三七对脓毒症急性肾损伤大鼠肾组织Toll样受体2/4和NOD样受体P1/3表达的干预作用  

Interference effect of Radix notoginseng on the expressions of Toll-like receptor 2/4 and NOD like receptor P1/3 in renal tissues of septic rats with acute kidney injury

作　　者：屠越兴[1] 俞云松[2] 韩芳[1] 倪银[1] 李曦[3] Tu Yuexing;Yu Yunsong;Han Fang;Ni Yin;Li Xi(Department of Intensive Care Unit,Zhejiang Provincial People's Hospital,People's Hospital of Hangzhou Medical College,Hangzhou 310014,Zhejiang,China;Department of Infection Disease,Sir Run Run Shaw Hospital,School of Medicine,Zhejiang University,Hangzhou 310016,Zhejiang,China;Centre of Laboratory Medicine,Zhejiang Provincial People's Hospital,People's Hospital of Hangzhou Medical College,Hangzhou 310014,Zhejiang,China)

机构地区：[1]浙江省人民医院,杭州医学院附属人民医院重症医学科,浙江杭州310014 [2]浙江大学医学院附属邵逸夫医院感染科,浙江杭州310016 [3]浙江省人民医院,杭州医学院附属人民医院检验科中心,浙江杭州310014

出　　处：《中国中西医结合急救杂志》2019年第1期5-10,共6页Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care

基　　金：浙江省自然科学基金(LQ17H190006).

摘　　要：目的探讨Toll样受体/NOD样受体(TLRs/NLRs)在脓毒症急性肾损伤(AKI)大鼠中的表达及三七的干预作用.方法选择150只雄性SD大鼠,将大鼠按随机数字表法分为假手术组、模型组、三七预处理组,每组50只.采用盲肠结扎穿刺术(CLP)复制脓毒症大鼠AKI模型;假手术组只开腹不结扎.三七预处理组预先给予三七3 g/kg灌胃、连续3 d,假手术组、脓毒症模型组给予等量生理盐水灌胃.各组大鼠于制模后6、12、24 h取10只采血后处死,并留取肾脏标本.采用鲎试剂检测内毒素水平;采用全自动生化分析仪检测血清肌酐(SCr)、血尿素氮(BUN)水平;采用酶联免疫吸附试验(ELISA)测定血清白细胞介素(IL-1β、IL-18、IL-6)、肿瘤坏死因子-α(TNF-α)水平;采用实时荧光定量聚合酶链反应(qPCR)及蛋白质免疫印迹试验(Western Blot)检测肾组织NLRP1/3、TLR2/4的mRNA及蛋白表达水平;光镜观察术后24 h肾组织病理学改变;每组剩余20只大鼠观察4 d、7 d存活率.结果脓毒症模型组和三七预处理组内毒素、SCr、BUN、IL-1β、IL-6、IL-18、TNF-α、NLRP3和TLR2/4的mRNA及蛋白表达水平均较假手术组明显升高,内毒素、IL-18、NLRP3、TLR2/4的mRNA及蛋白表达水平均于术后6 h即出现统计学差异〔内毒素(kU/L):61.3±25.7、56.9±18.6比0.2±0.1,IL-18(ng/L):16.7±5.0、13.8±2.9比10.6±2.8,NLRP3 mRNA(2^-ΔΔCt):6.3±1.9、4.0±1.2比1.1±0.4,TLR2 mRNA(2^-ΔΔCt):5.7±1.3、2.0±0.8比0.9±0.3,TLR4 mRNA(2^-ΔΔCt):4.4±1.2、2.3±0.7比0.6±0.2,NLRP3/β-actin:38.2±9.3、26.1±7.2比18.3±5.1,TLR2/β-actin:21.9±6.1、16.2±4.4比10.9±2.8,TLR4/β-actin:18.3±6.7、12.0±3.9比7.5±2.0,均P<0.05〕,SCr、BUN、IL-1β、TNF-α、IL-6均于术后12 h出现统计学差异〔SCr(μmol/L):62.3±21.6、38.1±13.9比36.0±11.9,BUN(mmol/L):16.5±7.2、6.9±2.6比6.8±2.5,IL-1β(ng/L):37.6±10.9、31.2±9.3比20.3±6.5,TNF-α(ng/L):15.6±3.9、10.2±2.8比7.3±2.1,IL-6(ng/L):9.3±2.5、6.8±1.7比5.0±1.3,均P<0.05�Objective To investigate the expressions of Toll-like receptors and NOD-like receptor(TLR/NLRP)in septic rats with acute kidney injury(AKI)and influence of Radix notoginseng.Methods One hundred and fifty male Sprague-Dawley(SD)rats were selected,and they were divided into three groups:sham operation group,model group and Radix notoginseng pretreatment group with 50 rats in each group.Sepsis rats mth AKI models were established by cecal ligation puncture(CLP);only was laparotomy performed in the sham operation group without ligation.The rats in Radix notoginseng pretreatment group were given Radix notoginseng(3 g/kg)for consecutive 3 days by gastric perfusion before treatment,and the sham operation group and sepsis model group rats were given equal amount of normal saline by gastric perfusion.After their blood was collected,at each following time points 6,12 and 24 hours after modeling,10 rats in each group were sacrificed,and the kidney specimens were collected.The endotoxin levels were detected by limulus assay;the levels of serum creatinine(SCr)and urea nitrogen(BUN)were detected by automatic biochemical analyzer;serum levels of interleukins(IL-1β,IL-18,IL-6)and tumor necrosis factor-α(TNF-α)were measured by enzyme-linked immunosorbent assay(ELISA);the expression levels of NLRP1/3 and TLR2/4 mRNA and protein in renal tissue were detected by real-time fluorescence quantitative polymerase chain reaction(qPCR)and Western Blot,respectively;the pathological changes of renal tissues were observed by optical microscope.The 4-day and 7-day survival rates were observed in the remaining 20 rats in each group.Results The levels of endotoxin,SCr,BUN,IL-1β,IL-6,IL-18,and TNF-αand the expressions of NLRP1/3,TLR2/4 mRNA and proteins in sepsis model group and Radix notoginseng pretreatment group were significantly higher than those in sham operation group,and the levels of endotoxin,IL-18,and the expressions of NLRP1/3,TLR2/4 mRNA and proteins showed statistically significant differences at 6-hour after operation[endotoxi

关 键 词：TOLL样受体 NOD样受体 脓毒症 肾损伤 急性 三七 

分 类 号：R392.11[医药卫生—免疫学]