调控RANKL/RANK/TLR4信号通路对脓毒症相关急性肾损伤的影响  

Impact of regulating the RANKL/RANK/TLR4 signaling pathway on sepsis-associated acute kidney injury

作　　者：牛新荣[1] 李辉 陈蔚霖 NIU Xinrong;LI Hui;CHEN Weilin(Second Department of Critical Care Medicine,People's Hospital of Xinjiang Uygur Autonomous Region,Xinjiang,Urumqi 830000,China;不详)

机构地区：[1]新疆维吾尔自治区人民医院重症医学二科,乌鲁木齐市830000 [2]新疆医科大学研究生院

出　　处：《河北医药》2025年第3期381-385,390,共6页Hebei Medical Journal

基　　金：新疆维吾尔自治区自然科学基金(编号:2022D01C604)。

摘　　要：目的探讨核因子-κB受体活化因子配体(RANKL)/核因子-κB受体激活因子(RANK)/toll样受体4(TLR4)信号通路在脓毒症相关急性肾损伤(SA-AKI)炎性反应中的作用及可能的机制。方法将24只雄性C57BL/6小鼠按随机数字表法分为假手术组(S组)、盲肠结扎穿孔术(CLP)诱导的脓毒症模型组(C组)、重组RANKL预处理组(R组)、抗RANKL预处理组(A组),每组6只。术前2 h CLP+重组RANKL组予以腹腔注射重组RANKL,CLP+抗RANKL组给予腹腔注射抗RANKL,制模后24 h采血并处死小鼠留取肾脏组织,用肌酐(Scr)及尿素氮(BUN)检测试剂盒检测血Scr、BUN水平,酶联免疫吸附测定(ELISA)法检测血清中IL-6、IL-1β、TNF-α水平,苏木精-伊红染色法(HE染色)观察肾脏组织病理学变化,反转录聚合酶链反应(RT-qPCR)及蛋白印迹法(Western blotting)检测肾脏组织中RANKL、RANK、TLR4 mRNA及蛋白表达水平。结果与假手术组比较,脓毒症组血清Scr、BUN、IL-1β、TNF-α、IL-6明显升高,肾脏组织病理学结果示肾小球部分缺血皱缩,球囊间隙扩大,肾小管上皮细胞坏死脱落于管腔中,管腔扩大,肾间质水肿,炎性细胞浸润,肾脏病理学评分明显升高,肾脏组织中RANKL的mRNA及蛋白表达均明显降低,而RANK和TLR4的mRNA及蛋白表达均明显升高,差异有统计学意义(均P<0.05);与脓毒症组比较,重组RANKL预处理组Scr、BUN、IL-1β、TNF-α、IL-6水平及病理学评分均明显降低,肾脏组织病理学结果示肾脏损害明显缓解,肾脏组织中RANKL的mRNA及蛋白表达均明显升高,而RANK和TLR4的mRNA及蛋白表达均明显下降,差异有统计学意义(均P<0.05);而抗RANKL预处理组肾功能及炎性因子水平有所升高,肾脏损伤程度加重,肾脏组织中RANKL的mRNA及蛋白表达均明显降低,RANK和TLR4的mRNA及蛋白表达均明显升高,差异有统计学意义(均P<0.05)。结论在SA-AKI的小鼠模型中,通过增加RANKL/RANK/TLR4信号通路的信号转导可减轻炎性反�Objective To investigate the role and possible mechanisms of the receptor activator of nuclear factor-κB ligand(RANKL)/receptor activator of nuclear factor-κB(RANK)/toll-like receptor 4(TLR4)signaling pathway in the inflammatory response of sepsis-associated acute kidney injury(SA-AKI).Methods Twenty-four male C57BL/6 mice were randomly assigned into the sham operation group(S group),cecal ligation and puncture(CLP)-induced sepsis model group(C group),recombinant RANKL pretreatment group(R group),and anti-RANKL pretreatment group(A group),with 6 mice in each group.Two hours before surgery,CLP-induced in R group and A group were intraperitoneally injected with recombinant RANKL and anti-RANKL antibody,respectively.Blood was collected 24 hours after modeling,and mice were sacrificed to harvest kidney tissue.Creatinine(Scr)and blood urea nitrogen(BUN)levels were measured using commercial kits.Enzyme-linked immunosorbent assay(ELISA)was performed to detect serum interleukin 6(IL-6),IL-1β,and tumour necrosis factor alpha(TNF-α)levels.Hematoxylin and eosin(H&E)staining was performed to observe the histopathological changes in mouse kidney tissue.Reverse transcription quantitative polymerase chain reaction(RT-qPCR)and Western blotting were used to detect the mRNA and protein expression levels of RANKL,RANK,and TLR4 in mouse kidney tissue,respectively.Results Compared with those of S group,mice in the C group had significantly higher serum Scr,BUN,IL-1β,TNF-α,and IL-6 levels.Histopathological assessment of kidney tissue showed partial ischemic shrinkage of renal glomeruli,widening of the glomerular interstitium,necrosis and shedding of renal tubular epithelial cells into the lumen,enlargement of tubular lumens,renal interstitial edema,inflammatory cell infiltration,and a significant increase in renal pathology scores in mice of C group.The mRNA and protein expressions of RANKL in kidney tissue were significantly downregulated,while RANK and TLR4 were significantly upregulated in mice of C group than S group(all P<0

关 键 词：脓毒症 急性肾损伤 RANKL/RANK/TLR4信号通路 炎性反应 

分 类 号：R692.5[医药卫生—泌尿科学]