Immunofluorescent detection of apoptosis in chronic viral hepatitis and other liver diseases

ABSTRACT

Chronic viral hepatitis is a common health problem in many countries including Egypt. The microscopic appearance of chronic viral hepatitis is usually associated with the appearance of eosinophilic structures called apoptotic bodies. Apoptosis is a type of programmed cell death. Abnormalities of apoptosis might contribute to many disease processes, the most important of which is neoplasia. The aim of this work is to study the frequency of apoptosis in cases of chronic viral hepatitis C [HCV], B [HB], pure Bilharzial liver affection, combined Bilharzial and chronic viral hepatitis C in addition to cases of hepatocellular carcinoma [HCC]. Liver biopsies [10 cases of HCV+ve patients, 6 cases HBs Ag +ve patients, 8 cases pure Bilharzial liver affection, 10 cases of combined Bilharzial and HCV, 10 cases of HCC and 6 cases of normal liver tissue obtained from areas adjacent to focal hepatic lesions were used as control]. A modified method of the terminal deoxynucleotidyl transferase- mediated dUTP-biotin nick end labeling [TUNEL] using immunofuorescence [FITC - dUTP] was used to detect apoptosis. Apoptotic index was determined for each case by counting apoptotic cells per 1000 cells in different fields. Apoptotic index for control cases was 0.1 +/- 0.08, cases of HCV 0.23 +/- 0.125, cases of HB 0.25 +/- 0.1, pure Bilharzial cases 0.125 +/- 0.08, combined Bilharzial and HCV cases 0.24 +/- 0.126 and for cases of HCC, it was 0.08 +/- 0.07.The difference between apoptotic index in cirrhotic and non-cirrhotic cases was statistically non significant [P = 0.896]. As a conclusion, we found that the modified TUNEL method using fluorescein isocyanate labeled dUTP was useful and sensitive for detecting apoptosis in the liver. The apoptotic index increases significantly than the control in cases of chronic viral hepatitis C [P = 0.044], B [P = 0.024] and combined HCV and Bilharzial liver affection [P = 0.033]. Apoptosis in pure Bilharzial liver affection does not significantly differ from the control [P = 0.636]. In cases of HCC, apoptosis decreases than the control in a non-significant manner [P = 0.647]. Apoptotic index does not differ significantly between cirrhotic and non- cirrhotic cases