| ObjectiveTo investigate the association between IL-32 gene polymorphisms and hepatic flare of chronic HBV infection,and the mechanism of IL-32 in liver fibrosis.Methods1.Patients who saw the Department of Infectious Diseases of the Third Affiliated Hospital of Guang Zhou Medical University from June 2016 to February 2019,were enrolled in the present study,including 151 chronic hepatitis B patients(hepatitis group)and 104 chronic HBV carriers(carrier group).Their peripheral blood samples were collected and blood DNA was extracted,respectively.Then alleles and genotypes of rs28732698 and rs12934561 were assayed by Sanger sequencing,and the association between different genotypes and alleles and hepatic flare of chronic HBV infection were analyzed,respectively.2.Hepatic tissue samples were collected from parts of patients with chronic HBV infection.According to pathological results,patients were divided into two groups: non or mild liver fibrosis group,moderate to severe liver fibrosis group.The expression levels of Integrin αⅤβ6 and FAK were detected by using immunohistochemistry.3.Different concentrations of rh IL-32γ were used to stimulate hepatic stellate cell(LX-2 cells),and the expression levels of FAK,paxillin,and α-SMA were detected by using Western blot,respectively.Results1.At the IL-32 rs28372698,the frequencies of AA,AT,and TT genotype were of35.1%,49.7%,15.2% in hepatitis group,and of 51%,40.3%,8.7% in carrier group,respectively.The frequency of TT genotype in hepatitis group was higher than that in carrier group(15.2% vs 8.7%,P=0.03).The frequencies of A and T allele were of 59.9%and 40.1% in hepatitis group,and of 71.2% and 28.8% in carrier group,respectively.The frequency of T allele in hepatitis group was higher than that in carrier group(40.1% vs28.8%,P=0.009).At the IL-32 rs12934561,the frequencies of CC,CT,and TT genotype were of 16.6%,55.6%,27.8% in hepatitis group,and of 10.6%,47.1%,42.3% in carrier group,respectively.The frequencies of CC+CT genotypes in hepatitis group were higher than that in carrier group(72.2% vs 57.7%,P=0.04).The frequencies of C and T allele were of 44.4% and 55.6% in hepatitis group,and of 34.1% and 65.9% in carrier group,respectively.The frequency of C allele in hepatitis group was higher than that in carrier group(44.4% vs 34.1%,P=0.02).2.The median of mean optical density of Integrin αⅤβ6 and FAK were of 0.263 and0.017 in the hepatic tissue of patients with moderate to severe liver fibrosis group,and of0.223 and 0.007 in non or mild liver fibrosis group,respectively(P=0.037,P=0.035).The difference of the two groups was statistically significant.3.Stimulated by rh IL-32γ at a concentration of 80ng/ml,the expression levels of FAK,paxillin,and α-SMA in LX-2 cells were of 1.37±0.57,0.16±0.01,and 0.16±0.02,respectively.However,the expression levels of FAK,paxillin,and α-SMA were of0.86±0.27,0.08±0.03,and 0.07±0.02 in control group,respectively(P=0.042,P=0.044,P=0.004).The difference of the two groups was statistically significant.Conclusions1.At the rs28372698 of IL-32 promoter,the TT genotype and T allele might be associated with hepatic flare of chronic HBV infection;2.At the rs12934561 of IL-32 intron,the CC+CT genotypes and C allele might be associated with hepatic flare of chronic HBV infection;3.IL-32 could stimulate the expression of α-SMA,FAK and paxillin by LX-2 cells,and might be participated in the progress of liver fibrosis through Integrin/FAK pathway. |
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