Effects of plumbagin on ROS and TGF-βl/Smad signaling pathway in hepatic stellate cells / 中国药理学通报

ABSTRACT

Aim To investigate the effects of plumbagin on reactive oxygen species (ROS) level and expressions of a-smooth muscle actin (ct-SMA), Smad2/ 3, p-Smad2/3, and Smad7 proteins. Methods We isolated primary hepatic stellate cell (HSC) of rat and inoculated the activated HSC. Then these cells would be divided into TGF-ßl treatment group,NADPH oxidase inhibitor (DPI) group,plumbagin treatment group, and control group. MIT was used to test cell proliferation and fluorescent probe was used to detect ROS level. Moreover, the protein expressions of Smad2/3, pSmad2/3, and Smad7 were determined using Western blot. Results Results of fluorescent probe showed that the positive rate of a-SMA was up to 91%. MTT test indicated the optimal concentrations of plumbagin were 2,4, and 8 u.mol • L " 1 . A decreased level of ROS could be found in DPI and plumbagin treatment groups. In addition, results of Western blot showed that a decreased expression of Smad2/3, p-Smad2/3 and an increased expression of Smad7 could be found in DPI and plumbagin treatment groups compared to control group. Conclusions Plumbagin may decrease the expression of smad2/3 and p-smad2/3 by down-regulating the level of ROS,and thus play an anti-HSC activation role.