还原型辅酶Ⅰ和还原型辅酶Ⅱ治疗缺血性脑卒中与抗氧化作用的比较
首发时间:2020-03-27
摘要:目的:研究还原型烟酰胺腺嘌呤二核苷酸(NADH)和还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)治疗缺血性脑卒中的疗效差异,并对其发挥神经保护作用的机制进行探讨。方法:采用体内小鼠短暂性大脑中动脉阻塞再灌注(t-MCAO/R)模型和体外小鼠海马神经元缺糖缺氧再复糖复氧(OGD/R)模型。在体内实验中,以缺血再灌注24h后小鼠脑梗死体积等指征进行NADH、NADPH的药效和时间窗研究。通过缺血再灌注28天后小鼠的生存率、脑萎缩程度和行为学测试评价各给药组缺血脑卒中小鼠的长期病理学变化。Western blot检测各给药组小鼠缺血皮层氧化损伤相关蛋白水平的变化。用ATP、MDA检测试剂盒检测小鼠缺血皮层组织的氧化应激和能量代谢水平。体外实验中,通过CCK-8法检测NADH、NADPH对OGO/R诱导神经元死亡的保护作用。用DCFH2-DA探针标记检测它们对活性氧水平的影响。结果:本实验成功建立了小鼠缺血性脑卒中的体内和体外模型。药效研究结果显示NADH、NADPH均可降低缺血性脑卒中小鼠的脑梗死体积,改善小鼠的神经行为学评分和降低脑水肿含量,其中NADPH的治疗效果优于NADH。NADPH治疗缺血性脑卒中小鼠在再灌注5小时内均有效,NADH治疗窗较窄,仅在再灌后2小时内有效。在缺血脑卒中小鼠的长期病理学观察中,无论是脑萎缩还是神经功能恢复,NADPH效果均优于NADH。在抗氧化能力和能量代谢上,NADPH抗氧化效果优于NADH,而在能量代谢方面,NADH效果明显优于NADPH。结论:NADH和NADPH均对小鼠脑缺血再灌注损伤具有神经保护作用。NADH对脑缺血小鼠的保护作用是通过对能量代谢的调节产生的。NADPH主要通过抗氧化应激抑制ROS从而抑制神经细胞的死亡。综合来说,NADPH提供了更好的保护作用。
关键词: 药理学 NADH NADPH 缺血性脑卒中 氧化应激 能量代谢
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Comparison of NADH and NADPH in the treatment of ischemic stroke and antioxidation
Abstract:Aim: To investigate the difference between NADH and NADPH in the treatment of ischemic stroke and explore the mechanism of their neuroprotective effect.Methods: tMCAO/Rwas produced by using an intraluminal filament technique in mice; in vitro the OGD/R model was established by deprivation of oxygen and glucose and reoxygenation. In vivo experiments, the efficacy and time window of NADH and NADPH were studied with indicators of cerebral infarct volume in mice after 24h of ischemia-reperfusion. The long-term pathological changes of ischemic stroke mice were evaluated by survival rate, the degree of brain atrophy and behavioral tests after 28 days of ischemia-reperfusion.Western blot was used to detect the changes of protein levels related to oxidative damage in ischemic cortex of mice. The ATP, MDA kits were used to detect the changes of oxidative stress and energy metabolism level in the ischemic cortex of mice. The protective effect of NADH and NADPH on OGO/R-induced neuronal death was detected by the CCK-8 method. DCFH2-DA probe labeling was used to detect their effect on active oxygen levels.Results: In this study, we have successfully established in vivo and in vitro models of stroke in mice. The results showed that NADH and NADPH can reduce the infarct volume of mice with ischemic stroke and improve the neurological deficient and water content, where NADPH is superior to NADH. NADPH had a longer therapeutic window for ischemic stroke mice than NADH. In the long-term pathological observation of ischemic stroke mice, NADPH is better than NADH, whether it is cerebral atrophy or nerve function recovery. NADPH had better antioxidant effect than NADH, and in terms of energy metabolism, NADH was better than NADPH. Conclusion: NADH and NADPH have protective effects on ischemia-reperfusion injury in mice. The protective effect of NADH on cerebral ischemia mice is produced by regulating energy metabolism while NADPH mainly inhibits ROS through antioxidant stress. Overall, NADPH provides better protection.
Keywords: Pharmacology NADH NADPH Ischemic stroke Oxidative stress Energy metabolism
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还原型辅酶Ⅰ和还原型辅酶Ⅱ治疗缺血性脑卒中与抗氧化作用的比较
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