清开灵注射液对神经细胞凋亡和线粒体膜电位的影响

Influences of Qingkailing Injection on neuron apoptosis and mitochondrial membrane potential

目的以缺氧、缺糖再给氧为模型，观察清开灵注射液对大鼠海马神经细胞凋亡和线粒体膜电位的影响。方法四唑盐比色实验（MTT）检测细胞线粒体活性，流式细胞术检测细胞凋亡百分率和线粒体膜电位，荧光显微镜观察细胞形态学变化和坏死百分率。结果缺氧、缺糖5h再给氧3、12、24h时，细胞凋亡率明显增高，形态学观察也发现大量神经细胞的染色质凝聚、核碎裂、凋亡小体产生；细胞线粒体膜电位和活性均显著降低，随再给氧时间的延长而进一步下降，清开灵注射液能显著降低神经细胞凋亡及坏死的百分率，提高线粒体膜电位和活性，与缺氧、缺糖再给氧组相比均有显著性差异（P〈0．05-0．01）。结论清开灵注射液可抑制缺氧、缺糖再给氧损伤所致的线粒体膜电位降低，从而稳定线粒体膜电位、抑制细胞凋亡的发生，而对海马神经细胞起到保护作用。

Objective To observe the influences of Qingkailing Injection （QI） on apoptosis of hippocampal neurons and mitochondrion membrane potential （MMP） in rat model of hypoxiahypoglycemia-rexyenation （HHR）. Methods Mitochondrion activity was detected by methyl thiazolyl tetrasolium test （MTI＇）. The apoptosis percentage and MMP were detected by flow cytometry, and the cytomorphological changes and neuron necrotic percentage were observed with fluorescence microscope. Results After hypoxia-hypoglycemia treating in rats for 5 hours and reoxygenation treating for 3,12,24 hours the apoptosis rate of neurons increased significantly. There were chromatinic coacervation, karyorrhexis and apoptotic bodies＂ in neurons were observed cytomorphologically. The MMP and mitochondrion activity decreased significantly after reoxygenation for 3,12,24 hours, and decreased further along with the prolongation of reoxygenation. QI could decrease obviously percentage of neuron apoptosis and necrosis and increase MMP and mitochondrion activity with a significant difference （P 〈 0.05 - 0.01 ） compared with the HHR group. Conclusion Qingkailing Injection could inhibit the decrease of MMP induced by injury of hypoxia-hypoglycemia-reoxygenation, slabilize MMP, inhibit cell apoptosis and protect hippocampal neurons.