尼氟灭酸对氯化钴诱导海马神经元凋亡的抑制作用

发布时间：2018-05-16 17:16

本文选题：尼氟灭酸 + 海马神经元　；参考：《吉林大学学报(医学版)》2015年05期

【摘要】：目的:探讨尼氟灭酸(NFA)对氯化钴(CoCl2)诱导的海马神经元凋亡的抑制作用,阐明其可能的机制。方法:原代培养乳鼠海马细胞,并在倒置显微镜下观察海马细胞的形态变化。将海马神经元随机分为对照组(含1%血清培养液)、CoCl2损伤组(含200μmol·L-1 NFA等体积培养液)和不同浓度NFA组(在CoCl2损伤组基础上加入20、40、80μmol·L-1 NFA)。MTT法检测各组海马神经元的存活率,流式细胞术分析海马神经元调亡率,ELISA法检测神经元培养液上清中相关凋亡蛋白Bcl-2、Bax、caspase-3和缺氧相关蛋白低氧诱导因子(HIF-1α)的表达水平。结果:海马神经元在培养7d后形态规则,突触相互交织形成网络。MTT检测,与对照组比较,CoCl2损伤组神经元存活率明显降低(P0.01);与CoCl2损伤组比较,20、40和80μmol·L-1 NFA组神经元存活率升高(P0.05或P0.01)。流式细胞术检测,与对照组比较,CoCl2损伤组神经元早期凋亡率、晚期凋亡率和总凋亡率升高(P0.05或P0.01);与CoCl2损伤组比较,20、40和80μmol·L-1 NFA组神经元早期凋亡率、晚期凋亡率及总凋亡率下降(P0.05或P0.01)。ELISA法检测,与对照组比较,CoCl2损伤组神经元培养液上清中HIF-1α、caspase-3和Bax表达水平升高(P0.01);与CoCl2损伤组比较,20和40μmol·L-1 NFA组神经元培养上清中HIF-1α、caspace-3和Bax表达水平下降,Bcl-2表达水平升高(P0.05或P0.01)。结论:NFA可以降低CoCl2对海马神经元的缺氧损伤作用,其机制与抑制HIF-1α的表达、下调Bax/Bcl-2及抑制神经元凋亡有关联。
[Abstract]:Aim: to investigate the inhibitory effect of NFAA on apoptosis of hippocampal neurons induced by cobalt chloride (CoCl2) and to elucidate its possible mechanism. Methods: the hippocampal cells were cultured in primary culture and the morphological changes of hippocampal cells were observed under inverted microscope. Hippocampal neurons were randomly divided into two groups: the control group (containing 1% serum culture medium) and the control group (containing 200 渭 mol L-1 NFA) and the NFA group with different concentrations (20 ~ 4080 渭 mol ~ (-1) NFA).MTT) was added to the CoCl2 injury group to detect the survival rate of hippocampal neurons in each group. Apoptosis rate of hippocampal neurons was analyzed by flow cytometry and the expression of apoptotic protein Bcl-2Caspase-3 and hypoxia inducible factor (HIF-1 伪) in the supernatant of cultured neurons was detected by Elisa. Results: the morphology of hippocampal neurons was regular and the synapses intertwined with each other to form a network. MTT assay showed that the survival rate of hippocampal neurons in the CoCl2 injured group was significantly lower than that in the control group, and the survival rate of neurons in the NFA group was significantly higher than that in the CoCl2 injury group (P0.05 or P0.01). Compared with the control group, the neuronal early apoptosis rate, late apoptosis rate and total apoptosis rate were increased by flow cytometry (P 0.05 or P 0.01), and the early apoptosis rate of neurons in the NFA group (20 40 渭 mol L-1) and 80 渭 mol L-1 NFA group (80 渭 mol L-1) was higher than that in the control group (P 0.05 or P 0.01). The late apoptosis rate and the total apoptotic rate were detected by P05 or P0.01).ELISA. Compared with the control group, the expression of HIF-1 伪 -caspase-3 and Bax in the supernatant of cultured neurons in the CoCl2 injured group was higher than that in the CoCl2 injury group, and the expression level of HIF-1 伪 -caspase-3 and Bax in the supernatant of the cultured supernatant of the CoCl2 group was significantly lower than that in the CoCl2 injury group (P 0.05 or P 0.01). Conclusion the effects of CoCl2 on hypoxic injury of hippocampal neurons can be reduced by CoCl2. The mechanism is related to the inhibition of the expression of HIF-1 伪, the down-regulation of Bax/Bcl-2 and the inhibition of neuronal apoptosis.
【作者单位】：吉林医药学院生理学教研室;吉林医药学院病理学教研室;吉林医药学院病原学教研室;
【基金】：国家自然科学基金资助课题(81201671) 吉林省教育厅科研项目资助课题(2014369)
【分类号】：R965

【参考文献】