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目的:研究白花丹醌(plumbagin)对转化生长因子-β1(TGF-β1)刺激的体外培养人肝星状细胞HSC-LX2α-平滑肌肌动蛋白(α-SMA)mRNA和蛋白表达的影响。方法:体外培养HSC-LX2,随机设立空白组,TGF-β1刺激的模型组,TGF-β1+白花丹醌2.0μmol·L-1高剂量组,TGF-β1+白花丹醌1.5μmol·L-1中剂量组和TGF-β1+白花丹醌1.0μmol·L-1低剂量组,各药物与细胞孵育72 h后,采用RT-PCR检测各组HSC-LX2α-SMA mRNA的表达,采用免疫细胞化学方法(ICC)检测HSCLX2中α-SMA蛋白的表达。结果:与正常组比较,模型组α-SMA mRNA及蛋白表达明显升高(P<0.01);与模型组比较,白花丹醌作用72 h后,高、中剂量组HSC-LX2细胞中α-SMA mRNA的表达明显降低(P<0.01);免疫细胞化学结果显示白花丹醌高、中剂量组对HSC-LX2细胞α-SMA蛋白的表达有显著地抑制作用(P<0.05),尤以高剂量组更明显(P<0.01)。结论:白花丹醌能抑制HSC-LX2的活化和增殖,其机制之一可能是从mRNA水平和蛋白水平抑制α-SMA的表达,从而发挥其抗肝纤维化作用。
AIM: To investigate the effect of Plumbagin on the expression of HSC-LX2α-SMA (smooth muscle actin) mRNA and protein in cultured human hepatic stellate cells (HSCs) stimulated by transforming growth factor-β1 (TGF-β1) Methods: The HSC-LX2 cells were cultured in vitro and were randomly divided into blank group, TGF-β1-stimulated model group, high dose TGF-β1 + plumbagin solution 2.0μmol·L-1, TGF- Dose group and TGF-β1 + plumbagin 1.0μmol·L-1 low dose group, each drug and cells incubated for 72 h, RT-PCR detection of HSC-LX2α-SMA mRNA expression in each group by immunocytochemistry ICC) to detect the expression of α-SMA protein in HSCLX2. Results: Compared with the normal group, the expression of α-SMA mRNA and protein in the model group was significantly increased (P <0.01). Compared with the model group, the expression of α- The expression of SMA mRNA in HSC-LX2 cells was significantly lower than that in HSC-LX2 cells (P <0.01). The results of immunocytochemistry showed that the high dose and middle dose of plumbagin had a significant inhibitory effect on the expression of α-SMA protein The dose group was more obvious (P <0.01). Conclusion: Plumbagin can inhibit the activation and proliferation of HSC-LX2. One of the mechanisms may be that the expression of α-SMA is inhibited by mRNA and protein levels, which may play an important role in anti-hepatic fibrosis.