文章摘要
许传先,凌卫明,张岳春,欧萌萌,方斌斌.姜黄素对冈田酸损伤的HT⁃22细胞的保护作用[J].南京医科大学学报,2021,(1):41~48
姜黄素对冈田酸损伤的HT⁃22细胞的保护作用
Protective effects of curcumin on HT⁃22 cells damaged by okadaic acid
投稿时间:2020-02-24  
DOI:10.7655/NYDXBNS20210108
中文关键词: 阿尔茨海默病  冈田酸  姜黄素  HT⁃22细胞  p⁃Tau
英文关键词: Alzheimer’s disease  okadaic acid  curcumin  HT⁃22 cell  p⁃Tau
基金项目:无锡市卫生计生科研青年项目(Q201755)
作者单位
许传先 无锡市儿童医院检验科江苏 无锡 214023 
凌卫明 南京医科大学附属无锡精神卫生中心检验科江苏 无锡 214151 
张岳春 南京医科大学附属无锡精神卫生中心检验科江苏 无锡 214151 
欧萌萌 南京医科大学附属无锡精神卫生中心检验科江苏 无锡 214151 
方斌斌 南京医科大学附属无锡精神卫生中心检验科江苏 无锡 214151 
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中文摘要:
      目的:探讨姜黄素(curcumin,Cur)对冈田酸(okadaic acid,OA)损伤的小鼠海马神经元细胞保护作用的机制,为阿尔茨海默病(Alzheimer’s disease,AD)的治疗提供新的理论基础。方法:小鼠海马神经元HT?22细胞经OA处理,分为对照组、Cur组、OA组、OA+Cur组,MTT检测各组细胞活力;Annexin V?FITC/PI双染色荧光显微镜及流式细胞仪检测各组细胞凋亡情况;DCFH?DA探针荧光显微镜及流式细胞仪测定细胞内活性氧(reactive oxygen species,ROS)水平;免疫印迹法检测Cleaved?caspase?3、Bcl?2、总微管相关蛋白Tau(t?Tau)、磷酸化的微管相关蛋白Tau(p?Tau)蛋白的水平。结果:与对照组相比,不同浓度的OA作用HT?22细胞24 h后,细胞活力呈剂量依赖性下降,差异有统计学意义。OA损伤持续一定时间后细胞内ROS水平升高,OA损伤后细胞凋亡增加,抗凋亡蛋白Bcl?2表达量降低、凋亡蛋白Cleaved?caspase?3蛋白表达升高且t?Tau、p?Tau蛋白表达异常增加。姜黄素作用OA损伤的HT?22细胞后,与OA组相比,OA+Cur组细胞活性增加,凋亡率降低,细胞内ROS生成减少,同时Bcl?2表达增加、Cleaved?caspase?3蛋白和t?Tau、p?Tau蛋白表达减少。结论:姜黄素可以保护OA损伤的小鼠海马神经元细胞,为阿尔茨海默病相关神经元细胞的保护提供了新的理论依据。
英文摘要:
      Objective:To explore the protective mechanism of curcumin(Cur)on the hippocampal neurons of mice injured by okadaic acid(OA),and to provide a new theoretical basis for the treatment of Alzheimer’s disease. Methods:Mouse hippocampal neuron HT?22 cells were treated with OA,and divided into four groups:control group,Cur group,OA group,and OA+Cur group. The cell viability was measured by MTT assay. The apoptosis was detected by using Annexin V?FITC/PI double staining fluorescence microscope and flow cytometry. DCFH?DA probe fluorescence microscopy and flow cytometry were used to determine the intracellular reactive oxygen species(ROS)production,and Western blot was used to detect the levels of cleaved?caspase?3,Bcl?2,total Tau(t?Tau),and phosphorylated Tau(p?Tau)proteins. Results:Compared with the control group,the cell viability decreased in a dose?dependent manner after HT?22 cells were treated with different concentrations of OA for 24 hours,and the difference was statistically significant. When OA injury lasted for a certain period of time,the level of ROS in the cells increased. OA injury promoted cell apoptosis,reduced expression of anti?apoptotic protein Bcl?2,increased expression of cleaved?caspase?3 protein,and the expression of t?Tau and p?Tau protein increased abnormally. Compared with the OA group,the cell viability increased,the apoptosis rate and the production of ROS decreased in the OA+Cur group,meanwhile,the expression of Bcl?2 protein increased,whereas the expressions of cleaved?caspase?3 protein,t?Tau and p?Tau decreased. Conclusion:Curcumin can protect hippocampal neurons from OA injury in mice. The present results provide a new theoretical basis for the protection of Alzheimer’s disease?related neurons.
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