] To make the objectives, technical solutions and advantages of the present invention more clear and understandable, the present invention will be described below through specific embodiments shown in the accompanying drawings. However, it should be understood that these descriptions are merely exemplary and are not intended to limit the scope of the present invention. In addition, in the following description, the descriptions of well-known structures and technologies are omitted to avoid unnecessarily confusing the concepts of the present invention.

[0046] Combined with the attached Figure 1-29 , the present invention discloses the application of Bifidobacterium animalis (children's probiotic powder, belonging to solid beverage) or foods and drugs containing Bifidobacterium animalis in the prevention and / or treatment of filial atrial fibrillation. The prevention and / or treatment of filial atrial fibrillation is filial atrial fibrillation caused by cold exposure during pregnancy. The food is any one of dietary supplements, prebiotics, solid beverages, and dairy products. The prebiotic is fructooligosaccharide or xylooligosaccharide or spirulina. The solid beverage is fruit juice. The dairy product is any one or several of yogurt, fermented milk with flavor, or cheese. The food is in the form of pills, capsules, tablets, powders, or liquids. The dose of Bifidobacterium animalis contained in the drug is viable count ≥ 4 billion / bag (Chr. Hansen Bb-12).

[0047] Example 1

[0048] The application of Bifidobacterium animalis (Chr. Hansen BB-12 children's probiotic powder, Chr. Hansen Bb-12) in the prevention and treatment of filial atrial fibrillation caused by cold exposure during pregnancy.

[0049] Example 2

[0050] The application of adding Bifidobacterium animalis to yogurt in the prevention and treatment of filial atrial fibrillation caused by cold exposure during pregnancy.

[0051] Example 3

[0052] The application of adding Bifidobacterium animalis to fruit juice in the prevention and treatment of filial atrial fibrillation.

[0053] Example 4

[0054] The combined use of Bifidobacterium and antispasmodics can treat adult inflammatory bowel disease (viable count ≥ 1.0 * 10 7 CFU / capsule, twice a day, 2-4 capsules each time).

[0055] Combined with Examples 1-3, to explore the effect of cold exposure during pregnancy on the susceptibility of offspring rats to atrial fibrillation, the present invention constructs a rat model of cold exposure during pregnancy. After the rats are pregnant, they are placed in a constant temperature incubator at 4±1°C for 18 days of cold exposure, and then placed at room temperature for delivery and feeding of the offspring. The offspring born to pregnant rats with cold exposure during pregnancy are cold offspring rats. After the offspring rats reach adulthood, electrophysiological detection of the atria is performed on room temperature offspring and cold offspring rats to observe the susceptibility to atrial fibrillation, and atrial pathological staining and Western blot are performed to observe the indicators related to atrial structural remodeling.

[0056] To explore the effect of Bifidobacterium on atrial fibrillation in cold offspring rats, the present invention randomly divides the cold offspring rats into two groups. One group is given a control solvent, and the other group is given intragastric administration of Bifidobacterium animalis (2×10 9 CFU / mL). The rats are given 100 μl of Bifidobacterium animalis solution by intragastric administration at 1-3 weeks of age and 500 μl of Bifidobacterium animalis solution by intragastric administration at 3-8 weeks of age, once a week. After the offspring rats reach adulthood, electrophysiological detection of the atria is performed to observe changes in the atrial fibrillation induction rate and duration; atrial pathological staining is used to observe the levels of atrial fibrosis and apoptosis; Western blot is used to detect the expression levels of the atrial fibrosis-related proteins TGF-β1 and α-SMA and the apoptosis-related indicators BAX and Bcl2; flow cytometry is used to detect the levels of CD4+, CD8+ and Treg cells in the spleen and circulation; ELISA is used to detect the levels of the cardiac and circulating inflammatory factors IL5, IL-6, TNF-α and IFN-γ.

[0057] The test results are as follows: Cold exposure during pregnancy leads to an increase in the susceptibility of offspring rats to atrial fibrillation.

[0058] Combined with the appendix Figure 1-4 , in the present invention, electrophysiological detection of the atria is performed on adult room temperature offspring and cold offspring rats. The results show that compared with room temperature offspring rats, the atrial fibrillation induction rate and duration of cold offspring rats are significantly increased (P = 0.0198), and the duration is significantly prolonged (P = 0.0399), and the atrial effective refractory period has no obvious change.

[0059] Effect of cold exposure during pregnancy on atrial myocardial fibrosis and apoptosis in offspring rats

[0060] Atrial myocardial fibrosis and apoptosis are important markers of atrial structural remodeling. In the present invention, Masson staining and Tunel staining are performed on the atrial tissues of cold offspring rats and room temperature offspring rats. The results of Masson staining show that compared with room temperature offspring rats, the collagen deposition in the atrial tissues of cold offspring rats is increased (P = 0.0011). Tunel staining shows that compared with room temperature offspring rats, the percentage of apoptotic cells in the atria of cold offspring rats is significantly increased (P = 0.0072). Combined with the appendix Figure 5-8, The results suggest that cold exposure during pregnancy increases the levels of atrial fibrosis and apoptosis in offspring rats.

[0061] Effect of Cold Exposure during Pregnancy on Atrial Fibrosis and Apoptosis-Related Proteins in Offspring Rats

[0062] Combined with Appendix Figure 9-12 , the present invention further detected atrial fibrosis and apoptosis-related proteins in room-temperature offspring and cold-exposed offspring rats. Compared with room-temperature offspring rats, the expressions of atrial fibrosis-related proteins TGF-β1 (P = 0.0053) and α-SMA (P = 0.0094), and the pro-apoptotic-related protein BAX (P = 0.0012) were significantly up-regulated in cold-exposed offspring rats, while the expression of the anti-apoptotic protein Bcl2 was significantly down-regulated (P = 0.0146).

[0063] Supplementation with Bifidobacterium animalis significantly reduces the susceptibility to atrial fibrillation in cold-exposed offspring rats.

[0064] Combined with Appendix Figure 13-15 , the present invention performed atrial electrophysiological detection on adult control rats and Bifidobacterium animalis group rats. The results showed that compared with control rats, the induction of atrial fibrillation was significantly reduced (P = 0.0406) and the duration of atrial fibrillation was significantly shortened (P = 0.0368) in the Bifidobacterium animalis group rats, suggesting that Bifidobacterium animalis can reduce the susceptibility to atrial fibrillation in cold-exposed offspring rats. Figure 15 In Figure 15 , Vehicle represents cold-exposed offspring rats in the control group given solvent, and B.animals represents cold-exposed offspring rats given Bifidobacterium; n = 8.

[0065] Supplementation with Bifidobacterium animalis improves atrial fibrosis and apoptosis levels in cold-exposed offspring

[0066] Combined with Appendix Figure 16-19 , after electrophysiological detection of adult control and Bifidobacterium animalis rats, atrial tissues were taken for pathological staining. The results of Masson staining showed that compared with control rats, the atrial collagen deposition was significantly reduced and the degree of fibrosis was alleviated (P = 0.0012) in the Bifidobacterium animalis group rats; the results of atrial Tunel staining showed that compared with control rats, the percentage of apoptotic cells in the atria of Bifidobacterium animalis group rats was significantly decreased (P = 0.0005). These results indicate that supplementation with Bifidobacterium animalis can significantly improve atrial fibrosis and apoptosis levels in cold-exposed offspring rats.

[0067] Supplementation with Bifidobacterium animalis reduces the expression of atrial fibrosis and apoptosis-related proteins in cold-exposed offspring rats

[0068] Combined with Appendix Figure 20-23, Western blot detection was performed on the atrial tissues of two groups of rats in this invention. The results showed that, compared with the solvent control group, the expressions of TGF-β1 (P < 0.0001) and α-SMA (P = 0.0002) in the atria of cold offspring rats in the group supplemented with Bifidobacterium animalis were significantly down-regulated, the expression of BAX (P = 0.0002) was decreased, and the expression of Bcl2 (P = 0.0356) was significantly increased. These results suggest that supplementing Bifidobacterium animalis can reduce the levels of atrial fibrosis and apoptosis in cold offspring rats.

[0069] Supplementation with Bifidobacterium animalis increases the CD4 / CD8 T cell ratio in cold offspring rats

[0070] Combined with the appendix Figure 24-27 , Bifidobacterium has an immunomodulatory effect, and immune-inflammatory disorders are one of the important mechanisms of atrial structural remodeling. To explore the effect of Bifidobacterium on immune T cells in cold offspring rats, flow cytometry was used in this invention to detect the spleens and blood of cold offspring rats in the solvent control group and the group supplemented with Bifidobacterium animalis. The results showed that, compared with the solvent control group, the proportion of CD4 / CD8 T cells in the spleens (P = 0.0116) and blood (P = 0.0202) of cold offspring rats in the group supplemented with Bifidobacterium animalis was significantly increased, suggesting that supplementing Bifidobacterium animalis reverses the immune imbalance in offspring rats caused by cold exposure during pregnancy.

[0071] Supplementation with Bifidobacterium animalis reduces the levels of plasma and atrial inflammatory factors in cold offspring rats

[0072] Combined with the appendix Figure 28-29 , to further verify whether supplementing Bifidobacterium animalis can improve the inflammatory level in rats, ELISA kits were used in this invention to detect inflammatory factors in the plasma and atrial tissues of rats. The results showed that, compared with the solvent control group, the levels of IFN-γ (P = 0.0004), TNF-α (P = 0.0015), and IL-6 (P < 0.0001) in the plasma of cold offspring rats in the group supplemented with Bifidobacterium were significantly reduced, and the levels of IFN-γ (P = 0.0061), IL-5 (P = 0.001), and IL-6 (P = 0.0002) in the atrial tissues were also significantly reduced. This suggests that supplementing Bifidobacterium animalis can reduce the systemic and local atrial inflammatory factor levels in cold offspring rats and improve the high-inflammatory state of the offspring rats caused by cold exposure during pregnancy.

[0073] The present invention demonstrates that Bifidobacterium animalis can prevent the occurrence of atrial fibrillation in offspring caused by cold exposure during pregnancy by regulating immune homeostasis, reducing circulatory and atrial inflammatory responses, suggesting that it can be used as a target for the prevention and treatment of atrial fibrillation. Previous studies have shown that Bifidobacterium can regulate the development of the neonatal immune system and reduce the risk of immune-related diseases. Bifidobacterium animalis is applicable to the occurrence of atrial fibrillation in offspring caused by cold exposure during pregnancy, regulates the body's immune homeostasis, and reduces the body's inflammatory level.

[0074] The above drawings and explanations are only a specific embodiment of the present invention, but the specific protection scope of the present invention is not limited to the above explanations. Any simple replacement or change within the technical concept disclosed by the present invention and based on the technical solution of the present invention should be within the protection scope of the present invention.

Claims

1. Use of Bifidobacterium animalis in the preparation of a medicament for preventing atrial fibrillation in offspring caused by cold exposure during pregnancy, characterized in that, The Bifidobacterium animalis is Bb-12 from Chr. Hansen, and the dosage of Bifidobacterium animalis contained in the drug is that the viable count is ≥ 4 billion per bag.

Images