高脂饮食小鼠脂肪组织胞外囊泡损伤海马神经元的研究  

Effect of extracellular vesicles derived from adipose on hippocampal neuronal damage in mice with a high-fat diet

作　　者：周迪 吕子阳 聂盛丹[2] 周高雅[3] 王珊[1] ZHOU Di;LYU Zi-yang;NIE Sheng-dan;ZHOU Gao-ya;WANG Shan(Dept of Pharmaceutical Engineering,School of Chemistry and Chemical Engineering,Central South University,Changsha 410083,China;Institute of chinical Medicine,Hunan Provincial People’s Hospital,Changsha 410081,China;Dept of Orthopedic Surgery,Brain Hospital of Hunan Province,Changsha 410007,China)

机构地区：[1]中南大学化学化工学院制药工程系,湖南长沙410083 [2]湖南省人民医院药物临床试验机构办,湖南长沙410081 [3]湖南省脑科医院神经内科,湖南长沙410007

出　　处：《中国药理学通报》2022年第1期79-85,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 82070857);湖南省自然科学基金资助项目(No 2019JJ50782)。

摘　　要：目的探讨高脂饮食小鼠脂肪组织分泌胞外囊泡对海马神经元以及小鼠认知行为的影响。方法C57BL/6J雄性小鼠20只,随机分为2组(n=10):正常饮食(normal diet,ND)组、高脂饮食(high-fat diet,HFD)组,分组喂养28周,每周记录小鼠体重。于第27周检测小鼠血糖、胰岛素水平,计算胰岛素抵抗指数(HOMA-IR),第28周用Morris水迷宫实验检测小鼠学习和记忆能力。取小鼠脂肪组织,HE染色观察形态学差异,并提取其分泌的胞外囊泡,采用TEM及NTA进行鉴定。将PKH 67标记的脂肪组织胞外囊泡经尾静脉注射正常小鼠,30 h后检测脑组织海马神经元摄取脂肪组织胞外囊泡的情况;用相同蛋白量的胞外囊泡处理原代海马神经元,观察其对神经元细胞活性及细胞形态的影响。结果与ND组相比,HFD组小鼠体重明显增加,血糖及胰岛素水平升高,并出现胰岛素抵抗状态。HFD组小鼠逃避潜伏期显著延长,目标象限停留时间明显缩短。HFD组小鼠脂肪细胞明显增大,脂肪组织分泌的胞外囊泡明显增多。脂肪组织分泌的胞外囊泡可被原代海马神经元及正常小鼠脑组织海马神经元摄取。与ND组相比,HFD组小鼠脂肪组织胞外囊泡处理可明显减少海马神经元树突长度、初级树突数和次级树突数,并降低神经元细胞活性。结论长期高脂饮食可能通过影响脂肪组织胞外囊泡,损伤海马神经元。Aim To investigate the effect of extracellular vesicles secreted by adipose tissue of mice on hippocampal neurons and cognitive behavior of mice with a high-fat diet. Methods Twenty C57 BL/6 J male mice were randomly divided into normal diet(ND) group(n=10) and high-fat diet(HFD) group(n=10), fed for 28 weeks. The weight of mice was recorded weekly. The level of fasting blood glucose, insulin and the insulin resistance index(HOMA-IR) of mice were tested at week 27. At week 28, the learning and memory abilities of mice were assessed by the Morris water maze. The morphological differences in adipose tissue were observed by HE staining, and the extracellular vesicles secreted from adipose tissue were quantified by TEM and NTA. Extracellular vesicles derived from adipose tissue labeled with PKH 67 were injected into normal mice via the tail vein, and after 30 h, the uptake of extracellular vesicles was detected in the hippocampal slice. The primary hippocampal neurons were treated with extracellular vesicles with the same amount of protein, and the effects of them on neuronal morphology and cell viability were observed. Results Compared with ND group, mice in HFD group were significantly heavier, with hyperglycemia, hyperinsulinemia and higher insulin resistance index. In the Morris watermaze test, the HFD group showed a longer escape la-tency and less swimming time in the target zone. Thevolume of adipocytes and the amount of extracellularvesicles secreted from them significantly increased inHFD group. Extracellular vesicles secreted by adiposetissue could be internalized by both the primary hipp-ocampal neurons and the hippocampal neurons in thenormal mice. Compared with ND group, extracellularvesicles secreted by adipose tissue of the HFD group significantly reduced the length of primary hippocampalneuronal dendrites, the number of primary and second-ary dendrites, and the cell viability of neuron cells.Conclusion Long-term high-fat diet could damage thehippocampal neurons by affecting the extracellular vesi-cles

关 键 词：高脂饮食 肥胖 脂肪组织 胞外囊泡 海马神经元损伤 认知功能 

分 类 号：R-332[医药卫生] R322.81