Effects of PI3K/PKB signaling pathway on expression of osteopontin in human hepatic stellate cells induced by transforming growth factor-β1 / 中国病理生理杂志

ABSTRACT

AIM:To investigate the regulatory effects of phosphatylinositol 3-kinase/protein kinase B (PI3K/PKB) signaling pathway on the expression of osteopontin ( OPN) in transforming growth factor-β1 ( TGF-β1 )-induced hu-man hepatic stellate cells .METHODS:Human hepatic stellate cell line LX-2 was cultured in DMEM and stimulated by TGF-β1 at the final concentration of 2.5, 5, 10 and 20 μg/L for 24 h or at final concentration of 10 μg/L for 12 h, 24 h and 48 h.LX-2 cells were pretreated with wortmannin , a specific inhibitor of PI3K/PKB signaling pathway , at final con-centration of 0.1 μmol/L for 1 h, followed by incubation with TGF-β1 at final concentration of 10μg/L for 24 h.The cells were collected.The expression of OPN was detected by real-time PCR and Western blotting .RESULTS: In LX-2 cells, the expression of OPN was apparently elevated when incubated with TGF-β1 .With the increase in TGF-β1 concentration or the extension of incubation hours , the expression of OPN was increased gradually in a dose-and time-dependent manner with certain limits.LX-2 cells pretreated with wortmannin and incubated with TGF-β1 had a significant decrease in the OPN expression as compared with control group (P<0.01).CONCLUSION:The expression of OPN in TGF-β1-induced LX-2 cells is regulated by the PI3K/PKB signaling pathway.