Resveratrol, a polyphenol phytoalexin, protects cardiomyocytes against anoxia/reoxygenation injury via the TLR4/NF-κB signaling pathway

被引:62
作者
Zhang, Cui
Lin, Guosheng [1 ]
Wan, Weiguo
Li, Xuyon
Zeng, Bin
Yang, Bo
Huang, Congxin
机构
[1] Wuhan Univ, Dept Cardiol, Renmin Hosp, Wuhan 430060, Peoples R China
关键词
resveratrol; anoxia/reoxygenation; cardiomyocytes; Toll-like receptor 4; nuclear factor-kappa B; MYOCARDIAL REPERFUSION; CARDIAC MYOCYTES; KAPPA-B; ISCHEMIA/REPERFUSION; ACTIVATION; APOPTOSIS; MECHANISMS; SURVIVAL; HEART; TLR4;
D O I
10.3892/ijmm.2012.885
中图分类号
R-3 [医学研究方法]; R3 [基础医学];
学科分类号
1001 ;
摘要
Previous studies indicate resveratrol pretreatment can protect cardiomyocytes. However, it is largely unknown whether resveratrol protects cardiomyocytes when applied at reperfusion. The purpose of this study was to investigate whether resveratrol given at reoxygenation could protect cardiomyocytes under the anoxia/reoxygenation (A/R) condition and to examine the underlying mechanism. In this study, primary cultures of neonatal rat cardiomyocytes were randomly distributed into three groups: control group, A/R group (cultured cardiomyocytes were subjected to 3 h anoxia followed by 2 h reoxygenation), and the resveratrol group (cardiomyocytes were subjected to 3 h anoxia/2 h reoxygenation, and 5, 10 or 20 mu M resveratrol was applied 5 min after reoxygenation). In order to evaluate cardiomyocyte damage, cell viability, lactate dehydrogenase (LDH) release, caspase-3 activity, and apoptosis were analyzed by the cell counting kit (CCK)-8 assay, colorimetric method and flow cytometry, respectively. The mRNA and protein expression of Toll-like receptor 4 (TLR4) were detected by quantitative real-time PCR and western blot analysis. Nuclear factor-kappa B (NF-kappa B) p65 protein and I-kappa B alpha. protein levels were also examined by western blot analysis. The levels of proinflammatory cytokines in the culture medium were assessed by enzyme-linked immunosorbent assay. We found that resveratrol prevented a reduction in cell viability, decreased the amount of LDH release, attenuated apoptotic cells and decreased caspase-3 activity induced by A/R in cardiomyocytes. Furthermore, resveratrol treatment significantly attenuated the TLR4 expression, inhibited NF-kappa B activation and reduced the levels of tumor necrosis factor (TNF)-alpha and interleukin (IL)-1 beta caused by A/R injury in the culture medium. Treatment with resveratrol shortly after the onset of reoxygenation improves cell survival and attenuates A/R-induced inflammatory response. This protection mechanism is possibly related to the TLR4/NF-kappa B signaling pathway.
引用
收藏
页码:557 / 563
页数:7
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