芦可替尼靶向抑制肝星状细胞活化的作用及机制

Role of ruxolitinib in the target inhibition of hepatic stellate cell activation and mechanism

目的探讨芦可替尼靶向抑制肝星状细胞活化的作用及机制。方法培养原代大鼠肝星状细胞,IL-6活化肝星状细胞后加入芦可替尼观察肝星状细胞内JAK1/2-STAT3通路及下游α-SMA、COL1A1、cyclin D1、cleaved caspase-3蛋白表达,通过CCK-8实验检测肝星状细胞增殖活性,Transwell实验检测肝星状细胞侵袭性。结果IL-6可明显激活JAK1/2-STAT3通路并活化肝星状细胞,芦可替尼可显著抑制肝星状细胞内JAK1/2、STAT3蛋白磷酸化水平,显著抑制肝星状细胞内α-SMA蛋白、COL1A1蛋白及cyclin D1蛋白表达,升高cleaved caspase-3蛋白表达,差异有统计学意义(均P<0.05)。芦可替尼能显著抑制肝星状细胞的细胞增殖能力及侵袭能力,差异有统计学意义(P<0.05)。结论芦可替尼可通过靶向抑制肝星状细胞中JAK1/2-STAT3通路激活进而降低肝星状细胞活化程度,抑制细胞增殖、侵袭能力,减少肝星状细胞细胞外基质合成能力。

Objective To explore the inhibitory effect of ruxolitinib on the hepatic stellate cell activation and its mechanism.Methods The primary rat hepatic stellate cells were cultured.The cultured rat hepatic stellate cells were activated by IL-6,and then given different concentrations of ruxolitinib.The effects of ruxolitinib on JAK1/2-STAT3 pathway and its downstream related molecules expression includingα-SMA,COL1A1,cyclin D1,cleaved caspase-3 in hepatic stellate cells were observed,the proliferation activity of hepatic stellate cells was detected by CCK-8 assay,and the invasive abilities of hepatic stellate cells were detected by Transwell assay.Results IL-6 significantly triggered the JAK1/2-STAT3 signaling pathway and the activation of hepatic stellate cells.Ruxolitinib significantly inhibited the phosphorylation of JAK1/2 and STAT3 protein,decreased the expression of alpha-SMA protein,collagenⅠand cyclin D1 protein and increased the expression of cleaved caspase-3 protein in hepatic stellate cells(all P<0.05).Ruxolitinib significantly reduced the proliferation and invasion abilities of hepatic stellate cells(P<0.05).Conclusion Ruxolitinib can reduce the activation of hepatic stellate cells,inhibit the cell proliferation and invasion,and reduce the synthesis of extracellular matrix by target inhibition of the JAK1/2-STAT3 pathway.