卡泊三醇抑制人肝星状细胞纤维化及其作用机制研究

The inhibitory effect of calcipotriol on fibrosis of human hepatic stellate cell and its mechanism

目的 探索卡泊三醇对人肝星状细胞纤维化的作用及机制。方法 以人肝星状细胞LX-2为研究对象,以转化生长因子β1(transforming growth factor-β1,TGF-β1)(5 ng/ml, 24 h)诱导,建立体外肝纤维化细胞模型,将细胞模型分成模型组和卡泊三醇组(100 nmol/L),以无处理的细胞为对照组。分别处理后,采用MTT法检测LX-2细胞的增殖;以qPCR的方法检测Ⅰ型胶原(type I collagen, Col-Ⅰ)、Col-Ⅲ、α-平滑肌肌动蛋白(αsmooth muscle actin,α-SMA)、丝裂原活化蛋白激酶(mitogen activated protein kinase, P38)及细胞外信号调节激酶(extracellular signal regulated kinase, ERK)的表达;Western blot检测p-P38、P38、p-ERK和ERK的表达;ELISA检测Col-Ⅰ、Col-Ⅲ、α-SMA和Smad3的含量。结果 与对照组比较,模型组LX-2细胞增殖能力增强,Col-Ⅰ、Col-Ⅲ、α-SMA、p-P38及p-ERK的表达均显著升高(P<0.05)。与模型组比较,卡泊三醇组细胞增殖能力下降,Col-Ⅰ、Col-Ⅲ、α-SMA、p-P38及p-ERK的表达均显著降低(P<0.05)。结论 卡泊三醇可以通过抑制MAPK通路,减少Col-Ⅰ、Col-Ⅲ的含量,抑制肝星状细胞纤维化。

Objective To explore the effect of calcipotriol on the fibrosis of human hepatic stellate cells and its mechanism.Methods Human hepatic stellate cells LX-2 were induced by transforming growth factor-β1(TGF-β 1)(5 ng/ml, 24 h) to establish the stereoscopic liver fibrosis cell model.The cell model was divided into the model group and the calcipotriol group(100 nmol/L),with untreated cells as the control group.After the treatment, the proliferation of LX-2 cells was detected by MTT,and the expression of type I collagen(Col-I),Col-III,α smooth muscle actin(α-SMA),mitogen activated protein kinase(P38) and extracellular signal regulated kinase(ERK) were detected by qPCR;Western blot was used to detect the expression of p-P38 and p-ERK.Elisa was used to detect the contents of Col-I,Col-III,α-SMA and Smad3.Results Compared with the control group, the proliferation ability of LX-2 cells in the model group was enhanced, and the expression levels of Col-I,Col-III,α-SMA,p-P38 and p-ERK were significantly increased(P<0.05).Compared with the model group, the proliferation ability of LX-2 cells in the calcipotriol group was decreased, and the expression levels of Col-I,Col-III,α-SMA,p-P38 and p-ERK were significantly decreased(P<0.05).Conclusion Calcipotriol can inhibit hepatic stellate cells fibrosis by inhibiting the MAPK pathway, and decreasing the contents of Col-I and Col-III.