IL-17A对人永生化角质形成细胞角蛋白17表达及STAT3信号通路的影响

摘要

Objective To observe the effects of interleukin-17A (IL-17A) on the secretion of keratin 17 (K17) from human immortalized keratinocyte cell line (HaCaT) cultured in vitro and the signal transducer and activator of transcription 3 (STAT3) signaling pathway,and to provide a theoretical basis for exploring the pathogenesis and targeted treatment of psoriasis. Methods HaCaT cells cultured in RPMI1640 culture medium were randomly divided into three groups: blank control group,induction group,and inhibitor group. The cells in the blank control group were treated with DMEM high-glucose medium only,cells in the induction group were treated with DMEM high-glucose medium containing 50 μg/L IL-17A, and cells in the inhibitor group were treated with a inhibitor (Piceatannol) containing 10 μmol/L STAT3 + the DMEM highglucose medium containing 50 μg/L IL-17A. The cultured HaCaT cells were collected,MTT assay was used to detect the proliferation of cells,flow cytometry (FCM) was used to detect the apoptosis rate,the expression of K17 mRNA was detected by RT-PCR,and Western blotting was used to detect the protein expression of K17 and phosphorylated STAT3 (p-STAT3). Results The A value of cell proliferation in the induction group was higher than that in the blank control group and the induction group,and the apoptosis rate was lower than that in the blank control group and the inhibitor group (both P < 0.01); the expression of K17 mRNA and the protein level of K17 and p-STAT3 in the induction group were both higher than those of the blank control group and the inhibitor group (all P < 0.01). Conclusion IL-17A can up-regulate the expression of K17 in HaCaT cells cultured in vitro,and its regulation mechanism may be achieved through the activation of STAT3,indicating that the role of IL-17A in psoriasis may be related to K17.%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(HaCaT)角蛋白17(K17)表达及信号转导和转录激活因子3(STAT3)信号通路的影响.方法 采用RPMI1640培养液培养HaCaT细胞,将细胞随机分为空白对照组、诱导组、抑制剂组,空白对照组仅加DMEM高糖培养基,诱导组加入含50 μg/L IL-17A的DMEM高糖培养基,抑制剂组加入含50 μg/L IL-17A的DMEM高糖培养基和10 μmol/L STAT3抑制剂Piceatannol.收集培养的HaCaT细胞,采用MTT比色法检测细胞增殖情况,流式细胞术检测细胞凋亡率,RT-PCR法检测细胞K17 mRNA表达水平,Western blotting法检测细胞K17和磷酸化STAT3(p-STAT3)蛋白表达水平.结果 诱导组细胞增殖A值高于空白对照组和抑制剂组,细胞凋亡率低于空白对照组和抑制剂组(P均<0.01);诱导组K17 mRNA及K17、p-STAT3蛋白相对表达量均高于空白对照组和抑制剂组(P均<0.01).结论 IL-17A能够上调体外培养的HaCaT细胞K17表达,其调控机制可能是通过激活STAT3来实现,表明IL-17A在银屑病中发挥的作用可能与K17有关.