摘要
目的探讨芒果苷(MF)在蛛网膜下腔出血(SAH)后早期阶段对神经细胞的保护作用及机制。方法采用小鼠来源神经瘤母细胞(Neuro-2a细胞)和氧合血红蛋白(OxyHb)建立体外SAH模型,分为对照组、蛛网膜下腔出血组(SAH)及芒果苷干预组(MF+SAH)3组,利用免疫荧光检测受损线粒体与溶酶体的结合;利用JC-1和DCFH-DA分别检测线粒体膜电位(Δψm)和活性氧(ROS);Western blot分别检测细胞内磷脂酰肌醇-3-激酶(PI3K)、蛋白激酶B(Akt)和雷帕霉素靶蛋白(mTOR)的表达,胞浆、线粒体中凋亡相关蛋白淋巴细胞瘤-2(Bcl-2)、Bcl2关联X蛋白(Bax)、细胞色素C(Cyt-C)和Caspase-3的表达。结果与对照组相比,SAH组线粒体膜电位(Δψm)明显去极化(P<0.05),细胞内ROS水平增多(P<0.01),p-PI3K、p-Akt、p-mTOR的表达水平显著减少(P<0.05),胞浆内Cyt-C表达水平显著增加(P<0.05),线粒体内Cyt-C表达水平显著减少(P<0.05),Bax和Caspase-3表达水平显著增加(P<0.05),Bcl-2表达水平显著减少(P<0.05);与SAH组相比较,MF+SAH组Δψm和ROS明显恢复(P<0.05),p-PI3K、p-Akt、p-mTOR的表达水平明显增加(P<0.05),胞浆内Cyt-C表达水平显著下降(P<0.05),线粒体内Cyt-C表达水平显著增加(P<0.05),Bax和Caspase-3表达显著减少(P<0.05),Bcl-2表达显著增加(P<0.05)。结论芒果苷可以通过稳定线粒体膜电位,减少ROS的产生,改善线粒体功能障碍,从而抑制SAH后早期阶段神经细胞的凋亡。
Objective To investigate the protective effect and mechanism of mangiferin(MF)on nerve cells in the early stage after subarachnoid hemorrhage(SAH).Methods The SAH model in vitro was established by using the Neuro-2a cells originated from the mouse neuroblastoma cells and OxyHb.The SAH models were divided into the control group(CON),SAH group and mangiferin intervention group(MF+SAH).The binding of damaged mitochondria to lysosome was detected by using the immunofluorescence;the mitochondrial membrane potential(Δψm)and reactive oxygen species(ROS)were measured by JC-1 and DCFH-DA,respectively;the expressions of phosphatidylinositol-3-kinase(PI3K)/protein kinase B(Akt),PKB)/rapamycin target protein(mTOR)were detected by Western blot;the expressions of apoptosis related proteins lymphoma-2(Bcl-2),BCL2 associated X protein(Bax),cytochrome C(Cyt-C)and cleaved-caspase-3(Caspase-3)in cytoplasm and mitochondria were detected.Results Compared with the control group,theΔψm in the SAH group was significantly depolarized(P<0.05),the intracellular ROS content was increased,the expression levels of p-PI3K,p-Akt and p-mTOR were significantly decreased(P<0.05),the expression of Cyt-C in cytoplasm was significantly increased(P<0.05),the expression of Cyt-C in mitochondria was significantly decreased(P<0.05),the expression of Bax and Caspase-3 was significantly increased(P<0.05),and the expression of Bcl-2 was significantly decreased(P<0.05);compared with SAH group,Δψm and ROS in the MF+SAH group were significantly recovered compared with before(P<0.05),the p-pi3k,p-Akt,p-mTOR expression levels were significantly increased(P<0.05),Cyt-C expression in cytoplasm was significantly decreased(P<0.05),Cyt-C expression in mitochondria was significantly increased(P<0.05),the Bax and Caspase-3 expression levels were significantly decreased(P<0.05),and the Bcl-2 expression level was significantly increased(P<0.05).Conclusion Mangiferin can inhibit the nerve cells apoptosis in the early stage after SAH by stabilizing the mitochondrial membrane potential,reducing the ROS production and improving the mitochondrial dysfunction.
作者
姬康祁
周文科
JI Kangqi;ZHOU Wenke(Department of Neurosurgery,First Affiliated Hospital of Xinxiang Medical University/Henan Provincial Key Laboratory of Neural Repair,Xinxiang,Henan 453000,China)
出处
《重庆医学》
CAS
2021年第13期2176-2181,共6页
Chongqing medicine
基金
国家自然科学基金项目(81541030)
河南省神经修复重点实验室开放课题项目(HNSJXF-2018-008)
新乡医学院研究生科研创新支持计划项目(YJSCX201947Y)。
关键词
蛛网膜下腔出血
脑损伤
细胞凋亡
线粒体疾病
芒果苷
subarachnoid hemorrhage
brain injuries
apoptosis
mitochondrial diseases
mangiferin