TLR4/NF-κB信号通路在肾缺血再灌注损伤中的作用机制  

Mechanism of TLR4/NF-κB Signaling Pathway in Renal Ischemia-reperfusion Injury

作　　者：李强[1] 靳书滨[1] 霍韶军[1] LI Qiang;JIN Shubin;HUO Shaojun(Second Department of Urology,Handan Central Hospital,Hebei 056000,China)

机构地区：[1]邯郸市中心医院泌尿外二科,056000

出　　处：《医学研究杂志》2023年第6期73-77,82,共6页Journal of Medical Research

基　　金：河北省医学科学研究课题计划(20191258)。

摘　　要：目的探讨Toll样受体4/核因子-κB(Toll-like receptor 4/nuclear factor-κB,TLR4/NF-κB)信号通路在肾缺血再灌注(renal ischemia-reperfusion,RIR)损伤中的作用机制。方法96只健康SD大鼠随机分为4组,即假手术(Sham)组、模型(Model)组、Model+TAK242(TLR4抑制剂,10mg/kg)组和Model+LPS(TLR4激活剂,5mg/kg)组,每组24只。除Sham组外,其他组采用夹闭双侧肾动脉45min的方法复制RIR大鼠模型;造模前7天开始,各组分别1次/天腹腔注射给药。再灌注6h后,检测血清肾功能指标,HE染色法行肾组织病理学检查;透射电子显微镜观察肾小管上皮细胞超微结构改变;RT-PCR法检测肾组织TLR4、NF-κB p65mRNA表达;Western blot法检测肾组织TNF-α、IL-1β、TLR4、NF-κB p65mRNA和蛋白表达。结果与Sham组比较,Model组血清BUN、SCr水平升高(P<0.05);肾组织可见肾小球体积增大、肾小管管腔扩张、炎性细胞浸润等病理学改变,病理评分升高(P<0.05);肾小管上皮细胞可见线粒体肿胀、膜破裂、嵴断裂,内质网扩张,核糖体减少等超微结构病变;肾组织TNF-α、IL-1β蛋白表达量升高,TLR4、NF-κB p65mRNA和蛋白表达量均升高(P<0.05)。与Model组比较,Model+TAK242组BUN、SCr水平降低(P<0.05),肾组织病变和肾小管上皮细胞超微结构改变均明显改善,TNF-α、IL-1β蛋白表达量降低,TLR4、NF-κB p65mRNA和蛋白表达量降低(P<0.05);Model+LPS组BUN、SCr水平升高(P<0.05),肾组织病变和肾小管上皮细胞超微结构改变均明显加重,TNF-α、IL-1β蛋白表达量升高,TLR4、NF-κB p65mRNA和蛋白表达量升高(P<0.05)。结论TLR4/NF-κB信号通路可能通过调节炎性反应参与RIR损伤过程。Objective To investigate the mechanism of TLR4/NF-κB signaling pathway in renal ischemia-reperfusion(RIR)injury.Methods 96healthy SD rats were randomly divided into Sham group,Model group,Model+TAK242(TLR4 inhibitor,10mg/kg)group and Model+LPS(TLR4 activator,5mg/kg)group,n=24.Except for the Sham group,the rats in other groups were prepared RIR rat model by clamping bilateral renal arteries for 45minutes.7days before modeling,the rats in each group was given intraperitoneal injection once a day.6hours after RIR,the level of renal function indexes in serum were detected,the renal histopathological was examined by HE staining;the ultrastructural changes of renal tubular epithelial cells was observed by transmission electron microscope;the mRNA expression of TLR4,NF-κB p65 were detected by RT-PCR;the protein expression of TNF-α,IL-1β,TLR4,NF-κB p65 were detected by Western blot.Results Compared with the Sham group,the level of BUN,SCr in serum of Model group were increased(P<0.05);the renal histopathological changes such as glomerular volume enlargement,renal tubule lumen expansion and inflammatory cell infiltration could be seen,and the pathological score was increased(P<0.05);the ultrastructural lesions such as mitochondrial swelling,membrane rupture,cristae fracture,endoplasmic reticulum expansion and ribosome reduction could be seen in renal tubular epithelial cells;the protein expression of TNF-α,IL-1βwere increased,the mRNA and protein expression of TLR4,NF-κB p65 were increased(P<0.05).Compared with the Model group,the level of BUN,SCr in Model+TAK242group were decreased(P<0.05);the renal tissue lesions and renal tubular epithelial cell ultrastructural changes were significantly improved;the expression of TNF-α,IL-1βwere decreased,the mRNA and protein expression of TLR4,NF-κB p65 were decreased(P<0.05).The level of BUN,SCr in Model+LPS group were increased(P<0.05);the renal tissue lesions and renal tubular epithelial cell ultrastructural changes were significantly worse;the expression of TNF-α,IL-1

关 键 词：TLR4/NF-κB信号通路 肾缺血再灌注 肾小管 炎性反应 

分 类 号：R363.1[医药卫生—病理学]