Triptolide Inhibits the Proliferation of HaCaT Cells Induced by IL22 via Upregulating miR-181b-5p
Triptolide Inhibits the Proliferation of HaCaT Cells Induced by IL22 via Upregulating miR-181b-5p
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DOI：
10.2147/dddt.s254466
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发表时间：
2020-01-01
影响因子：
4.8
通讯作者：
Li, Jiawen
中科院分区：
文献类型：
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作者：
He, Qi;Zhang, Bo;Li, Jiawen
背景：有证据表明雷公藤内酯醇可有效治疗牛皮癣；然而，人们对这些机制仍知之甚少。因此，本研究旨在探讨雷公藤甲素对用IL22处理的HaCaT细胞增殖和分化的作用，以模拟银屑病角质形成细胞的异常增殖/分化。材料与方法：用miR-181b-5p antagomir转染HaCaT细胞24小时，然后暴露于10μM雷公藤甲素24小时，随后加入100ng/mL的IL22 24小时。此外，分别通过免疫荧光或流式细胞术检测HaCaT细胞的增殖和细胞周期分布。结果：雷公藤甲素明显上调HaCaT细胞中miR-181b-5p的水平。此外，雷公藤甲素通过诱导细胞周期停滞显着抑制 IL22 诱导的 HaCaT 细胞增殖。此外，IL22显着抑制HaCaT细胞的分化，而雷公藤甲素处理可逆转这种现象。相比之下，雷公藤甲素对 IL22 刺激的 HaCaT 细胞增殖和分化的影响被 miR-181b-5p antagomir 显着逆转。此外，双荧光素酶测定显示 E2F5 是 HaCaT 细胞中 miR-181b-5p 的直接靶标。同时，miR-181b-5p的上调明显降低了HaCaT细胞中E2F5的水平。结论：在本研究中，我们发现雷公藤甲素可以通过上调miR-181b-5p来抑制IL22刺激的角质形成细胞的增殖并促进其分化。这些数据表明雷公藤甲素可能是治疗牛皮癣的潜在药物。
Background: Evidence has been shown that triptolide was effective in the treatment of psoriasis; however, the mechanisms remain poorly understood. Thus, this study aimed to investigate the role of triptolide on the proliferation and differentiation of HaCaT cells which are treated with IL22 to mimic abnormal proliferation/differentiation in keratinocyte of psoriasis.Materials and Methods: HaCaT cells were transfected with miR-181b-5p antagomir for 24 h, and then exposed to 10 mu M Triptolide for 24 h, following by 100 ng/mL of IL22 for 24 h. In addition, the proliferation and cell cycle distribution in HaCaT cells were assessed by immunofluorescence or flow cytometry assays, respectively.Results: Triptolide obviously upregulated the level of miR-181b-5p in HaCaT cells. In addition, triptolide significantly inhibited IL22-induced proliferation of HaCaT cells via inducing cell cycle arrest. Moreover, IL22 markedly inhibited the differentiation of HaCaT cells, and this phenomenon was reversed by triptolide treatment. In contrast, the effects of triptolide on the proliferation and differentiation in IL22-stimulated HaCaT cells were notably reversed by miR-181b-5p antagomir. Moreover, dual-luciferase assay showed that E2F5 was the direct target of miR-181b-5p in HaCaT cells. Meanwhile, upregulation of miR-181b-5p obviously decreased the level of E2F5 in HaCaT cells.Conclusion: In this study, we found that triptolide could inhibit the proliferation and promote the differentiation in IL22-stimulated keratinocytes via upregulating miR-181b-5p. These data indicated that triptolide may be a potential agent for the treatment of psoriasis.