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目的:以缺氧/缺糖再给氧为模型,观察清开灵注射液对神经细胞线粒体膜电位的保护作用。方法:四唑盐比色实验(MTT)检测细胞线粒体活性,激光共聚焦扫描显微镜检测细胞内游离钙离子含量([Ca2+]i),流式细胞术检测细胞凋亡百分率和线粒体膜电位。结果:缺氧-缺糖5 h再给氧3 h时,细胞内[Ca2+]i和细胞凋亡率明显增加,并随再给氧时间的延长而明显增高,线粒体膜电位和活性明显降低,并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细胞内[Ca2+]i浓度,抑制细胞凋亡的发生,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的降低,从而具有稳定线粒体膜电位的作用,抑制细胞凋亡的发生,而对海马神经细胞起到保护作用,这种作用可能与其能抑制神经细胞内钙超载有关。
OBJECTIVE: To observe the protective effect of Qingkailing injection on mitochondrial membrane potential of nerve cells by hypoxia/glucose reoxygenation model. Methods: Cell mitochondrial activity was measured by tetrazolium salt colorimetric assay (MTT). Intracellular free calcium ion content ([Ca2+]i) was detected by confocal laser scanning microscopy. Apoptosis percentage and mitochondrial membrane potential were measured by flow cytometry. RESULTS: During hypoxia-hypoglycemia for 5 h and then for 3 h, the intracellular [Ca 2+ ] i and apoptosis rate were significantly increased, and the mitochondrial membrane potential and activity were significantly decreased with prolonged reoxygenation time. With the further extension of oxygen supply time, Qingkailing injection can significantly reduce intracellular [Ca2+]i concentration, inhibit the occurrence of apoptosis, increase mitochondrial membrane potential and activity, and hypoxia-glucose re-supply There was a significant difference between the oxygen group (P<0.05, <0.01). Conclusion: Qingkailing injection can inhibit the decrease of mitochondrial membrane potential caused by hypoxia-hypoglycemia and reoxygenation injury, which can stabilize the mitochondrial membrane potential, inhibit the occurrence of apoptosis, and play a role in hippocampal neuronal cells. The protective effect may be related to its ability to inhibit calcium overload in nerve cells.