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

Abstract

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 μM 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-κB (NF-κB) p65 protein and I-κBα 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-κB activation and reduced the levels of tumor necrosis factor (TNF)-α and interleukin (IL)-1β 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-κB signaling pathway.

Resveratrol Attenuates Ischemia/Reperfusion Injury in Neonatal Cardiomyocytes and Its Underlying Mechanism

“…Previous studies have suggested that Res has a cardioprotective effect [11], [23]. Our present study focused on the underlying mechanisms of the effect, suggesting that several pathways are involved in the process, such as anti-apoptosis and anti-oxygen free radical.…”

“…Res has extensive pharmacological effects, including anticancer [12]–[15], modulating inflammation response [16]–[18], improving ifosfamide-induced Fanconi syndrome in rats [19], treating diabetic nephropathy [20] and protecting neurons [21], [22]. Moreover, there have been additional studies implicating the cardioprotective role of Res [23]. However, the underlying mechanism involved in the protective role of Res is still not completely clear.…”

Resveratrol Regulates Activated Hepatic Stellate Cells by Modulating NF‐κB and the PI3K/Akt Signaling Pathway

“…And resveratrol contents in peanut varieties are up to 1.92 μg/g (Tokuşoglu and others ). Preclinical and clinical studies have demonstrated that resveratrol possesses several pharmacological effects including anticancer, antioxidant, anti‐inflammatory, and antiasthmatic activity, and has an effective role in glaucoma, pancreatitis, and osteoarthritis (Lyons and others ; Bishayee and others ; Al‐Dissi and Weber ; Zhang and others ; Luo and others ; Wang and others ). In addition, resveratrol administration reduces portal pressure, HSC activation, and hepatic fibrosis in vivo (Di Pascoli and others ).…”

Resveratrol attenuates hypoxia/reoxygenation-induced Ca2+ overload by inhibiting the Wnt5a/Frizzled-2 pathway in rat H9c2 cells

“…Similarly, resveratrol treatment also decreased the LDH levels in H/R cardiomyocytes. Therefore, as previous studies have demonstrated, it was proven that resveratrol exerted a protective effect on H/R cardiomyocytes (18). It is believed that the cardioprotection induced by resveratrol is achieved through a preconditioning effect; however, it is difficult to restrict its administration as a pretreatment (24,25).…”

“…The absorbance of each well at 450 nm related to the reference absorbance at 630 nm was measured using a microplate reader (Bio-Rad, Hercules, CA, USA). The cell viability percentage was calculated using the following formula: % Cell viability = (mean absorbance in the test wells)/(mean absorbance in the control well) x 100 (18).…”