陈运转;吕爱红;王军杰;辛凯;赵曼;

• 1:郑州人民医院
• 2:河南中医药大学第三附属医院

摘要(Abstract)：

目的通过构建大鼠缺血再灌注损伤模型,观察再灌注后自噬现象的发生,探讨自噬在脑缺血再灌注损伤中的作用。方法选用健康Sprague-Dawley(SD)大鼠(n=108),随机分为3组各36只:(1)假手术组;(2)对照组;(3)三-甲基腺嘌呤(3-MA)组,然后根据再灌注时间(6 h、12 h和24 h)将各组随机分为3个亚组各12只。根据神经行为学评分、TTC染色、脑组织含水量、透射电子显微镜几方面结果进行统计学分析。结果假手术组无症状,TTC染色未见梗死灶,脑组织含水量最少,电镜下海马神经细胞细胞器正常,无自噬现象。对照组症状较3-MA组减轻(P<0.05),梗死灶占比较3-MA组降低(P<0.05),脑组织含水量较3-MA组减少(P<0.05),细胞器破坏较3-MA组减轻,自噬溶酶体较3-MA组增多。结论缺血再灌注可致神经细胞损伤,细胞出现自噬现象,表现为大鼠神经功能缺失,缺血侧脑组织见梗死灶,梗死灶内海马神经细胞受损,细胞内自噬溶酶体显著增多。在脑缺血再灌注损伤中,抑制细胞自噬可致神经元损伤加重,自噬的激活可能对神经细胞发挥保护作用。

关键词(KeyWords)： 自噬;缺血再灌注;3-甲基腺嘌呤;神经细胞;Sprague-Dawley大鼠

Abstract:

Keywords:

基金项目(Foundation): 河南省中医药科学研究专项课题(编号:2018ZY2057)

作者(Author): 陈运转;吕爱红;王军杰;辛凯;赵曼;

Email:

DOI:

参考文献(References)：

• [1] LIU C Y,CHEN C Q.Intra-and extracranial atherosclerotic stenosis in China:epidemiology,diagnosis,treatment and risk factors[J].Eur RMed Pharmaco Sci,2014,18(22):3368-3379.
• [2] BROUNS R,DE DEYN P P.The complexity of neurobiological processes in acute ischemic stroke[J].Clin Neurol Neurosurg,2009,11(6):483-495.DOI:10.1016/j.clineuro.2009.04.001.
• [3] KANG Y,SUN Y,LI T,et al.Garcinol protects against cerebral ischemia-reperfusion injury in vivo and in vitro by inhibiting inflammation and oxidative stress[J].Mol Cell Probes,2020,10:101672.DOI:10.1016/j.mcp.2020.101672.
• [4] YANG Y,HU F,YANG G,et al.Lack of sphingomyelin synthase 2 reduces cerebral ischemia/reperfusion injury by inhibiting microglial inflammation in mice[J].Exp Ther Med,2020,20(6):241.DOI:10.3892/etm.2020.9371.
• [5] LIU D,GU Y,WANG W,et al.Astragalin alleviates ischemia/reperfusioninduced brain injury via suppression of endoplasmic reticulum stress[J].Mol Med Rep,2020,22(5):4070-4078.DOI:10.3892/mmr.2020.11448.
• [6] MIZUSHIMA N,KLIONSKY D J.Protein turnover via autophagy:implications for metabolism[J].Annu Rev Nutr,2007,27:19-40.DOI:10.1146/annurev.nutr.27.061406.093749.
• [7] RYTER S W,BHATIA D,CHOI M E.Autophagy:A Lysosome-Dependent Process with Implications in Cellular Redox Homeostasis and Human Disease[J].Antioxid Redox Signal,2019,30(1):138-159.DOI:10.1089/ars.2018.7518.
• [8] HUANG Y G,TAO W,YANG S B,et al.Autophagy:novel insights into therapeutic target of electroacupuncture against cerebral ischemia/ reperfusion injury[J].Neural Regen Res,2019,14(6):954-961.DOI:10.4103/1673-5374.250569.
• [9] LONGA E Z,WEINSTEIN P R,CARLSON S,et al.Reversible middle cerebral artery occlusion without craniectomy in rats[J].Stroke,1989,20(1):84-91.DOI:10.1161/01.str.20.1.84.
• [10] MELONI B P,MEADE A J,KITIKOMOLSUK D,et al.Characterisation of neuronalcell death in acute and delayed in vitro ischemia (oxygen-glucose deprivation) models[J].Neurosci Methods,2011,195(1):67-74.
• [11] DEGTEREV A,HUANG Z H,BOYCE M,et al.Chemical inhibitor of nonapoptotic cell death with therapeutic potential for ischemic brain injury[J].Nat Chem Biol,2005,1(2):112-119.DOI:10.1038/nchembio711.
• [12] HJELDE A,HJELSTUEN M,HARALDSETH O,et al.Hyperbaric oxygen and neutrophil accumulation/tissue damage during permanent focal cerebral ischaemia in rats[J].Eur J Appl Physiol,2002,86(5):401-405.DOI:10.1007/s00421-001-0573-1.
• [13] ALIREZAEI M,KEMBALL C C,WHITTON J L.Autophagy,inflammation and neurodegenerative disease[J].Eur J Neurosci,2011,33(2):197-204.DOI:10.1111/j.1460-9568.2010.07500.x.
• [14] DORIA A,GATTO M,PUNZI L.Autophagy in human health and disease[J].N Engl J Med,2013,368(19):1845.DOI:10.1056/NEJMc1303158.
• [15] CLARKE A J,SIMON A K.Autophagy in the renewal,differentiation and homeostasis of immune cells[J].Nat Rev Immunol,2019,19(3):170-183.DOI:10.1038/s41577-018-0095-2.
• [16] HUANG X,DING J,LI Y,et al.Exosomes derived from PEDF modified adipose-derived mesenchymal stem cells ameliorate cerebral ischemia-reperfusion injury by regulation of autophagy and apoptosis[J].Exp Cell Res,2018,371(1):269-277.DOI:10.1016/j.yexcr.2018.08.021.
• [17] TAO J,SHEN C,SUN Y,et al.Neuroprotective effects of pinocembrin on ischemia/reperfusion-induced brain injury by inhibiting autophagy[J].Biomed Pharmacother,2018(10),106:1003-1010.DOI:10.1016/j.biopha.2018.07.026.
• [18] WANG P,SHAO B Z,DENG Z,et al.Autophagy in ischemic stroke[J].Prog Neurobiol,2018,163-164:98-117.DOI:10.1016/j.pneurobio.2018.01.001.
• [19] LEVINE B,PACKER M,CODOGNO P.Development of autophagy inducers in clinical medicine[J].J Clin Invest,2015,125(1):14-24.DOI:10.1172/JCI73938.
• [20] WEI K,WANG P,MIAO C Y.A double-edged sword with therapeutic potential:an updated role of autophagy in ischemic cerebral injury[J].CNS Neurosci Ther,2012,18(11):879-886.DOI:10.1111/cns.12005.
• [21] ZHU J,WANG L,ZHANG J.Galuteolin Inhibited Autophagy for Neuroprotection Against Transient Focal Cerebral Ischemia in Rats[J].Neuromolecular Med,2020 Oct 21.DOI:10.1007/s12017-020-08606-2.
• [22] YU S,YU M,BU Z.FKBP5 Exacerbates Impairments in Cerebral Ischemic Stroke by Inducing Autophagy via the AKT/FOXO3 Pathway[J].Front Cell Neurosci,2020,14:193.DOI:10.3389/fncel.2020.00193.
• [23] SUN X,WANG D,ZHANG T,et al.Eugenol Attenuates Cerebral Ischemia-Reperfusion Injury by Enhancing Autophagy via AMPK-mTOR-P70S6K Pathway[J].Front Pharmacol,2020,11:84.DOI:10.3389/fphar.2020.00084.
• [24] CARLONI S,BUONOCORE G,BALDUINI W.Protective role of autophagy in neonatal hypoxia-ischemia induced brain injury[J].Neurobiol Dis,2008,32(3):329-339.DOI:10.1016/j.nbd.2008.07.022.
• [25] ZHANG X,YAN H,YUAN Y,et al.Cerebral ischemia-reperfusion-induced autophagy protects against neuronal injury by mitochondrial clearance[J].Autophagy,2013,9(9):1321-1333.DOI:10.4161/auto.25132.
• [26] ADHAMI F,SCHLOEMER A,KUAN C Y.The roles of atuophagy in cerebral ischemia[J].Autophagy,2007,3(1):42-44.
• [27] PENG C,RAO W,ZHANG L,et al.Mitofusin 2 exerts a protective role in ischemia reperfusion injury through increasing autophagy[J].Cell Physiol Biochem,2018,46(46):2311-2324.DOI:10.1159/000489621.
• [28] CARLONI S,GIRELLI S,SCOPA C,et al.Activation of autophagy and Akt/CREB signaling play an equivalent role in the neuroprotective effect of rapamycin in neonatal hypoxia-ischemia[J].Autophagy,2010,6(3):366-377.DOI:10.4161/auto.6.3.11261.
• [29] CAI C C,ZHU J H,YE L X,et al.Glycine protects against hypoxic-ischemic brain injury by regulating mitochondria-mediated autophagy via the AMPK pathway[J].Oxid Med Cell Longev,2019:4248529.DOI:10.1155/2019/4248529.