基于NF-κB通路研究槲皮素对心肌缺血再灌注损伤大鼠的影响

The Effect of Quercetin on Myocardial Ischemia-Reperfusion Injury in Rats Based on NF-κB Pathway

目的:探讨槲皮素对心肌缺血再灌注损伤(myocardial ischemia-reperfusion injury,MIRI)大鼠的保护作用及其对核转录因子kappa B(nuclear factor of kappa B,NF-kB)通路的影响。方法:将SD雄性大鼠按体质量随机均分为5组,即假手术组、模型组及槲皮素低、中、高剂量(25 mg·kg^(-1)、50 mg·kg^(-1)、100 mg·kg^(-1))组。除假手术组,其余各组均采用结扎大鼠左冠前降支的方法建立MIRI模型,连续给予相应药物7 d。于末次给药12 h后,采用超声心动仪检测大鼠左心功能;HE染色观察大鼠心肌组织病理学;TTC染色观察心肌梗死面积变化;检测血清乳酸脱氢酶(lactate dehydrogenase,LDH)、肌酸激酶同工酶MB(creatine Kinase-MB,CK-MB)、白细胞介素-1β(interleukin-1β,IL-1β)、IL-6、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)水平,同时检测心肌组织中丙二醛(malondialdehyde,MDA)、超化物歧化酶(superoxide dismutase,SOD)及Na^(+)-K^(+)-ATP酶、Ca^(2+)-ATP酶活力;Western Blot法检测大鼠心肌组织κB抑制蛋白α(inhibitor of NF-κBα,IκBα)、κB抑制蛋白激酶(inhibitor of nuclear factorκB kinase,IKK)和NF-κB p65蛋白及其磷酸化蛋白表达。结果:槲皮素低、中、高剂量组大鼠左室射血分数(left ventricular ejection fraction,LVEF)、左室短轴缩短率(left ventricular fractional shortening,LVFS)水平、心肌梗死面积及心肌组织Na^(+)-K^(+)-ATP酶、Ca^(2+)-ATP酶、SOD酶活力及p-IκBα、p-IKK、p-NF-κB p65蛋白表达显著增加(P<0.05,P<0.01),血清中TNF-α、IL-6、LDH、CK-MB水平显著降低(P<0.05),同时可改善心肌组织损伤。结论:槲皮素可显著减轻MIRI引起的心肌组织损伤,改善心脏功能,抑制心肌组织炎症反应和氧化应激反应,其作用机制可能与增强超微量ATP酶活力及抑制NF-κB通路有关。

Objective:To investigate the protective effect of quercetin on myocardial ischemia-reperfusion injury(MIRI)rats and its effect on the nuclear factor kappa B(NF-κB)pathway.Methods:The SD male rats were randomly divided into five groups according to body mass,namely sham operation group,model group and quercetin low,medium and high dose(25 mg·kg^(-1),50 mg·kg^(-1),100 mg·kg^(-1))group.In addition to the sham operation group,the other groups were ligated the left anterior descending branch of the crown to establish MIRI model.The corresponding drugs were given for 7 days.12 hours after the last administration,the left ventricular function was detected by echocardiography;HE staining was used to observe myocardial histopathology;TTC staining was used to observe the change of infarct size;serum lactate dehydrogenase(LDH),creatine Kinase-MB(CK-MB)and interleukin-1β(IL-1β),IL-6,tumor necrosis factor-α(TNF-α)level were detected;meanwhile,the activities of malondialdehyde(MDA),superoxide dismutase(SOD),Na^(+)-K^(+)-ATPase and Ca^(2+)-ATPase were detected;Western Blot method was used to detect the expression of inhibitor of NF-κBα(IκBα),inhibitor of nuclear factorκB kinase(IKK),NF-κB p65 protein and its phosphorylated protein in rat myocardial tissue.Results:Left ventricular ejection fraction(LVEF),left ventricular fractional shortening(LVFS)levels,myocardial infarction area and myocardial tissue Na^(+)-K^(+)-ATPase,Ca^(2+)-ATPase,SOD enzyme activity and p-IκBα,p-IKK,p-NF-κB p65 protein expression increased significantly(P<0.05;P<0.01).Serum levels of TNF-α,IL-6,LDH,and CK-MB were significantly reduced(P<0.05),and at the same time,it could improve myocardial tissue damage.Conclusion:Quercetin can significantly reduce myocardial tissue damage caused by MIRI,improve cardiac function,and inhibit myocardial tissue inflammation and oxidative stress.Its mechanism of action may be related to enhancing the activity of ultra-micro ATPase and inhibiting the NF-κB pathway.