IL-17a通过STAT3信号通路调节HaCaT细胞中IL-6、IL-22、IL-23的表达

IL-17a Regulates the Expression of IL-6,IL-22 and IL-23 in HaCaT Cells Through the STAT3 Signaling Pathway

目的:探讨IL-17a刺激人角质形成(HaCaT)细胞对分泌白介素-6(IL-6)、白介素-22(IL-22)、白介素-23(IL-23)的影响以及IL-17a调控角质形成细胞中IL-6、IL-22、IL-23表达的可能机制。方法:将HaCaT细胞随机分为空白对照组(DMEM高糖培养基)、IL-17a诱导组(含80μg/L IL-17a的DMEM高糖培养基)、抑制剂组(含80μg/L IL-17a的DMEM高糖培养基和10μmol/L STAT3抑制剂白皮衫醇),48 h后用ELISA法检测各组上清液的IL-6、IL-22、IL-23浓度,实时荧光免疫定量-聚合酶链式反应法(RT-PCR)检测各组HaCaT细胞中IL-6、IL-22、IL-23 mRNA表达水平,同时用Western Blot检测STAT3的表达水平。结果:与空白对照组和抑制剂组比较,诱导组的HaCaT细胞IL-6、IL-22、IL-23表达水平及mRNA均明显升高(P<0.05);与空白对照组和抑制剂组比较,诱导组的HaCaT细胞STAT3蛋白表达水平明显升高(P<0.05)。结论:IL-17a可促进HaCaT细胞分泌IL-6、IL-22、IL-23,其调控机制可能通过STAT3来实现。

Objective:To explore the effect of interleukin-17a(IL-17a)on the expression of IL-6,IL-22,IL-23 in human immortalized keratinocytes(HaCaT)cells and possible mechamism of IL-17a regulating IL-6,IL-22,IL-23 in keratinocytes.Method:HaCaT cells were divided into blank control group(DMEM high-glucose medium only),induction group(DMEM high-glucose medium containing 80μg/L IL-17a)and inhibitor group(DMEM high-glucose medium containing 80μg/L IL-17a and 10μg/L STAT3 inhibitor Piceatannol).After 48 h,we examined the expression of IL-6,IL-22,IL-23 by double antibody sandwich enzyme-1inked immunosorbent assay(ELISA)in the cells supernatant and realtime polymerase chain reaction(RT-PCR)was used to detect the mRNA expression levels of IL-6,IL-22,IL-23 in HaCaT cell of all group.Protein expression of STAT3 was detected by Western Blot.Result:Compared with blank control group and inhibitor group,the expression levels and mRNA of IL-6,IL-22 and IL-23 in HaCaT cells in induction group were significantly increased(P<0.05).Compared with the blank control group and the inhibitor group,the expression level of STAT3 protein in HaCaT cells in the induction group was significantly increased(P<0.05).Conclusion:Il-17a can promote the secretion of IL-6,IL-22 and IL-23 by HaCaT cells,which may be regulated by STAT3.