干扰STAT3基因对哮喘气道损伤的影响

发布时间：2018-04-02 15:38

本文选题：信号转导及转录激活因子　切入点：哮喘　出处：《中国病理生理杂志》2017年12期

【摘要】：目的:探讨信号转导及转录激活因子3(STAT3)在哮喘气道损伤中的作用及相关机制。方法:首先,通过Western blot检测哮喘患者的支气管黏膜组织中STAT3的表达及活化情况;其次,以支气管上皮细胞为研究对象,检测屋尘螨抗原P1(Derp1)刺激对STAT3的表达及活化的影响;再次,采用shRNA干扰支气管上皮细胞中STAT3的表达,CCK-8实验检测细胞活力,同时检测细胞培养液中肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)和IL-6的含量来反映细胞的炎症反应,检测细胞中丙二醛(MDA)的含量及超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)的活性来反映细胞氧化应激水平;流式细胞术检测细胞凋亡情况。结果:哮喘患者的支气管黏膜组织中p-STAT3的蛋白水平显著上升,Derp1刺激可显著上调支气管上皮细胞中p-STAT3的蛋白水平。沉默STAT3可抑制TNF-α、IL-1β和IL-6释放,降低MDA的含量,升高SOD和GSH-Px的活性,并抑制细胞凋亡,提高细胞活力(P0.05)。结论:沉默STAT3可减轻尘螨诱导的气道损伤,其作用机制可能是通过抑制JAK/STAT信号通路的激活,抑制支气管上皮细胞分泌炎症细胞因子及细胞内氧化应激反应,进而减少细胞凋亡。
[Abstract]:Objective: To investigate the signal transduction and transcription factor 3 (STAT3) in airway injury and the related mechanism. Methods: first, through the expression of STAT3 Western blot detection in patients with asthma and bronchial mucosa tissues and activation; secondly, the bronchial epithelial cells as the research object, the detection of house dust mite antigen P1 (Derp1) effect of stimulation on the expression of STAT3 and activation of STAT3 shRNA expression; again, the interference of bronchial epithelial cells, cell viability by CCK-8 assay, cell culture and detection of tumor necrosis factor alpha (TNF- alpha) solution, interleukin 1 beta (IL-1 beta) content and IL-6 can reflect the inflammation cell detection of cell, malondialdehyde (MDA) content and superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activity to reflect the stress level of cell oxidation; cell apoptosis was detected by flow cytometry: The protein level of p-STAT3 increased significantly in patients with asthma and bronchial mucosa tissues, Derp1 stimulation can significantly increase the p-STAT3 protein level of bronchial epithelial cells. Silencing of STAT3 can inhibit the release of TNF- alpha, IL-1 beta and IL-6, decrease the content of MDA, increase SOD and GSH-Px activity, and inhibit cell apoptosis, increase cell viability (P0.05). Conclusion: the silencing of STAT3 can reduce airway injury induced by dust mites, and its mechanism may be through inhibiting activation of JAK/STAT signaling pathway, inhibiting the secretion of bronchial epithelial cells in response to oxidative stress inflammatory cytokines and cells, and reduce cell apoptosis.

【作者单位】：郑州大学第一附属医院呼吸与危重症医学科;
【基金】：河南省卫生厅科技攻关项目(No.201503057) 河南省高等学校重点科研项目(No.18A320056) 河南省卫生科技创新型人才工程(豫卫科2010-52号)
【分类号】：R562.25

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