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第41卷第1期 南京医科大学学报（自然科学版）
2021年1月 Journal of Nanjing Medical University（Natural Sciences） · 41 ·

·基础研究·

姜黄素对冈田酸损伤的HT⁃22细胞的保护作用

许传先 ，凌卫明 ，张岳春 ，欧萌萌 ，方斌斌 2*
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无锡市儿童医院检验科，江苏 无锡 214023；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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［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⁃cas⁃
pase⁃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。
［关键词］ 阿尔茨海默病；冈田酸；姜黄素；HT⁃22细胞；p⁃Tau
［中图分类号］ R749.1 ［文献标志码］ A ［文章编号］ 1007⁃4368（2021）01⁃041⁃08
doi：10.7655/NYDXBNS20210108

Protective effects of curcumin on HT⁃22 cells damaged by okadaic acid

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XU Chuanxian ，LING Weiming ，ZHANG Yuechun ，OU Mengmeng ，FANG Binbin 2*
1 Department of Clinical Laboratory，Wuxi Children’s Hospital，Wuxi 214023；Department of Clinical Laboratory，
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Wuxi Mental Health Center Affiliated with Nanjing Medical University，Wuxi 214151，China
［Abstract］ 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.
［Key words］ Alzheimer’s disease；okadaic acid；curcumin；HT⁃22 cell；p⁃Tau
［J Nanjing Med Univ，2021，41（01）：041⁃048］

［基金项目］ 无锡市卫生计生科研青年项目（Q201755）
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通信作者（Corresponding author），E⁃mail：65645419@qq.com