竹黄颗粒介导IL-22/miR-330调控角质形成细胞增殖的机制研究

Mechanism of Zhuhuang Granule Intervenes IL-22/miR-330 in Regulating Keratinocyte Proliferation

目的研究竹黄颗粒干预IL-22/miR-330调控角质形成细胞增殖的分子机制。方法 IL-22处理Ha Ca T和HKCs细胞,应用QPCR技术检测细胞中miR-330的表达变化;之后慢病毒感染Ha Ca T和HKCs细胞,实现miR-330的过表达和干扰,通过CCK8和Brd U技术检测细胞增殖情况;采用CCK8、Western blot、QPCR、ELISA实验分别检测不同浓度竹黄颗粒处理Ha Ca T、HKCs后细胞增殖能力,IL-22蛋白表达水平,miR-330表达水平及细胞分泌IL-22含量的变化。结果 IL-22处理后的Ha Ca T和HKCs细胞中miR-330的表达水平显著下调(P<0.01),而细胞增殖能力显著提升(P<0.01);过表达miR-330后Ha Ca T和HKCs细胞增殖能力显著下降(P<0.05),干扰miR-330后Ha Ca T和HKCs细胞增殖能力显著上升(P<0.05);竹黄颗粒处理Ha Ca T、HKCs后,细胞增殖能力下降(P<0.01),IL-22蛋白表达水平显著下降(P<0.01),miR-330表达水平显著上升(P<0.01),细胞分泌IL-22含量显著下降(P<0.01)。结论竹黄颗粒可以通过抑制IL-22的表达,促进miR-330的表达,从而抑制角质形成细胞增殖。

Objective To study the molecular mechanism of IL-22/miR-330 intervened by Zhuhuang granule in regulating the proliferation of keratinocytes. Methods The expression of miR-330 in Ha Ca T and HKCs cells after IL-22 intervention was detected by QPCR technique. Overexpression and interference of miR-330 in Ha Ca T and HKCs cells were achieved by lentivirus infection, their cell proliferation was detected by CCK8 and Brd U. The cell proliferation ability, the expression of IL-22 protein, the expression level of miR-330 and the secretion of IL-22 in Ha Ca T and HKCs were determined by CCK8,West ern blot, QPCR and ELISA, respectively. Results The expression of miR-330 in Ha Ca T and HKCs cells significantly decreased after treatment with IL-22(P<0.01), and the cell proliferation ability of Ha Ca T and HKCs cells was significantly improved(P<0.01). The cell proliferation ability of Ha Ca T and HKCs cells significantly decreased after miR-330 overexpression(P<0.05). After interference of miR-330 expression, cell proliferation ability of Ha Ca T and HKCs had a significant increase(P<0.05).Cell proliferation ability of Ha Ca T and HKCs treated by different concentrations of Zhuhuang granule decreased(P<0.01). The expression of IL-22 protein decreased significantly(P<0.01), the expression level of miR-330 increased significantly(P<0.01),and the content of IL-22 decreased significantly(P<0.01). Conclusion Zhuhuang granule could inhibit the expression of IL-22, promote the expression of miR-330 and inhibit the proliferation of keratinocytes.