Effect of Cordyceps polysaccharides on hepatocyte apoptosis induced by tumor necrosis factor-α

Abstract

Objective To investigate the effect of Cordyceps polysaccharides on hepatocyte apoptosis and its mechanism of action. Methods Normal human L02 hepatocytes without any drug treatment was established as normal group. The L02 hepatocytes were treated with different concentrations of tumor necrosis factor-α (TNF-α) (5, 10, 20, and 40 ng/ml) for 24 hours to screen out the modeling conditions for the model of hepatocyte apoptosis, which were established as model group. According to the experimental design, L02 hepatocytes pretreated with three different concentrations of Cordyceps polysaccharides (50, 100, and 200 μg/ml) for 12 hours, with or without TNF-α treatment for 24 hours at the selected concentration, were established as experimental group. Related samples were collected for analysis. CCK8 assay was used to measure cell proliferation; Annexin V/PI double staining was used to measure the number of apoptotic hepatocytes; RT-PCR was used to measure the mRNA expression of cell apoptosis genes (Bax, caspase-3, caspase-8, caspase-9, and death receptor Fas); Western blotting was used to measure the protein expression of cleaved-caspase-3 and cleaved-caspase-8. A one-way analysis of variance or the Kruskal-Wallis H test was used for comparison of continuous data between multiple groups, and the least significant difference t-test or the Dunnett-t test was used for further comparison between two groups. Results CCK8 assay showed that compared with the normal group, the L02 hepatocytes induced by 40 ng/ml TNF-α for 24 hours had a significant reduction in proliferation (73.54%±14.19% vs 100.00%±23.61%, P Conclusion Cordyceps polysaccharides can effectively inhibit the apoptosis of normal L02 hepatocytes induced by TNF-α.