Study On The Toxicity Of Three Anti-virus Drugs Of Nucleosides To Primary Hepatocytes Of Rats

The hepatocytes were primarily cultured and used to evaluate the toxicity of three anti-virus agent of nucleosides on the hepatocytes.The hepatocytes were isolated and harvested by collagenase and trypsin from Wistar rats. The viability of the hepatocytes was tested by trypan blue exclusion and the quantity was calculated at the same time. The morphological change of primarily cultured hepatocytes was observed. The proliferative hepatocytes were tested by MTT experiment. The results showed that the quantity, isolated by collagenase, was 2.4 times as much as trypsin, the viability was three-tenths of trypsin, the activity and the adherence effect was inferior to trypsin. The results showed that although the quantity isolated by collagenase was so much, the viability was bad, the quantity isolated by trypsin was inferior to collagenase, but the viability was high, and used to evaluate the hepatocytes toxicity of three anti-virus drugs.The density of the hepatocytes were adjusted to 3×10⁵cells/mL by DMEM medium, and the hepatocytes were devided into the drug group, the positive control group, the negative control group and the blank control group. The hepatocytes were cultured in 37℃, 5%CO₂ incubator for 4h, then the cultural supernatants were discarded. The hepatocytes were exposed to 10, 5, 3.3, 2, 0.4 and 0.08mmol/L AZT; 5, 2.5, 1.25, 0.625, 0.125 and 0.025mmol/L ACV; 10, 4, 2, 1, 0.2 and 0.04mmol/L GCV; 10,2 and 0.5mmol/L CCl₄; DMEM medium containing 1% DMSO and DMEM medium. The activity of the hepatocytes was detected at 6, 12 and 24h by MTT experiment, the content of AST, ALT, LDH and BUN was detected at 6 and 12h, the hepatocytes were stained with HE at 12h to observe the morphological changes.