摘要
目的观察缺氧预处理后骨髓间充质干细胞(BMSCs)来源外泌体对心肌缺血再灌注(I/R)大鼠心肌损伤的影响,并探讨其可能的机制。方法分离培养大鼠BMSCs,采用ExoQuick试剂法分离提纯BMSCs来源外泌体。将50只SD大鼠随机分为正常组(常规饲养未接受任何干预的大鼠)、假手术组(开胸后仅打开心包而不结扎左冠状动脉前降支)、I/R组(建立心肌I/R模型)、缺氧干预组(建立心肌I/R模型并采用缺氧预处理后BMSCs来源外泌体400μg/g进行干预)和常氧干预组(建立心肌I/R模型并采用常氧预处理后BMSCs来源外泌体400μg/g进行干预),每组10只。各组干预1周后处死,取心肌组织,HE染色观察心肌病理变化,TTC染色后测定心肌梗死面积比,ELISA法检测心肌组织氧化应激指标[超氧化物歧化酶(SOD)、丙二醛(MDA)、总抗氧化力(T-AOC)]。采用实时荧光定量PCR法和Western blotting法检测缺氧及常氧预处理BMSCs来源外泌体中缺氧诱导因子1α(HIF-1α)mRNA和蛋白表达。结果正常组和假手术组大鼠心肌纤维排列整齐,结构清楚,细胞和间质无明显充血水肿及炎症细胞浸润。I/R组心肌纤维断裂、排列紊乱,存在细胞间质充血水肿及炎性细胞浸润。与I/R组相比,缺氧干预组、常氧干预组大鼠心肌纤维断裂、排列紊乱程度及细胞间质充血水肿程度均有所减轻。与正常组、假手术组比较,I/R组、缺氧干预组、常氧干预组大鼠心肌组织SOD、T-AOC表达均降低,心肌梗死面积比、MDA表达均升高;与I/R组比较,缺氧干预组、常氧干预组大鼠心肌组织SOD、T-AOC表达均升高,心肌梗死面积比、MDA表达均降低,且缺氧干预组变化更明显(P均<0.05)。缺氧预处理BMSCs来源外泌体中HIF-1αmRNA和蛋白相对表达量分别为6.84±0.62、7.35±0.86,均高于常氧预处理BMSCs来源外泌体的4.27±0.39、5.14±4.76(P均<0.05)。结论 BMSCs来源外泌体可减轻I/R大鼠的心肌损伤,且低氧预处
Objective To observe the effects of exosomes derived from bone marrow mesenchymal stem cells(BM-SCs)after hypoxic preconditioning on myocardial ischemia/reperfusion(I/R)rats with myocardial injury and to investi-gate the possible mechanism.Methods BMSCs were isolated and cultured from normal SD rats.Exosomes were isolated and purified by ExoQuick reagent method from BMSCs.Fifty SD rats were randomly divided into 5 groups after weighing:the normal group(without any intervention),sham operation group(without ligation of left anterior descending coronary ar-tery after thoracotomy),I/R group(myocardial I/R rat model without any intervention),hypoxic intervention group(es-tablishing the I/R models,using BMSCs-derived exosomes 400μg/g after hypoxic preconditioning to intervene the myocar-dial I/R rat models),and normoxic intervention group(using normoxic cultured BMSCs-derived exosomes 400μg/g to in-tervene the myocardial I/R rat models),with 10 rats in each group.Rats in each group were sacrificed after 1 week of in-tervention.The myocardial tissues were taken and the pathological changes were observed by HE staining.The myocardial infarction area was measured by TTC staining.The oxidative stress indexes[superoxide dismutase(SOD),malondialde-hyde(MDA),total antioxidant capacity(T-AOC)]of myocardial tissues were detected by ELISA.Real-time PCR and Western blotting were used to detect the expression levels of HIF-1αmRNA and protein in exosomes derived from BMSCs.Results In the normal group and the sham group,the myocardial fibers were neatly arranged and the structure was clear,and there were no obvious hyperemia and edema or inflammatory cell infiltration in the cells and stroma.In the I/R group,myocardial fibers were broken and disordered,with intercellular hyperemia and edema and inflammatory cell infil-tration.Compared with the normal group and sham group,the expression levels of SOD and T-AOC decreased in the myo-cardial tissues of the I/R group,hypoxia intervention group and normoxia intervention group,while th
作者
王玉梅
韩炜
李惠
张媛
郭艳丹
乔玲玲
许文平
WANG Yumei;HAN Wei;LI Hui;ZHANG Yuan;GUO Yandan;QIAO Lingling;XU Wenping(Outpatient Department,General Hospital of Southern Theater Command of the Chinese PLA,Guangzhou 510010,China)
出处
《山东医药》
CAS
2021年第23期37-41,46,共6页
Shandong Medical Journal
基金
广东省公益研究与能力建设专项(2015A030302062)。
关键词
心肌缺血再灌注损伤
缺氧预处理
骨髓间充质干细胞
外泌体
氧化应激
大鼠
myocardial ischemia-reperfusion injury
hypoxia preconditioning
bone marrow mesenchymal stem cells
exosome
oxidative stress
rats
