Expression And Significance Of INOS,CD206,CD163 And Arg1 In Chronic Hepatitis B And Hepatitis B-related Liver Cirrhosis

Objective To study the expression differences of iNOS,CD206,CD163 and Arg1 in the peripheral blood and hepatic tissue between chronic hepatitis B and hepatitis B-related liver cirrhosis,understanding the polarization type of mononuclear macrophages and analyzing their relationships with the development of the disease.Methods In peripheral blood,61 cases with chronic hepatitis B,including 31 cases of HBeAg negative chronic hepatitis B,30 cases of HBeAg positive chronic hepatitis B;63 cases with hepatitis B-related liver cirrhosis,including 33 cases with compensated liver cirrhosis,30 cases with decompensated liver cirrhosis;36 normal people were included in the study.Collect peripheral venous blood.Total RNA was extracted from the whole blood for measurement of iNOS,CD206,CD163 and Arg1 mRNA expression levels using real-time PCR.The relationships between mRNA expression levels of the target above and clinical indicators of chronic hepatitis B and hepatitis B-related liver cirrhosis were analyzed.Hepatic tissue sections:Choose 19 cases tissue samples(? 5cm from the tumor or away from tumor)taken from surgical removal hepatic carcinoma with hepatitis B-relatedliver cirrhosis patients and 1 case tissue sample taken from hepatitis B-related liver cirrhosis patient except liver cancer as hepatitis B-related liver cirrhosis group,a total of 20 cases;Choose 5 cases adjacent normal tissue taken from surgical removal hepatic benign tumor or liver cyst patients as a normal control group.Extract total RNA from part of liver tissue and detect mRNA expression of iNOS,CD206,CD163 and Arg1 in the liver of hepatitis B-related liver cirrhosis;Another part of the liver tissue was made into wax blocks and HE staining and Masson staining were proceeded.Detect protein expression levels of iNOS,CD206 with immunohistochemical method,and use the IPP image analysis software to detect the average Optical Density value.Results The mRNA expression of iNOS,CD206,CD163 and Arg1 in chronic hepatitis B and liver cirrhosis groups increased when compared with normal control group(all P < 0.05),and iNOS mRNA rose mainly in hepatitis group,with CD206,CD163 and Arg1 mRNA rising mainly in cirrhosis group.Besides the expression levels of CD206,CD163 in decompensated hepatitis B-related liver cirrhosis were higher than compensated cirrhosis(all P < 0.01).INOS mRNA expression increased in HBeAg positive chronic hepatitis B when compared with HBeAg negative hepatitis(P < 0.05).In addition the expression level of CD163 was positively related with the level of total bilirubin,with negative correlation with prothrombin activity in liver cirrhosis group.Liver tissue sections: The mRNA expression of iNOS,CD206,CD163 and Arg1 in hepatitis B-related liver cirrhosis group increased when compared with normal control group(all P < 0.01).The mRNA expression of CD206 and CD163 in liver cirrhosis group were higher than iNOS mRNA expression(all P < 0.05).It had no statistical difference when comparing the mRNA expression quantity of iNOS with those of CD206,CD163 and Arg1 in normal group.HE stainingresults of hepatitis B-related liver cirrhosis group and normal group showed that the liver tissue structure more disorder and inflammatory cells and necrotic cells increased obviously in liver cirrhosis group compared with normal group.Masson staining showed that amount of collagen expression in hepatitis B-related liver cirrhosis group was much more than that in the normal group(P< 0.01),and pseudolobule formed in hepatitis B-related liver cirrhosis.The expression of iNOS and CD206 in liver of hepatitis B-related liver cirrhosis group was higher than normal group(P < 0.01).In liver cirrhosis group the expression level of CD206 was higher than iNOS(P < 0.01)and it had no statistical difference when comparing the expression quantity of iNOS with CD206 in normal group.Conclusions Peripheral blood mononuclear cells in chronic hepatitis B group may be mainly in the polarization direction of M1,and iNOS may participate in the process of chronic development in chronic hepatitis B;In hepatitis B-related liver cirrhosis group,macrophages may be in the polarization directions of M1 / M2 at the same time,but may be given priority to the polarization direction of the M2.CD163 is closely related to the occurrence and development of hepatitis B-related liver cirrhosis,and it may be used as one of the important indices to predict prognosis of hepatitis B-related liver cirrhosis and evaluate the severity of disease.