The expression changes in IL-6,GP130,JAK1 and STAT1 genes in PC12 cell after combined soman and hypoxia injury / 解放军医学杂志

ABSTRACT

Objective To investigate the complex effect of soman and hypoxia on PC12 cell line as shown by the expression changes in IL-6/GP130, JAK1 and STAT1. Methods A cell model of intoxication by combining effect of hypoxia and soman was reproduced, and it was divided into four groups control, soman intoxication, hypoxia combined with soman intoxication, and Genistein inhibition groups. PC12 cells were cultured in RPMI 1640 and were treated with NGF (50ng/ml) for seven days. The differentiated PC12 cells were then exposed to hypoxia in an incubator containing 5% CO2, 95% N2 and (or) incubated with soman (20?mol/L) for 2, 6, 12 or 24 hours. The expression levels of IL-6/GP130, JAK1, STAT1 mRNA and protein were assessed by RT-PCR and Western blot in PC12 cells. The products were sequenced by Sanger's double strand DNA sequence determination. Results In soman intoxicated group, the expression levels of IL-6/GP130, JAK1, STAT1 mRNA and protein were elevated in PC12 cell, reaching the peak level at 12 hours, and then lowered at 24 hours, but remaining higher than that of control group. In combined soman intoxicated and hypoxia group, the expression levels of IL-6/GP130, JAK1, STAT1 mRNA and protein reached the peak value at 6 hours, being higher than that of control group, soman intoxicated group and Genistein inhibition group. It was shown that the sequences of the products as amplified by RT-PCR were the same as that found in the GenBank. Conclusion Soman intoxication or (and) hypoxia up-regulate the expression of IL-6/GP130. Both hypoxic condition and soman treatment can up-regulate the expression of JAK1/STAT1 mRNA and protein in PC12 cells. JAK-STAT pathway may play a role in the mechanisms of brain injury resulted from hypoxia and soman poisoning.