The Affect Of Serum Substances On Growth-supressor Rate Of Bel-7402 Cell And Expression Of ALB MRNA In Patrents Suffered From Chronic Servere Hepatitis

ObjectivesSecondary liver damage plays an important role in pathogenesis of fulminant hepatic failure. Secondary Kver damage happens after primary liver damage. Secondary liver damage results in accumulation of a number of poisonous substances including large molecular substance, middle molecular substance and small molecular substance. The pathogenesis of these substances in secondary liver damage are complex, which will lead to hepatic cell damage finally. The present study was to investigate the affect of serum substances on growth - suppressor rate of Bel - 7402 cell and expression of ALB mRNA in vitro in patients suffered from chronic severe hepatitis complicated with hepatic encepha lopathy (II).MethodThe case group consist of 10 patients suffered from fulminant hepatic failure complicated with HE (II) and the control group consist of 10 patients suffered from chronic hepatitis B. After 12h fasting 10ml blood was taken . Strum was filtrated sterilly and added to Bel - 7402 cells cultivated 72 hours. According to MTT , the OD value was detected and then cell suppressor ratio was obtained. The damage condition of cell membrane was deduced by detecting the ALT and LDH releasing ratio of cell cultivative solution. We cultured Bel - 7402 cell in 25% of serum concentration for 24 hours and then extracted total mRNA of cells. It was reversely translated to cDNA, getted specific electrophoresis band of ALB mRNA, measuring the density and area of the band , the relative quantity of ALB mRNA was calculated.Resluts1. Through MTT examination, the growing suppress ratio of Bel - 7402 cells increases with serum concentration in the test group . When the serum concentration is up to 50% , growth suppressor ratio is 73% after cultured for 48 hours. There is significant difference between the test group and control group.2. Checking releasing ratio of ALT and LDH in cell culture solution in 25% serum and during different time such as Oh. 8h. 24h. 48h, there had no significant difference between examination group and control group.3. ALB mRNA expression in examination group decreased significantly in 25% serum concentration cultured after 24 hours for using RT - PCR .DiscussionA mass of different kinds of molecules accumulated in severe hepatitis patient , but the mechanism of how these materials injured hepatic cells was uncertain. In this examination, we conclude that growth suppress ratio of Bel - 7402 cell increased with the serum concentration, and the expression of ALB mRNA was decrease when the duration of culture prolonged, which maybe result from the death of a great deal of cells. In the examination group, the substance of serum restrained growth of cell, but had no effect on cell membrane, there maybe exist other mechanisms such as apoptosis, which need further reseach.ConclusionThe serum of patient with serve hepatitis complicated with hepatic encepha-lopathy (II) inhibit the growth of hepatic cell and make the expression of ALB mRNA decrease, but it has no effect on cell membrane during growth of hepatic cell.