n-6和n-3多不饱和脂肪酸对心肌缺血/再灌注损伤炎症因子TNF-α和IL-1释放的影响

Effects of n-6 and n-3 polyunsaturated fatty acids on the release of inflammatory cytokines TNF-α and IL-1 after myocardial ischemia-reperfusion injury

目的探讨n-6和n-3多不饱和脂肪酸对心肌缺血/再灌注损伤炎症因子TNF-α和IL-1释放的抑制作用及其抗炎作用机制。方法构建健康成年SD大鼠4组模型(sham、IR、n-3PUFA+IR和n-6PUFA+IR),通过肌酸激酶(CK)、肌酸激酶同工酶(CK-MB)和乳酸脱氢酶(LDH)活性检测缺血/再灌注的损伤程度;通过酶联免疫吸附测定促炎细胞因子TNF-α、IL-1的表达水平;通过qPCR和Westernblot检测TLR4/NF-κB的表达水平。结果与IR组相比,n-3PUFA+IR和n-6PUFA+IR组的CK、CK-MB和LDH显著降低,心肌梗死面积减少,心肌组织形态学结构改善;ELISA检测发现n-6和n-3PUFAs处理组,TNF-α、IL-1的表达水平显著降低;TLR4、NF-κB的mRNA和蛋白的表达也显著降低。结论n-3PUFA和n-6PUFA通过TLR4/NF-κB途径抑制促炎因子TNF-α和IL-1的表达,从而保护心肌缺血/再灌注损伤,TLR4介导的NF-κB信号传导途径可能成为心肌缺血/再灌注损伤的新策略。

Objective To investigate the inhibitory effects of n-6 and n-3 polyunsaturated fatty acids on the release of inflammatory cytokines TNF-αand IL-1 in myocardial ischemia/reperfusion injury and its anti-inflammatory mechanism.Methods Four sets of models of healthy adult SD rats(sham,IR,n-3 PUFA+IR,and n-6 PUFA+IR)were constructed.The degree of ischemia-reperfusion injury was measured by activity of creatine kinase(CK),creatine kinase isoenzyme(CK-MB)and lactate dehydrogenase(LDH);The expression levels of pro-inflammatory cytokines TNF-αand IL-1 were determined by enzyme-linked immunosorbent assay.The expression levels of TLR4/NF-κB were detected by qPCR and Western blot.Results Compared with the IR group,CK,CK-MB and LDH were significantly lower in the n-3 PUFA+IR and n-6 PUFA+IR groups,the myocardial infarct size was reduced,and the myocardial histomorphology was improved.ELISA showed that the expression levels of TNF-αand IL-1 in n-6 and n-3 PUFAs groups were significantly decreased;the mRNA and protein expression of TLR4 and NF-κB were also significantly decreased.Conclusion n-3 PUFA and n-6 PUFA inhibit the expression of pro-inflammatory factors TNF-αand IL-1 through the TLR4/NF-κB pathway,thereby protecting myocardial ischemia/reperfusion injury.TLR4-mediated NF-κB signaling pathway may be a new strategy for myocardial ischemia/reperfusion injury.