Application of probiotics in treatment of primary biliary cholangitis

Abstract

The invention discloses an application of probiotics in preparation of a medicine for preventing and / or treating cholestatic liver diseases. The probiotics are selected from one or more of plant lactobacillus Lp-G18, bifidobacterium longum subsp. Longum BL-G301, lactobacillus reuteri LR-G100 and lactobacillus johnsonii LJ-G55. Preferably, the cholestatic liver disease is primary biliary cholangitis.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to the use of probiotics in treating cholestatic liver diseases, in particular the use of Lactobacillus johnsonii or Bifidobacterium longum subsp. longum in preventing and / or treating primary biliary cholangitis. Background Art

[0002] Primary biliary cholangitis (PBC), formerly known as primary biliary cirrhosis, is a chronic autoimmune intrahepatic cholestatic disease of unknown etiology. Its main pathological manifestations are intrahepatic cholestasis, the presence of circulating antimitochondrial antibodies, degeneration and necrosis of interlobular bile duct epithelial cells in the portal tract, and inflammatory cell infiltration. Continued infiltration of inflammatory cells leads to progressive destruction of the small intrahepatic bile ducts, ultimately leading to cirrhosis. Ursodeoxycholic acid (UDCA) is currently the first-line treatment for PBC, with a favorable safety profile and few adverse effects. The only FDA-approved second-line treatment is obeticholic acid (OCA), a 6-ethylchenodeoxycholic acid, which can be used for patients who have an inadequate response to UDCA, but only 50% of patients respond. Case reports have shown that OCA can lead to severe hepatic compensatory events. Other drugs, including glucocorticoids, still require extensive clinical research to verify, and have adverse reactions such as immunosuppression and drug-induced liver injury. So far, UDCA is still the most effective treatment for PBC, but 30%-40% of patients do not respond well to UDCA treatment, and there is a risk of progression to cirrhosis or even liver failure. Therefore, it is of practical significance to develop a treatment method with high safety, no side effects, and good effects. Probiotics have been widely clinically proven to improve intestinal health, relieve diarrhea, and improve functional gastrointestinal disorders, and there have also been some explorations in improving liver damage. Patent document CN117320736A discloses that Leuconostoc citreum can be used to prevent and treat cholestatic liver disease, but Leuconostoc citreum is not on the list of edible fungi in my country. Summary of the Invention

[0003] In order to solve the problems existing in the prior art, the object of the present invention is to provide the use of probiotics in the preparation of a medicament for preventing and / or treating cholestatic liver disease, especially primary biliary cholangitis.

[0004] In one aspect, the present invention provides use of probiotics in the preparation of a medicament for preventing and / or treating cholestatic liver disease.

[0005] Wherein, the prevention and / or treatment of cholestatic liver disease includes the prevention and / or treatment of liver function damage, cholestasis, inflammatory lesions and / or liver fibrosis caused by cholestatic liver disease.

[0006] Preferably, the prevention and / or treatment of cholestatic liver disease includes: reducing the levels of alanine aminotransferase, aspartate aminotransferase and / or total bile acid in serum, reducing liver inflammatory cell infiltration, reducing liver fibrosis, and reducing the expression of aging-related secretory factors.

[0007] More preferably, the cholestatic liver disease is primary biliary cholangitis.

[0008] Wherein, the probiotics are one or more selected from Lactiplantibacillus plantarum, Bifidobacterium longum subsp. longum, Limosilactobacillus reuteri and Lactobacillus johnsonii.

[0009] Preferably, the name of the Lactobacillus plantarum is Lp-G18, and its preservation number is CCTCC NO: M2013690; the name of the Bifidobacterium longum subspecies longum is BL-G301, and its preservation number is CCTCC NO: M2013689; the name of the Lactobacillus reuteri is LR-G100, and its preservation number is CCTCC NO: M2013692; the name of the Lactobacillus johnsonii is LJ-G55, and its preservation number is CCTCC NO: M2017116.

[0010] More preferably, the probiotics are Bifidobacterium longum subsp. longum BL-G301 and / or Lactobacillus johnsonii LJ-G55.

[0011] In the medicine, the content of the probiotics is not less than 1×10 9 CFU / mL or 1×10 9 CFU / g, for example: 1×10 9 CFU / mL or 1×10 9 CFU / g, 2×10 9 CFU / mL or 2×10 9 CFU / g, 3×10 9 CFU / mL or 3×10 9 CFU / g, 4×10 9 CFU / mL or 4×10 9 CFU / g, 5×109 CFU / mL or 5×10 9 CFU / g, 6×10 9 CFU / mL or 6×10 9 CFU / g, 7×10 9 CFU / mL or 7×10 9 CFU / g, 8×10 9 CFU / mL or 8×10 9 CFU / g, 9×10 9 CFU / mL or 9×10 9 CFU / g, 1×10 10 CFU / mL or 1×10 10 CFU / g, or higher.

[0012] In another aspect, the present invention provides a pharmaceutical composition comprising a microbial agent comprising one or more selected from the group consisting of Lactobacillus plantarum Lp-G18 with a deposit number of CCTCC NO: M 2013690, Bifidobacterium longum subsp. longum BL-G301 with a deposit number of CCTCC NO: M2013689, Lactobacillus reuteri LR-G100 with a deposit number of CCTCC NO: M2013692, and Lactobacillus johnsonii LJ-G55 with a deposit number of CCTCC NO: M 2017116. More preferably, the microbial agent comprises Bifidobacterium longum subsp. longum BL-G301 and / or Lactobacillus johnsonii LJ-G55.

[0013] Furthermore, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient. The dosage form of the microbial agent includes but is not limited to lyophilized powder, tablets, capsules or granules.

[0014] Furthermore, the pharmaceutical composition also contains active bacteria of other species.

[0015] In another aspect, the present invention provides a method for preventing and / or treating cholestatic liver disease, comprising administering to a subject in need thereof a preventively or therapeutically effective amount of a probiotic, wherein the probiotic is one or more selected from the group consisting of Lactobacillus plantarum Lp-G18, deposited with CCTCC NO: M 2013690, Bifidobacterium longum subsp. longum BL-G301, deposited with CCTCC NO: M2013689, Lactobacillus reuteri LR-G100, deposited with CCTCC NO: M2013692, and Lactobacillus johnsonii LJ-G55, deposited with CCTCC NO: M 2017116. More preferably, the probiotic is Bifidobacterium longum subsp. longum BL-G301 and / or Lactobacillus johnsonii LJ-G55.

[0016] Preferably, the cholestatic liver disease is primary biliary cholangitis.

[0017] Terminology Notes:

[0018] “And / or” will be taken as a specific disclosure of each of the two specified features or components with or without the other. Thus, the term “and / or” used in phrases such as “A and / or B” is intended to include “A and B,” “A or B,” “A” (alone), and “B” (alone). Likewise, the term “and / or” used in phrases such as “A, B, and / or C” is intended to cover each of the following: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0019] "Comprising" and "including" have the same meaning and are intended to be open ended and permit, but not require, the inclusion of additional elements or steps. When the terms "comprising" or "including" are used herein, the terms "consisting of" and / or "consisting essentially of" are also included and disclosed.

[0020] "Pharmaceutically acceptable excipient" refers to any ingredient other than the probiotics described herein and having substantially non-toxic and non-inflammatory properties in patients, including but not limited to any and all solvents, dispersion media or other liquid carriers, dispersion or suspension aids, diluents, granulation and / or dispersing agents, surfactants, isotonicity agents, thickeners or emulsifiers, preservatives, binders, lubricants, colorants, sweeteners or flavorings, stabilizers, antioxidants, osmolarity regulators, pH regulators, buffers, chelating agents, cryoprotectants and / or fillers, as appropriate for the particular dosage form desired. Various excipients for formulating pharmaceutical compositions and techniques for preparing compositions are known in the art. Exemplary preservatives include, but are not limited to, vitamin A, vitamin C, vitamin E, β-carotene, citric acid, ascorbic acid, butylated hydroxyanisole, ethylenediamine, sodium lauryl sulfate (SLS), sodium lauryl ether sulfate (SLES), and the like, and combinations thereof. Exemplary buffers for controlling pH may include, but are not limited to, sodium phosphate, sodium citrate, sodium succinate, histidine (or histidine-HCl), sodium malate, sodium carbonate, and / or combinations thereof. Exemplary cryoprotectants include, but are not limited to, mannitol, sucrose, trehalose, lactose, glycerol, dextrose, and combinations thereof. Exemplary fillers include, but are not limited to, sucrose, trehalose, mannitol, glycine, lactose, raffinose, and combinations thereof. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1General evaluation of probiotic-treated primary cholangitis mice: After successful cholangitis model establishment, mice were given probiotics by oral gavage for 12 days. Thirty-five mice were randomly divided into seven groups, each containing five mice: a blank control group (control), a modeling group (ANIT group), a positive control group (ANIT + UDCA group), and four probiotic groups (ANIT + Lp-G18; ANIT + BL-G301; ANIT + LR-G100; and ANIT + LJ-G55). A represents the survival curve; B represents the body weight curve; C represents the liver index; and D represents the spleen index. Data are expressed as mean ± standard deviation. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

[0022] Figure 2 Serum liver function assessment in mice treated with probiotics for primary cholangitis: A represents aspartate aminotransferase (AST); B represents alanine aminotransferase (ALT); C represents alkaline phosphatase (ALP); and D represents total bile acid (TBA). Data are expressed as mean ± standard deviation. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

[0023] Figure 3 .Hematoxylin and eosin staining of the liver in mice with primary cholangitis model treated with probiotics.

[0024] Figure 4 Assessment of liver fibrosis in mice with probiotic intervention for primary cholangitis: A shows Sirius red staining of liver tissue from each group of mice; B shows Masson staining of liver tissue from each group of mice; C shows quantitative statistical graph of Sirius red staining-positive area; D shows quantitative statistical graph of Masson staining-positive area. Data are expressed as mean ± standard deviation. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

[0025] Figure 5 Evaluation of probiotic-induced liver inflammation in mice with primary cholangitis: A shows CD68 immunohistochemical staining of liver tissue from each group of mice; B shows quantitative statistical analysis of CD68-positive areas. Data are presented as mean ± standard deviation. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

[0026] Figure 6Probiotic intervention evaluated the aging-associated secretory phenotype in mice with primary cholangitis. A represents tumor necrosis factor-α (TNF-α); B represents interleukin-6 (IL-6); C represents interleukin-1β (IL-1β); D represents transforming growth factor-β (TGF-β); and E represents tissue inhibitor of metalloproteinase-1 (TIMP-1). Data are expressed as mean ± standard deviation. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. DETAILED DESCRIPTION

[0027] Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiments are only used to illustrate the present invention and are not used in limiting the scope of the present invention.In addition, should be understood that after reading the content of the present invention being recorded, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms fall within the limited scope of the present invention equally.

[0028] Example 1 Materials and Methods

[0029] 1.1 Preparation of probiotic suspension

[0030] The probiotics to be tested were Lactiplantibacillus plantarum Lp-G18 (deposit number: CCTCC NO: M 2013690), Bifidobacterium longum subsp. longum BL-G301 (deposit number: CCTCC NO: M2013689), Limosilactobacillus reuteri LR-G100 (deposit number: CCTCC NO: M 2013692) and Lactobacillus johnsonii LJ-G55 (deposit number: CCTCC NO: M 2017116). All of them were freeze-dried bacterial powders and stored at 4°C. The total number of viable bacteria in each bacterial powder was tested before the experiment. The above-mentioned probiotics are all probiotic strains that have been sold to the outside world by the applicant (JuneYao Runying Biotechnology (Shanghai) Co., Ltd.) and have been recorded in Chinese applications CN11134235A and CN119331778A.

[0031] Before feeding mice every day, fresh bacterial suspension was prepared by weighing an appropriate amount of bacterial powder and dissolving it in sterile saline to a concentration of 5×10 9 CFU / mL of bacterial solution.

[0032] 1.2 Construction of animal model and experimental grouping

[0033] Female C57BL6 / J mice (n=35), 6-8 weeks old, were housed at (25±2)°C with a 12-hour light and dark cycle, and allowed to acclimate to a diet ad libitum for one week. Thirty-five mice were randomly divided into seven groups of five. Oral gavage was performed according to the grouping schedule. After successful model establishment 48 hours later, intervention was continued for 12 consecutive days.

[0034] Blank control group (control group): The mice were gavaged with olive oil on the first day, and 48 hours later, 0.2 mL of sterile saline was gavaged every day.

[0035] Modeling group (ANIT group): On day 1, a single oral gavage of 60 mg / kg of α-naphthyl isothiocyanate (ANIT, dissolved in olive oil) was administered to establish a primary biliary cholangitis model. After 48 hours of successful model establishment, 0.2 mL of sterile saline was administered daily.

[0036] Positive control group (ANIT + UDCA group): On day 1, a single oral gavage of 60 mg / kg of α-naphthyl isothiocyanate (ANIT, dissolved in olive oil) was administered to establish a primary biliary cholangitis model. Forty-eight hours after successful model establishment, 15 mg / kg of ursodeoxycholic acid (UDCA) was administered daily.

[0037] Four probiotic intervention groups (ANIT+probiotic group): On the first day, α-naphthyl isothiocyanate (ANIT, dissolved in olive oil) was given by oral gavage at 60 mg / kg to establish a primary biliary cholangitis model. After 48 hours of successful model establishment, 0.2 mL of the corresponding probiotic solution was given by oral gavage daily (the probiotic intervention dose was 1×10 9 CFU / animal / day).

[0038] During the modeling and intervention period, mice were weighed daily. Serum and liver tissue samples were collected after the intervention for subsequent analysis.

[0039] 1.3 Serum biochemical test

[0040] Mouse blood samples were stored at room temperature for no more than 1 hour and centrifuged at 3000 rpm for 10 minutes to obtain serum. Serum levels of alanine aminotransferase (ALT / GPT), aspartate aminotransferase (AST / GOT), alkaline phosphatase (ALP), and total bile acid (TBA) were measured.

[0041] 1.4 Liver tissue sections and pathological observation

[0042] The liver tissue was fixed with 4% paraformaldehyde for 24 hours, embedded in paraffin, and mouse liver sections were prepared and stained with HE. Finally, the prepared sections were scanned with a PRECICE 500 digital slicer to evaluate the pathological and fibrotic changes of the liver tissue.

[0043] 1.5 Sirius red staining, Masson staining, and liver fibrosis assessment

[0044] Liver tissue was fixed in 10% formalin, dehydrated, embedded, sectioned, dewaxed, and hydrated according to conventional histological methods. Sirius red and Masson staining were performed, and liver collagen deposition and liver fibrosis were analyzed using Image-Pro Plus 6.0.

[0045] 1.6 Immunohistochemical analysis

[0046] Liver tissue was preserved in 4% paraformaldehyde and embedded in paraffin. Paraffin sections (5 μm) were dewaxed and dehydrated, and heat-induced antigen retrieval was performed. Sections were then incubated with 3% hydrogen peroxide to inhibit endogenous enzymes. After blocking with 5% goat serum, sections were incubated with anti-CD68 antibody (1:4000) overnight and further incubated with HRP-conjugated antibody for 1 hour. Sections were then incubated with streptavidin-biotin-peroxidase for 10 minutes. Representative images captured using a microscope were analyzed using Image-Pro Plus 6.0 software to count positive cells.

[0047] 1.7 RT-PCR detection of liver aging-related secretory factors

[0048] Total RNA was extracted from liver tissue using the standard Trizol (ThermoFisher, Massachusetts, USA) protocol. Expression levels of the liver aging-associated secretory phenotype factors interleukin-6 (IL-6), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), transforming growth factor-β (TGF-β), and tissue inhibitor of metalloproteinase 1 (TIMP-1) were measured. Relative mRNA expression levels were determined using the 2-ΔΔCt method. Primer sequences are shown in Table 1.

[0049] Table 1 Primer sequences for RT-PCR

[0050]

[0051] 1.8 Data Analysis

[0052] Data are expressed as mean ± standard deviation. Differences between groups were analyzed using one-way analysis of variance (ANOVA) followed by comparison using the Benjamini, Krieger, and Yekutieli two-stage step-up method. All statistical analyses were performed using GraphPad Prism software (version 8.0.1). A p value of < 0.05 was considered statistically significant.

[0053] Example 2: Effect of probiotics on the improvement of primary biliary cholangitis

[0054] 2.1 Effects of probiotics on the general condition of mice with primary biliary cholangitis

[0055] After the primary biliary cholangitis model in mice was successfully established using α-naphthyl isothiocyanate (ANIT, dissolved in olive oil), the mice were given probiotics by gavage for 12 days, and the survival status, body weight, and liver and spleen weight of the mice on the day of sacrifice were recorded.

[0056] The survival curve of mice is as follows Figure 1 As shown in A, the results showed that no mice died in each group during the experiment.

[0057] Weight is an important indicator of the health status of mice. Being underweight or losing weight too quickly indicates that the mice are in poor general condition and have a high probability of death. Figure 1 As shown in B, the results showed that ANIT model mice lost weight on the second day after ANIT administration, and their weight began to recover after UDCA or probiotics administration. Among them, the weight of mice in the Lactobacillus plantarum Lp-G18 group returned to the initial weight on the fifth day, which was better than that of other groups. On the seventh day, the weight of mice in the UDCA and probiotics administration groups had returned to the initial weight and slowly increased over time.

[0058] Compared with the blank control group, the liver and spleen indexes of mice in the ANIT group (biliary cholangitis model) were significantly increased (p < 0.05) (organ index = organ weight (g) / mouse weight (g) × 100%). Intervention with Lactobacillus plantarum Lp-G18, Bifidobacterium longum subsp. longum BL-G301 or Lactobacillus johnsonii LJ-G55 significantly reduced the liver index, and the effect was comparable to that of the positive drug UDCA intervention ( Figure 1 C); intervention with probiotics Bifidobacterium longum subsp. longum BL-G301 or Lactobacillus johnsonii LJ-G55 significantly reduced spleen index, and the effect was comparable to that of the positive control ANIT+UDCA group ( Figure 1 D). These results suggest that Lactobacillus plantarum Lp-G18, Bifidobacterium longum subsp. longum BL-G301, or Lactobacillus johnsonii LJ-G55 may improve the phenotype of primary cholangitis mice by alleviating hepatic and systemic inflammatory responses.

[0059] 2.2 Effects of probiotics on serological liver function indices in mice with primary biliary cholangitis

[0060] Liver function is an important indicator that reflects the liver's synthesis, metabolism and detoxification functions, as well as whether there is cholestasis and liver cell damage. Figure 2Compared with the blank control group, ANIT (cholangitis model) mice showed significant increases in aspartate aminotransferase (AST), alanine aminotransferase (ALT), and total bile acid (TBA) (p < 0.05), with alkaline phosphatase (ALP) also showing a slight upward trend. Intervention with Lactobacillus plantarum Lp-G18, Bifidobacterium longum subsp. longum BL-G301, or Lactobacillus johnsonii LJ-G55 significantly reduced serum AST, ALT, and TBA levels (p < 0.05). Lactobacillus reuteri LR-G100 only significantly reduced ALT and TBA levels (p < 0.05). The intervention effects of Bifidobacterium longum subsp. longum BL-G301 and Lactobacillus johnsonii LJ-G55 were comparable to those of the active drug UDCA. In fact, Lactobacillus johnsonii LJ-G55 showed a more significant reduction in TBA than UDCA.

[0061] 2.3 Probiotics alleviate liver histological damage in mice with primary biliary cholangitis

[0062] The liver tissue of mice was pathologically analyzed by HE staining. Figure 3 Compared with the blank control group, mice in the ANIT group (cholangitis model) showed a large number of neutrophil and lymphocyte infiltration around the bile ducts, especially in the portal area, accompanied by bile duct hyperplasia, increased fibroblasts, collagen fiber deposition in the portal area, and fibrous tissue hyperplasia. Intervention with the positive drug UDCA or the probiotics Lp-G18, BLG301, LR-G100, or LJ-G55 all alleviated cholangitis to varying degrees. The LJ-G55 group had the most significant alleviating effect, as demonstrated by reduced neutrophil and lymphocyte infiltration around the bile ducts, reduced bile duct hyperplasia, and no collagen fiber deposition and fibrous tissue hyperplasia in the portal area. BL-G301, Lp-G18, and LR-G100 had the second-best alleviating effects.

[0063] 2.4 Probiotics alleviate liver fibrosis in mice with primary biliary cholangitis

[0064] Cholestasis caused by cholangitis is one of the important causes of liver fibrosis. Sirius red staining and MASSON staining can be used to show the degree of fibrosis in liver tissue. In this experiment, the liver tissues of each group of mice were stained with Sirius red ( Figure 4 A) and MASSON staining ( Figure 4 B), and the collagen fibers were quantified. The quantitative results were as follows: Figure 4As shown in Figures C and D, the experimental results showed that compared with the blank control group, the ANIT group (cholangitis model) showed a significant proliferation of collagen fibers in the liver tissue. Intervention with the probiotics Lp-G18, BL-G301, LR-G100, or LJ-G55 significantly reduced the area of ​​collagen fiber-positive areas in the liver tissue. BL-G301 and LJ-G55 were comparable in alleviating liver fibrosis to the positive drug UDCA.

[0065] 2.5 Effect of probiotics on the alleviation of liver inflammation in mice with primary biliary cholangitis

[0066] To evaluate the effect of probiotics on inflammation in mice with primary cholangitis, CD68 macrophage immunohistochemical staining was performed on liver tissue sections of mice in each group. Figure 5 Compared with the blank control group, the number of CD68-positive macrophages in the liver tissue of mice in the ANIT group (cholangitis model) was significantly increased (p < 0.0001). After intervention with probiotics Lp-G18, BL-G301, LR-G100, or LJ-G55, the number of CD68-positive macrophages was significantly reduced, and the reduction effect was more significant than that of the positive drug UDCA.

[0067] Effects of probiotics on the aging-related secretory phenotype in mice with primary biliary cholangitis

[0068] The mRNA expression levels of the five most common molecules of SASP (senescence-associated secretory phenotype) (IL-6, IL-1β, TNF-α, TGF-β and TIMP-1) in liver tissue were detected. Figure 6 As shown in the results, compared with the blank control group, the mRNA expression levels of TNF-α and IL-6 in the liver tissue of mice in the ANIT group (cholangitis model) were significantly increased (p < 0.05), and the mRNA expression levels of IL-1β, TGF-β, and TIMP-1 also showed a certain upward trend. After intervention with the probiotics Lp-G18, BL-G301, LR-G100, or LJ-G55, the mRNA expression levels of TNF-α and IL-6 were significantly downregulated (p < 0.05), with an effect comparable to that of the positive drug UDCA. At the same time, the mRNA expression levels of IL-1β, TGF-β, and TIMP-1 also showed a certain downward trend. The combined effects of downregulating TNF-α and IL-6 showed that the intervention effects of the probiotics LR-G100 and LJ-G55 were the most significant.

[0069] In summary, the present invention explores the therapeutic and ameliorative effects of four probiotics (Lactiplantibacillus plantarum Lp-G18, Bifidobacterium longum subsp. longum BL-G301, Limosilactobacillus reuteri LR-G100, and Lactobacillus johnsonii LJ-G55) on primary cholangitis in mice. The effects were evaluated from the aspects of general status, serological liver function indicators, histopathology, liver fibrosis, inflammation, and aging-related secretory factors. The results are summarized in Table 2. Comprehensive evaluation results from different dimensions show that the therapeutic effects of Bifidobacterium longum subsp. longum BL-G301 and Lactobacillus johnsonii LJ-G55 are comparable to those of UDCA, the first-line treatment for primary biliary cholangitis; Lactobacillus plantarum Lp-G18 and Lactobacillus reuteri LR-G100 also have certain therapeutic effects.

[0070] Table 2 The alleviating effects of four probiotics on primary cholangitis

[0071]

[0072] Culture collection:

[0073] (1) Lactiplantibacillus plantarum Lp-G18, deposited in China Center for Type Culture Collection, Wuhan, China, with a CCTCC NO: M2013690 and a deposit date of December 23, 2013;

[0074] (2) Bifidobacterium longum subsp. Longum BL-G301, deposited in China Center for Type Culture Collection, Wuhan, China, with a CCTCC NO: M2013689 and a deposit date of December 23, 2013;

[0075] (3) Limosilactobacillus reuteri LR-G100, deposited in China Center for Type Culture Collection, Wuhan, China, with a CCTCC NO: M2013692 and a deposit date of December 23, 2013;

[0076] (4) Lactobacillus johnsonii LJ-G55, deposited at China Center for Type Culture Collection, Wuhan, China, with a CCTCC NO: M2017116 and a deposited date of March 15, 2017.

[0077] The above describes the embodiments of the present invention. However, the present invention is not limited to the above embodiments. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included in the scope of protection of the present invention.

Claims

1. Use of probiotics in the preparation of a medicament for preventing and / or treating cholestatic liver disease, characterized in that: The probiotics are one or more selected from Lactiplantibacillus plantarum, Bifidobacterium longum subsp. longum, Limosilactobacillus reuteri and Lactobacillus johnsonii.

2. The use according to claim 1, characterized in that: The name of the plant lactobacillus is Lp-G18, and its preservation number is CCTCC NO: M 2013690; the name of the Bifidobacterium longum subspecies longum is BL-G301, and its preservation number is CCTCC NO: M2013689; the name of the Lactobacillus reuteri is LR-G100, and its preservation number is CCTCC NO: M 2013692; and the name of the Lactobacillus johnsonii is LJ-G55, and its preservation number is CCTCC NO: M 2017116.

3. The use according to claim 1, characterized in that The probiotics are Bifidobacterium longum subspecies longum BL-G3011, whose preservation number is CCTCC NO: M2013689, and / or Lactobacillus johnsonii LJ-G55, whose preservation number is CCTCC NO: M 2017116.

4. The use according to any one of claims 1 to 3, characterized in that: The cholestatic liver disease is primary biliary cholangitis.

5. The use according to any one of claims 1 to 4, characterized in that The medicine prevents and / or treats liver function damage, cholestasis, inflammatory lesions and / or liver fibrosis caused by cholestatic liver disease.

6. The use according to any one of claims 1 to 4, characterized in that The drug reduces the levels of alanine aminotransferase, aspartate aminotransferase and / or total bile acid in serum, reduces liver inflammatory cell infiltration, and reduces liver fibrosis.

7. The use according to any one of claims 1 to 4, characterized in that The content of the probiotics in the medicine is not less than 1×10 9 CFU / mL or 1×10 9 CFU / g.

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