Effects of Curc-OEG on primary rat hepatic stellate cells in vitro

Abstract

Abstract: AIM: To investigate the effects and mechanisms of Curc-OEG on the proliferation, activation and apoptosis of hepatic stellate cells (HSCs). METHODS: The HSCs were isolated from male SD rats by collagenase IV and DNase digestion of liver, and further purified by percoll gradient centrifugation. Freshly isolated cells at day 2 were treated with Curc-OEG at concentrations of 0, 6.25 and 12.5 μg/mL for 7 days. The expression of α-SMA, TGF-β₁ and Smad2 was confirmed by RT-PCR and Western blot. Activated HSCs at day 14 were treated with Curc-OEG at concentrations of 0, 6.25, 12.5, 25, 50 and 75 μg/mL for 24 h. The expression of Bax and Bcl-2 and other genes associated with fibrogenesis including TGF-β₁, collagen I, NF-κB and TIMP-1 was observed with RT-PCR. RESULTS: After 7 days cultivation, Curc-OEG caused a significant concentration-dependent reduction in cell numbers of 56% and 86% at 6.25 and 12.5 μg/mL respectively compared with the control. At 12.5 μg/mL, Curc-OEG reduced α-SMA, TGF-β₁ and Smad2 mRNA levels by 83%, 85% and 75%, and protein levels by 94%, 92% and 73%, respectively (P<0.05). Cell apoptosis was significantly increased at a concentration of 25 μg/mL Curc-OEG. At 50 μg/mL, Curc-OEG significantly upregulated the mRNA levels of the proapoptotic Bax by 2.3-fold,downregulated the antiapoptotic Bcl-2, TGF-β₁, collagen I, NF-κB and TIMP-1 by 5.6-fold, 90%, 83%, 74% and 65%, respectively (P<0.05). CONCLUSION: Curc-OEG not only significantly inhibited the proliferation and activation of primary HSCs, but also induced apoptosis of activated HSCs and reduced ECM accumulation.

Key words: hepatic fibrosis, curcumin, hepatic stellate cell, activation, apoptosis

CLC Number:

R965.2

SUN Yong, HUANG Xiao-hua, SHEN Neng, TANG Hua-dong, REN Hong, PENG Ming-li. Effects of Curc-OEG on primary rat hepatic stellate cells in vitro[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2014, 19(10): 1086-1092.

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