姜黄素后处理肢体缺血再灌注肾损伤模型大鼠基质金属蛋白酶9及基质金属蛋白酶组织抑制因子1平衡的变化  

Effects of curcumin post-conditioning on the balance of matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 in a rat model of renal injury induced by limb ischemia/reperfusion

作　　者：邹海波[1] 孙晓峰[1] Zou Haibo;Sun Xiaofeng(Department of Anesthesiology,Affiliated Central Hospital of Shenyang Medical College,Shenyang 110024,Liaoning Province,China)

机构地区：[1]沈阳医学院附属中心医院麻醉科

出　　处：《中国组织工程研究》2019年第23期3643-3648,共6页Chinese Journal of Tissue Engineering Research

基　　金：辽宁省教育厅一般项目(L2015540),项目负责人:孙晓峰;沈阳医学院青年基金项目(20162029),项目负责人:邹海波~~

摘　　要：背景:姜黄素预处理可减轻肢体缺血再灌注对肾脏的损伤,但姜黄素后处理是否对肾脏损伤有保护作用及其机制目前研究甚少。目的:探讨大鼠肢体缺血再灌注肾组织损伤时姜黄素后处理对基质金属蛋白酶9/基质金属蛋白酶组织抑制因子1平衡的影响。方法:成年雄性SD大鼠80只由中国医科大学实验室中心提供。实验方案经沈阳医学院附属中心医院动物实验伦理委员会批准。采用随机数字表法,将80只SD大鼠分为4组(n=20):假手术组、缺血再灌注组、姜黄素后处理组和地塞米松组。除假手术组外,其余大鼠采用夹闭双侧股动脉4h,再灌注4h的方法制备肢体缺血再灌注模型。在大鼠肢体缺血后4h,姜黄素后处理组经腹腔注射姜黄素200mg/kg,地塞米松组经腹腔注射地塞米松1.5mg/kg,假手术组、缺血再灌注组以等量生理盐水替代。于再灌注4h后,经颈动脉采集血样,检测肌酐、尿素氮;麻醉下取大鼠肾脏组织检测丙二醛水平,苏木精-伊红染色检测病理变化和肾小管间质半定量评分(SSRT);应用Westen blot法和RT-PCR法分别测定基质金属蛋白酶9、基质金属蛋白酶组织抑制因子1蛋白和核因子κB p65 mRNA含量,计算基质金属蛋白酶9/基质金属蛋白酶组织抑制因子1比值;应用ELESA法检测肺组织肿瘤坏死因子α和白细胞介素1β等炎性因子的水平。结果与结论:①与假手术组比较,缺血再灌注组血浆肌酐、尿素氮和丙二醛均升高(P<0.05);肾苏木精-伊红染色示肾小管扩张,管腔内可见管型,刷状缘消失、肾间质炎症细胞浸润等病变;SSRT升高(P<0.05);基质金属蛋白酶9、基质金属蛋白酶9/基质金属蛋白酶组织抑制因子1比值和核因子κB p65 mRNA含量均升高(P<0.05);肿瘤坏死因子α和白细胞介素1β炎性因子释放增加(P<0.05);②与缺血再灌注组比较,姜黄素后处理组血浆肌酐、尿素氮和丙二醛降低(P<0.05);肾组织炎性细胞浸BACKGROUND:Curcumin preconditioning can alleviate renal injury induced by limb ischemia/reperfusion,but whether curcumin preconditioning has protective effect on renal injury and its mechanisms are still poorly studied.OBJECTIVE:To investigate the effects of the balance of matrix metalloproteinase-9/tissue inhibitor of metalloproteinase-1 when curcumin post-conditioning towards renal injury induced by limb ischemia/reperfusion in rats.METHODS:Eighty adult male Sprague-Dawley rats were provided by Laboratory Center of China Medical University,and the study was approved by the Laboratory Animal Ethics Committee of Affiliated Central Hospital of Shenyang Medical College.The rats were randomly divided into four groups(n=20/group)using a random number table:sham,ischemia/reperfusion,curcumin post-conditioning and dexamethasone groups.Limb ischemia was induced by occlusion of bilateral femoral arteries of rats except for sham group for 4 hours,followed by reperfusion for 4 hours.At 4 hours after ischemia,200 mg/kg curcumin and 1.5 mg/kg dexamethasone was immediately injected by intraperitoneal pathway into curcumin post-conditioning and dexamethasone groups,respectively.The same volume of normal saline was injected into the rats in the other two groups.Arteries blood samples were taken at 4 hours of reperfusion for examining biochemical indexes such as creatinine and blood urea nitrogen.Kidney was removed to detect the level of malondialdehyde.Hematoxylin-eosin staining was used to detect pathological changes and semi-quantitative score of renal tubulointerstitia.The expression levels of matrix metalloproteinase-9,tissue inhibitor of metalloproteinase-1 and nuclear factor-κB p65 mRNA were detected by western blot assay and RT-PCR,respectively.The levels of tumor necrosis factor-α and interleukin-1β in lung tissue were detected by ELISA.RESULTS AND CONCLUSION:(1)Compared with sham group,the levels of creatinine,blood urea nitrogen and malondialdehyde in the ischemia/reperfusion group were significantly increased(P<0.

关 键 词：姜黄素 缺血再灌注 姜黄素后处理 肾 基质金属蛋白酶9 基质金属蛋白组织抑制因子1 肢体缺血再灌注 肾损伤模型 

分 类 号：R453[医药卫生—治疗学] R318[医药卫生—临床医学]