| Objectives:Continued viral replication and immune response is the big problem of HBV treatment. A recent study showed that HMGB1 is seemed to been an important cytokine in infectious disease, previous studies had found that HMGB1 related with the pathogenesis of many viral diseases. This research was to elucidate the HMGB1,sRAGE and IL-6 levels in blood of patients with HBV, and the expression of HMGB1,RAGE mRNA in liver tissues of patients with various types of HBV infection. All types of patients were compared with clinical indicators, and explore the function of HMGB1 in HBV replication and liver tissue injury. Aboved research provided the theoretical basis for the pathogenesis of HBV infection.Methods:1.Collected the blood of patients with various types of HBV infection. ELISA and Real-Time fluorescence quantitative PCR were used to detect the expression of HMGB1,sRAGE and IL-6 protein and HMGB1 mRNA, then analysis related clinical indicators. 2. Collected the liver tissue of patients with various types of HBV infection. Real-Time fluorescence quantitative PCR were used to detecte the HMGB1 mRNA. The distribution and expression of HMGB1 protein were detected by Western blot and Immunofluorescence, then analysis related clinical indicators.Results:1.Real-Time fluorescence quantitative PCR showed the expression levels of HMGB1 mRNA in the peripheral blood of patients with HBV, AsC,CHB-1,CHB-2,CHB-3,SH,LC-1,LC-2 groups 2-△△CT>2,the HMGB1 mRNA in AsC,CHB-1,CHB-2,CHB-3,SH groups were gradually increased, there was a significant difference between all group; LC-1,LC-2 groups 2-△△CT>2,the HMGB1 mRNA in LC-1,LC-2 groups were gradually increased, there was a significant difference between all group .2.ELISA showed that the concentrations of HMGB1 protein in the serum of patients with HBV, it was gradually increased in AsC,CHB-1,CHB-2,CHB-3,SH groups. AsC group was no significant difference with control group(P >0.05);CHB-1 group had a significant difference with control group (P<0.05);there was a significant difference between CHB-2,CHB-3,SH groups and control group (p<0.01);HMGB1 protein gradually increased in LC-1,LC-2 groups, there is a significant difference with control group.(p<0.01) 3.ELISA showed that the concentrations of sRAGE protein in the serum of patients with HBV, it was gradually decreased in AsC,CHB-1,CHB-2 groups, was significant difference (p<0.01); it was gradually increased in CHB-3,SH groups, there was a significant difference with control group(p<0.01); it was gradually increased in LC-1,LC-2groups, there was a significant difference with control group(p< 0.01).4. ELISA showed that the concentrations of IL-6 protein in the serum of patients with HBV, it was gradually increased in AsC,CHB-1,CHB-2,CHB-3,SH groups;there was no significant difference between AsC group with control group(P >0.05);there was a significant difference between CHB-1 group with control group (P<0.05);there was a significanct difference between CHB-2,CHB-3,SH groups and control group (p<0.01);it was gradually increased in LC-1,LC-2 groups and control group, there was a significant difference with control group(p<0.01).5. The correlation analysis of HMGB1 with each indexs in the peripheral serum of patients with HBV, there were positive relation between HMGB1 with TBIL,IL-6 (r=0.495,0.886,0.511,P<0.01), There were negative relation between HMGB1 with PTA,A,CHE (r=-0.727,-0.848,-0.849, P<0.001);There was a negative correlation between HMGB1 and sRAGE in control,AsC,CHB-1,CHB-2 groups(r=-0.873,P<0.01); there was a positive correlation between HMGB1 and sRAGE in CHB-3,SH,LC-1, LC-2 groups and control group(r=0.832,P<0.01).6. Real-Time fluorescence quantitative PCR showed that the expression levels of HMGB1 mRNA in liver tissue of patients with HBV. AsC,CHB-1,CHB-2,LC-1 groups were significant higher than that in control group, and 2-△△CT>2; it was gradually increased in AsC,CHB-1,CHB-2groups.7. Western blot showed that the expressions of HMGB1 protein in liver tissue of patients with HBV, it was significantly higher in AsC,CHB-1,CHB-2 groups than in control group(P<0.01);the expressions of RAGE protein in AsC,CHB-1,CHB-2,LC-1 groups were significantly higher than that in control group(P<0.01).8.Western blot showed that the expressions of HMGB1 protein in hepatocytic cytoplasm of patients with HBV, it was significantly higher in AsC,CHB-1,CHB-2 groups than that in control group(P<0.01);the expressions of RAGE protein in AsC,CHB-1,CHB-2,LC-1 groups were significantly higher than that in control group(P<0.01)9. Western blot showed that the expressions of HMGB1 protein in hepatocytic nuclear of patients with HBV, it was significantly higher in AsC,CHB-1,CHB-2 groups compare with control group(P<0.01),LC-1 group was no significant difference with control group .10. The correlation analysis of HMGB1 protein in liver tissue with each index in patients with HBV, there was a positive relation between HMGB1 with TBIL (r=0.725,P<0.01); there were a negative relation between HMGB1 with PTA,A,CHE (r= -0.736,-0.684,-0.861, P<0.01); there was no relation between HMGB1 with HBV– DNA (r =0.008,P>0.05) .11.Immunofluorescence results showed that the expression of HMGB1 in liver tissue of patients with HBV, the HMGB1 was distribution in hepatocellular nuclear and cytoplasm, expression in hepatocellular cytoplasm especially, the HMGB1 expression in nuclear and cytoplasm were gradually increased in AsC,CHB-1,CHB-2 groups, expression enhanced in cytoplasm especially; the HMGB1 expression enhanced in LC-1 group, especially in cytoplasm .12. Immunofluorescence results showed that the expression of RAGE in liver tissue of patients with HBV, the RAGE was distribution in hepatocellular cytoplasm but not in nuclear in all groups, it was gradually enhanced in AsC,CHB-1,CHB-2,LC-1 groups . .Conclusions:1. The contents of HMGB1 protein and HMGB1 mRNA in the peripheral blood of patients with HBV were significantly higher than that in healthy control, the severity of patients has a positive correlated with expression of HMGB1, which indicated that the HMGB1 was closely related to the occurrence and development of HBV infection.2. Concentration of HMGB1,tissue inflammation and fibrosis affecting the expression of sRAGE, indicated that HMGB1 coordination sRAGE may affect the liver inflammatory response and the pathological process of liver cirrhosis.3. There was a negative correlated with HMGB1 and clinical indicators(CHE,A,PTA), HMGB1 in serum of patients with HBV, indicating that HMGB1 may affected liver metabolism, also showed that HMGB1 may as a new index of liver function.4. The levels of HMGB1 mRNA in liver tissue of patients with HBV, further illustrated that the expression of HMGB1 mRNA is related with severity of HBV disease.5. There was a positive correlation with HMGB1 and HBV-DNA in peripheral blood , but the liver HMGB1 protein was no correlation with HBV-DNA, the HMGB1 functional status in the nucleus and cytoplasm of liver cells is unknown, the relation with HMGB1 and HBV replication need further research to demonstrate.6. The distribution of HMGB1 protein in the nucleus and cytoplasm changed with the different types of diseases, indicated that the distribution of HMGB1 in the liver cells affects its function.7. The expression of RAGE and HMGB1 in the liver tissue gradually increased with the severity of liver disease, especially in the cirrhosis group, suggested that HMGB1 may play an important role in liver cirrhosis through RAGE receptor.
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