siRNA沉默JAK/STAT相关基因对COPD大鼠肺泡上皮细胞的影响

Effects of siRNA Silenced JAK/STAT Relevant Gene on Alveolar Epithelial Cells of COPD Rats

目的:观察JAK/STAT基因表达对慢性阻塞性肺疾病(COPD)大鼠肺泡上皮细胞增殖和炎症反应的影响。方法:实验分为对照组、模型组及实验组,提取正常肺泡上皮细胞作为对照组,构建COPD大鼠模型,提取肺泡上皮细胞作为模型组,实验组在模型组基础上利用siRNA体外沉默肺泡上皮细胞JAK2/STAT3基因转录,24 h后利用Western blot法检测细胞内JAK2和STAT3 mRNA和蛋白表达变化,MTT法检测肺泡上皮细胞增殖情况,ELISA法检测炎症因子TNF-α及IL-1β表达。结果:与对照组比较,模型组与实验组肺泡上皮细胞pJAK2和p-STAT3基因及蛋白表达升高,细胞增殖能力降低,TNF-α及IL-1β表达上调(P<0.05);与模型组比较,实验组肺泡上皮细胞p-JAK2和p-STAT3基因及蛋白表达降低,细胞增殖能力提高,TNF-α及IL-1β表达下调(P<0.05)。结论:阻断JAK/STAT信号通路可促进COPD模型大鼠肺泡上皮细胞增殖,抑制炎症反应,从而发挥保护作用。

Objective： To observe the effect of JAK/STAT gene expression on the proliferation and inflammatory response of alveolar epithelial cells in COPD rats. Methods： The rat model of COPD was constructed,the alveolar epithelial cells were extracted and siRNA was used to silence COPD model alveolar epithelial cell gene transcription. After 24 h,Western blot and RT-PCR were adopted to detect mRNA and protein expression of JAK / STAT3 and ELISA method was used to detect the inflammatory factor TNF alpha,IL-1 beta expression. Results： The proliferation capacity of alveolar epithelial cells increased after blocking the JAK / STAT3 signaling pathway in the COPD model（ P〈 0. 05）. RTPCR method result showed that mRNA expression of JAK / STAT3 was inhibited after blocking the JAK /STAT3 signaling pathway and Western blot result also showed that p-JAK2 and p-ATST3 protein expression were inhibited（ P〈 0. 05）. Simultaneously,the content of TNF-α and IL-1β were down-regulated after blocking JAK / STAT signal pathway（ P 〈0. 05）. Conclusion： Blocking the JAK / STAT signaling pathway may enhance the proliferation of alveolar epithelial cells in the COPD model,inhibit inflammatory response,and thus play a protective role.