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Aim To investigate whether salvianolic acid B ( Sal B) has inhibitory effect on hepatoma HuH- 7 cells and explore whether it works via Hippo/YAP signaling pathway. Methods HuH-7 cells were induced by TGF-β1 (9 pmol · L
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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.
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;Aim To investigate the role of periostin in PDGF-induced proliferation and extracellular matrix accumulation of mouse mesangial cells. Methods The effects of PDGF on periostin protein, cell proliferation and extracellular matrix accumulation were detected. The cells were collected at 0, 2,4, 6, 12 h after stimulation with PDGF(10 (ig • L " 1 ) to detect the expression of periostin, PCNA, FN and TGF-ßl by Western blot. The silencing effect of sh-periostin vector on periostin protein in mouse mesangial cells was identified by Western blot. Cells were randomly divided into control group, PDGF group, PDGF + sh-nc group and PDGF + sh-periostin group to detect the role of periostin in PDGF-induced proliferation and extracellular matrix accumulation of mouse mesangial cells. Results PDGF could elevate periostin protein expression. Western blot result showed that periostin protein expression in PDGF-stimulated groups was significantly higher than that in Oh group, which was consistent with the result of immunofluorescence. Positive expression of periostin was located in cytoplasm. Western blot result showed that PCNA, FN and TGF-ßl protein in PDGF-stimulated groups increased as compared with Oh group. shRNA vector aimed at periostin (sh-periostin vector) could partially reverse PDGF-induced mesangial cell proliferation and extracellular matrix expression. PCNA, Fn and TGF-ßl expressions were attenuated significantly. Conclusions PDGF can enhance periostin protein expression and increase mouse mesangial cell proliferation and extracellular matrix accumulation. Periostin shRNA vector can partially reverse PDGF-induced mesangial cell proliferation and extracellular matrix generation.