| 摘要: |
| 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、15滋mol/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；Westernblot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10滋mol/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10滋mol/L紫草素能够显著诱导HaCaT细胞凋亡（P＜0.01）。Westernbolt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。 |
| 关键词: 紫草素 HaCaT Notch-1 Jagged1 Hes1 Hes5 细胞凋亡 |
| DOI：10.12056/j.issn.1006-2785.2017.39.03.2016-1917 |
| 分类号: |
| 基金项目:浙江省自然科学基金（Y2100759） |
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| Effects of Shikonin on proliferation of HaCaT Cells |
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CHEN Wei, CAO Yi, YANG Xiaohong
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the Third Affiliated Hospital of Zhejiang Chinese Medical University
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| Abstract: |
| 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 and15滋mol/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 ＞10滋mol/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. |
| Key words: Shikonin HaCaT Notch-1 Jagged1 Hes1 Hes5 Cell apoptosis |
