Application of KLB protein in preparation of medicine for treating schistosome infection

Abstract

The invention relates to the technical field of medicines, in particular to application of KLB protein in preparation of a medicine for treating schistosome infection. The invention verifies the treatment effect of the KLB protein on hepatic tissue EMT and fibrosis caused by schistosoma japonicum infection, and proves that the KLB can be used as a medicine for treating schistosoma japonicum hepatic fibrosis.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of pharmaceutical technology, and in particular to the application of KLB protein in the preparation of a drug for treating schistosome infection. BACKGROUND

[0002] Schistosomiasis is a global distribution of zoonosis caused by schistosome of schistosoma. In the world, the disease can affect 78 countries in the world, about 240 million people are infected with schistosome, and more than 800 million people live in the risk area of infection. The main schistosome infection of human is Japanese blood fluke, mansoni and egyptian blood fluke, and the main schistosome infection of human is Japanese blood fluke, mansoni and egyptian blood fluke. The main epidemic area is yunnan, hubei, anhui, jiangxi and hunan province. Although the schistosomiasis in our country is low prevalence at present, but there are still a large number of schistosomiasis liver fibrosis patients.

[0003] Klotho is a single transmembrane protein, which is a co-receptor of fibroblast growth factor receptor (FGFR). Klotho was first discovered as an anti-aging gene, which can be divided into α-Klotho, β-Klotho and γ-Klotho according to its homology, and the main expression in liver is β-Klotho, namely KLB. In recent years, researchers have gradually deepened the research on KLB. Studies have shown that FGF21-KLB-FGFR signaling pathway plays an important role in regulating liver metabolism. Mice lacking KLB have elevated AST / ALT ratio and increased triglyceride accumulation in liver. The KLB gene polymorphism of obese people is significantly higher than that of non-obese people. In obese population, the secondary G allele of rs7674434 and the T allele of rs12152703 in KLB are risk alleles of NAFLD, and the decrease of KLB expression may be the fundamental reason for FGF21 resistance and the development of NAFLD. HSCs were cultured in vitro with different concentrations of FBS, and the expression of KLB mRNA and protein in HSCs cultured with 10% FBS was significantly lower than that in low FBS culture group. Compared with 0.5% FBS cultured HSCs, the mRNA expression levels of a-SMA, collagen I, collagen III and other fibrosis indicators in 10% FBS cultured HSCs were increased, which was negatively correlated with the expression level of KLB. The down-regulation of KLB expression plays an important role in inducing fibrosis genes, promotes the activation of hepatic stellate cells, and aggravates the liver damage of metabolic associated fatty liver (MAFLD) adult patients. Current studies have shown that KLB plays an important role in the regulation of liver diseases, but there is no research on the treatment effect of KLB on liver damage caused by schistosome infection. SUMMARY

[0004] The application aims to provide application of KLB protein in preparation of a medicine for treating schistosome infection.

[0005] The technical problem of the application is solved by the following technical scheme:

[0006] In a first aspect of the application, the application of KLB protein in preparation of a medicine for treating schistosome infection is provided.

[0007] In some embodiments of the application, the application of KLB protein is in preparation of a medicine for treating Japanese schistosome infection.

[0008] In some embodiments of the application, the application of KLB protein is in preparation of a medicine for treating liver fibrosis caused by Japanese schistosome infection.

[0009] In a second aspect of the application, the application of Klotho recombinant protein in preparation of a medicine for treating schistosome infection is provided.

[0010] In some embodiments of the application, the application of Klotho recombinant protein is in preparation of a medicine for treating Japanese schistosome infection.

[0011] In some embodiments of the application, the application of Klotho recombinant protein is in preparation of a medicine for treating liver fibrosis caused by Japanese schistosome infection.

[0012] In a third aspect of the application, the application of AAV8 adeno-associated virus loaded with KLB gene in preparation of a medicine for treating schistosome infection is provided.

[0013] In some embodiments of the application, the AAV8 adeno-associated virus is used as a carrier to load KLB gene, and the AAV8 adeno-associated virus is used to direct infection of liver so as to make liver tissue highly express KLB protein.

[0014] In some embodiments of the application, the application of AAV8 adeno-associated virus loaded with KLB gene is in preparation of a medicine for treating Japanese schistosome infection.

[0015] In some embodiments of the application, the application of AAV8 adeno-associated virus loaded with KLB gene is in preparation of a medicine for treating liver fibrosis caused by Japanese schistosome infection.

[0016] The present application provides two methods for supplementing KLB protein in mice. The first method is intraperitoneal injection of klotho recombinant protein; the second method is tail vein injection of AAV8-KLB (AAV8 adeno-associated virus encapsulating KLB gene).

[0017] The present application verifies the overexpression effect of KLB in liver tissue. The present application verifies that the mice supplemented with KLB protein have reduced liver tissue EMT and fibrosis levels after Schistosoma japonicum infection.

[0018] The present application includes the following beneficial effects: the present application verifies the therapeutic effect of KLB protein on liver tissue EMT and liver fibrosis caused by Schistosoma japonicum infection, and proves that KLB protein can be used as a drug to treat liver fibrosis caused by Schistosoma japonica. BRIEF DESCRIPTION OF DRAWINGS

[0019] Figure 1 It is a column chart of the worm load and egg load in Example 3.

[0020] Figure 2 It is the location and expression level of GFP in the tissue of the AAV8-GFP injected mouse detected by the frozen section in Example 3.

[0021] Figure 3 It is the qPCR detection of the fibrosis index of the liver tissue of the mouse in Example 2.

[0022] Figure 4 It is the qPCR detection of the fibrosis index of the liver tissue of the mouse in Example 3.

[0023] Figure 5 It is the result of western blot detection of the EMT and liver fibrosis level of the liver tissue of the mouse in Example 2.

[0024] Figure 6 It is the result of western blot detection of the KLB expression of the liver tissue of the mouse in Example 3.

[0025] Figure 7 It is the result of western blot detection of the EMT and liver fibrosis level of the liver tissue of the mouse in Example 3.

[0026] Figure 8 It is the HE and Masson staining chart of Example 3.

[0027] Figure 9 It is the percentage of the liver tissue and spleen tissue of the mouse in the total body weight of the mouse in Example 3.

[0028] Unless there is a contrary statement, the terms used in the specification and claims have the following meanings.

[0029] As described herein, the term "prevention" refers to preventing the occurrence of a disease and / or preventing the recurrence of a disease. DETAILED DESCRIPTION

[0030] The present specification describes in detail the specific embodiments, and those skilled in the art should recognize that the following embodiments are exemplary and cannot be understood as a limitation of the present application. For those skilled in the art, several improvements and modifications of the present application are made without departing from the principles of the present application, and the technical solutions obtained by the improvements and modifications also fall within the protection scope of the claims of the present application. The beneficial effects of the present application are specifically illustrated by the following examples.

[0031] Klotho recombinant protein is KLB protein, and the full name is Beta-klotho. The klotho recombinant protein is purchased from R&D Systems Company. The klotho protein has three subtypes, namely α-klotho, β-klotho and γ-klotho, and the main expression in liver tissue is β-klotho, that is, KLB. Klotho is not equal to KLB. AAV8-KLB: AAV8-KLB is constructed by Jikai Company, that is, AAV8 adeno-associated virus loaded with KLB gene.

[0032] AAV8-ctrol: AAV8-ctrol is constructed by Jikai Gene Company, that is, the control adeno-associated virus of AAV8-KLB, and the main function is to prove that after injection of AAV8-KLB, the KLB of mouse liver tissue is significantly increased, and after injection of AAV8-ctrol, there is no significant change.

[0033] Example 1

[0034] Establishment of Schistosoma japonicum infected mouse model

[0035] The abdominal hair of SPF level 6-8 week old female C57BL / 6 mice was removed, and the mice were infected with Schistosoma japonicum cercaria by abdominal patch method, and the infection dose of Schistosoma japonicum cercaria was (20±1) pieces per mouse, that is, the Schistosoma japonicum infected mouse model was obtained.

[0036] Example 2

[0037] Therapeutic efficacy experiment of klotho intraperitoneal injection

[0038] 1. Experimental grouping

[0039] The mice infected with Schistosoma japonicum for 4 weeks in Example 1 were randomly divided into PBS group and klotho group, and each group had 6 mice.

[0040] 2. Drug treatment

[0041] Klotho group mice were intraperitoneally injected with 0.8 μg of klotho recombinant protein, and PBS group mice were intraperitoneally injected with an equal amount of PBS, once every 2 days. After 6 weeks of infection, the mice were euthanized after anesthesia, and the mouse liver tissues were taken.

[0042] Example 3

[0043] Therapeutic efficacy experiment of AAV8-KLB targeting liver

[0044] 1. Experimental grouping

[0045] 6-8 week old C57BL / 6 female mice were randomly divided into AAV8-ctrol group (control group) and AAV8-KLB group (experimental group: AAV8 adeno-associated virus as a carrier to encapsulate KLB gene, AAV8 adeno-associated virus to direct infection of liver, making liver tissue highly express KLB protein), each group of 6. In addition, 3 6-8 week old C57BL / 6 female mice were set as AAV8-GFP positive reference group.

[0046] 2. Drug treatment

[0047] The AAV8-KLB group of mice was injected with 2x10 11 vg AAV8-KLB, and the AAV8-ctrol group of mice was injected with an equal amount of AAV8-ctrol; the positive reference group of AAV8-GFP group of mice was injected with 2x10 11 vg AAV8-GFP.

[0048] 3. Mouse infection

[0049] After 3 weeks of injection, GFP was highly expressed in liver tissue, and the AAV8-ctrol and AAV8-KLB mice were infected with Schistosoma japonicum according to the method of Example 1 for 6 weeks after infection. After anesthesia, the mice were euthanized, and the mouse liver tissues were taken. Figure 2 The results showed that the AAV8-GFP 2x10 11 vg virus was injected into the tail vein, and after three weeks of injection, the results of frozen section showed that GFP was more significantly expressed in liver tissue, suggesting that AAV8 was mainly directed to liver tissue, rather than spleen and intestine. Figure 6 It was shown that KLB protein was highly expressed in liver tissue of AAV8-KLB group of mice after 6 weeks of Schistosoma japonicum infection.

[0050] Figure 9The percentage of liver tissue and spleen tissue of mice in Example 3 to the total weight of mice, respectively. The mice in Example 3 were injected 6 weeks after being infected with Schistosoma japonicum, and the liver tissue and spleen tissue of the mice were taken to calculate the liver index and spleen index, respectively. The results show that the liver index and spleen index of the AAV8-KLB mice are significantly lower than those of the AAV8-ctrl group, indicating that supplementing the KLB protein in the liver tissue of mice can reduce the damage to the liver tissue and reduce the immune response level of the spleen tissue.

[0051] Example 4

[0052] Calculate the worm load of mice after being infected with Schistosoma japonicum

[0053] ① After the mice in Example 3 infected with Schistosoma japonicum for 6 weeks were anesthetized and euthanized, the abdominal cavity of the mice was opened to fully expose the internal organs, and the intestinal tube and fat were gently pushed to the right side of the mouse abdomen;

[0054] ② The perfusion needle was inserted into the inferior vena cava, and the peristaltic pump was used to perfuse PBS preheated to 37°C into the blood vessels of the mice at a speed of 70 rpm per minute. When the liver tissue of the mice was filled, the hepatic portal vein was cut open, the adult worms were flushed out, and collected in a dish for counting.

[0055] Figure 1 The bar chart of the worm load and egg load in Example 3, and the statistical chart ns represents non-significant difference. As can be seen from the figure, there is no significant difference in the worm load between the AAV8-ctrl group of mice and the AAV8-KLB group of mice, indicating that the infection of Schistosoma japonicum is random infection, and there is no difference in group infection.

[0056] Example 5

[0057] Calculate the egg load of liver tissue of mice after being infected with Schistosoma japonicum

[0058] ① After the mice in Example 3 infected with Schistosoma japonicum for 6 weeks were anesthetized and euthanized, the abdominal cavity of the mice was opened to fully expose the internal organs, and the intestinal tube and fat were gently pushed to the right side of the mouse abdomen; 100 mg of liver tissue of the mice was taken and cut into tissue fragments, and placed in 15 mL of 1% sodium hydroxide solution;

[0059] ② Shake on a 37°C shaking table for 2 hours until the liver tissue fragments completely disappear.

[0060] ③ Take 20 μL of the digestion solution containing Schistosoma japonicum eggs, and count under a microscope. Each mouse liver tissue was counted three times independently and the average value was taken.

[0061] Figure 1For the bar graphs of the worm burden and egg burden in Example 3, ns in the statistical chart represents non-significant difference. As can be seen from the figure, there is no significant difference in the worm burden between the AAV8-ctrol group mice and the AAV8-KLB group mice, indicating that the infection of Schistosoma japonicum is random infection, and there is no difference in the infection between groups.

[0062] Example 6

[0063] Freeze section detection of the location and expression level of AAV8 in mouse tissues

[0064] ①Take the liver tissue, spleen tissue and small intestine tissue of the positive reference group of mice injected with AAV8-GFP in Example 3, and place them in 4% paraformaldehyde. Since AAV8-GFP has spontaneous fluorescence, it is stored in the dark;

[0065] ②Paraffin section deparaffinization to water: sequentially place the section in the environmentally friendly deparaffinization liquid for 10 min, the environmentally friendly deparaffinization liquid for 10 min, the environmentally friendly deparaffinization liquid for 10 min, anhydrous ethanol I for 5 min, anhydrous ethanol II for 5 min, anhydrous ethanol III for 5 min, and distilled water for washing;

[0066] ③Antigen repair: place the tissue section in a repair box filled with EDTA antigen repair buffer (PH 8.0) in a microwave oven for antigen repair. Medium heat for 8 min, stop heating for 8 min, and then medium-low heat for 7 min. During this process, prevent the buffer from evaporating too much, and do not dry the section. After natural cooling, place the slide in PBS (PH 7.4) and shake for 3 times, 5 min each time.

[0067] ④Since AAV8-GFP has spontaneous fluorescence, only DAPI needs to be used to restain the nucleus. After slightly shaking the section dry, add DAPI staining solution in the circle, and incubate at room temperature for 10 min in the dark.

[0068] ⑤Mounting: place the slide in PBS (PH 7.4) and shake for 3 times, 5 min each time on a decolorizing shaker. After slightly shaking the section dry, mount the section with an anti-fluorescence quenching mounting agent.

[0069] ⑥Microscopy and photography: place the section under a scanner to collect images or under a fluorescence microscope to take photographs. (DAPI ultraviolet excitation wavelength 330-380 nm, emission wavelength 420 nm, blue light; AAV8-GFP has spontaneous fluorescence of GFP, green light).

[0070] Figure 2To implement the frozen section detection of the protein in Example 3 with AAV8 as the carrier to locate and express the level of the protein in the mouse tissue, the frozen section shows the location of AAV8 in the mouse tissue (liver tissue, spleen tissue, small intestine tissue) after 3 weeks of AAV8-GFP tail vein injection (blue represents the nucleus, green represents GFP, and the color area represents the expression level). The picture result shows that the green fluorescence mainly occupies a large area in the liver tissue, indicating that the protein with AAV8 as the carrier will mainly express in the mouse liver tissue after injection. The use of AAV8 as the viral carrier to encapsulate KLB can achieve the purpose of high expression of KLB in the liver tissue of the mouse.

[0071] Example 7

[0072] qPCR detection of EMT and liver fibrosis level of mouse liver tissue

[0073] Part of the liver tissue of the mouse infected with Schistosoma japonicum for 6 weeks in Example 2 and Example 3 was taken and placed in a 2 mL EP tube, and the Trizol method was used to extract the liver tissue RNA. After determining the RNA concentration and purity by the microplate reader, the RNA was reversely transcribed into cDNA by using the reverse transcription kit (Xinbei Biological, R202-02). The cDNA after reverse transcription was used as a template, and the qPCR mix (Xinbei Biological, Q204-01) was used to perform the qPCR reaction to detect the gene expression of EMT and liver fibrosis indicators Collagen I, Collagen III, Collagen IV, α-SMA, and vimentin in the liver tissue (primers are shown in Table 1).

[0074] Table 1 Primer sequences of liver fibrosis indicators

[0075]

[0076]

[0077] Figure 3 The mouse liver tissue fibrosis indicators in Example 2. In Example 2, the mouse was infected with Schistosoma japonicum for 4 weeks, and then klotho recombinant protein was injected intraperitoneally for 2 weeks. The qPCR was used to detect the gene expression levels of the mouse liver fibrosis indicators, including α-SMA, Collagen 1, Collagen 3, and Collagen 4. As can be seen from the figure, after the mouse infected with Schistosoma japonicum was injected intraperitoneally with klotho recombinant protein, the expression levels of the mouse liver fibrosis indicators α-SMA, Collagen 1, Collagen 3, and Collagen 4 were significantly lower than those of the mouse injected intraperitoneally with PBS.

[0078] Figure 4is the fibrosis index of mouse liver tissue of Example 3. Example 3 detected the gene expression level of EMT and fibrosis related indicators vimentin, Collagen 1 and Collagen 3 in liver tissue by qPCR at 6 weeks after the AAV8-KLB injected mice were infected with Japanese blood flukes. It can be seen from the figure that the expression level of EMT indicator vimentin and fibrosis indicators Collagen 1 and Collagen 3 in the liver tissue of AAV8-KLB injected mice after Japanese blood fluke infection was significantly lower than that of AAV8-ctrl group mice.

[0079] Figure 3 and Figure 4 The results all suggest that supplementing the content of KLB in mouse liver tissue can alleviate the liver fibrosis caused by Japanese blood fluke infection.

[0080] Example 8

[0081] Western blot detection of KLB, EMT and liver fibrosis level in mouse liver tissue

[0082] ①Take part of the liver tissue of the mice infected with Japanese blood flukes for 6 weeks in Example 2 and Example 3, add appropriate amount of RIPA lysis buffer to fully lyse the tissue, and measure the protein concentration by BCA method;

[0083] ②Add 5xloading, mix thoroughly, and then place in 100℃ metal bath for oscillation boiling for 10 min to fully denature the protein.

[0084] ③After SDS-PAGE electrophoresis, use a rapid wet transfer instrument to transfer the membrane.

[0085] ④After transferring the membrane, block at room temperature for 1 h, and wash with TBST for 3 times;

[0086] ⑤KLB antibody (R&D, AF2619), EMT related antibodies E-cadherin (CST, 3195s), N-cadherin (CST, 13116s), vimentin (CST, 5741s), and fibrosis related antibodies α-SMA (CST, 19245s), COL1 (bioss, bs-10423R) as primary antibodies, incubate at 4℃ overnight, and wash with TBST for 5 times;

[0087] ⑥Add secondary antibody, incubate at room temperature for 1 h, wash with TBST for 3 times;

[0088] ⑦Spread the color developing solution evenly on the membrane, select appropriate exposure time for exposure and take pictures for preservation. Use Image J software to analyze the gray value of the band.

[0089] Figure 5Figure 2 is the result of western blot detection of EMT and liver fibrosis levels in mouse liver tissue of Example 2. Example 2 detected EMT and fibrosis indicators in mouse liver tissue by western blot after 4 weeks of infection with Schistosoma japonicum and 2 weeks of intraperitoneal injection of Klotho recombinant protein. The figure shows that the protein expression levels of E-cadherin, a-SMA and Collagen 1 in the liver tissue of mice injected with Klotho recombinant protein were significantly lower than those of mice injected with PBS.

[0090] Figure 6 Figure 3 is the result of western blot detection of KLB expression in mouse liver tissue of Example 3. Example 3 detected the KLB protein expression level in the liver tissue of AAV8-KLB group mice and AAV8-ctrol group mice 6 weeks after infection with Schistosoma japonicum. The number of * in the statistical chart represents the degree of significant difference. The results in the figure show that the KLB protein expression level in the liver tissue of AAV8-KLB group mice was significantly higher than that of AAV8-ctrol group mice, indicating that the overexpression of KLB in the liver tissue was successful.

[0091] Figure 7 Figure 4 is the result of western blot detection of EMT and liver fibrosis levels in mouse liver tissue of Example 3. Example 3 detected the protein expression levels of EMT and fibrosis-related indicators E-cadherin, N-cadherin, vimentin and a-SMA in the liver tissue 6 weeks after infection with Schistosoma japonicum. The results in the figure show that the protein expression of E-cadherin, an epithelial marker in the liver of AAV8-KLB group mice, was significantly higher than that of AAV8-ctrol group mice, and the protein expression levels of N-cadherin, vimentin and a-SMA, mesenchymal markers and fibrosis markers, were significantly lower than those of AAV8-ctrol group mice.

[0092] Figure 5 and Figure 7 The results all show that supplementing KLB protein expression in mouse liver tissue can reduce the EMT and liver fibrosis changes caused by Schistosoma japonicum infection.

[0093] Example 9

[0094] Detection of egg granuloma size by hematoxylin-eosin staining (HE staining)

[0095] ① Prepare paraffin sections of the liver tissue of mice infected with Schistosoma japonicum for 6 weeks in Example 2 and Example 3;

[0096] ② Dewaxing to water-sudan staining-erythrosin staining

[0097] ③ Dehydration, mounting, and microscopic examination, the size of individual granulomas and granulomas around the eggs can be observed.

[0098] Figure 8 HE and Masson staining figures of Example 3. The HE staining figure shows the egg granuloma size of liver tissue of AAV8-KLB group mice and AAV8-ctrl group mice at 6 weeks after Schistosoma japonicum infection. The number of * in the statistical chart represents the significant difference, and the number represents the degree of significant difference. The HE results in the figure show that the area of egg granuloma of AAV8-KLB group mice is significantly smaller than that of AAV8-ctrl group mice, which indicates that supplementing KLB in mouse liver tissue can reduce the formation of egg granuloma caused by Schistosoma japonicum infection and reduce the inflammatory response,

[0099] Example 10

[0100] Collagen fiber area was detected by ponceau acid fuchsin-aniline blue staining (Masson staining)

[0101] ① The liver tissue of mice infected with Schistosoma japonicum for 6 weeks in Example 2 and Example 3 was made into paraffin sections;

[0102] ② Dewaxing to water-potassium dichromate staining-iron hematoxylin staining-ponceau acid fuchsin staining-phosphomolybdate acid staining-aniline blue staining;

[0103] ③ Dehydration, mounting, and microscopic examination, the collagen fibers are blue;

[0104] ④ The area of collagen fibers was detected to evaluate the fibrosis level of liver tissue.

[0105] Figure 8 HE and Masson staining figures of Example 3. The Masson staining figure shows the area of collagen fibers of liver tissue of AAV8-KLB group mice and AAV8-ctrl group mice at 6 weeks after Schistosoma japonicum infection. The number of * in the statistical chart represents the significant difference, and the number represents the degree of significant difference. The Masson results in the figure show that the area of collagen fibers of AAV8-KLB group mice is significantly smaller than that of AAV8-ctrl group mice. It indicates that supplementing KLB in mouse liver tissue can reduce the collagen fibers formed by Schistosoma egg deposition, and the fibrosis level is reduced.

[0106] The present application verifies the therapeutic effect of KLB protein on EMT and liver fibrosis caused by Schistosoma japonicum infection, and proves that KLB can be used as a drug to treat liver fibrosis of Schistosomiasis.

[0107] The detailed description of the application is described in detail, and those skilled in the art should recognize that the above embodiments are exemplary and cannot be understood as limiting the application, and those skilled in the art can make several improvements and modifications to the application without departing from the principles of the application, and the technical solutions obtained by the improvements and modifications also fall within the protection scope of the claims of the application.

Claims

1. Use of KLB protein in the preparation of a drug for treating schistosome infection.

2. Use according to claim 1, characterized in that, Use of the KLB protein in the preparation of a drug for treating Schistosoma japonicum infection.

3. Use according to claim 1, characterized in that, Use of the KLB protein in the preparation of a drug for treating liver fibrosis caused by Schistosoma japonicum infection.

4. Use of Klotho recombinant protein in the preparation of a drug for treating schistosome infection.

5. Use according to claim 4, characterized in that, Use of the Klotho recombinant protein in the preparation of a drug for treating Schistosoma japonicum infection.

6. Use according to claim 4, characterized in that, Use of the Klotho recombinant protein in the preparation of a drug for treating liver fibrosis caused by Schistosoma japonicum infection.

7. Use of AAV8 adeno-associated virus loaded with KLB gene in the preparation of a drug for treating schistosome infection.

8. Use according to claim 7, characterized in that, The AAV8 adeno-associated virus as a carrier loaded with KLB gene, and the AAV8 adeno-associated virus targets the liver for infection, so that the liver tissue highly expresses KLB protein.

9. Use according to claim 7, characterized in that, Use of the AAV8 adeno-associated virus loaded with KLB gene in the preparation of a drug for treating Schistosoma japonicum infection.

10. Use according to claim 7, characterized in that, Use of the AAV8 adeno-associated virus loaded with KLB gene in the preparation of a drug for treating liver fibrosis caused by Schistosoma japonicum infection.

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