The Effects Of Plasma From Patients With Severe Hepatitis On The Growth And Metabolism Of HepG2 And Human Fetus Hepatocytes,Porcine Hepatocytes

The highly differentiated hepatocytes are the most important constituents in bioartificial liver which have many special liver functions like the synthesis and biotransformation. However, the same time as efficient removal of xenobiotics and reactive compounds from the liver failure plasm, function and viability of hepatocytes in bioreactors will be influenced by the interaction of liver failure patient plasma. In China there are a lot of patients with severe viral hepatitis accepting the BAL therapy. Therefore, it would be important to investigate the direct interactions between severe hepatitis plasma(SHP) and hepatocytes in bioreactor.Here I have this study to investigate the effects of SHP on Hep G2 cells and Human fetus, porcine hepatocytes. The methods and results are following.1. The effects on viability of HepG2 cellsThe HepG2 cell viability was inhibited in 10%, 30%, 50%, 100% SHP during the 5 day culture, a significant difference in viability occurred after 3 days of culture in medium containing 100% SHP compared to the controls. Although the HepG2 cell viability was inhibited in both activation and deactivation SHP, there was no significant difference between themselves. 2. The effects on cell membrane injury of HepG2 cells After 5h exposure to the SHP, the leakages of LDH and γ-GT were significantly increased in the HepG2 cells, at the end of the experient they are approximately 2 times higher than that of media containing 10% NCS. The bullas were observed from SEM pictures of HepG2 cells cultured in SHP for 24h. The membranes of the cells became indistinct. Exposure to SHP resulted in a signifiant increase in apoptosis compared to control cultures.3. The effects on function of HepG2 cellsThe protein content of HepG2 cells cultured in medium supplemented with 10%SHP was less than that of the controls in 10%NCS. The protein content of cultures grown in 100%SHP was significantly less than that of those grown in medium containing 10%SHP. Exposure to SHP resulted in no decrease in AFP compared to control cultures. When confluent HepG2 cells were challenged with medium containing SHP, the GSH content was significantly increased after 24h compared to control cultures in 10% NCS. After 3 days the GSH level declined dramatically to below the levels in control cultures.4. The effects on morphology and function of human fetus and pocine hepatocytesLDH, AST in medium containing SHP is higher than that of media containing 10%NCS, which indicates the injury of cell membrane of human fetal and pocine hepatocytes. AFP declined in both human fetal hepatocytes and pocine hepatocytes exposed to SHP. The morphology of human fetus and pocine hepatocytes underwent a dramatic change once they were put into SHP. The results of this study imply: SHP inhibits the growth rate and the synthesis of protein; disturbs glutathione homeostasis; and induces apoptosis and morphological changes in cultured HepG2 cells. SHP also inhibits the growth rate and the synthesis of protein; damages cell membrane in cultured human fetal and pocine hepatocytes.