Effects of phenylethanol glycosides from Cistanche tubulosa on proliferation of rat HSC induced by PDGF-BB and its mechanism / 中国药理学通报

Aim To investigate the effect of phenyle-thanol glycosides from Cistanche tubulosa(CPhGs) on the proliferation and activation of rrPDGF-BB induced HSC and their target points for resisting hepatic fibro-sis,to elucidate the molecular mechanism in molecular level, and provide basic data for the further develop-ment of new drugs. Methods HSCs were cultivated by CPhGs with different concentrations ( 0 , 3. 91 , 7. 81 , 15. 63 , 31. 25 , 62. 50 , 125. 00 , 250. 00 , and 500 mg ·L-1 ) and IC50 of CPhGs was determined. CPhGs with different concentrations ( 25 , 50 , 75 , 100 mg · L-1 ) were selected, and after the cells were stimulated with rrPDGF-BB, cell proliferation was determined by MTT. ERK1/2 ,α-SMA, c-fos, c-jun and Collagen I mRNA and Erk1/2 ,P-Erk1/2 and CollagenⅠprotein ex-pressions were assayed by RT-PCR and Western blot. Results CPhGs of ( 50 ~100 ) mg · L-1 concentra-tions groups could effectively inhibit rrPDGF-BB-medi-ated proliferation(P0. 05 ) . CPhGs of ( 25 ~100 ) mg · L-1 concentrations groups could inhibit ERK1/2 ,α-SMA,c-fos, c-jun and CollagenⅠmRNA levels, and also ob-viously inhibited Erk1/2 ,P-Erk1/2 and Collagen Ⅰ pro-tein expression on HSC. Conclusions CPhGs has the protective effect against hepatic fibrosis. The mecha-nism of this process may involve the interference with PDGF/ERK1/2 signaling pathway and inhibiting the activation and proliferation of HSC.

The Effects of Autophagy Inhibitor on Activation of Alcohol induced Hepatic Stellate Cells / 天津医药

Objective To observe the effect of autophagy inhibitor on the activation of alcohol induced hepatic stel-late cells, and the mechanisms thereof. Methods HSC-T6 cells were cultured in vitro and divided into four groups, includ-ing blank control group, alcohol group, 5 mmol/L 3-MA+alcohol group (low alcohol group) and 10 mmol/L 3-MA+alcohol group (high alcohol group). RT-PCR was used to detect the expression levels ofα-smooth muscle actin (α-SMA) and typeⅠcollagen. The levels of LC3Ⅱ,α-SMA and typeⅠcollagen were detected by Western blot assay. The cell viability of HSC-T6 was detected by MTT assay. Results The mRNA expressions ofα-SMA, typeⅠcollagen and the protein of expressionsα-SMA, typeⅠcollagen and LC3Ⅱwere significantly up-regulated in alcohol group compared with those of control group (P<0.05), while the expressions of those parameters were significantly down-regulated in 10 mmol/L 3-MA+alcohol group (P<0.01). The mRNA and protein levels ofα-SMA and typeⅠcollagen were significantly decreased in two 3-MA-treated groups compared with those in alcohol group (P<0.05). Meanwhile, compared with the 5 mmol/L 3-MA+alcohol group,the protein expressions ofα-SMA, typeⅠcollagen and LC3Ⅱwere significantly decreased in10 mmol/L 3-MA+alcohol group (P < 0.05 ). Compared with the alcohol group,there was significantly lower proliferation activity in all two 3-MA-treated groups (P<0.05). Conclusion 3-MA can inhibit the protein expression of LC3Ⅱ,α-SMA and typeⅠcollagen induced by alcohol in HSC-T6 cells, and inhibit the proliferation of HSC cells.