白细胞介素17A体外刺激角质形成细胞株对角蛋白17表达的影响  

Effect of interleukin-17A on keratin 17 in keratinocyte and its molecular mechanism in vitro

作　　者：时磊[1] 魏明[1] 

机构地区：[1]郑州大学第五附属医院药学部,450052

出　　处：《中国医药生物技术》2017年第6期526-531,共6页Chinese Medicinal Biotechnology

摘　　要：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HaCaT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HaCaT细胞随机分为空白对照组、IL-17A(50μg/L)刺激孔(诱导组)和IL-17A(50μg/L)+10μmol/L STAT3抑制剂白皮杉醇(抑制剂组)。收集培养的HaCaT细胞,采用四甲基偶氮唑盐(MTT)检测HaCaT细胞增殖;采用流式细胞术(FCM)检测HaCaT细胞凋亡;采用RT-PCR检测HaCaT细胞K17mRNA表达水平;采用Western blot法检测HaCaT细胞K17和磷酸化STAT3(p-STAT3)蛋白表达水平。结果诱导组、抑制剂组和空白对照组HaCaT细胞增殖差异有统计学意义(F=8.47,P=0.006),诱导组HaCaT细胞增殖高于空白对照组和抑制剂组(均P<0.05);诱导组、抑制剂组和空白对照组HaCaT细胞凋亡率差异有统计学意义(F=26.48,P=0.001),诱导组HaCaT细胞凋亡率(5.96%±0.68%)低于空白对照组(15.34%±1.32%)和抑制剂组(15.62%±1.26%)(均P<0.05);诱导组、抑制剂组和空白对照组HaCaT细胞K17 mRNA表达差异有统计学意义(F=10.25,P=0.001),与空白对照组和抑制剂组比较,诱导组的HaCaT细胞K17 mRNA表达明显升高(均P<0.05);3组HaCaT细胞K17和p-STAT3蛋白表达差异有统计学意义(F分别为11.62和9.86,P=0.001和0.003)。与空白对照组和抑制剂组比较,诱导组的HaCaT细胞K17和p-STAT3蛋白表达明显升高(均P<0.05)。结论 IL-17A能够上调体外培养的HaCaT细胞K17的表达,其调控机制可能是通过激活STAT3来实现,表明IL-17A在银屑病中所发挥的作用可能与K17有关,为银屑病的发病机制研究和治疗提供了新的思路。Objective To explore the effect of IL-17A on the expression of K17 in keratinocytes（KCs） and its molecular mechanism. Methods IL-17A（50 μg/L） was used to stimulate the KCs for 12 to 48 h, and three groups were divided that included control group, induction group and both induction and inhibitor（abbreviated as inhibitor） group. MTT assay was used to detect the proliferation. Flow cytometry was used to test apoptosis. Real-time PCR and Western blot were used to detect K17 mRNA and protein levels, as well as STAT3 signaling pathway. Results HaCaT cell proliferation was different among the blank control group, induction group and inhibitor group, as tested by MTT（F = 8.47, P = 0.006）. The apoptosis rate of HaCaT cells was（15.34% ± 1.32%）,（5.96% ± 0.68%） and（15.62% ± 1.26%） in the blank control group, induction group and inhibitor group, respectively（F = 26.48, P = 0.001）. K17 mRNA and protein expression levels were statistically significant among the three groups（F = 10.25, P = 0.001 for mRNA and F = 11.62, P = 0.001 for protein, respectively）. The p-STAT3 protein expression was statistically significant as well（F = 9.86, P = 0.003）. Conclusions IL-17A can up-regulate the expression of K17 in KC, and its specific regulation mechanism is by the activation of STAT3. Our experimental results prove that IL-17A takes part in the pathogenesis of psoriasis on the level of keratin. This work provides a new strategy toward pathogenesis and remedy in psoriasis.

关 键 词：白细胞介素17 银屑病 Th17细胞 角蛋白17 STAT3转录因子 

分 类 号：R758.63[医药卫生—皮肤病学与性病学]