氧糖剥离预处理通过保护线粒体减轻PC12细胞氧糖剥离/再灌注损伤  

Oxygen-glucose deprivation preconditioning alleviates oxygen-glucose deprivation/reperfusion injury in PC12 cells by protecting mitochondria

作　　者：黄丽 许娜 杨朝鲜 HUANG Li;XU Na;YANG Chaoxian(Department of Anesthesiology,Second Affiliated Hospital,Army Medical University(Third Military Medical University),Chongqing,400037;Department of Anatomy,School of Basic Medicine,Southwest Medical University,Luzhou,Sichuan Province,646000,China)

机构地区：[1]陆军军医大学(第三军医大学)第二附属医院麻醉科,重庆400037 [2]西南医科大学基础医学院解剖教研室,四川泸州646000

出　　处：《陆军军医大学学报》2024年第15期1763-1771,共9页Journal of Army Medical University

基　　金：四川省自然科学基金(2022NSFSC0718);西南医科大学应用基础面上项目(2021ZKMS011)。

摘　　要：目的研究氧糖剥离预处理(oxygen-glucose deprivation preconditioning,OGDPC)对氧糖剥离/再灌注(oxygen-glucose deprivation/reperfusion,OGD/R)损伤PC12细胞的保护作用及机制。方法简单随机抽样将PC12细胞分为Control、OGDPC、OGD/R和OGDPC+OGD/R共4组。透射电镜观察线粒体的超微结构,Mito-tracker染色观察线粒体数量,Western blot检测OGDH、PDHA1、COX4、HSP60的表达,JC-1和ATP试剂盒评估线粒体膜电位及ATP水平,TUNEL染色观察细胞凋亡情况,并与AIF、EndoG的免疫荧光染色进行共定位。结果与OGD/R组相比,OGDPC+OGD/R组PC12细胞线粒体超微结构损伤较轻;OGDH、PDHA1和COX4的表达增多(P<0.05),HSP60的表达减少(P<0.05);线粒体膜电位及ATP水平提高(P<0.05),细胞凋亡率降低(P<0.05),且AIF、EndoG向TUNEL染色阳性的细胞核聚集减少(P<0.05)。结论OGDPC减轻OGD/R引起的PC12细胞损伤,其机制可能与改善线粒体结构和功能、减少线粒体途径引起的细胞凋亡有关。Objective To investigate the protective effect of oxygen-glucose deprivation preconditioning(OGDPC)on PC12 cells damaged with oxygen-glucose deprivation/reperfusion(OGD/R)and its underlying mechanism.Methods PC12 cells were randomly divided into Control,OGDPC,OGD/R and OGDPC+OGD/R groups.The morphology and quantity of mitochondria were observed by transmission electron microscopy(TEM)and Mito-tracker staining.The expression of OGDH,PDHA1,COX4 and HSP60 was detected by Western blotting.Mitochondrial membrane potential and ATP production were evaluated by JC-1 and ATP assay kit.TUNEL staining was used to assess cell apoptosis,and translocation of AIF and EnodoG was observed by immunofluorescence staining and TUNEL staining.Results Compared with the OGD/R group,the mitochondrial ultrastructure damage were alleviated,the expression levels of OGDH,PDHA1 and COX4 were enhanced(P<0.05),that of HSP60 was decreased(P<0.05),the mitochondrial membrane potential and ATP level were increased(P<0.05),the cell apoptotic rate was reduced(P<0.05),and AIF and EndoG were less aggregated to the TUNEL-positive nucleus in OGDPC+OGD/R group(P<0.05).Conclusion OGDPC can alleviate the injury of PC12 cells caused by OGD/R,and the potential mechanism may be related to improving mitochondrial structure and function,and reducing cell apoptosis via mitochondrial pathway.

关 键 词：氧糖剥离预处理 氧糖剥离/再灌注 线粒体 PC12细胞 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R363.22[医药卫生—基础医学] R741.02