姜黄素对小鼠肾脏再灌注损伤的保护作用及其机制研究  

Curcumin attenuates renal ischemia reperfusion injury in mice by inhibiting the activation of toll-like receptor 4 signaling

作　　者：吴迪英[1] 郭闻渊[2] 张豪杰[1] 蔡旭东[1] 徐鹏杰[1] 

机构地区：[1]宁波大学附属宁波市李惠利医院肾内科,浙江宁波315040 [2]第二军医大学长征医院器官移植科,上海200003

出　　处：《解放军医学杂志》2012年第7期671-675,共5页Medical Journal of Chinese People's Liberation Army

基　　金：上海市科委医学引导项目(114119a7900)~~

摘　　要：目的探讨姜黄素对小鼠肾脏缺血再灌注(IR)损伤的保护作用及其可能机制。方法选择健康雄性C57BL/6小鼠20只,随机均分为4组。假手术(SO)组小鼠仅接受腹中线开腹、游离双侧肾蒂及关腹操作;对照组小鼠制成肾脏IR模型,并于肾脏缺血同时经尾静脉注射生理盐水;姜黄素低剂量(CM-L)及高剂量(CM-H)组小鼠建立肾脏IR模型,并经尾静脉分别注射5mg/kg及20mg/kg姜黄素。再灌注6h后检测各组血清肌酐(Scr)、尿素氮(BUN)水平,观察肾脏组织形态学变化;Western blotting检测肾脏组织内Toll样受体4(TLR-4)、高迁移率族蛋白B1(HMGB1)、透明质烷(HA)蛋白的表达,实时荧光定量RT-PCR检测透明质烷合成酶(HAS)1、HAS2、HAS3以及白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)mRNA的表达;免疫组化检测肾组织内巨噬细胞浸润情况;同时采用ELISA法检测血清IL-6、TNF-α水平。结果与对照组相比,CM-L及CM-H组小鼠再灌注6h血清Scr、BUN水平明显降低(Scr:对照组235.4±28.7μmol/L,CM-L组167.8±17.2μmol/L,CM-H组125.8±13.3μmol/L;BUN:对照组21.6±2.7mmol/L,CM-L组18.1±2.4mmol/L,CM-H组12.5±1.7mmol/L;P<0.05或P<0.01),肾小管上皮细胞损伤情况明显改善(Jablonski评分:对照组2.15±0.28分,CM-L组1.72±0.21分,CM-H组1.42±0.15分,P<0.01);肾脏组织内TLR-4、HMGB1的mRNA及蛋白水平,HA、HAS1、HAS2、HAS3以及IL-6、TNF-αmRNA水平均明显降低(P<0.05或P<0.01);巨噬细胞浸润减轻(对照组32.4±4.2个/高倍视野,CM-L组24.8±3.6个/高倍视野,CM-H组18.9±3.1个/高倍视野,P<0.01);血清IL-6、TNF-α含量亦显著降低(IL-6:对照组1411.2±150.6pg/ml,CM-L组918.7±113.4pg/ml,CM-H组809.6±108.3pg/ml;TNF-α:对照组154.7±21.1pg/ml,CM-L组135.8±15.2pg/ml,CM-H组125.6±14.6pg/ml;P<0.05或P<0.01)。结论姜黄素对小鼠肾脏IR损伤具有明显的保护作用,其机制可能与抑制肾脏再灌注后TLR-4信号活化有关。Objective To investigate the protective effect of curcumin on renal ischemia reperfusion（IR） injury and its related mechanism in mice.Methods C57BL/6 mice were randomly divided into 4 groups： sham-operated（SO） group,low-dose and high-dose curcumin（CM-L and CM-H） groups（the mice were injected with 5mg/kg or 20mg/kg curcumin just before ischemia induction.） and control group（the mice were injected with equal volume of physiological saline at the same time.）.At a 6-hour reperfusion,the serum and renal samples were collected.The serum creatinine（Scr） and blood urea nitrogen（BUN） levels in serum were determined and renal patho-histological changes were also examined.Expressions of Toll-like receptor 4（TLR-4）,high mobility group protein B1（HMGB1）,hyaluronan（HA）,hyaluronan synthase（HAS） 1,HAS2,HAS3 and interleukin-6（IL-6）,tumor necrosis factor-α（TNF-α） and TLR-4 protein were assessed using quantitative fluorescence RT-PCR and Western blotting.The infiltration of F4/80-positive macrophage cells was detected using immunohistochemical analysis and the IL-6 and TNF-α levels in serum were determined by enzyme linked immunosorbent assay（ELISA）.Results Compared with control group,CM-L and CM-H group exhibited lower levels of Scr and BUN（Scr： Control 235.4±28.7,CM-L 167.8±17.2,CM-H 125.8±13.3μmol/L;BUN： Control 21.6±2.7,CM-L 18.1±2.4,CM-H 12.5±1.7mmol/L;P0.05 or P0.01）,less pathological changes（Jablonski scores： Control 2.15±0.28,CM-L 1.72±0.21,CM-H 1.42±0.15;P0.01）,lower mRNA levels of TLR-4,HMGB1,HA,HAS1,HAS2,HAS3,IL-6 and TNF-α,lower levels of TLR-4 protein and HMGB1,less infiltration of macrophage [Control 32.4±4.2/high power field（HPF）,CM-L 24.8±3.6/HPF,CM-H 18.9±3.1/HPF;P0.01].Moreover,the serum levels of IL-6 and TNF-α were lower in CM-L and CM-H groups than those in control group（IL-6： Control 1411.2±150.6,CM-L 918.7±113.4,CM-H 809.6±108.3pg/ml;TNF-α： Control 154.7±21.1,CM-L 135.8±15.2,CM-H 125.6±14.6pg/ml;P0.05 or

关 键 词：姜黄素 再灌注损伤 TOLL样受体4 高迁移率族蛋白B 透明质烷 

分 类 号：R692.9[医药卫生—泌尿科学]