摘要
目的 探讨微小RNA-20a（microRNA-20a,miR-20a）在脂多糖诱导肺泡上皮细胞A549炎症反应中的作用及可能机制.方法 首先将miR-20a类似物（mimic）/抑制剂（inhibitor）转染至A549细胞,再用脂多糖共同刺激24 h后,用实时定量聚合酶链反应检测白细胞介素6（interleukin-6,IL-6）和IL-8 mRNA的表达,酶联免疫吸附法检测IL-6和IL-8蛋白的表达情况,并用Western blot检测细胞凋亡信号调节激酶1（apoptosis signal regulating kinase 1,ASK1）、P38、P-P38、JNK及P-JNK蛋白的表达.结果 mimic组IL-6、IL-8 mRNA和蛋白表达水平低于mimic阴性对照组,inhibitor组IL-6、IL-8 mRNA和蛋白表达水平高于inhibitor阴性对照组,差异均有统计学意义（P〈0.05）;mimic组ASK1、P-P38及P-JNK蛋白表达低于mimic阴性对照组,inhibitor组ASK1、P-P38、P-JNK蛋白表达高于inhibitor阴性对照组,差异均有统计学意义（P〈0.05）.结论 miR-20a对ASK1的靶向调控可能在急性呼吸窘迫综合征的炎症发生过程中发挥作用.
Objective To study the roles of miR-20a in lipopolysaccharide induced inflammation of A549 cells and the possible mechanisms.Method The miR-20a mimic/inhibitor were transfected into A549 cells, and the cells were stimulated using lipopolysaccharide for 24 h.Interleukin-6 （ IL-6） and IL-8 were detected at mRNA level and protein level using real-time PCR and ELISA method , respectively.Protein expression of apoptosis signal regulating kinase 1 （ASK1）、P38、P-P38、JNK and P-JNK were detected using Western blot. Result Compared to mimic negative control group , the levels of mRNA and protein expression of IL-6 and IL-8 in the mimic group were all significantly decreased （ P〈0.05）.Compared to inhibitor negative control group , the levels of mRNA and protein expression of IL-6 and IL-8 in the inhibitor group were all significantly increased （P〈0.05）.The levels of ASK1, P-P38 and P-JNK protein in the mimic group were significantly lower than the mimic negative control group （P〈0.05）;the level of protein expression of ASK1, P-P38 and P-JNK in the inhibitor group were all higher than the inhibitor negative control group （P〈0.05）.Conclusion The regulation of ASK1 by miR-20a may play an important role in the inflammation process of acute respiratory distress syndrome .
基金
国家自然科学基金（81571476）
江苏省自然科学基金（BK20141080）
湖北省自然科学基金（2015CFB626）
