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许永劼,许 雯,陈 钢,林海容,黄昶煜东,朱丽英,潘 卫.2 种高糖诱导海马神经元模型应用及优势比较[J].中国比较医学杂志,2021,31(8):1~8.
2 种高糖诱导海马神经元模型应用及优势比较
Application and comparison of the advantages of two high-glucose-induced hippocampal neuron models
投稿时间:2020-11-04  
DOI:10. 3969 / j.issn.1671-7856. 2021. 08. 001
中文关键词:  糖尿病脑病  海马神经元  细胞模型  应用优势
英文关键词:diabetic encephalopathy  hippocampal neurons  cell model  application advantages
基金项目:
作者单位E-mail
许永劼 1.贵州医科大学附属医院产前诊断中心,贵阳 550025
3.贵州医科大学公共卫生学院,贵阳 550025 
422526543@ qq.com 
许 雯 贵州医科大学医学检验学院,贵阳 550025  
陈 钢 贵州医科大学医学检验学院,贵阳 550025  
林海容 贵州医科大学医学检验学院,贵阳 550025  
黄昶煜东 贵州医科大学医学检验学院,贵阳 550025  
朱丽英 贵州医科大学医学检验学院,贵阳 550025  
潘 卫 1.贵州医科大学附属医院产前诊断中心,贵阳 550025
2.贵州医科大学医学检验学院,贵阳 550025
3.贵州医科大学公共卫生学院,贵阳 550025 
pw_6602@ sina.com 
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中文摘要:
       目的 比较 2 种高糖诱导海马神经元模型优缺点,探讨 2 种细胞模型在不同情况下应用效果。 方法 体外培养原代海马神经元和 HT-22 海马神经元细胞系,分为对照组和高糖组。神经元纯度鉴定,细胞活力检测,转染试剂对细胞活性和转染效率的影响,流式细胞术观察细胞凋亡情况,Western blot 检测凋亡蛋白表达情况。 结果 原代海马神经元纯度 85%以上;成功复制 2 种高糖诱导神经元细胞模型;高糖作用时间 48 h,原代海马神经元细胞活性稳定在 80%, HT-22 细胞活性达 95% 以上; 不同种脂质体转染试剂均能损伤 2 种细胞模型, LipoRNAiMax 毒性较小,其中 HT-22 细胞受损小,HT-22 细胞转染效率较高;2 种模型细胞凋亡显著(P<0. 05),其中原代神经元凋亡率更高,凋亡蛋白 Bcl-2 均表达下降、原代神经元 Bax 表达增高(P<0. 05),HT-22 细胞中 Bax 表达无差异。 结论 2 种高糖诱导海马神经元细胞模型具备不同的优缺点,细胞模型的选择应根据实际情况考虑。
英文摘要:
       Objective To compare the advantages and disadvantages of two high-glucose-induced hippocampal neuron models, and explore their application in different situations. Methods Primary hippocampal neurons and HT-22 hippocampal neuron cell lines were cultured in vitro and divided into control and high glucose groups. Neurons were purified and cell viability was measured. The effect of transfection reagents on cell viability and transfection efficiency was determined and flow cytometry was used to observe cell apoptosis. Western blot was used to detect apoptotic protein expression. Results The purity of primary hippocampal neurons was > 85%. Two high-glucose-induced neuronal cell models were successfully modeled. The high-glucose action time was 48 hours, the activity of primary neurons was stable at 80%, and the activity of HT-22 cells was >95%. The liposome transfection reagents caused damage in the two cell models. LipoRNAiMax was less toxic to HT-22 cells, which also had a high transfection efficiency. The two models showed significant apoptosis, compared with the primary neuron control group, the apoptosis rate of the primary neuron high glucose group increased, compared with the HT-22 control group, the HT-22 high glucose group increased cell apoptosis ( P< 0. 05). Compared with HT-22 high glucose group, the apoptotic rate of primary neurons high glucose group was higher, the expression of apoptotic protein Bcl-2 was decreased, and the expression of Bax was increased in primary neurons ( P< 0. 05). Compared with HT-22 control group, there was no difference in the expression of Bax in HT-22 high glucose group . Conclusions The two high-glucose-induced hippocampal neuron cell models had different advantages and disadvantages. The choice of cell model should be considered based on actual conditions.
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