十六烷酸对谷氨酸致PC12细胞凋亡的保护作用

发布时间：2018-03-15 03:25

本文选题：十六烷酸　切入点：PC细胞　出处：《中国药学杂志》2015年13期 　论文类型：期刊论文

【摘要】：目的研究十六烷酸对谷氨酸损伤PC12细胞凋亡的保护作用。方法将PC12细胞分为7组:正常对照组,二甲基亚砜溶剂对照组,模型对照组(25 mmol·L-1谷氨酸),阳性药物对照组(7μmol·L-1尼莫地平或10μmol·L-1MK801)以及十六烷酸组(1、10、100μmol·L-1)。噻唑蓝法验证十六烷酸对谷氨酸损伤PC12细胞保护作用;乳酸脱氢酶比色法、台盼蓝染色法检测不同浓度十六烷酸对谷氨酸损伤PC12细胞膜通透性变化的影响;AO/EB、Hoechst荧光染色法观察PC12细胞凋亡的形态。FITC-Annexin V/PI荧光标记流式细胞仪检测细胞凋亡率。Western blot法检测细胞内Bcl-2、Bax、GluR1蛋白表达。结果噻唑蓝、台盼蓝染色计数、细胞上清液中乳酸脱氢酶含量显示:与谷氨酸模型组相比,十六烷酸能明显改善谷氨酸损伤PC12细胞的活力,减少细胞膜通透性;AO/EB荧光染色显示谷氨酸损伤后,模型对照组可见大部分细胞经AO染色后核染色质部分呈现浓染,十六烷酸各组仅出现少数被EB染成橘红色的细胞;Hocehst荧光染色显示,与模型组相比十六烷酸组细胞核发出蓝色荧光较弱。十六烷酸能上调Bcl-2蛋白表达,抑制Bax蛋白表达;并能抑制谷氨酸导致的谷氨酸受体的过度表达。结论十六烷酸对谷氨酸损伤PC12细胞凋亡具有保护作用,该保护与抑制谷氨酸受体蛋白NMDA、Bax蛋白的表达的增加,上调Bcl-2蛋白表达等有关。
[Abstract]:Objective to study the protective effect of hexadecanoic acid on apoptosis of PC12 cells injured by glutamate. Methods PC12 cells were divided into 7 groups: normal control group and dimethyl sulfoxide solvent control group. Model control group (25 mmol 路L -1 glutamate), positive drug control group (7 渭 mol 路L -1 nimodipine or 10 渭 mol 路L -1 MK801) and cetanoic acid group 100 渭 mol 路L -1. Thiazolyl blue assay was used to verify the protective effect of hexadecanoic acid on PC12 cells injured by glutamate. Effects of different concentrations of hexadecanoic acid on the permeability of PC12 cell membrane induced by glutamic acid by trypan blue staining the morphology of PC12 cell apoptosis was observed by AOP / EBX Hoechst fluorescence staining. FITC-Annexin V / Pi fluorescence flow cytometry was used to detect apoptosis rate. Western blot. Methods to detect the expression of Bcl-2OBax-GluR1 protein in cells. Results thiazolyl blue, thiazolyl blue, and thiazolyl blue were detected. Trypan blue staining and lactate dehydrogenase content in cell supernatant showed that cetanoic acid could significantly improve the viability of glutamate injured PC12 cells and reduce cell membrane permeability after glutamate injury. In the model control group, most of the cells showed strong staining after AO staining, and only a few cells stained with EB in the cetanoic acid group showed Hocehst fluorescence staining. Compared with the model group, the cetanoic acid group had weaker blue fluorescence. Cetane acid could up-regulate the expression of Bcl-2 protein and inhibit the expression of Bax protein. Conclusion hexadecanoic acid has a protective effect on the apoptosis of PC12 cells injured by glutamate, which protects and inhibits the increase of the expression of glutamate receptor protein NMDA-Bax protein. Upregulation of Bcl-2 protein expression and so on.
【作者单位】：新安医学省部共建教育部重点实验室;安徽省中医药科学院中药药效与安全性评价研究所;安徽中医药大学药学院;
【基金】：国家自然科学基金资助项目(81374005) 十二五科技支撑计划资助项目(2012BAI26B03) 安徽省第五批优秀青年科技基金资助项目(10040606Y17) 2013年度安徽省学术及技术带头人及后备人选科研活动经费资助项目
【分类号】：R96

【参考文献】