miR-216a调控NF-κB信号通路参与脓毒症急性肾损伤的机制

Mechanism of involvement of miR-216a insepsis-inducedacute kidney injury by regulating NF-κB signaling pathway

目的探讨miR-216a通过NF-κB信号通路对脓毒症大鼠急性肾损伤的作用机制。方法选取SD大鼠40只,采用盲肠结扎穿孔法进行造模,随机分为假手术组、模型组、miR-NC组、miR-216a组,各组10只。观察肾脏病理形态学变化;采用RT-PCR检测各组大鼠肾组织中miR-216a表达情况;采用全自动生化分析仪检测大鼠血清中生化指标水平;采用ELISA法检测各组大鼠炎症因子水平;采用Western blot检测鼠NF-κB通路相关蛋白表达。结果假手术组大鼠肾组织结构无异常;模型组和miR-NC组肾组织伴有炎性浸润、肾间质加宽;miR-216a组炎性浸润现象以及肾小球病变情况有所缓解;与假手术组相比,模型组、miR-NC组肾组织miR-216a表达水平较低,与miR-NC组相比,miR-216a组肾组织的miR-216a表达水平较高(P<0.05);与假手术组相比,模型组、miR-NC组大鼠血清Scr、BUN、KIM-1水平较高,与miR-NC组相比miR-216a组大鼠血清Scr、BUN、KIM-1水平较低(P<0.05);与假手术组相比,模型组、miR-NC组大鼠血清TNF-α、IL-6、IL-1β水平较高,与miR-NC组相比miR-216a组大鼠血清TNF-α、IL-6、IL-1β水平较低(P<0.05);与假手术组相比,模型组、miR-NC组大鼠肾组织中NF-κB p65、p-NF-κB p65表达较高,与miR-NC组相比miR-216a组大鼠肾组织中NF-κB p65、p-NF-κB p65表达较低(P<0.05)。结论miR-216a过表达可以降低脓毒症大鼠炎症水平,改善肾损伤,可能是通过调控NF-κB信号通路实现的。

Objective To investigate the mechanism ofaction ofmiR-216a on acute kidney injury in septic ratsvia NF-κB signaling pathway.Methods Forty SD rats were selected,and a rat model was established by cecal ligation and puncture.The rates were randomly divided into asham-operation group,model group,miR-NC group,and miR-216a group,10 for each group.The pathologicalchangesin thekidneys were observed.RT-PCR was used to detect expression of miR-216a in the ratkidney tissues in each group;serum levels of biochemical indexes were detected by automatic biochemical analyzer;levels of inflammatory factors in each group were detected by ELISA;and expression of murine NF-κB pathway-related protein was detected by WB.Results In the sham-operation group,no abnormal kidney tissue structure developed;in the model and the miR-NC groups,the kidney tissues wereaccompanied by inflammatory infiltration and widened interstitium;andin the miR-216a group,inflammatory infiltration and glomerular lesions were alleviated.As compared with the sham-operationgroup,expression levels of miR-216a inthe kidney tissues were lower in the model group andthemiR-NC group,and as compared with the miR-NC group,expression levels of miR-216a inthe kidney tissue were higher in the miR-216a group.As compared with the sham-operationgroup,serum levels of Scr,BUN and KIM-1 were higher in the model group and the miR-NC group while lower in the miR-216a groupascompared with the miR-NC group(P<0.05).As compared to the shamoperation group,serum levels of TNF-α,IL-6,and IL-1βwere higher in the model groupand miR-NC group,butlower in themiR-216a group ascompared to miR-NC group(P<0.05).In comparison withthe sham-operation group,expressions of NF-κB p65,p-NF-κB and p65 were higher inthemodel groupand miR-NC group,while those expressionswerelower in the miR-216a groupthan in the miR-NC group(P<0.05).Conclusions miR-216a overexpression reduced inflammation levels and ameliorated renal injury in septic rats,possibly through regulation of the NF-κB signaling pathway.