miR-29a靶向信号转导及转录激活因子3调控炎症反应在脂多糖诱导大鼠肾小管上皮细胞损伤中的保护作用

The protective effect of miR-29a on LPS induced rat renal tubular epithelial cell injury by targeting STAT3

目的研究miR-29a靶向信号转导及转录激活因子3(signal transduction and activator of transcription 3,STAT3)调控炎症反应在脂多糖(LPS)诱导大鼠肾小管上皮细胞损伤中的保护作用。方法培养大鼠肾小管上皮细胞株NRK-52E,LPS刺激建立脓毒症细胞模型、诱导细胞损伤,转染miR-NC、miR-29a、pcDNA3.1质粒、pcDNA3.1-STAT3质粒或用STAT3抑制剂AG490处理,比较细胞活力A490、STAT3、p-STAT3、bcl-2表达量,TNF-α、IL-1β含量的差异;双荧光素酶报告基因验证miR-29a靶向STAT3。结果LPS组的miR-29a表达水平、A490水平、Bcl-2的表达水平均低于对照组,STAT3、p-STAT3的表达水平、TNF-α、IL-1β的含量均高于对照组;LPS+miR-29a组的miR-29a表达水平、A490水平、Bcl-2的表达水平均高于LPS组,STAT3、p-STAT3的表达水平、TNF-α、IL-1β的含量均低于LPS组;pcDNA3.1-STAT3+miR-29a组的A490水平及Bcl-2的表达水平均低于pcDNA3.1+miR-29a组,STAT3、p-STAT3的表达水平、TNF-α、IL-1β的含量均高于pcDNA3.1+miR-29a组;miR-29a组STAT3双荧光素酶报告基因的荧光活力低于miR-NC组。结论STAT3通路激活与LPS诱导的肾小管上皮细胞炎症反应及凋亡有关,miR-29a通过靶向STAT3减轻LPS引起的炎症反应及凋亡。

Objective To study the protective effect of miR-29a on LPS induced rat renal tubular epithelial cell injury by targeting signal transduction and activator of transcription 3(STAT3).Methods Rat renal tubular epithelial cell line NRK-52E was cultured.LPS stimulation was used to establish sepsis cell model and induce cell injury,then transfected with miR-nc,miR-29a,pcDNA3.1 plasmid,pcDNA3.1-STAT3 plasmid or treated with STAT3 inhibitor AG490.Then cell viability A490,the expression of STAT3,p-STAT3 and Bcl-2,TNF-αand IL-1βcontents were compared,and miR-29a targeting STAT3 was confirmed by double luciferase reporter gene.Result The expression levels of miR-29a,Bcl-2,the level of A490 of LPS group were lower than those of the control group,while the expression of STAT3,p-STAT3,the contents of TNF-αand IL-1βwere higher than those in control group;the expression levels of miR-29a,Bcl-2,the level of A490 of LPS+miR-29a group were higher than those of the LPS group,while the expression of STAT3,p-STAT3,the contents of TNF-αand IL-1βwere lower than those in LPS group;the expression levels of Bcl-2,the level of A490 of pcDNA3.1-STAT3+miR-29a were lower than pcDNA3.1+miR-29a group,while the expression of STAT3,p-STAT3,the contents of TNF-αand IL-1βwere higher than those in LPS group;the fluorescence activity of STAT3 double luciferase reporter gene of miR-29a group was lower than that of miR-nc group.Conclusion STAT3 pathway activation is related to LPS induced inflammatory response and apoptosis of renal tubular epithelial cells,miR-29a can reduce LPS induced inflammatory response and apoptosis by targeting STAT3.