SPINK7 对IL-22 介导的角质细胞异常增殖及炎症应答的影响

目的:探讨SPINK7对IL-22刺激后角质形成细胞HaCaT的过度增殖及炎症应答的影响及其可能的机制。方法:采用IL-22刺激体外培养的HaCaT细胞。采用RT-PCR检测SPINK7和细胞炎症因子的mRNA表达;Western blot检测SPINK7,cyclin D1、survivin和Wnt/β-catenin通路相关蛋白的表达;MTT法检测细胞增殖能力的变化;ELISA法检测上清液中IL-23、TNF-α和IL-17A的表达水平。结果:IL-22处理的HaCaT细胞中,SPINK7的mRNA和蛋白表达量均显著升高。此外,沉默SPINK7明显抑制IL-22诱导的细胞增殖,同时抑制凋亡相关蛋白cyclin D1及survivin的表达水平。SPINK7沉默显著降低IL-22刺激诱导的促炎性细胞因子IL-23、TNF-α和IL-17A的mRNA表达水平和分泌量。机制研究发现SPINK7沉默能够抑制IL-22激活的Wnt/β-catenin信号通路。结论:沉默SPINK7能够抑制IL-22诱导的细胞增殖和炎症应答反应,其机制可能与抑制Wnt/β-catenin信号通路有关,这为银屑病的诊断提供了新的标志物和治疗靶点。

Effects of SPINK7 on IL-22-induced hyperproliferation and inflammatory response of keratinocytes

Objective:To investigate the potential role and molecular mechanism of SPINK7 in IL-22-stimulated hyperproliferation and inflammatory response in HaCaT cells.Methods:The keratinocytes HaCaT were triggered by activation of IL-22.The mRNA expression of SPINK7 and inflammatory cytokines was determined using RT-PCR assay.Western blot analysis was performed to evaluate the protein expression of SPINK7,cyclin D1,survivin and associated protein of Wnt/β-catenin signaling.MTT assay was conducted to estimate cell proliferation.The secretion of inflammatory factors IL-23,TNF-αand IL-17A was measured using ELISA assay.Results:The mRNA and protein expressions of SPINK7 were elevated in IL-22-treated HaCaT cells.In addition,silencing of SPINK7 obviously restrained IL-22-triggered cell proliferation,as well as attenuated the expression of apoptosis associated proteins cyclin D1 and survivin.Moreover,siliencing of SPINK7 remarkably prohibited the production of inflammatory cytokines IL-23,TNF-αand IL-17A induced by IL-22 in HaCaT cells.Mechanically,siliencing of SPINK7 could inhibit the activation of Wnt/β-catenin signaling pathway in IL-22-stimulated HaCaT cells.Conclusion:These findings manifested that siliencing of SPINK7 restrained IL-22-stimulated abnormal proliferation and inflammatory response of keratinocyte via Wnt/β-catenin signaling,and provide diagnostic marker and a novel target for psoriasis treatment.