瞬时受体电位M7通道蛋白对大鼠肝星状细胞凋亡的影响

目的通过转染特异性siRNA,下调瞬时受体电位M7通道蛋白(TRPM7)在大鼠肝星状细胞系HSC-T6中的表达,研究其对HSC-T6凋亡的影响及内在机制。方法利用Li-pofectamineTM2000将特异性TRPM7-siRNA转染到HSC-T6内,分别运用流式细胞术检测HSCs凋亡变化;Western blot检测TRPM7、Caspase-3蛋白的表达;RT-PCR检测TRPM7,Caspase-3、Bid mRNA的表达。结果 TRPM7-siRNA明显抑制TRPM7 mRNA和蛋白的表达。与正常组或对照组相比,转染组细胞凋亡率、凋亡蛋白Caspase-3及凋亡基因Bid的表达均明显上调。结论特异性TRPM7-siRNA能明显抑制TRPM7表达,诱导HSCs凋亡,其机制可能与其诱导凋亡基因Bid表达上调有关。

The effects of transient receptor potential melastaln 7 channel protein on apoptosis of rat hepatic stellate cells

Aim To investigate the effects of downexpression of transient receptor potential melastatin 7(TRPM7) and its internal mechanisms on apoptosis through transfecting specific siRNA in rat hepatic stellate cell lines HSC-T6.Methods Specific TRPM7-siRNA was transfected by LipofectamineTM 2000.Cell apoptosis was measured by Flow Cytometry.TRPM7 and Caspase-3 protein expression were analyzed by Western blot;TRPM7,Caspase-3 and Bid mRNA expression were evaluated by RT-PCR.Results Suppression TRPM7 expression by siRNA significantly increased the apoptosis of HSCs,the expression of Caspase-3 protein and Bid mRNA compared with normal or control.Conclusions Specific TRPM7-siRNA not only inhibits the expression of TRPM7 but also induces apoptosis of HSCs.The enhanced gene expression of Bid may contribute to HSCs apoptosis.