地奥司明通过线粒体途径保护SH-SY5Y细胞免受氧糖剥夺损伤  

Diosmin protects SH-SY5Y cells from oxygen-glucose deprivation/reoxygenation injury via mitochondrial pathway

作　　者：梁东冉 张新超 任凯 刘芳芳 赵琳[1] 武明媚 LIANG Dongran;ZHANG Xinchao;REN Kai;LIU Fangfang;ZHAO Lin;WU Mingmei(Department of Anatomy,Medical School of Yan'an University,Yan'an 716099,China;Department of Neurobiology,School of Basic Medical Sciences,Air Force Medical University,Xi'an 710032,China)

机构地区：[1]延安大学医学院解剖学教研室,陕西延安716099 [2]空军军医大学基础医学院神经生物学教研室,陕西西安710032

出　　处：《空军军医大学学报》2024年第12期1406-1411,共6页Journal of Air Force Medical University

基　　金：国家自然科学基金(82471069);西安市神经免疫疾病创新转化重点实验室开放课题(2024001)。

摘　　要：目的探讨地奥司明(DSM)通过线粒体途径对氧糖剥夺/复氧(OGD/R)处理的人神经母细胞瘤SH-SY5Y细胞的保护效果。方法体外培养SH-SY5Y细胞,建立OGD/R模型,通过CCK-8细胞活性检测,确定DSM最佳作用浓度并进行后续实验。将细胞分为空白对照组(Control组)、OGD/R组和DSM组(DSM浓度分别为0.01、0.02、0.04 g/L)。CCK-8法检测细胞存活率,使用相差显微镜来观察细胞的形态,并通过分光光度测量法来测定细胞内的ATP水平和线粒体复合体Ⅱ活性,JC-1法检测线粒体膜电位,TUNEL法检测细胞凋亡状况。结果浓度为0.01、0.02、0.04 g/L的DSM均能促进OGD/R损伤的细胞存活,且药物浓度为0.02 g/L时SH-SY5Y细胞的存活率最高。与Control组相比,OGD/R组细胞形态皱缩,细胞ATP含量和线粒体复合体Ⅱ活性显著下降,线粒体膜电位降低,细胞凋亡率升高;而0.02 g/L DSM预先孵育3 h,再进行OGD/R损伤的细胞存活率增高,细胞形态基本正常,ATP含量和线粒体复合体Ⅱ活性显著提升,且线粒体膜电位水平显著提高,凋亡细胞减少。结论DSM对OGD/R的细胞具有显著保护作用,且在0.02 g/L时发挥最佳保护作用,其作用机制与维持线粒体正常功能和减少细胞凋亡状态密切相关。Objective To investigate the protective effects of diosmin(DSM)on human neuroblastoma SH-SY5Y cells treated with oxygen-glucose deprivation/reoxygenation(OGD/R)via mitochondrial pathway.Methods SH-SY5Y cells were cultured in vitro,OGD/R model was established,and the optimal concentration of DSM was determined by CCK-8 assay and subsequent experiments were conducted.The cells were divided into blank control group(Control group),OGD/R group,and DSM group(DSM concentration was 0.01,0.02,and 0.04 g/L,respectively).Cell survival rate was detected by CCK-8 assay,cell morphology was observed by phase-contrast microscope,intracellular ATP levels and mitochondrial complexⅡactivity were measured by spectrophotometry,mitochondrial membrane potential was detected by JC-1 staining method,and apoptosis was detected via TUNEL assay.Results DSM at three different concentrations(0.01,0.02,and 0.04 g/L)promoted the survival of SH-SY5Y cells damaged by OGD/R,with 0.02 g/L concentration exhibiting the most significant effect on cell survival.Compared with Control group,OGD/R group was characterized by shrunken cell morphology,significantly decreased ATP content and mitochondrial complexⅡactivity,decreased mitochondrial membrane potential,and increased apoptosis rate.After pre-incubation with 0.02 g/L DSM for 3 h,the survival rate of OGD/R damaged cells was increased,cell morphology was basically normal,ATP content and mitochondrial complexⅡactivity were significantly increased,mitochondrial membrane potential was significantly increased,and cell apoptosis was decreased.Conclusion DSM has a significant protective effect on SH-SY5Y cells damaged by OGD/R and the best protective effect is displayed at 0.02 g/L,and its mechanism of action is closely related to maintaining the normal function of mitochondria and reducing the state of apoptosis.

关 键 词：地奥司明 氧糖剥夺/复氧模型 神经保护 线粒体 细胞凋亡 

分 类 号：R965[医药卫生—药理学]