干扰STAT3基因对哮喘气道损伤的影响  

Effect of STAT3 down-regulation on airway injury in asthma

作　　者：杨远舰 张桢[1] 程哲[1] 

机构地区：[1]郑州大学第一附属医院呼吸与危重症医学科,河南郑州450052

出　　处：《中国病理生理杂志》2017年第12期2269-2273,共5页Chinese Journal of Pathophysiology

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)

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(P<0.05)。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。AIM ： To investigate the effect of signal transducer and activator of transcription 3 （STAT3） on air-way injury in asthma and its mechanism. METHODS ： The expression and activation of STAT3 in bronchial mucosa tissues of asthmatic patients were measured by Western blot. The expression and activation of STAT3 in bronchial epithelial cells pretreated with Dermatophagoides pteronyssinus antigen PI （ Derpl ） were estimated. Bronchial epithelial cells were trans-fected with STAT3 shRNA. STAT3 expression was measured by Western blot. The cell viability was detected by CCK-8 as-say. The concentrations of TNF-a, IL-lp and IL-6 were detected by ELISA. The content of malondialdehyde （ MDA） and the activity of superoxide dismutase （SOD） and glutathione peroxidase （ GSH-Px） were also determined for evaluating the status of oxidative stress. The cell apoptosis was analyzed by flow cytometry. RESULTS ： The protein level of p-STAT3 was significantly up-regulated in both bronchial mucosa of asthmatic tissues and bronchial epithelial cells pretreated with Derpl. Knockdown of STAT3 inhibited the releases of TNF-a, IL-1 p and IL-6, decreased the content of MDA and enhanced the activity of SOD and GSH-Px with the suppression of cell apoptosis （ P 〈 0. 05 ） . CONCLUSION ： Down-regulation of STAT3 attenuates Derpl-induced the airway injury. The mechanism may involve that knockdown of STAT3 suppresses the activation of JAK/STAT signaling pathway, the release of asthma-related inflammatory cytokines and oxidative stress in bronchial epithelial cells, thus inhibiting cell apoptosis.

关 键 词：信号转导及转录激活因子3 哮喘 支气管上皮细胞 炎症细胞因子 氧化应激 

分 类 号：R0[医药卫生] R562.25