Medicine for preventing and relieving toxicity of alternariol and application thereof

Abstract

The invention provides a medicine for preventing or relieving toxicity of alternariol and application of the medicine. The medicine at least comprises tauroursodeoxycholic acid or pharmaceutically acceptable salt of tauroursodeoxycholic acid. The invention also provides application of tauroursodeoxycholic acid or pharmaceutically acceptable salts thereof in preparation of drugs for preventing or relieving toxicity of alternariol. The application disclosed by the invention has the beneficial effects that the TUDCA treatment recovers a part of damaged liver functions after chronic liver injury caused by alternariol toxicity, inhibits the expression level of endoplasmic reticulum stress markers, relieves liver cell apoptosis, reduces the liver coefficient, reduces the fibrosis degree of liver tissues, inhibits inflammatory response and reduces the serum transaminase level; the TUDCA and the resveratrol are combined for use, so that the use amount of the TUDCA can be reduced, and the production cost is remarkably reduced; the feed additive can also be applied to animal husbandry and has the effect of protecting the health of livestock and poultry as a feed additive.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, relates to the field of biomedical technology, and more specifically to a drug for preventing and alleviating the toxicity of cross-linked spore phenol and an application thereof. Background Art

[0002] Alternariol (AOH), also known as Alternaria, is a secondary toxic metabolite mainly produced by Alternaria, and is widely found in agricultural products such as grains, fruits, vegetables and nuts around the world. Studies have shown that the proportion of AOH contamination in wheat in my country is 47%, with a median contamination concentration of 7.9 µg / kg, the average contamination level of AOH in apples is 935.96 µg / kg, and the AOH contamination level in wolfberries in the northwest region is as high as 2607 μg / kg. In addition, 78% of food samples such as ketchup, tomatoes, sunflower oil and flour in the European market are contaminated with AOH, with average and maximum contamination concentrations of 6.7 µg / kg and 20.8 µg / kg, respectively. The European Union (EU) has set indicative levels (2-30 μg / kg) for AOH in grains, tomato products, sunflower oil and sesame. Although the indicative level is not a safe level, 22.2% of food samples still exceed this level.

[0003] At present, AOH has become one of the "emerging mycotoxins" that the world focuses on. The contamination of food is a serious problem faced by the whole world and has a high exposure risk. The European Food Safety Authority (EFSA) has put forward scientific opinions on the risks of AOH in food to public health, and set the threshold of toxicological concern (TTC) of AOH at 2.5 ng / kg bw / d based on genotoxicity. According to recent statistics from EFSA, the population most exposed to AOH is young children, with an average exposure level of 3.8 to 71.6 ng / kg bw / d. This level is higher than the TTC (2.5 ng / kg bw / d). A study on human exposure in Beijing in 2012 in my country showed that the exposure level of AOH was LOQ to 32.9 μg / L. Due to the high stability of AOH, it is widely present in the entire food processing chain and is difficult to remove. Once ingested by humans and animals, it will have toxic effects on the digestive system, immune system and reproductive system, leading to acute and chronic poisoning and disease. However, no method has been found to alleviate the toxicity of cross-linked spore phenol. As the main metabolic and detoxification site for toxic substances, the liver is very vulnerable to the harm of toxic substances. The liver is one of the main organs affected by AOH. Previous studies have shown that AOH can reduce the activity of liver cells, increase the level of cell apoptosis and H2A phosphorylation. Animal experimental results also show that AOH can cause liver damage manifestations such as mononuclear infiltration around the liver portal, hepatocyte degeneration, focal inflammation of hepatocytes, and elevated transaminases.

[0004] Tauroursodeoxycholic acid (TUDCA) is chemically named 2-[[(3α, 5β, 7β)-3,7-dihydroxy-24-oxocholestane-24-yl]amino]ethanesulfonic acid dihydrate, which is a hydrophilic bile acid that has the effect of inhibiting endoplasmic reticulum stress induced by unfolded protein reaction, and has therapeutic effects on cholestatic liver disease, obesity and neurodegenerative diseases. At present, it is mainly used in the clinic to treat gallbladder cholesterol stones, primary sclerosing cholangitis, primary biliary cirrhosis and chronic viral hepatitis C. In addition, the prior art also reports the use of TUDCA in neuroprotective drugs, avoiding secondary damage after spinal cord injury, and treating neurodegenerative diseases (such as amyotrophic lateral sclerosis, Alzheimer's disease, Parkinson's disease, Huntington's disease and / or retinitis pigmentosa). In addition, in a mouse model, tauroursodeoxycholic acid can reduce cyclosporine-induced renal fibrosis. Tauroursodeoxycholic acid can also alleviate cisplatin-induced hearing loss in rats, and tauroursodeoxycholic acid can also alleviate fumonisin B1-induced cytotoxicity by inhibiting endoplasmic reticulum stress in gastric epithelial cells. However, tauroursodeoxycholic acid has not yet been found to alleviate the liver toxicity caused by cross-linked spore phenol.

[0005] Therefore, how to take effective measures to alleviate the liver toxicity induced by spore necrosis is an urgent problem to be solved by those skilled in the art. Summary of the invention

[0006] In view of the above-mentioned shortcomings of the prior art, the object of the present invention is to provide a drug for preventing and alleviating the toxicity of spore necrosis and its application, so as to solve the problem of the current lack of drugs for preventing or alleviating the toxicity of spore necrosis.

[0007] To achieve the above-mentioned object and other related objects, the present invention provides a use of tauroursodeoxycholic acid or a pharmaceutically acceptable salt thereof in the preparation of a drug for preventing or alleviating the toxicity of spore necrosis.

[0008] As a preferred embodiment of the present application, the structural formula of tauroursodeoxycholic acid is as shown in formula (I): (I).

[0009] As a preferred embodiment of the present application, the pharmaceutically acceptable salt of tauroursodeoxycholic acid is tauroursodeoxycholic acid sodium salt, and its structural formula is as described in formula (II): (II).

[0010] As a preferred embodiment of the present application, the toxicity is liver toxicity induced by spore necrosis.

[0011] As a preferred embodiment of the present application, the application includes any one or more of 1) to 5): 1) Alleviate liver damage induced by cross-linked spore phenol; 2) Reduce the degree of liver tissue fibrosis induced by cross-linked spore phenol; 3) Inhibit the inflammatory response induced by cross-linked spore phenol; 4) Alleviate liver cell apoptosis induced by cross-linked spore phenol; 5) Inhibit the expression level of endoplasmic reticulum stress markers.

[0012] As a preferred embodiment of the present application, the application 1) includes: Reduce liver function; and / or Reduce serum transaminase levels.

[0013] As a preference of the present application, the dosage of tauroursodeoxycholic acid or a pharmaceutically acceptable salt thereof in the application is 300-600 mg / kg bw.

[0014] As a preference of the present application, the dosage of tauroursodeoxycholic acid or a pharmaceutically acceptable salt thereof in the application is 500 mg / kg bw.

[0015] The present invention also provides a drug for preventing or alleviating the toxicity of spore necrosis, characterized in that the drug at least comprises tauroursodeoxycholic acid or a pharmaceutically acceptable salt thereof.

[0016] As a preference of the present application, the drug comprises at least 70 wt % of tauroursodeoxycholic acid or a pharmaceutically acceptable salt thereof.

[0017] As a preferred embodiment of the present application, the active ingredient of the drug at least includes tauroursodeoxycholic acid or a pharmaceutically acceptable salt thereof.

[0018] As a preferred embodiment of the present application, the dosage of tauroursodeoxycholic acid or a pharmaceutically acceptable salt thereof in the drug for preventing or alleviating the toxicity of spore necrosis is 300-600 mg / kg bw.

[0019] As a preference of the present application, the dosage of tauroursodeoxycholic acid or a pharmaceutically acceptable salt thereof in the drug is 500 mg / kg bw.

[0020] As a preferred embodiment of the present application, the drug, tauroursodeoxycholic acid or a pharmaceutically acceptable salt thereof is administered by a route selected from the following: oral, injection, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intraventricular, transdermal, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual and rectal administration.

[0021] As a preference of the present application, tauroursodeoxycholic acid or a pharmaceutically acceptable salt thereof is administered orally or by injection.

[0022] As a preferred embodiment of the present application, the drug is administered orally or by injection, and the daily oral or injection dose of tauroursodeoxycholic acid or its pharmaceutically acceptable salt or the active ingredient in the drug is 500 mg / kg bw.

[0023] The present invention also provides a drug for preventing or alleviating the toxicity of spore necrosis, characterized in that the drug comprises resveratrol, tauroursodeoxycholic acid or pharmaceutically acceptable salts thereof, either separately or together.

[0024] As a preferred embodiment of the present application, the dosage of tauroursodeoxycholic acid or a pharmaceutically acceptable salt thereof in the drug is 100-300 mg / kg bw; the dosage of resveratrol is 30-60 mg / kg bw.

[0025] As a preferred embodiment of the present application, the dosage of tauroursodeoxycholic acid or a pharmaceutically acceptable salt thereof in the drug is 200 mg / kg bw; the dosage of resveratrol is 50 mg / kg bw.

[0026] As a preferred embodiment of the present application, tauroursodeoxycholic acid or a pharmaceutically acceptable salt thereof and resveratrol are used simultaneously or sequentially.

[0027] As a preference of the present application, the drug further comprises at least one pharmaceutically acceptable carrier or excipient.

[0028] As preferred in the present application, the drug is in the form of tablets, pills, capsules, powders, granules, suppositories, dragees, ointments, sprays, inhalants, emulsions, solutions, suspensions, syrups and elixirs.

[0029] The present invention also provides the use of tauroursodeoxycholic acid in in vitro screening for preparing drugs for preventing or alleviating the toxicity of spore phenol.

[0030] The present invention also provides an application of a drug for preventing or alleviating the toxicity of spore necrosis in preventing or alleviating the toxicity of spore necrosis.

[0031] The present invention also provides the use of tauroursodeoxycholic acid or a pharmaceutically acceptable salt thereof in establishing an animal model of spore toxicity injury.

[0032] The present invention also provides a method for establishing an animal model of cross-linked spore toxicity injury, which is characterized by comprising the following steps: randomly grouping the animal models into a control group, an AOH group and an AOH+TUDCA group, wherein the control group is fed with normal feed; the AOH group is fed with feed containing 10 μg / kg AOH; the AOH+TUDCA group is fed with feed containing 10 μg / kg AOH + daily oral administration of 500 mg / kg bw TUDCA for 90 days, and finally obtaining a toxic injury animal model.

[0033] As a preferred embodiment of the present application, when establishing an animal model of spore toxicity injury caused by spore phenol, the dosage of spore phenol is 10 μg / kg.

[0034] As a preferred embodiment of the present application, the animal model of spore toxicity injury is a mouse model, specifically a C57BL / 6 mouse.

[0035] The invention adopts tauroursodeoxycholic acid or a pharmaceutically acceptable salt thereof to prepare a method for preventing or alleviating the toxicity of cross-sporin.

[0036] The present invention provides a method for preventing or alleviating the toxicity of spore phenol, characterized in that the method comprises: An effective amount of tauroursodeoxycholic acid or a pharmaceutically acceptable salt thereof is administered to the subject to reduce the toxicity of the spore necrosis factor.

[0037] The test subject may be a living animal in an animal model of spore damage caused by the spore toxicity, or may be an ex vivo liver tissue obtained from the animal model of spore damage caused by the spore toxicity.

[0038] The present invention also provides a method for preventing or alleviating the toxicity of spore necrosis, characterized in that the method comprises: An effective amount of tauroursodeoxycholic acid or a pharmaceutically acceptable salt thereof and resveratrol are administered to the subject simultaneously or sequentially to reduce the toxicity of spore necrosis.

[0039] The present invention also provides a method for preventing or alleviating the toxicity of spore necrosis, characterized in that the method comprises: An effective amount of a drug for preventing or alleviating the toxicity of spores is administered to the subject to reduce the toxicity of spores; the drug comprises resveratrol, tauroursodeoxycholic acid or a pharmaceutically acceptable salt thereof, either separately or together.

[0040] The present invention also provides a feed additive for preventing or alleviating the toxicity of spore necrosis, characterized in that the feed additive comprises a therapeutically effective amount of tauroursodeoxycholic acid or a pharmaceutically acceptable salt thereof.

[0041] The present invention also provides a feed additive for preventing or alleviating the toxicity of spore necrosis and its application in preventing or alleviating the toxicity of spore necrosis.

[0042] As a preferred embodiment of the present application, the amount of tauroursodeoxycholic acid or a pharmaceutically acceptable salt thereof added to the feed is 400-1000 mg / kg.

[0043] The present invention has the following beneficial effects: (1) TUDCA treatment restored some of the liver functions damaged by chronic liver damage caused by spore toxicity, inhibited the expression of endoplasmic reticulum stress markers, alleviated liver cell apoptosis, reduced the liver coefficient and the degree of liver tissue fibrosis, inhibited inflammatory response and reduced serum transaminase levels; (2) The combined use of TUDCA and resveratrol can reduce the dosage of TUDCA. Since tauroursodeoxycholic acid is relatively expensive (12 yuan / mg), and the cheap resveratrol (RET, 2.5 yuan / mg) has a synergistic effect on the effect of tauroursodeoxycholic acid on the alleviation of spore-induced liver toxicity, the combined use of the two substances can significantly reduce the dosage of TUDCA by at least 2 times, thereby greatly reducing the economic cost of alleviating spore-induced liver toxicity. (3) The present invention can also be used in animal husbandry as a feed additive to protect the health of livestock and poultry. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] Figure 1The effect of tauroursodeoxycholic acid on liver injury induced by spore spore, including (A) experimental schematic diagram. (B) Changes in liver coefficient. (CD) Changes in serum transaminase levels. (E) Changes in tissue structure after H&E staining.

[0045] Figure 2 The effect of tauroursodeoxycholic acid on liver fibrosis induced by spore necrosis factor, including (A) the change of Sirius red staining area and (BE) the change of mRNA level of liver fibrosis related genes.

[0046] Figure 3 The effect of tauroursodeoxycholic acid on the inflammatory response of the liver induced by spores, including (A) the change of F4 / 80 staining area, (BE) the change of mRNA level of proinflammatory factors in the liver, and (FG) the change of proinflammatory factor content in the liver.

[0047] Figure 4 The alleviating effect of tauroursodeoxycholic acid on apoptosis of liver tissue cells induced by spore spore. The changes of protein expression level (A) and mRNA expression level (BD) of apoptosis markers.

[0048] Figure 5 Figure 3 The alleviating effect of tauroursodeoxycholic acid on apoptosis of human liver cells induced by spore necrosis factor. After the cells were treated with 40 μM AOH and 300 μM TUDCA alone or in combination, the changes in cell activity (A), cell apoptosis rate (B), protein expression level (C) and mRNA expression level (DF) of apoptosis markers were observed.

[0049] Figure 6 The figure shows the alleviating effect of tauroursodeoxycholic acid on apoptosis of primary mouse liver cells induced by spore spore. After the cells were treated with 40 μM AOH and 300 μM TUDCA alone or in combination, the changes in cell activity (A), cell apoptosis rate (B), protein expression level (C) and mRNA expression level (DF) of apoptosis markers were observed.

[0050] Figure 7 The effect of tauroursodeoxycholic acid on the expression level of endoplasmic reticulum stress markers in liver cells induced by spore spore. (A) Changes in the expression level of endoplasmic reticulum stress marker proteins in liver tissue. (B) Changes in the expression level of endoplasmic reticulum stress marker mRNA in liver.

[0051] Figure 8The synergistic effect of resveratrol on the mitigation of tauroursodeoxycholic acid-induced hepatotoxicity. Changes in cell viability, apoptosis rate, and expression levels of proinflammatory factors in mouse primary liver cells (AD) and human liver cells (EH) after cells were treated with 40 μM AOH, 150 μM TUDCA, and 10 μM resveratrol (RET) alone or in combination. DETAILED DESCRIPTION

[0052] The following describes the embodiments of the present invention through 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.

[0053] It should be noted that the process equipment or devices not specifically specified in the following embodiments are all conventional equipment or devices in the art.

[0054] In addition, it should be understood that one or more method steps mentioned in the present invention do not exclude the existence of other method steps before or after the combination step or the insertion of other method steps between these explicitly mentioned steps, unless otherwise specified; it should also be understood that the combination connection relationship between one or more devices / apparatuses mentioned in the present invention does not exclude the existence of other devices / apparatuses before or after the combination device / apparatus or the insertion of other devices / apparatuses between these explicitly mentioned two devices / apparatuses, unless otherwise specified. Moreover, unless otherwise specified, the numbering of each method step is only a convenient tool for identifying each method step, and is not intended to limit the order of arrangement of each method step or the scope of the present invention. Changes or adjustments in their relative relationships should also be regarded as the scope of the present invention without substantially changing the technical content.

[0055] In the following examples of the present application, among the reagents: tauroursodeoxycholic acid was purchased from MCE, USA, with the product number HY-19696; resveratrol was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd., with the product number R107315; and spore spore was purchased from Qingdao Puruibang Biotechnology Co., Ltd., with the product number MSS1030.

[0056] Example 1 Effects of tauroursodeoxycholic acid on liver damage induced by spore spore.

[0057] C57BL / 6 mice were used as the animal model and divided into 3 groups, with 6 animals in each group: control group: fed with normal feed; AOH group: fed with feed containing 10 μg / kg AOH; AOH+TUDCA group: fed with feed containing 10 μg / kg AOH + daily oral administration of 500 mg / kg bw TUDCA. After 90 days of feeding, the animals were weighed and serum was collected. They were then slaughtered, and the liver tissue was obtained and weighed. Part of the liver tissue was quickly frozen in liquid nitrogen and stored in a -80°C refrigerator, and the other part was stored in a 4% paraformaldehyde solution ( Figure 1 A). H&E staining and blood biochemical tests were used to analyze the extent of liver damage. The experimental results showed that AOH could increase the liver coefficient ( Figure 1 B) and serum transaminase levels ( Figure 1 CD). After AOH exposure, pathological changes such as cell swelling and necrosis appeared in liver tissue ( Figure 1 E). When mice were treated with tauroursodeoxycholic acid and AOH simultaneously, the liver coefficient, serum transaminase levels, and histopathological changes were not significantly different from those in the control group. These results suggest that tauroursodeoxycholic acid can alleviate AOH-induced liver damage.

[0058] Example 2 Effects of tauroursodeoxycholic acid on hepatic fibrosis induced by spore necrosis factor.

[0059] Sirius red staining and real-time fluorescence quantitative PCR were used to detect changes in liver fibrosis markers. The results of the Sirius red staining experiment showed that AOH could increase the deposition of collagen in liver tissue, while tauroursodeoxycholic acid could reverse this abnormal phenomenon ( Figure 2 A). Real-time fluorescence quantitative PCR results also confirmed that AOH treatment significantly increased the mRNA levels of profibrotic genes (collagen type I / III α1 (Col1a1, Col3a1)), connective tissue growth factor (Ctgf), and tissue inhibitor of metalloproteinase 1 (Timp1) in liver tissues. However, the expression levels of these fibrosis-related genes in the AOH+TUDCA group were not significantly different from those in the control group ( Figure 2 BE). These results suggest that tauroursodeoxycholic acid can improve the hepatic fibrosis induced by spore necrosis.

[0060] Example 3 Effect of tauroursodeoxycholic acid on inflammatory response in liver induced by spore necrosis factor.

[0061] The results of F4 / 80 staining experiments showed that AOH could promote the infiltration of macrophages in liver tissue, while in the AOH+TUDCA group, the infiltration of macrophages was significantly reduced, with no significant difference compared with the control group ( Figure 3A). RNA was extracted from liver tissue using an RNA extraction kit, and then the expression levels of pro-inflammatory factors (IL6, Tnfα, Cxcl10, Ccl2) were detected using real-time fluorescence quantitative PCR. At the same time, IL6 and Tnfα in liver tissue were detected using Elisa kits. The results showed that AOH could increase the expression levels of pro-inflammatory factors ( Figure 3 BE), and the contents of IL6 and Tnfα were significantly increased ( Figure 3 FG), however, tauroursodeoxycholic acid could inhibit the AOH-induced increase in macrophage infiltration and proinflammatory cytokine levels. This indicates that tauroursodeoxycholic acid can inhibit the AOH-induced inflammatory response.

[0062] Example 4 Effect of tauroursodeoxycholic acid on apoptosis of liver cells induced by spore spore.

[0063] RNA extraction kit and cell lysis buffer were used to extract RNA and protein from liver tissue and cells, respectively. Then, real-time fluorescence quantitative PCR and immunoblotting were used to detect the expression levels of apoptosis markers (Caspase3, Bax and Bcl2). CCK kit and apoptosis kit were used to analyze the cell activity and apoptosis of mouse primary liver cells and human liver cells (THLE-2). The results of animal experiments showed that after AOH treatment, the protein expression levels of the pro-apoptotic genes Cleaved Caspase3 and Bax and the mRNA expression levels of Caspase3 and Bax genes in liver tissue were significantly increased, while the expression level of the anti-apoptotic gene Bcl2 was significantly decreased ( Figure 4 The results of cell experiments showed that after 48 h of treatment, 40 μM AOH could reduce cell activity ( Figure 5 A and Figure 6 A) and increased apoptosis rate ( Figure 5 B and Figure 6 B), the abnormal change trend of apoptosis marker expression levels is similar to the results of animal experiments ( Figure 5 CF and Figure 6 CF). After simultaneous treatment with 300 μM tauroursodeoxycholic acid and 40 μM AOH, there was no significant difference in cell activity, cell apoptosis rate and expression level of apoptosis markers compared with the control group. This means that tauroursodeoxycholic acid can alleviate the apoptosis of liver cells induced by cross-linked spore phenol. In the cell experiment, the cell models were primary mouse hepatocytes and human liver cells, and the effective concentration of TUDCA was 100~350 μM.

[0064] Example 5 Effect of tauroursodeoxycholic acid on the expression level of endoplasmic reticulum stress markers in liver cells induced by spore spore.

[0065] RNA and protein were extracted from liver tissue using RNA extraction kit and cell lysis buffer, respectively. Then, the expression levels of endoplasmic reticulum stress markers (Bip, Phospho-PERK, PERK, ATF4 and CHOP) were detected using real-time fluorescence quantitative PCR and immunoblotting. The results of immunoblotting experiments showed that crosslinking spore phenol can significantly increase the protein expression level of Bip, promote the phosphorylation of PERK protein, and increase the expression levels of downstream signaling proteins ATF4 and CHOP. The results of real-time fluorescence quantitative PCR experiments also showed similar trends ( Figure 7 ). When mice were treated with tauroursodeoxycholic acid, the expression levels of these ER stress markers returned to normal levels, with no significant difference from the control group, indicating that tauroursodeoxycholic acid can inhibit the occurrence of ER stress.

[0066] Example 6 Synergistic effect of combined use of resveratrol and tauroursodeoxycholic acid on the alleviation of hepatotoxicity induced by spore necrosis.

[0067] In this example, the CCK kit and the apoptosis kit were used to analyze the cell activity and apoptosis of mouse primary liver cells and human liver cells (THLE-2). RNA was extracted from liver cells using an RNA extraction kit, and then the expression levels of proinflammatory factors (IL6 and Tnfα) were detected using real-time fluorescence quantitative PCR. The results showed that after 48 h of cell treatment, 150 μM tauroursodeoxycholic acid and 10 μM resveratrol (RET) alone could only partially alleviate the cell activity inhibition, cell apoptosis and increased expression levels of proinflammatory factors induced by 40 μM AOH. However, when the cells were treated with 150 μM tauroursodeoxycholic acid and 10 μM resveratrol at the same time, there was no significant difference in cell activity, cell apoptosis level, and proinflammatory factor expression level compared with the control group ( Figure 8 ), to achieve the mitigation effect of 300 μM tauroursodeoxycholic acid (this is the optimal concentration). Since tauroursodeoxycholic acid is expensive (12 yuan / mg), this experiment explored the synergistic effect of the relatively cheap resveratrol (RET, 2.5 yuan / mg) on ​​the mitigation effect of tauroursodeoxycholic acid on the liver toxicity induced by cross-linked spores, and reduced the economic cost of mitigating the liver toxicity of cross-linked spores by using the two substances in combination. This shows that resveratrol can enhance the mitigation effect of tauroursodeoxycholic acid on the liver toxicity induced by cross-linked spores, and the two show a synergistic effect. The combined use of the two ingredients can significantly reduce the economic cost compared with the use of tauroursodeoxycholic acid alone.

[0068] Example 7 Synergistic effect of resveratrol and tauroursodeoxycholic acid on the alleviation of hepatotoxicity induced by spore necrosis.

[0069] C57BL / 6 mice were used as animal models and divided into 5 groups, with 6 animals in each group: control group: fed with normal feed; AOH group: fed with feed containing 10 μg / kg AOH; AOH+TUDCA group: fed with feed containing 10 μg / kg AOH + daily oral administration of 200 mg / kg bw TUDCA; AOH+RET group: fed with feed containing 10 μg / kg AOH + daily oral administration of 50 mg / kg bw resveratrol; AOH+TUDCA+RET group: fed with feed containing 10 μg / kg AOH + daily oral administration of 200 mg / kg bw TUDCA + 50 mg / kg bw resveratrol. After 90 days of feeding, the animals were weighed and serum was collected. Afterwards, they were slaughtered, and liver tissue was obtained and weighed. Part of the liver tissue was quickly frozen in liquid nitrogen and stored in a -80℃ refrigerator, and the other part was stored in a 4% paraformaldehyde solution, and the liver damage, liver fibrosis, liver inflammation and other indicators were analyzed. The experimental results showed that the use of resveratrol alone had a poor effect on the alleviation of liver toxicity induced by cross-sporin, while the AOH+TUDCA group and the AOH+TUDCA+RET group partially or completely restored some of the liver functions damaged by chronic liver injury induced by cross-sporin, reduced the liver coefficient and the degree of fibrosis of liver tissue, inhibited the inflammatory response and reduced the serum transaminase level. Among them, the liver coefficient, serum transaminase level and tissue pathological changes of mice in the AOH+TUDCA+RET group were not significantly different from those in the control group, indicating that resveratrol can enhance the alleviation of tauroursodeoxycholic acid on liver toxicity induced by cross-sporin, and the two showed a synergistic effect. Moreover, the dosage of tauroursodeoxycholic acid or a pharmaceutically acceptable salt thereof used alone for preventing or alleviating the toxicity of cross-linked spore phenol is 500 mg / kg bw, but when used in combination with resveratrol, the dosage of tauroursodeoxycholic acid or a pharmaceutically acceptable salt thereof is 200 mg / kg bw; the dosage of resveratrol is 50 mg / kg bw. From the above data, it can be seen that the usage of tauroursodeoxycholic acid or a pharmaceutically acceptable salt thereof can be reduced by 2 times or more, that is, the combined use of the two ingredients can significantly reduce the economic cost compared with the use of tauroursodeoxycholic acid alone.

[0070] In summary, TUDCA treats liver toxicity induced by spore spore, restores part of the liver function damaged after chronic liver damage induced by spore spore, inhibits the expression level of endoplasmic reticulum stress markers, alleviates liver cell apoptosis, reduces the liver coefficient and the degree of fibrosis of liver tissue, inhibits inflammatory response and reduces serum transaminase levels; in order to reduce costs, TUDCA is used in combination with resveratrol, which can reduce the amount of TUDCA by at least 2 times, thereby greatly reducing the economic cost of alleviating spore spore liver toxicity; in addition, the present invention can also be used in animal husbandry, as a feed additive to protect the health of livestock and poultry.

[0071] The above examples are for the purpose of illustrating the embodiments disclosed by the present invention and are not to be construed as limitations of the present invention. In addition, the various modifications listed herein and the variations of methods and compositions in the invention are obvious to those skilled in the art without departing from the scope and spirit of the present invention. Although the present invention has been specifically described in conjunction with various specific preferred embodiments of the present invention, it should be understood that the present invention should not be limited to these specific embodiments. In fact, various modifications obvious to those skilled in the art as described above to obtain the invention should be included within the scope of the present invention.

Claims

1. Use of tauroursodeoxycholic acid or a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing or alleviating the toxicity of spore necrosis.

2. The use according to claim 1, characterized in that: The structural formula of the tauroursodeoxycholic acid is shown in formula (I): （I）。 3. The use according to claim 1, characterized in that: The pharmaceutically acceptable salt of tauroursodeoxycholic acid is tauroursodeoxycholic acid sodium salt, and its structural formula is as described in formula (II): (II).

4. The use according to claim 1, characterized in that: The toxicity of the cross-linked spore is the liver toxicity induced by the cross-linked spore.

5. The use according to claim 4, characterized in that: The application includes any one or more of 1) to 5): 1) Alleviate liver damage induced by spore necrosis; 2) Reduce the degree of liver tissue fibrosis induced by cross-linked spore phenol; 3) Inhibit the inflammatory response induced by cross-linked spore phenol; 4) Alleviate liver cell apoptosis induced by cross-linked spore phenol; 5) Inhibit the expression level of endoplasmic reticulum stress markers.

6. The use according to claim 1, characterized in that: The dosage of tauroursodeoxycholic acid or a pharmaceutically acceptable salt thereof in the application is 300-600 mg / kg bw.

7. The use according to claim 6, characterized in that: The dosage of tauroursodeoxycholic acid or a pharmaceutically acceptable salt thereof in the application is 500 mg / kg bw.

8. A drug for preventing or alleviating the toxicity of spore necrosis, characterized in that: The drug at least comprises tauroursodeoxycholic acid or a pharmaceutically acceptable salt thereof for use in any one of claims 1 to 7.

9. The drug according to claim 8, characterized in that: The drug comprises resveratrol, tauroursodeoxycholic acid or pharmaceutically acceptable salts thereof, separately or together.

10. The drug according to claim 9, characterized in that: The dosage of tauroursodeoxycholic acid or a pharmaceutically acceptable salt thereof in the drug is 100-300 mg / kg bw; the dosage of resveratrol is 30-60 mg / kg bw.

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