姜黄素对冈田酸损伤的HT⁃22细胞的保护作用
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无锡市卫生计生科研青年项目(Q201755)


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

    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.

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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(自然科学版),2021,(1):41-48

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  • 收稿日期:2020-02-24
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  • 在线发布日期: 2021-02-04
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