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目的:探讨姜黄素体外对氧化应激致肝星状细胞(HSC-T6)增殖、表达转化生长因子-β1(TGF-β1)的抑制作用。方法:以100μmol.L-1过氧化氢(H2O2)致HSC氧应激,将细胞分成对照组、H2O2组和H2O2+姜素素干预组(终质量浓度分别为25,50,100μmol.L-1)5组,用四甲基偶氮唑(MTT)比色法检测HSC细胞增殖;用流式细胞术检测姜黄素对HSC细胞周期的影响;硫代巴比妥酸法、黄嘌呤氧化酶法和消化法分别检测细胞培养液丙二醛(MDA)、超氧化物歧化酶(SOD)和羟脯氨酸(HyP)的含量;用逆转录聚合酶链式反应(RT-PCR)检测HSC表达TGF-β1的水平。结果:HSC经H2O2诱导后,细胞增殖显著(P<0.01),细胞培养液上清中MDA,Hyp含量明显上升,SOD活力下降(P<0.01);经姜黄素各剂量干预后,MDA,Hyp含量下降,SOD活力升高,以50,100μmol.L-1姜黄素浓度作用明显(P<0.01);H2O2刺激HSC后,TGF-β1 mRNA表达被上调(P<0.001),经姜黄素干预后,TGF-β1 mRNA表达水平下降(P<0.01)。结论:姜黄素可显著抑制氧化应激致HSC的增殖,该作用可能与其抗氧化、抑制TGF-β1的表达有关,表明姜黄素作为抗氧化剂在肝纤维化防治中具有重要价值。
Objective: To investigate the inhibitory effect of curcumin on proliferation and expression of transforming growth factor-β1 (TGF-β1) induced by oxidative stress in hepatic stellate cells (HSC-T6) in vitro. Methods: The cells were divided into control group, H2O2 group and H2O2 + gingerin intervention group (final concentration of 25, 50, 100μmol.L-1) in 5 groups with HSC oxygen stress induced by 100μmol.L-1 H2O2 , The proliferation of HSC cells was detected by MTT colorimetric assay; the effect of curcumin on the cell cycle of HSC was detected by flow cytometry; thiobarbituric acid method, xanthine oxidase method and digestion The contents of malondialdehyde (MDA), superoxide dismutase (SOD) and hydroxyproline (HyP) in the cell culture medium were detected respectively. The expression of TGF-β1 in HSC was detected by reverse transcriptase-polymerase chain reaction s level. Results: The proliferation of HSC induced by H2O2 was significant (P <0.01). The content of MDA and Hyp in the supernatant of cell culture increased significantly and the activity of SOD decreased (P <0.01). After the intervention of curcumin, the contents of MDA and Hyp (P <0.01). After the stimulation of HSC with H2O2, the expression of TGF-β1 mRNA was up-regulated (P <0.001). After the intervention of curcumin, TGF-β1 mRNA expression decreased (P <0.01). CONCLUSION: Curcumin can significantly inhibit the proliferation of HSC induced by oxidative stress, which may be related to its anti-oxidation and inhibition of TGF-β1 expression, indicating that curcumin has important value as an antioxidant in the prevention and treatment of hepatic fibrosis.