神经干细胞源性外泌体对脑缺血再灌注损伤后大鼠氧化应激和细胞凋亡的影响
Effects of neural stem cell-derived exosomes on oxidative stress and apoptosis of rats with after cerebral ischemia reperfusion injuries at different time points
陈珊 1赵雪 2刘若静 2周璐 2龙婷婷 2朱俊德2
作者信息
- 1. 贵阳康养职业大学基础医学院,贵州贵阳 550081;贵州医科大学基础医学院人体解剖学教研室,贵州贵安 561113
- 2. 贵州医科大学基础医学院人体解剖学教研室,贵州贵安 561113;贵州省高等学校功能与疾病人脑组织库重点实验室,贵州贵安 561113
- 折叠
摘要
目的 探讨神经干细胞源性外泌体(NSC-Exos)对脑缺血再灌注损伤(MCAO)模型大鼠氧化应激和神经细胞凋亡的影响.方法 135只雄性SD大鼠随机分为Sham组(手术、不缺血、不微量注射)、MCAO组(手术、缺血、不微量注射)及NSC-Exos组(手术、缺血、侧脑室微量注射),于微量注射后大鼠脑缺血再灌注第1、7及14天时行神经功能缺陷评分,2,3,5-三苯基氯化四氮唑(TTC)检测脑组织相对梗死体积,HE染色和透射电子显微镜观察海马CA3区神经细胞形态及亚显微结构,比色法检测海马组织匀浆中谷胱甘肽过氧化物酶(GSH-PX)、超氧化物歧化酶(SOD)和丙二醛(MDA)的含量,Western blot检测海马组织中Bcl-2、Bax蛋白的表达.结果 与Sham组相比,MCAO组大鼠神经功能缺陷评分增高(P＜0.05),脑组织相对梗死体积显著增大(P＜0.05),神经细胞形态及超微结构受损严重,海马组织匀浆中GSH-PX、SOD酶活性降低,MAD含量增高(P＜0.05),Bcl-2含量降低、Bax蛋白含量增加(P＜0.05);与MCAO组相比,NSC-Exos组大鼠神经功能缺陷评分降低(P＜0.05),脑组织相对梗死体积减小(P＜0.05),存活神经细胞形态较为完好,超微结构较为清晰,海马组织匀浆中GSH-PX、SOD酶活性均增高,MAD含量减小(P＜0.05),Bcl-2蛋白随缺血再灌注时间的延长表达增高,蛋白Bax表达减少(P＜0.05).结论 NSC-Exos可改善MCAO大鼠的神经功能,减轻神经元的病理损伤,其机理可能与抑制海马组织的氧化应激和细胞凋亡有关.
Abstract
Objective To explore the effects of neural stem cell-derived exosomes(NSC-Exos)on oxidative stress and the influence of nerve cell apoptosis of middle cerebral artery occlusion(MCAO)model rats with cerebral ischemia reperfusion(CIR)injury.Methods One hundred and thirty-five male SD rats were randomly divided into Sham group(operation,no ischemia,no microinjection),MCAO group(with operation of CIR injury,ischemia,no microinjection),and NSC-Exos group(operation,ischemia,microinjection into the lateral ventricle).After microinjection,the rats were tested for neurological deficit scores on days 1,7 and 14 of CIR.Relative infarct volume of brain tissues was detected by 2,3,5-triphenyltetrazolium chloride(TTC).Morphology and ultrastructure of hippocampal CA3 region were observed by HE staining and transmission electron microscopy.The contents of glutathione peroxidase(GSH-PX),superoxide dismutase(SOD)and malondialdehyde(MDA)in hippocampus homogenate were detected by colorimetric method.The changes of Bcl-2 and Bax protein expression in hippocampus were detected by Western blot.Results Compared with Sham group,the neurological deficit score of MCAO group increased(P＜0.05),relative infarct volume of brain tissues increased significantly(P＜0.05).Morphology and ultrastructure of nerve cells were severely damaged.The activities of GSH-PX and SOD decreased,while the content of MAD increased in the hippocampus homogenate(P＜0.05).The expression of Bcl-2 protein decreased,while the expression of Bax protein increased(P＜0.05).Compared with MCAO group,the neurological function score of NSC-Exos group significantly decreased(P＜0.05),relative infarct volume of brain tissue was reduced(P＜0.05).Morphology of surviving nerve cells was relatively intact,and the ultrastructure was relatively clear.The activities of GSH-PX and SOD in hippocampus homogenate increased,while the content of MAD decreased(P＜0.05).The expression of Bcl-2 protein increased with extension of reperfusion time,while the expression of Bax protein was opposite(P＜0.05).Conclusion NSC-Exos can improve the neurological function and reduce the pathological injury of neurons in MCAO rats,and its mechanism may be related to the inhibition of oxidative stress and cell apoptosis in hippocampus.
关键词
脑卒中/神经干细胞/外泌体/细胞凋亡/大鼠Key words
cerebral ischemia/neural stem cells/exosome/apoptosis/rat引用本文复制引用
基金项目
贵州省自然科学基金项目(黔科合基础-ZK[2023]一般323)
贵州省卫生计生委自然科学基金项目(gzwkj2022-512)
贵州医科大学国家自然基金培育项目(20NSP006)
贵阳康养职业大学校级科研项目(贵康大K2024-21)
出版年
2024
NETL
NSTL
万方数据