摘要
目的:脑缺血/再灌注损伤(ischemia/reperfusion,I/R)是缺血性卒中发生后的一种复杂的病理过程。目前,治疗脑I/R损伤的药物疗效尚不理想。我国Ⅰ类新药丁苯酞(dl-3-N-butylphthalide,NBP)被批准用于治疗缺血性脑卒中,有研究报道,NBP具有减轻脑I/R损伤的作用,但其确切机制仍有待进一步研究。本研究从线粒体融合与裂变的角度探讨NBP抗小鼠脑I/R损伤的作用及机制。方法:将ICR小鼠随机分为假手术组(Sham)、I/R组、I/R+NBP组、I/R+NBP+dorsomorphin组。采用线栓法制备小鼠大脑中动脉栓塞(middle cerebral artery occlusion,MCAO)模型。Longa五分法评定小鼠神经功能损伤情况;TTC染色法检测脑梗死体积;透射电镜观察线粒体形态;Western blot检测腺苷酸活化蛋白激酶(AMPK)、磷酸化腺苷酸活化蛋白激酶(p-AMPK)、线粒体融合蛋白2(Mfn2)、线粒体裂变相关蛋白(Drp1,p-Drp1)的表达。结果:与Sham组相比,I/R组小鼠发生线粒体损伤,表现出线粒体裂变的典型特征,小鼠神经行为学评分、脑梗死体积增加(均P<0.01),p-Drp1/Drp1比值增加(P<0.05),p-AMPK/AMPK比值、Mfn2蛋白表达降低(均P<0.01);在I/R组的基础上使用NBP后,小鼠线粒体损伤有所改善,小鼠神经行为学评分、脑梗死体积降低(均P<0.01),p-Drp1/Drp1比值降低(P<0.01),p-AMPK/AMPK比值和Mfn2蛋白表达增加(均P<0.01),而dorsomorphin(AMPK抑制剂)可明显降低NBP的上述保护作用。结论:NBP具有减轻小鼠脑I/R损伤的作用,其作用机制可能与通过AMPK调节线粒体融合与裂变有关。
Objective:Cerebral ischemia/reperfusion(I/R)injury is a complex pathophysiological process which occurs during ischemic stroke.At present,the efficacy of currently available drugs in the treatment of cerebral I/R injury is not ideal.Dl-3-N-butylphthalide(NBP),a new class I drug in China,has been approved for the treatment of ischemic stroke.It has been reported that NBP can reduce cerebral I/R damage,but its exact mechanism remains to be further studied.This study investigated the effect and mechanism of NBP on cerebral I/R damage in mice from mitochondrial fusion and fission.Methods:Male ICR mice were randomly divided into four groups,i.e.,Sham operation group(sham group),cerebral ischemia/reperfusion group(I/R group),I/R+NBP group,I/R+NBP+dorsomorphin group.Mice models of cerebral I/R injury were established by middle cerebral artery occlusion/reperfusion(MCAO/R),and the neurobehavioral deficit was evaluated by Longa score.The infract volume was detected by TTC staining.Mitochondrial morphology of brain cells was assessed by transmission electron microscopy(TEM).And the expression levels of AMP-activated protein kinase(AMPK),p-AMPK,mitochondrial fusion protein 2(Mfn2),mitochondrial fission protein(Drp1 and p-Drp1)were determined by western blot.Results:Compared with sham group,the mice in I/R group showed mitochondrial damage and typical changes of mitochondrial fission,with increased neurological dysfunction,infarct volume and neurobehavioral score(all P<0.01),the ratio of p-Drp1/Drp1(P<0.05),decreased ratio of p-AMPK/AMPK(P<0.01)and decreased the expression of Mfn2 protein(P<0.01).Moreover,based on I/R group,NBP reduced the damage of mitochondrial,decreased the neurobehavioral score and infarct volume(all P<0.01),decreased the radio of p-Drp1/Drp1(P<0.01),increased the radio of p-AMPK/AMPK(P<0.01),and increased the expression of Mfn2 protein(P<0.01).However,these protect effects were attenuated by dorsomorphin(an inhibitor of AMPK).Conclusion:NBP has protective effects against cerebral I/R injury in mice and the underlying mechanisms may be related to regulation of mitochondrial fusion and fission through AMPK signaling pathway.
作者
陈蕾
王雪如
黄杰
胡淼
童旭辉
董淑英
CHEN Lei;WANG Xue-ru;HUANG Jie;HU Miao;TONG Xu-hui;DONG Shu-ying(Department of Pharmacology,School of Pharmacy,Bengbu Medical College,Bengbu 233030,China;Anhui Engineering Technology Research Center of Biochemical Pharmaceutical,Bengbu 233030,China;Basic and Clinical Key Laboratory of Cardiovascular and Cerebrovascular Diseases,Bengbu Medical College,Bengbu 233030,China)
出处
《中国新药杂志》
CAS
CSCD
北大核心
2022年第19期1929-1935,共7页
Chinese Journal of New Drugs
基金
国家自然科学基金资助项目(81402930)
蚌埠医学院512人才培养计划项目(BY51201104)
蚌埠医学院研究生科研创新训练项目(Byycx21030)。
关键词
脑缺血/再灌注
丁苯酞
线粒体融合与裂变
腺苷酸活化蛋白激酶
cerebral ischemia/reperfusion
dl-3-N-butylphthalide
mitochondrial fusion and fission
AMP-activated protein kinase
