白藜芦醇对脂多糖诱导的H9c2细胞损伤的保护作用及其机制

Protective effect of resveratrol on lipopolysaccharide-induced damage of H9c2 cells and relevant mechanisms

目的研究白藜芦醇(RSV)对脂多糖(LPS)诱导的H9c2细胞损伤的保护作用及其机制。方法用LPS 10μg/m L处理大鼠H9c2心肌细胞0、5、10、20、30、60 min,评估LPS在不同时间点对H9c2细胞核因子κB抑制蛋白(IκBα)、p-p65、p65和β-肌动蛋白(β-actin)表达的诱导作用;用不同浓度RSV预处理H9c2细胞后再用LPS刺激,观察RSV对LPS诱导的H9c2细胞损伤的影响。MTT法检测H9c2细胞增殖能力;荧光法检测细胞内活性氧(ROS)水平;Western blotting检测IκBα、p-p65、p65和β-actin的表达。结果 LPS抑制H9c2细胞增殖,使细胞内ROS产生量明显增加;LPS使IκBα表达水平显著降低,使p65磷酸化水平显著升高。RSV预处理能明显降低LPS对H9c2细胞的损伤,使细胞内ROS产生明显减少;RSV可上调IκBα表达,下调p-p65的表达。结论 RSV可能通过下调p-p65及上调IκBα抑制LPS诱导的H9c2细胞损伤。

Objective To investigate the protective effect of resveratrol （RSV） on lipopolysaccharide （LPS）- induced damage of H9c2 cells and relevant mechanisms. Methods Mouse myocardial cells H9c2 were treated by LPS of 10 μg/mL for 0, 5, 10, 20, 30, and 60 min. The inductive effect of LPS on the expressions of nuclear factor κB inhibitory protein （IκBα）, p-p65, p65, and β-actin at different time points was evaluated. H9c2 cells were stimulated by LPS after being pre-treated by different concentrations of RSV and the effects of RSV on the LPS-induced damage of H9c2 cells were observed. The proliferation of H9c2 cells was detected by MTT. The intracellular oxygen species reactive （ROS） level was detected by fluorescence method. The expressions of IκBα p-p65, p65, and β-actin were detected by Western blotting. Results LPS inhibited the proliferation of H9c2 cells and the intracellular ROS level significantly elevated. LPS significandy decreased the expression level of IκBα and signilicandy increased the phosphorylation of p65. Pretreatment by RSV remarkably reduced the LPS-induced damage of H9c2 cells and significantly decreased the intracellular ROS level. RSV up- regulated the expression of IκBα and down-regulated the expression of p-p65. Conclusion RSV may inhibit the LPS-induced damage of H9c2 cells via down-regulating the expression of p-p65 and up-regulating the expression of IκBα.