Application of lactobacillus rhamnosus in preparation of medicine for treating intestinal barrier and drug-induced liver injury
By using Lactobacillus rhamnosus preparations, the treatment challenges of drug-induced liver injury and intestinal barrier damage have been solved, resulting in improved liver function and restored intestinal health, significantly reducing the risk of inflammatory diseases and enhancing intestinal barrier function.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-11-19
- Publication Date
- 2026-04-03
AI Technical Summary
There is a lack of effective methods in the current technology to treat drug-induced liver injury and intestinal barrier damage, especially the application of improving liver function and intestinal health by modulating the gut microbiota has not been reported.
Using Lactobacillus rhamnosus as the active ingredient, an oral drug formulation was prepared for the treatment of drug-induced liver injury and intestinal barrier damage. It reduces liver damage and improves intestinal barrier integrity by improving intestinal flora diversity and restoring intestinal flora dysbiosis.
Lactobacillus rhamnosus can significantly reduce drug-induced liver injury and intestinal barrier damage, improve liver function indicators, restore intestinal flora diversity, reduce the risk of inflammatory bowel disease and liver disease, increase intestinal butyrate content, and enhance intestinal barrier function.
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Figure CN121775016A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of microbial technology, specifically to the application of Lactobacillus rhamnosus in the preparation of drugs for treating intestinal barrier and drug-induced liver injury. Background Technology
[0002] Drug-induced liver injury (DILI) refers to liver damage caused by drugs or their metabolites, and has become a significant global public health issue. Recent surveys show that DILI is a major cause of acute liver failure worldwide, and its incidence is on the rise. In recent years, the proportion of herb-induced liver injury (HILI) among DILI cases has continued to rise, and it remains one of the main reasons for drug development termination, post-marketing warnings and restrictions, and market withdrawal.
[0003] The composition of the gut microbiota influences all aspects of the gut-hepatic interaction, from the intestinal epithelium to the intestinal vascular barrier, to the hepatic sinusoids, and back to the gut via the biliary tree and systemic circulation. In a healthy state, the gut microbiota is contained within the intestinal lumen, but its metabolites act locally and systemically to maintain gut-hepatic axis homeostasis; disruption of homeostasis at any of these interfaces can lead to liver disease. Probiotics have demonstrated numerous health benefits in gut health and immunology research, including modulating the gut microbiota, stimulating immune cells, synthesizing immunomodulatory molecules, and improving mucosal barrier function; they also translocate across the intestinal barrier and survive in the mesenteric lymph nodes and liver, influencing both gut and liver health.
[0004] Currently, most research on the functions of probiotics focuses on improving gastrointestinal function, regulating nutrient metabolism, and immunity. Probiotics have significant effects on constipation, enteritis, lactose intolerance, infection control, inflammation, allergies, and disorders of glucose and lipid metabolism. With the gradual maturation of the "gut-liver axis" theory, regulating the gut microbiota through probiotics to improve liver function has become a new approach to treating drug-induced liver injury. The beneficial effects of certain lactobacilli on colitis have also been confirmed by animal studies and clinical research.
[0005] However, there are currently no reports on the application of a type of Lactobacillus rhamnosus in the preparation of drugs for treating intestinal barrier and drug-induced liver injury. Summary of the Invention
[0006] The purpose of this invention is to provide the application of *Lactobacillus rhamnosus* in the preparation of drugs for treating intestinal barrier and drug-induced liver injury. This invention uses probiotics as an adjunct to treat drug-induced liver injury and intestinal barrier damage. The strain can alleviate liver function indicators in mice with drug-induced liver injury, improve intestinal barrier damage associated with drug-induced liver injury, improve intestinal flora diversity, and restore drug-induced intestinal flora imbalance.
[0007] In a first aspect, the present invention provides the use of *Lactobacillus rhamnosus* in the preparation of drugs for the prevention and / or treatment of intestinal barrier and drug-induced liver injury; wherein the *Lactobacillus rhamnosus* is *Lactobacillus casei*. Lacticaseibacillus rhamnosus .
[0008] The aforementioned Lactobacillus rhamnosus Lacticaseibacillus rhamnosus It can be purchased from the China Industrial Microbial Culture Collection Center (CICC), with accession number CICC 6001.
[0009] Furthermore, the aforementioned drug-induced liver injury refers to liver damage caused by drugs managed as prescription or over-the-counter drugs, such as chemical drugs, biological products, and traditional Chinese medicines, as well as products such as Chinese medicinal materials, natural medicines, health products, and dietary supplements, or their metabolites, excipients, pollutants, and impurities.
[0010] Furthermore, the intestinal barrier injury mentioned above is intestinal barrier injury accompanying drug-induced liver injury.
[0011] In a specific embodiment of the present invention, the drug-induced liver injury is either drug-induced liver injury caused by genipin or drug-induced liver injury caused by acetaminophen.
[0012] Furthermore, the aforementioned Lactobacillus rhamnosus can reduce liver damage caused by hepatotoxic drugs, improve intestinal barrier damage, improve intestinal flora diversity, restore intestinal flora imbalance caused by stress, increase the abundance of fecal cocci, increase intestinal butyrate content, and reduce the risk of inflammatory bowel disease and inflammatory liver disease.
[0013] Furthermore, the drug is a pharmaceutical preparation with Lactobacillus rhamnosus as the active ingredient.
[0014] Furthermore, the pharmaceutical preparation contains 1×10 8 ~2×10 8 CFU of live Lactobacillus rhamnosus. The Lactobacillus rhamnosus is a bacterial suspension.
[0015] Furthermore, the pharmaceutical preparation is an oral pharmaceutical preparation. The dosage form of the pharmaceutical preparation is selected from one or more of the following: powder, tablet, granule, capsule, solution, and suspension.
[0016] Furthermore, the pharmaceutical formulation may also include other pharmaceutically acceptable carriers or excipients.
[0017] Furthermore, the pharmaceutical preparation further includes one or more of the following: excipients, binders, diluents, disintegrants, fillers, wetting agents, absorption promoters, surfactants, adsorbent carriers, and lubricants.
[0018] Furthermore, the excipient is selected from one or more of lactose, microcrystalline cellulose, mannitol, and starch;
[0019] The adhesive is selected from one or more of the following: methylcellulose, ethylcellulose, polyvinylpyrrolidone, polyvinyl alcohol, hydroxypropylcellulose, and hydroxypropyl methylcellulose;
[0020] The diluent is selected from one or more of the following: calcium sulfate, glucose binder, dextrin, monosaccharide or polysaccharide, kaolin, sugar alcohol, and microcrystalline cellulose;
[0021] The disintegrant is selected from one or more of the following: dry starch, sodium carboxymethyl starch, low-substituted cellulose, polyvinylpyrrolidone, sodium carboxymethyl cellulose, microcrystalline cellulose, potassium polyacrylaldehyde, sodium glycolate starch, and low-substituted hydroxypropyl cellulose.
[0022] In a second aspect, the present invention provides the use of Lactobacillus rhamnosus in the preparation of medicaments that improve the intestinal mucosal barrier, improve the intestinal mucosal mechanical barrier, improve the intestinal mucosal immune barrier, improve the intestinal mucosal biological barrier, and improve the intestinal mucosal chemical barrier.
[0023] A third aspect of the present invention provides a pharmaceutical preparation for preventing and / or treating intestinal barrier and drug-induced liver injury, wherein the active ingredient is *Lactobacillus rhamnosus*; wherein the *Lactobacillus rhamnosus* is *Lactobacillus casei*. Lacticaseibacillus rhamnosus .
[0024] The advantages of this invention are:
[0025] 1. The *Lactobacillus rhamnosus* used in this invention can restore intestinal function in mice, improve intestinal flora diversity in mice, and have a hepatoprotective effect on mice with drug-induced liver injury. The *Lactobacillus rhamnosus* can reduce liver damage caused by hepatotoxic drugs in mice, has a hepatoprotective effect on mice with drug-induced liver injury, effectively improve intestinal barrier damage in mice, enhance the integrity of the intestinal barrier, improve intestinal flora diversity, restore intestinal flora imbalance caused by stress, increase the abundance of *Femcoccus*, increase intestinal butyrate content, and reduce the risk of inflammatory bowel disease and inflammatory liver disease.
[0026] 2. This invention provides a novel use of Lactobacillus rhamnosus as an adjunct to the treatment of drug-induced liver injury and intestinal barrier damage, which can reduce liver function indicators of drug-induced liver injury, improve intestinal barrier damage associated with drug-induced liver injury, improve intestinal flora diversity, and restore drug-induced intestinal flora disorder. Attached Figure Description
[0027] Figure 1 The effect of each experimental group of mice on liver biochemical indicators (Mean ± SEM, n=6). P < 0.0001, compared with the blank control group, #### P < 0.0001, compared with the model group.
[0028] Figure 2 The results of HE staining of mouse liver sections in each experimental group (×20) are shown. The upper left is the normal group, the upper right is the model group, the lower left is the inactivated probiotic group, and the lower right is the probiotic group.
[0029] Figure 3 The effect of each experimental group of mice on liver biochemical indicators (Mean ± SEM, n=6). P < 0.0001, compared with the blank control group, #### P < 0.0001, compared with the model group, ### P < 0.001, compared with the model group.
[0030] Figure 4 The results of HE staining of mouse liver sections (×20) in each experimental group are shown. The left group is the normal group, the middle group is the model group, and the right group is the probiotic group.
[0031] Figure 5 This is the effect of intestinal permeability on mice in each experimental group. P < 0.05, compared with the blank control group, # P < 0.05, compared with the model group.
[0032] Figure 6 The effect of each experimental group of mice on the expression of tight junction protein in the colon (Mean ± SEM, n=6). P < 0.01, compared with the blank control group, P < 0.001, compared with the blank control group, #### P < 0.0001, compared with the model group.
[0033] Figure 7 The results of AB-PAS staining of colon sections (×10) of mice in each experimental group are shown. The left group is the normal group, the middle group is the model group, and the right group is the probiotic group. Detailed Implementation
[0034] The specific implementation methods provided by the present invention will be described in detail below with reference to the embodiments.
[0035] Example 1: Lactobacillus rhamnosus can restore genipin-induced liver damage in mice:
[0036] 1. Model establishment and animal grouping
[0037] Twenty-four male C57 mice, each weighing approximately 20g, were used as experimental animals. After one week of acclimatization, the animals were randomly divided into four groups of six mice each: a control group, a model group, a probiotic group, and an inactivated probiotic group. The control group received PBS solution via gavage. The model group received genipin at a dose of 180 mg / kg via gavage for three consecutive days. The probiotic group received 2 × 10⁻⁶ mg / kg of genipin via gavage. 8 CFU of Lactobacillus rhamnosus ( Lacticaseibacillus rhamnosus The probiotics were purchased from the China Industrial Microbial Culture Collection Center (accession number CICC 6001) at a dose of 0.2 mL. The probiotics were administered continuously for 14 days, followed by 3 days of concurrent administration of genipin at a dose of 180 mg / kg. The inactivated probiotic group received 2 × 10⁻⁶ probiotics via gavage. 8 0.2 mL of CFU was inactivated and cooled at 121°C and administered continuously for 14 days, followed by administration of genipin at a dose of 180 mg / kg for the last 3 days.
[0038] 2. Methods for evaluating liver injury
[0039] (1) Measurement of liver function biochemical indicators: mouse serum samples were taken and the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the serum were detected to monitor liver damage in mice.
[0040] (2) HE staining observation of liver tissue: The liver tissue was removed from the fixative, dehydrated, embedded in paraffin, and cut into sections with a thickness of about 4 μm. The sections were then soaked in xylene I for 20 min, xylene I for 20 min, anhydrous ethanol for 15 min, anhydrous ethanol II for 5 min, and 75% ethanol for 5 min, and washed with tap water. The sections were then stained in hematoxylin staining solution for 3-5 min, washed with tap water, differentiated with differentiation solution, washed again with tap water, and then stained with blue solution and rinsed with running water. The sections were then dehydrated in 85% and 95% graded ethanol solutions for 5 min, and then stained in eosin staining solution for 5 min. The sections were then soaked in anhydrous ethanol I for 5 min, anhydrous ethanol I for 5 min, anhydrous ethanol III for 5 min, xylene I for 5 min, and xylene for 15 min until transparent, and then mounted with neutral resin. The sections were then subjected to 3D panoramic scanning.
[0041] 3. Results show
[0042] (1) such as Figure 1 As shown, the serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in the model group were significantly higher than those in the normal group, indicating the success of the liver injury model. The serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in the probiotic group were significantly lower than those in the model group, basically at the same level as those in the normal group. Moreover, there was no significant decrease in biochemical indicators in the inactivated probiotic group, suggesting that Lactobacillus rhamnosus has a protective effect against traditional Chinese medicine-induced liver injury.
[0043] (2) For example Figure 2 As shown, no obvious abnormalities were observed in the blank control group and the probiotic group. In the model group, a large number of hepatocytes showed uneven cytoplasmic staining, with lighter staining at the edges and darker staining in the center. Nuclei and kernels were aggregated. Extensive oval cell proliferation was observed around the portal vein, accompanied by a small amount of connective tissue proliferation and inflammatory cell infiltration. A small number of hepatocytes within the lobules showed focal caseous necrosis with a small amount of inflammatory cell infiltration. In the inactivated probiotic group, oval cell proliferation was observed around the portal vein, accompanied by a small amount of connective tissue proliferation and inflammatory cell infiltration. Hepatocytes in the inactivated probiotic group showed mild swelling, loose and lightly stained cytoplasm, and a small amount of focal inflammatory cell infiltration within the lobules. This suggests that *Lactobacillus rhamnosus* has a protective effect against drug-induced liver injury.
[0044] Example 2: Lactobacillus rhamnosus can restore acetaminophen-induced liver damage in mice:
[0045] 1. Model establishment and animal grouping
[0046] Eighteen male C57 mice, each weighing approximately 20g, were used as experimental animals. After one week of acclimatization, the animals were randomly divided into three groups of six each: a control group, a model group, and a probiotic group. The control group was administered PBS solution by gavage; the probiotic group was administered 2×10⁻⁶ PBS solution by gavage. 8 CFU / mL Lactobacillus rhamnosus ( Lacticaseibacillus rhamnosus The mice were administered 0.2 mL of acetaminophen (accession number CICC6001) for 2 weeks. After fasting overnight on day 13, the mice in the model group and probiotic group were given acetaminophen by gavage at a dose of 300 mg / kg. All mice were sacrificed 24 hours after administration of acetaminophen.
[0047] 2. Methods for evaluating liver injury
[0048] (1) Measurement of liver function biochemical indicators: mouse serum samples were taken and the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the serum were detected to monitor liver damage in mice.
[0049] (2) HE staining observation of liver tissue: The liver tissue was removed from the fixative, dehydrated, embedded in paraffin, and cut into sections with a thickness of about 4 μm. The sections were then soaked in xylene I for 20 min, xylene I for 20 min, anhydrous ethanol for 15 min, anhydrous ethanol II for 5 min, and 75% ethanol for 5 min, and washed with tap water. The sections were then stained in hematoxylin staining solution for 3-5 min, washed with tap water, differentiated with differentiation solution, washed again with tap water, and then stained with blue solution and rinsed with running water. The sections were then dehydrated in 85% and 95% graded ethanol solutions for 5 min, and then stained in eosin staining solution for 5 min. The sections were then soaked in anhydrous ethanol I for 5 min, anhydrous ethanol I for 5 min, anhydrous ethanol III for 5 min, xylene I for 5 min, and xylene for 15 min until transparent, and then mounted with neutral resin. The sections were then subjected to 3D panoramic scanning.
[0050] 3. Results show
[0051] (1) such as Figure 3 As shown, the serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in the model group were significantly higher than those in the normal group, indicating the success of the liver injury model; the serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in the probiotic group were significantly lower than those in the model group, suggesting that Lactobacillus rhamnosus has a protective effect against drug-induced liver injury.
[0052] (2) For example Figure 4 As shown, no obvious abnormalities were observed in the blank control group and the probiotic group; in the model group, a large number of hepatocytes showed uneven cytoplasmic staining, with lighter staining at the edges and darker staining in the center, and aggregation of nuclei and kernels. Extensive oval cell proliferation was observed around the portal vein, accompanied by a small amount of connective tissue proliferation and a small amount of inflammatory cell infiltration. A small number of hepatocytes within the lobules showed focal caseous necrosis with a small amount of inflammatory cell infiltration. This suggests that *Lactobacillus rhamnosus* has a protective effect against drug-induced liver injury.
[0053] Example 3: Lactobacillus rhamnosus can improve the intestinal barrier in mice:
[0054] 1. Model establishment and animal grouping
[0055] Eighteen male C57 mice, each weighing approximately 20g, were used as experimental animals. After one week of acclimatization, the animals were randomly divided into three groups of six each: a control group, a model group, and a probiotic group. The control group received PBS solution via gavage. The model group received genipin at a dose of 180 mg / kg via gavage for three consecutive days. The probiotic group received 2 × 10⁻⁶ mg / kg of genipin via gavage. 8 CFU of Lactobacillus rhamnosus ( Lacticaseibacillus rhamnosus(Accession number CICC 6001) 0.2 mL, administered continuously for 14 days, followed by 3 days of simultaneous administration of genipin at a dose of 180 mg / kg.
[0056] 2. Intestinal barrier evaluation methods
[0057] (1) Intestinal permeability test: Mice were given FITC-glucan at a dose of 600 mg / kg by gavage, and the serum fluorescence value was measured 4 hours later to monitor the intestinal permeability of mice.
[0058] (2) Detection of key proteins in the intestinal barrier using molecular biotechnology: The expression levels of tight junction proteins ZO-1 and Occludin in mouse colon tissue were detected by Western blot method to monitor the integrity of the mouse intestinal mucosal barrier.
[0059] (3) AB-PAS staining observation of colon tissue: The colon tissue was removed from the fixative, dehydrated, embedded in paraffin, and cut into sections with a thickness of about 4 μm. The sections were then soaked in xylene I for 20 min, xylene II for 20 min, anhydrous ethanol for 15 min, anhydrous ethanol II for 5 min, and 75% alcohol for 5 min, and washed with tap water. The sections were then stained in alexandrite blue solution for 5 min, rinsed with tap water, stained in periodic acid solution for 15 min, rinsed twice with tap water and ultrapure water, stained with Scheffer solution in the dark for 30 min, rinsed with tap water for 5 min, stained with hematoxylin solution for 5 min, rinsed with tap water, differentiated with 1% hydrochloric acid alcohol for 10 s, rinsed with tap water, blued with 1% ammonia water, and rinsed with running water.
[0060] 3. Results show
[0061] (1) such as Figure 5 As shown, the fluorescence value of FITC-glucan in the serum of the model group was significantly higher than that of the normal group, indicating impaired intestinal permeability; the fluorescence value of FITC-glucan in the serum of the probiotic group was significantly lower than that of the model group, and basically the same as that of the normal group, suggesting that Lactobacillus rhamnosus improves the intestinal barrier and reduces intestinal permeability.
[0062] (2) For example Figure 6 As shown, the expression levels of the colonic tight junction proteins ZO-1 and Occludin in the model group were significantly lower than those in the normal group, indicating impaired intestinal mucosal barrier. The expression levels of the colonic tight junction proteins ZO-1 and Occludin in the probiotic group were significantly higher than those in the normal group, even higher than those in the normal group, suggesting that Lactobacillus rhamnosus improves the integrity of the intestinal mucosal barrier and improves intestinal barrier function by promoting the expression of intestinal tight junction proteins.
[0063] (3) such as Figure 7As shown, compared with the normal group, the number of goblet cells in the colon of mice in the model group was significantly reduced; compared with the model group, the number of goblet cells in the colon of mice in the probiotic group was increased. The results indicate that drug-induced liver injury can cause a decrease in the number of goblet cells in the colon of mice. After two weeks of probiotic administration, the number of goblet cells in the colon of mice was restored. This suggests that *Lactobacillus rhamnosus* improves the integrity of the intestinal mucosal barrier and enhances intestinal barrier function by increasing the number of goblet cells in the colon of mice.
[0064] The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited to the embodiments described. Those skilled in the art can make various equivalent modifications or substitutions without departing from the spirit of the present invention, and these equivalent modifications or substitutions are all included within the scope defined by the claims of this application.
Claims
1. The use of *Lactobacillus rhamnosus* in the preparation of drugs for the prevention and / or treatment of intestinal barrier and drug-induced liver injury, characterized in that... The Lactobacillus rhamnosus mentioned is Lactobacillus casei. Lacticaseibacillus rhamnosus .
2. The application according to claim 1, characterized in that, Drug-induced liver injury refers to liver damage caused by chemical drugs, biological products, traditional Chinese medicines managed as prescription or over-the-counter drugs, as well as Chinese medicinal materials, natural drugs, health products, dietary supplements, or their metabolites, excipients, pollutants, and impurities.
3. The application according to claim 1, characterized in that, The intestinal barrier injury mentioned is intestinal barrier injury accompanying drug-induced liver injury.
4. The application according to claim 1, characterized in that, The drug is a pharmaceutical preparation with Lactobacillus rhamnosus as the active ingredient.
5. The application according to claim 4, characterized in that, The pharmaceutical preparation mentioned is an oral pharmaceutical preparation.
6. The application according to claim 5, characterized in that, The pharmaceutical preparation further includes one or more of the following: excipients, binders, diluents, disintegrants, fillers, wetting agents, absorption promoters, surfactants, adsorbent carriers, and lubricants.
7. The application according to claim 6, characterized in that, The excipients are selected from one or more of lactose, microcrystalline cellulose, mannitol, and starch; The adhesive is selected from one or more of the following: methylcellulose, ethylcellulose, polyvinylpyrrolidone, polyvinyl alcohol, hydroxypropylcellulose, and hydroxypropyl methylcellulose; The diluent is selected from one or more of the following: calcium sulfate, glucose binder, dextrin, monosaccharide or polysaccharide, kaolin, sugar alcohol, and microcrystalline cellulose; The disintegrant is selected from one or more of the following: dry starch, sodium carboxymethyl starch, low-substituted cellulose, polyvinylpyrrolidone, sodium carboxymethyl cellulose, microcrystalline cellulose, potassium polyacrylaldehyde, sodium glycolate starch, and low-substituted hydroxypropyl cellulose.
8. Application of Lactobacillus rhamnosus in the preparation of drugs that improve the intestinal mucosal barrier, improve the intestinal mucosal mechanical barrier, improve the intestinal mucosal immune barrier, improve the intestinal mucosal biological barrier, and improve the intestinal mucosal chemical barrier.
9. A pharmaceutical formulation for the prevention and / or treatment of intestinal barrier and drug-induced liver injury, characterized in that, Its active ingredient is *Lactobacillus rhamnosus*; the *Lactobacillus rhamnosus* is *Lactobacillus casei*. Lacticaseibacillus rhamnosus .