迷迭香酸通过抑制NF-κB活化对抗谷氨酸诱导PC12细胞凋亡

发布时间：2018-04-27 00:07

本文选题：迷迭香酸 + NF-κB　；参考：《南华大学》2012年硕士论文

【摘要】：目的：观察迷迭香酸（RA）对抗谷氨酸诱导PC12细胞凋亡发挥神经保护作用，探讨作用机制与核转录因子-κB（NF-κB）信号通路的关系。方法：体外培养PC12细胞，建立谷氨酸诱导细胞凋亡模型。采用MTT法检测细胞存活率；Hoechst33258荧光染色观察细胞凋亡形态的改变，碘化丙啶（PI）染色流式细胞仪检测分析细胞凋亡率；Western blot检测IκBα、p-IκBα蛋白、NF-κBp65蛋白在细胞浆和细胞核中表达变化情况。结果：不同浓度谷氨酸（0、4、8、12、16、20mmol/L）作用PC12细胞24h后，细胞存活率呈剂量依赖性下降，其IC75大约为16mmol/L。因此，本实验以16mmol/L谷氨酸作为PC12细胞损伤模型的工作浓度，，将后续实验分组为：对照组，损伤组（16mmol/L谷氨酸），RA1组（30μmol/L RA+16mmol/L谷氨酸），RA2组（60μmol/L RA+16mmol/L谷氨酸）。16mmol/L谷氨酸孵育PC12细胞24h后，细胞核形态出现凋亡小体、细胞核浓缩或碎块状等典型凋亡样改变，细胞凋亡率为(28.6±2.5)%；与损伤组比较，RA1组和RA2组细胞存活率随RA浓度增加明显升高（p 0.05），并能减少细胞凋亡发生，使其凋亡率分别下降至(18.3±3.7)%、(7.8±1.9)%。16mmol/L谷氨酸作用PC12细胞1h后，细胞浆中IκBα蛋白和NF-κB p65蛋白减少，p-IκBα蛋白表达增加，NF-κB p65蛋白在细胞核中表达增加，2h达到高峰；给予30、60μmol/L RA预处理1h后，能抑制NF-κBp65蛋白活化进入细胞核。结论： 1、在PC12细胞模型中，迷迭香酸有抗谷氨酸诱导细胞凋亡的作用； 2、 RA对抗谷氨酸毒性发挥神经保护作用的机制，可能与抑制NF-κB活性有关。
[Abstract]:Objective: To observe the neuroprotective effect of rosmarinic acid (RAA) on glutamate induced apoptosis of PC12 cells, and to explore the relationship between the mechanism and the signal pathway of nuclear transcription factor-魏 B (NF- 魏 B) NF- 魏 B. Methods: PC12 cells were cultured in vitro, and glutamate induced apoptosis model was established. The morphology of cell apoptosis was observed by MTT assay. The apoptosis rate was detected by flow cytometry. The expression of NF- 魏 Bp65 in cytoplasm and nucleus of I 魏 B 伪 -p-I 魏 B 伪 protein was detected by Western blot. Results: The survival rate of PC12 cells decreased in a dose-dependent manner after 24 h treatment with different concentrations of Glutamic acid. The IC75 was about 16 mmol 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) ~ (-1). Therefore, in this experiment, 16mmol/L glutamate was used as the working concentration of PC12 cell injury model. The following experiments were divided into two groups: control group. The injured group was divided into two groups: the injured group was treated with 60 渭 mol/L RA 16mmol/L glutamate, 16 mmol / L glutamate / L glutamic acid for 24 hours, and the injured group was treated with 60 渭 mol/L RA 16mmol/L glutamate, 16 mmol / L glutamic acid for 24 hours after incubating PC12 cells with 60 渭 mol/L RA 16mmol/L glutamate or 16 mmol / L glutamate. The apoptotic bodies appeared in nuclear morphology, the apoptotic rate was 28.6 卤2.5g, the cell survival rate of RA1 group and RA2 group increased significantly with the increase of RA concentration, and the apoptosis rate decreased with the increase of RA concentration. After treated with glutamate of 7.8 卤1.9)%.16mmol/L for 1 h, I 魏 B 伪 protein and NF- 魏 B p65 protein in the cytoplasm decreased the expression of p-I 魏 B 伪 protein and increased the expression of NF- 魏 B p65 protein in the nucleus of PC12 cells. The expression of NF- 魏 B p65 protein increased to a peak at 2 h after pretreatment with 3060 渭 mol/L RA for 1 h. It can inhibit the activation of NF- 魏 Bp65 protein into the nucleus. Conclusion: 1. In PC12 cell model, rosemary acid could inhibit the apoptosis induced by glutamate. 2. The neuroprotective mechanism of RA against glutamate toxicity may be related to the inhibition of NF- 魏 B activity.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363

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