线粒体损伤在高糖高脂培养的多巴胺能神经元损伤中的作用研究  

Role of mitochondrial damage in dopaminergic neuron damage induced by high glucose and high-fat culture

作　　者：刘明之 张倬清 张雨鑫 苏建云 杨洋 杨晓春[1] LIU Mingzhi;ZHANG Zhuoqing;ZHANG Yuxin;SU Jianyun;YANG Yang;YANG Xiaochun(The First People's Hospital of Yunnan Province(the Affiliated Hospital of Kunming University of Science and Technology),Kunming,Yunnan 650032,China;Qingdao Academy of Chinese Medical Sciences,Shandong University of Traditional Chinese Medicine,Qingdao,Shandong 266114,China)

机构地区：[1]云南省第一人民医院/昆明理工大学附属医院,云南昆明650032 [2]山东中医药大学青岛中医药科学院,山东青岛266114

出　　处：《中国医学工程》2025年第4期17-24,共8页China Medical Engineering

基　　金：云南省应用基础研究计划项目(202101AY070001-275);云南省兴滇英才项目(YNWR-QNBJ-2018-315);云南省科技人才和平台计划项目(2019HB050);内分泌国家级临床重点专科培育项目(2024NMKFKT-02)。

摘　　要：目的研究体外高糖高脂培养的SN4741多巴胺神经元细胞的增殖和细胞显微结构的变化,探索糖尿病多巴胺能神经元损伤的机制。方法将体外培养的SN4741细胞株分为对照(control)和糖尿病(DM)两组,分别给予单纯胎牛血清培养基和含浓度30 mmol/L高糖、0.3 mmol/L棕榈酸、10%胎牛血清的DEME培养基培养24 h。CCK8检测细胞的存活率,流式细胞术检测细胞凋亡,透射电镜观察细胞显微结构的变化,并检测细胞线粒体膜电位。结果加入不同浓度高糖培养后,CCK8检测显示DM组SN4741细胞存活率明显下降,且下降的程度与高糖浓度和孵育时间呈正比(P<0.001)。流式细胞术检测显示,DM组细胞凋亡现象较control组明显增加(P<0.05)。透射电镜显示,DM组线粒体的体积增大,线粒体峭模糊、断裂,部分线粒体发生空泡化。免疫荧光显示,与control组相比,高糖培养的SN4741细胞线粒体膜电位降低。结论糖尿病导致多巴胺能无长突细胞损伤,线粒体结构和功能异常可能参与了该细胞损伤的发生。【Objective】To investigate the proliferation and cell microstructure changes of SN4741 dopamine neuron cells cultured in vitro with high glucose and high fat,and to explore the mechanism of diabetic dopaminergic neuron damage.【Methods】The SN4741 cell line cultured in vitro was divided into two groups:control group and diabetes mellitus(DM)group.The cells were cultured in pure fetal bovine serum medium and DEME medium containing 30 mmol/L high glucose,0.3 mmol/L palmitic acid,and 10%fetal bovine serum for 24 hours,respectively.CCK8 was used to detect cell survival rate,flow cytometry was used to detect cell apoptosis,transmission electron microscopy was used to observe cell microstructure changes,and cell mitochondrial membrane potential was detected.【Results】After adding different concentrations of high glucose,CCK8 detection showed that the survival rate of SN4741 cells in the DM group was significantly decreased,and the degree of decrease was proportional to the high glucose concentration and incubation time,with statistically significant differences(P<0.001).Flow cytometry showed that the apoptosis of cells in the DM group was significantly increased compared with the control group,with statistically significant differences(P<0.05).Transmission electron microscopy showed that the volume of mitochondria in the DM group was enlarged,the mitochondrial ridges were blurred and broken,and some mitochondria were vacuolated.Immunofluorescence showed that compared with the control group,the mitochondrial membrane potential of SN4741 cells cultured in high glucose was reduced.【Conclusion】Diabetes leads to the damage of retinal dopaminergic amacrine cells,and abnormal mitochondrial structure and function may be involved in the occurrence of this cell damage.

关 键 词：糖尿病 多巴胺能无长突细胞 凋亡 增殖 线粒体 

分 类 号：R816.97[医药卫生—放射医学]