摘要
目的探究miRNA-21调节JAK2/STAT3通路及其关键因子在脂多糖(LPS)诱导急性肺损伤(ALI)/急性呼吸窘迫综合征(ARDS)中的作用。方法构建LPS诱导ALI/ARDS细胞模型。通过实时荧光定量聚合酶链反应检测miRNA-21表达;蛋白质印迹技术检测JAK2、STAT3、BAX、BCL-2、NF-κB表达,细胞计数实验检测细胞活力,流式细胞仪检测细胞凋亡。结果不同时间点内,1 mg/L LPS干预促进细胞凋亡,促进JAK2、STAT3表达,使miRNA-21表达下调。抑制miRNA-21表达,细胞活力下降,凋亡表达增加。抑制JAK2、STAT3表达,细胞活力上升,凋亡表达减少;抑制miRNA-21表达,促进JAK2/STAT3激活,进而增加BAX、NF-κB表达;反之,增加miRNA-21表达,抑制JAK2/STAT3激活,降低BAX、NF-κB表达。结论LPS诱导ALI/ARDS致miRNA-21表达下调,JAK2/STAT3激活,BAX、NF-κB表达增加。JAK2/STAT3抑制剂干预可在一定程度上逆转由ALI/ARDS诱导促炎、促凋亡反应。抑制miRNA-21表达下调可在LPS诱导ALI/ARDS中发挥保护作用。
Objective To explore the role of miRNA-21 in modulating the JAK2/STAT3 signaling pathway and its critical factors in lipopolysaccharide(LPS)-induced acute lung injury(ALI)/acute respiratory distress syndrome(ARDS).Methods The LPS-induced ALI/ARDS cell model was established.The expression of miRNA-21 was detected by real-time fluorescent quantitative polymerase chain reaction.The expressions of JAK2,STAT3,BAX,BCL-2,NF-κB were detected by Western Blotting.Cell viability was measured by Cell Counting Kit-8.Cell apoptosis was detected by flow cytometry.Results miRNA-21 was down-regulated and JAK2/STAT3 were up-regulated in ALI/ARDS induced by 1 mg/L LPS.Inhibition of miRNA-21 resulted in decreased cell activity and pro-apoptotic expression.When miRNA-21 was down-regulated,and JAK2/STAT3 pathway was activated,BAX and NF-κB increased.Otherwise,when miRNA-21 was up-regulated and JAK2/STAT3 pathway was inactivated,BAX and NF-κB decreased.Conclusion miRNA-21 was down-regulated,JAK2/STAT3 pathway was activated,BAX and NF-κB were up-regulated in LPS-induced ALI/ARDS.Inhibition of JAK2/STAT3 could reverse the pro-inflammatory and pro-apoptotic responses induced by ALI/ARDS.Inhibiting the down-regulation of miRNA-21 might exert a protective role in LPS-induced ALI/ARDS.
作者
周璐
冯建国
贾静
伍春林
章卓
姜鲜
Zhou Lu;Feng Jianguo;Jia Jing;Wu Chunlin;Zhang Zhuo;Jiang Xian(Department of Anesthesiology, Affiliated Hospital of Southwest Medical University, Luzhou 646000, China;Laboratory of Anesthesiology, Southwest Medical University, Luzhou 646000, China;School of Pharmacy, Southwest Medical University, Luzhou 646000, China)
出处
《成都医学院学报》
CAS
2021年第4期424-430,436,共8页
Journal of Chengdu Medical College
