川芎嗪对脑缺血再灌注小鼠脑损伤及NADPH氧化酶表达的影响

TMP Improves Oxidative Stress and Inflammatory Injury After Transient Global Brain Ischemia by Inhibition of NADPH Oxidase and Myeloperoxidase in Mice

目的研究川芎嗪对脑缺血再灌注小鼠大脑氧化应激损伤的保护作用和还原型烟酰胺腺嘌呤二核苷酸磷酸(nicotinamide adenine dinucleotide phosphate,NADPH)氧化酶表达的影响。方法采用线栓法建立大脑中动脉闭塞/再灌注模型。造模后对小鼠进行Longa神经功能评分;采用HE和氯化三苯基四氮唑染色法(triphenyltetrazolium chloride staining,TTC)分别观察脑组织的病理改变和梗死情况;检测脑组织中超氧化物歧化酶(total antioxidative capacity,T-AOC)、过氧化物酶(superoxide dismutase,SOD)、乳酸脱氢酶(malondialdehyde,MDA)、过氧化氢(hydrogen peroxide,H_(2)O_(2))和髓过氧化物酶(myeloperoxidase,MPO)的变化,Western blot检测NADPH氧化酶活化蛋白p47抗体(NADPH oxidase activated protein p47 antibody,P47phox)和NADPH氧化酶(NADPH oxidase,Nox)4蛋白的表达,免疫组化检测肿瘤坏死因子(tumor necrosis factor,TNF)-α的表达。结果川芎嗪能显著减轻脑缺血再灌注小鼠的脑组织损伤、缩小脑梗死体积,升高脑缺血再灌注小鼠脑组织中T-AOC、SOD水平而降低MDA、H_(2)O_(2)和MPO的含量(P<0.05),同时降低其脑组织中P47phox、Nox4和TNF-α的蛋白表达(P<0.05)。结论川芎嗪对脑缺血再灌注小鼠大脑损伤具有明显的保护作用,其机制与降低NADPH氧化酶的表达进而抑制氧化应激及减轻炎症反应有关。

Objective To investigate the effects of TMP on oxidative stress injury and NADPH oxidase expression in transient global brain ischemia mice.Methods The model of middle cerebral artery occlusion/reperfusion was established by means of thread embolism.Longa nerve function was evaluated after modeling.HE and triphenyltetrazolium chloride staining(TTC)were used to observe the pathological changes and infarction of brain tissue.The changes of total antioxidative capacity(T-AOC),superoxide dismutase(SOD),malondialdehyde(MDA),hydrogen peroxide(H_(2)O_(2)),and myeloperoxidase(MPO)in brain tissue were detected.The expression of NADPH oxidase activated P47phox,NADPH Nox4 proteins and TNF-αwere detected by Western blot.Results TMP can significantly reduce the brain tissue injury of cerebral ischemia-reperfusion mice,reduce the volume of cerebral infarction,increase the levels of T-AOC and SOD,and decrease the contents of MDA,H_(2)O_(2)and MPO(P<0.05).Meanwhile,TMP can reduce the expression of P47phox,Nox4 and TNF-αin cerebral tissue(P<0.05).Conclusion TMP has obvious protective effect on brain injury in cerebral ischemia-reperfusion mice,and the mechanism is related to reducing the expression of NADPH oxidase,inhibiting oxidative stress and alleviating inflammation.