IL-17A对体外培养角质形成细胞角蛋白17的影响及作用机制

Effects of Interleukin-17A on Cytokeratin 17 in Keratinocytes Cultured in Vitro and Its Molecular Mechanism

目的观察白细胞介素-17A(IL-17A)对体外培养的人永生化角质形成细胞株(Ha Ca T)分泌角蛋白17(K17)及信号转导和信号转录激活因子3(STAT3)信号通路的影响。方法 RPMI1640培养液培养的Ha Ca T细胞随机分为空白对照组、IL-17A(50μg/L)刺激(诱导组)和IL-17A(50μg/L)+10μmol/L STAT3抑制剂Piceatannol(抑制剂组)。收集培养后Ha Ca T细胞,四甲基偶氮唑蓝(MTT)检测Ha Ca T细胞增殖,流式细胞术(FCM)检测Ha Ca T细胞凋亡,RT-PCR检测Ha Ca T细胞K17 mRNA水平;Western印迹法检测Ha Ca T细胞K17和磷酸化STAT3(pSTAT3)蛋白水平。结果三组Ha Ca T细胞增殖和凋亡差异有统计学意义(P<0.05);三组Ha Ca T细胞K17 mRNA和Ha Ca T细胞K17和p-STAT3蛋白表达差异均有统计学意义(P<0.05)。结论 IL-17A能够上调Ha Ca T细胞K17的表达,表明IL-17A在银屑病中所发挥的作用可能与K17有关,为研究银屑病的发病机理和治疗提供了新的策略。

ObjectiveTo explore the effect of Interleukin-17A（IL-17A） on the expression of keratin 17（K17） in keratinocytes（HaCaT） cultured in vitro and on STAT3 signaling pathway. MethodsHaCaT cells cultured in RPMI1640 medium were randomly divided into blank control group,IL-17A（50μg/L） stimulation in KCs group（induction group） and IL-17A（50μg/L） ＋10μmol/L STAT3 inhibitor Picestanol group（inhibition group）. After HaCaT cells were harvested,proliferation and apoptosis of HaCaT cells were detected by MTT assay and flow cytometry（FCM）,respectively. Additionally,K17 mRNA level in HaCaT cells was detected by RT-PCR,and protein levels of K17 of HaCaT cells and phosphorylated STAT3（p-STAT3） were detected by Western blotting. ResultsProliferation and apoptosis rate of HaCaT cells were significantly different in blank control group,induction group and inhibition group. In addition,there were significant differences in expression of K17 mRNA,K17 and p-STAT3 in HaCaT in three groups. ConclusionIL-17A can up regulate the expression of K17 in KC,which indicates the association between IL-17A and K17 in the pathogenesis of psoriasis,potentially providing a novel strategy for pathogenesis and treatment of psoriasis.