目的 研究紫草素对HaCaT细胞抑制增殖的作用及其作用机制.方法 倒置显微镜下观察不同浓度紫草素(0、2、5、10、15μmol/L)作用后HaCaT细胞的形态学改变;MTT法检测紫草素对HaCaT细胞抑制增殖的作用;流式细胞术检测细胞凋亡率变化;Western blot法检测Notch-1、Jagged1和Hes5蛋白的表达情况.RT-PCR检测下游信号分子Hes1和Hes5的mRNA表达.结果 10μmol/L紫草素即可显著抑制HaCaT细胞的增殖,且随着浓度的增加,紫草素呈剂量依赖性地抑制HaCaT细胞的增殖(均P＜0.01).流式细胞术检测结果显示101μmol/L紫草素能够显著诱导HaCaT细胞凋亡(P＜0.01).Western bolt法检测结果显示,Notch-1、Jagged1和Hes5蛋白表达逐渐降低.RT-PCR法检测结果显示,Notch信号通路下游信号分子Hes1和Hes5的mRNA表达水平亦呈剂量依赖性降低.结论 紫草素呈剂量依赖性地抑制HaCaT细胞的增殖,并诱导细胞凋亡,Notch-1信号通路在其中起了重要作用.%Objective To investigate effects of Shikonin on proliferation of HaCaT cells and its mechanism.Methods Human immortal keratinocyte HaCaT cells were treated with 0,2,5,10 and15μmol/L of Shikonin.The morphology of cells was observed with inverted microscopy,the growth inhibition rate of HaCaT cells was determined with MTT method,the apoptosis rate was detected by flow cytometer(FCM),the expression of Notch-1,Jagged1 and Hes5 proteins was detected by Western blot;the expression of Hes1 and Hes5 mRNA was detected by RT-PCR.Results MTT assay showed that cell growth was significantly inhibited by shitonin in a dose-dependent manner (P ＜0.01).FCM showed that the percentage of apoptotic cells was significantly increased and the cell viability was significantly decreased after HaCaT cells were treated with ＞ 10μmol/L shikonin (P＜0.01).Western blot showed that the expression of Notch-1,Jagged1 and Hes5 was decreased;and RT-PCR showed that the mRNA expression of Hes1 and Hes5 was also decreased.Conclusion Shikonin can inhibit proliferation and induce apoptosis of HaCaT cells,which may be regulated by Notch-1 signal pathway.
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