Expression And Significance Of IL-17,IL-23 And IL-8 In HBV Chronic Infection Tree Shrew Model And Clinical HBV-Related Liver Diseases

1.Expression and significance of IL-17,IL-23,IL-8 in HBV chronically infected tree shrew modelObjective:To inoculate newborn tree shrews with high-titer human serum HBV to construct a model of HBV chronic infection of tree shrews,and to observe the changes of IL-17,IL-23 and IL-8 in tree shrews after infection.Methods:(1)Establishment of a tree shrew model of HBV chronic infection:25new-born tree shrews were randomly divided into two groups:18 in the experimental group and 7 in the control group.The experimental group was subcutaneously inoculated with human HBV(10~7IU/ml)serum on both thighs at the 1st and 3rd days,and the control group was replaced with normal saline.Each inoculation volume was 300μL,a total of 600μL.After vaccination,blood was collected once through the femoral vein at weeks 8,12,16,20,and24,and liver biopsy was performed at 12 and 24 weeks.ELISA to detect the HBs Ag in serum.Quantitative real-time PCR(probe method)to detect HBV DNA in serum and liver tissue.To observe the HBs Ag in liver tissue by immunohistochemistry.The pathological conditions in liver tissue were observed by HE staining.(2)Expression of IL-17,IL-23 and IL-8 in the tree shrew model of HBV chronic infection:According to the test results,the experimental group were divided into a chronic infection group and a suspected chronic infection group.quantitative Real-time PCR(q PCR)was used to detect IL-17,IL-23 and IL-8m RNA in the liver tissues of three groups.ELISA was used to detect the expression of IL-17,IL-23 and IL-8 protein in the serum of three groups of tree shrews.And IL-17,IL-23 and IL-8 protein in liver tissue of three tree shrews was observed by immunohistochemistry.Results:(1)Validation of the tree shrew model of HBV chronic infection:Serum HBs Ag or HBV-DNA continued to be positive for 24 weeks was defined as the chronic infection group,and serum HBs Ag or HBV-DNA intermittently appeared twice or more positively as the suspected chronic infection group.During the observation period of 8 to 24 weeks,the HBs Ag and HBV-DNA test results of 25 tree shrews defined 8 tree shrews as chronic infection group.10tree shrews were defined as suspected chronic infection groups.The results of detection of HBs Ag and HBV-DNA in the serum of the control group were negative.2 of the chronic infection groups had HBs Ag positive expression at 12and 24 weeks,while no suspected chronic infection group and control group had no HBs Ag positive expression.HBs Ag positive expression is brownish yellow,mainly expressed in liver cytoplasm.The pathological and morphological changes of the liver tissue of the suspected chronic infection group and the chronic infection group were lighter than those of the control group.In the infection group,there were a small amount of monocytes and lymphocytes infiltration,and the cytoplasm of the liver cells became loose.(2)Expression of IL-17,IL-23 and IL-8 in the tree shrew model of HBV chronic infection:At the 12th and 24th weeks,the IL-17,IL-23,and IL-8m RNA in the liver tissue of tree shrews in the chronic infection group were significantly higher than those in the control group(P<0.05).The IL-17,IL-23,and IL-8 m RNA in liver tissue of tree shrews in the chronic infection group were slightly higher than those in the control group,but there was no significant difference(P>0.05).The serum IL-17,IL-23 and IL-8 in the chronic infection group were significantly higher than those in the control group at five detection points(8 weeks,12 weeks,16 weeks,20 weeks,and 24 weeks)(P<0.05).Compared with the control group,the levels of IL-17,IL-23 and IL-8 in the serum of the suspected chronic infection group were slightly higher,and only IL-17 had a significant difference at week 16(P<0.05).The IL-17,IL-23 and IL-8 proteins were mainly expressed in the cytoplasm of tree shrew liver tissues,and a few were expressed on the cell membrane.Positive staining showed mostly brown or brown.The IL-17,IL-23 and IL-8 proteins in tree shrew liver tissues in the chronic infection group were higher than those in the control group and suspected chronic infection group.There were significant differences(P<0.05).Conclusions:(1)High titer(107 IU/ml)of human serum HBV can successfully construct HBV chronic infection tree shrew model.(2)IL-17,IL-23 and IL-8 in the serum and liver tissues of tree shrews in HBV chronic infection group were higher than those in control group,suggesting that IL-17,IL-23 and IL-8 may be related to the chronicization of HBV infection and the development of hepatitis B.2.Expression and significance of IL-17,IL-23,IL-8 in clinical HBV-related liver diseasesObjective:To investigate the role of IL-17,IL-23 and IL-8 in the pathogenesis of chronic HBV infection.Collect serum from patients with HBV-related liver disease,and further detect changes in the expression of IL-17,IL-23 and IL-8 in patients.The role of IL-17,IL-23,and IL-8 in the progress of HBV-related liver disease was explored.Methods:From May 2019 to July 2019,a total of 240 patients were treated in the outpatient department of Infectious Diseases of the First Affiliated Hospital of Guangxi Medical University,including 48 asymptomatic carriers(As C),47patients with chronic hepatitis B(CHB),51 patients with HBV-related liver cirrhosis(HBV-LC),62 patients with HBV-related liver cancer(HBV-HCC),and 32 healthy patients during the same period.Quantitative quantitative PCR(probe method)to detect HBV DNA in the serum of all patients.ELISA was used to detect the IL-17,IL-23 and IL-8 protein in the serum of all patients.Results:The average level of HBV-DNA in selected patients with HBV liver-related diseases was at of 10~5to 10~6IU/ml.The HBV-DNA in healthy controls were lower than clinically positive standard(<500 IU/ml).The serum protein IL-17,IL-23 and IL-8 in As C group,CHB group,HBV-LC group and HBV-HCC group were higher than those in HC group(control group).The serum protein IL-17,IL-23 and IL-8 increases with the severity of HBV-related liver disease.Conclusions:During the process of HBV infection,it may develop into hepatitis B carriers,chronic hepatitis B,liver fibrosis and liver cancer.With the aggravation of the disease,the expression levels of IL-17,IL-23 and IL-8 are higher,suggesting that IL-17,IL-23 and IL-8 may be involved in the progression of HBV-related liver diseases.