姜黄素对小鼠心肌缺血再灌注损伤的保护作用及其与Toll样受体4/核因子-κB信号通路的相关性

Protecitve effect of curcumin onf myocardial ischemia-reperfusion injury in mice as well as its relationship to Toll-like receptor 4/nuclear factor kappa-light-chain-enhance of activated B cell signaling pathway

目的探讨姜黄素对小鼠心肌缺血再灌注损伤(myocardial ischemia-reperfusion injury,MIRI)的保护作用及其与Toll样受体4(Toll-like receptor 4,TLR4)/核因子-κB(nuclear factor kappa-light-chain-enhancer of activated B cells,NF-κB)信号通路的相关性。方法取健康C57BL/6小鼠,采用随机区组法分为假手术组(Sham组)、MIRI组及姜黄素预处理的MIRI组(Cur组):使用丝线暂时阻断小鼠冠状动脉前降支(left anterior descending,LAD)血流30 min,再灌注4 h,建立小鼠MIRI模型;假手术组接受相同的手术过程,但未阻断LAD;Cur组在缺血前30 min腹腔注射姜黄素(100 mg/kg)。采用伊文思蓝和氯化三苯基四氮唑(2,3,5-triphenyltetrazolium chloride,TTC)染色评估小鼠心肌梗死面积;电化学发光法检测小鼠血清肌钙蛋白(cardiac troponin T,cTnT)水平;RT-PCR法检测小鼠心肌组织TLR4及肿瘤坏死因子-α(tumor nectosis factor-alpha,TNF-α,)mRNA水平;Western blot法检测小鼠心肌组织NF-κB蛋白表达水平;ELISA法检测小鼠心肌组织TNF-α蛋白表达水平。结果 Cur组小鼠心肌梗死面积及血清cTnT均低于MIRI组(P均<0.01);Cur组小鼠心肌组织TLR4、TNF-αm RNA水平及NF-κB、TNF-α蛋白表达水平均低于MIRI组(P均<0.05)。结论姜黄素对小鼠MIRI的心肌具有保护作用,可能是通过TLR4/NF-κB信号通路实现的。

Objective To investigate the protective effect of curcumin（Cur） on the mice with myocardial ischemiareperfusion injury（MIRI）as well as its relationship to the Toll-like receptor 4 / nuclear factor kappa-light-chain-enhancer of activated B cell（TLR4 / NF-κB） signaling pathway. Methods Healthy C57BL / 6 mice were randomly divided into sham operation（Sham）, MIRI and Cur groups. The left anterior descending（LAD） of mice was ligated with a thread for 30 min, then subjected to reperfusion for 4 h to establish a model of MIRI. The mice in Sham group received the same operation process as that in MIRI group, while the LAD was not ligated. However, the mice in Cur group were injected i.p.with Cur at a dosage of 100 mg / kg 30 min before MIRI. The area of myocardial infarction（MI）was measured by Evans blue and 2, 3, 5-triphenyltetrazolium chloride（TTC） staining. The serum cardiac troponin T（c Tn T） level of mice was determined by electrochemiluminescence assay. The transcription levels of TLR4 and tumor necrosis factor-alpha（TNF-α）m RNAs in myocardial tissue of mice were determined by RT-PCR, while the expression levels of NF-κB and TNF-αproteins by Western blot and ELISA respectively. Results The area of MI of mice in Cur group was smaller, while the serum c Tn T level was lower, than those in MIRI group（both P 〈 0. 01）. However, the transcription levels of TLR4 and TNF-α m RNAs as well as expression levels of NF-κB and TNF-α proteins in Cur group were significantly lower than those in MIRI group（each P 〈0. 05）. Conclusion Cur showed protective effect on the myocardial tissue of mice with MIRI by a possible TLR4 / NF-κB signaling pathway.