Toll样受体2/4在糖尿病大鼠肾缺血再灌注损伤时的表达及肾康注射液的保护作用  

Expression of Toll-like receptors 2/4 and the protective effect of Shenkang injection on renal ischemia-reperfusion injury in diabetic rats

作　　者：刘洋[1] 马强[1] 杨光[1] 敖强国[1] 刘胜[1] 赵佳慧[1] 王晓华[1] 程庆砾[1] Liu Yang;Ma Qiang;Yang Guang;Ao Qiangguo;Liu Sheng;Zhao Jiahui;Wang Xiaohua;Cheng Qingli.(Department of Geriatric Nephrology, Chinese PLA General Hospital, Beijing, Chin)

机构地区：[1]解放军总医院南楼临床部肾脏病科,北京100853

出　　处：《中华肾病研究电子杂志》2018年第2期59-64,共6页Chinese Journal of Kidney Disease Investigation（Electronic Edition）

基　　金：国家自然科学基金青年项目(81600655);军队保健专项科研基金(16BJZ12)

摘　　要：目的探讨糖尿病(DM)大鼠肾缺血再灌注损伤(IRI)的可能机制及肾康注射液(SKI)的保护作用。方法采用高糖高脂饮食诱导6周联合小剂量链脲佐菌素的方法建立DM大鼠模型,将造模成功的SD大鼠随机分为糖尿病肾缺血再灌注组(DMIR)、糖尿病肾缺血再灌注SKI治疗组(DMIR+SKI)、糖尿病假手术组(DMS)和正常对照组(NCS)。DMIR+SKI组大鼠应用SKI[6.0 g/(kg·d)]干预8周,其它组用等量生理盐水。应用右肾切除联合左肾动脉夹闭45 min后再灌注24 h的方法诱导肾IRI。最后,抽血、留取肾组织。ELISA法检测大鼠血清高迁移率族蛋白(HMGB1)水平;免疫组化方法检测肾组织HMGB1、Toll样受体(TLR)2和TLR4的表达;免疫印迹法和实时定量PCR法检测肾组织TLR2、TLR4、髓样分化因子88(MyD88),核因子-κBp56(NF-κBp56)的表达。结果 DMIR组大鼠血清HMGB1水平,肾小管HMGB1、TLR2和TLR4的表达以及肾组织TLR2、TLR4、MyD88和NF-κBp56的蛋白及mRNA水平均明显高于DMS组(P<0.01);经过SKI干预治疗8周,DMIR+SKI组大鼠的上述指标均明显低于DMIR组(P<0.01)。结论 SKI可通过抑制TLR2/4炎症反应通路,减轻DM大鼠肾IRI炎症反应而起到肾脏保护作用。Objective To investigate the mechanisms and therapeutic effects of Shenkang injection（ SKI） on renal ischemia-reperfusion injury（ IRI） in the rats with diabetes mellitus（ DM）. Methods The DM rat model was induced by high-sugar,high-fat diet plus low-dose intraperitoneal injection of streptozotocin for 6 weeks. Then,the Sparague-Dawley rats were divided into four groups： DM ischemia-reperfusion（ DMIR） group,DMIR plus treatment with SKI（ DMIR ＋ SKI） group,DM sham（ DMS） group,and normal control sham（ NCS） group. The DMIR ＋ SKI group were treated with SKI（ 6. 0 g/kg/d） for eight weeks after DM model established,while rats of the other groups were treated with normal saline. Afterwards,the renal IRI was induced by a right nephrectomy plus clamping the left renal artery for 45 minutes and reperfusion for24 hours. Finally,blood drawing and renal tissues collection were made. The serum high mobility group box 1（ HMGB1） was detected with ELISA. The renal expression of HMGB1,Toll-like receptor（ TLR） 2,TLR4,myeloid differentiation factor 88（ MyD88）,and nuclear factor κBp56（ NF-κBp56） were detected with immunohistochemistry,Western blot,and real-time polymerase chain reaction. Results The level of serum HMGB1 and renal expression of HMGB1,TLR2,TLR4,MyD88,and NF-κBp56 were all significantly higher in DMIR group than in DMS group（ P〈0. 01）,but were significantly decreased in the DMIR ＋ SKI group.Conclusions Our results indicated that TLR2/4 pathway activation might play an important role in renal IRI of DM rats. The renal protective effects of SKI may be related with the TLR2/4 pathway inhibition.

关 键 词：糖尿病 缺血再灌注肾损伤 TOLL样受体2 TOLL样受体4 肾康注射液 

分 类 号：R285.5[医药卫生—中药学]