摘要
目的探讨7,8-二羟基-4-甲基香豆素(Dhmc)对谷氨酸诱导的PC12细胞损伤的神经保护作用。方法采用谷氨酸诱导PC12细胞损伤凋亡模型,给予Dhmc处理后,MTT法检测细胞增殖;化学检测法测定乳酸脱氢酶(LDH)漏出量、细胞上清液中丙二醛(MDA)含量及超氧化物歧化酶(SOD)水平;采用Annexin-V-FITC双染法检测细胞凋亡率;采用荧光法检测活性氧(ROS)的生成;采用流式细胞仪检测线粒体膜电位(MMP);Western blot法检测Bcl-2、Bax、Cyt C、cleaved caspase-3蛋白表达。结果 Dhmc可显著降低由谷氨酸诱导的细胞损伤,提高了细胞存活率,减少LDH漏出量,降低MDA含量,增加SOD活性,抑制ROS的生成,维持线粒体膜电位水平,下调Bcl-2/Bax比率,减少Cyt C的释放,降低Caspase-3活性。结论 Dhmc具有减轻由谷氨酸诱导的PC12细胞损伤的作用,其机制可能与抗氧化应激和减轻线粒体损伤有关。
Objective To determine the neuroprotective effect of 7, 8-dihydroxy-4-methylo-umarin against glutamate induced PC12 cell injury. Methods PC12 cells were treated with glutamate to establish the cell model. After Dhmc treatment, MTT assay was used to analyze the PC 12 cell viability. The contents of LDH, MDA and the level of SOD in the cells were determined by kit respectively. Cell apoptosis detection was performed with an Annexin-V-FITC Apoptosis Detection Kit. The production of ROS was determined by fluorescence method. The mitochondrial membrane potential （MMP） was measured by flow cytometer. Western blot was carried out to detect the protein expression. Results Dhmc significantly decreased the contents of LDH and MDA, enhanced the activity of SOD induced by glutamate. In addition, Dhmc treatment decreased the intracellular ROS generation and maintained the level of MMP. Finally, Dhmc treatment increased the ratio of Bcl-2/Bax, reduced the release of Cyt C, decreased the caspase-3 activity. Conclusion Dhmc can significantly ameliorate glutamate induced PC 12 cell injury. The mechanism may be partly through anti-oxidative stress and alleviating mitochondrial damage.
出处
《中南药学》
CAS
2017年第4期450-454,共5页
Central South Pharmacy
