三七总皂苷抗缺氧缺糖再给氧诱导大鼠海马神经细胞凋亡的研究

目的 :以原代培养的大鼠海马神经细胞缺氧 /缺糖再给氧为模型 ,观察三七总皂苷对神经细胞凋亡的抑制作用。方法 :流式细胞仪检测凋亡细胞百分率 ,激光共聚焦扫描显微镜检测细胞内Ca²⁺浓度 ,荧光显微镜观察细胞形态学变化和坏死细胞百分率 ,同时 ,测定细胞乳酸脱氢酶 (LDH)的释放。结果 :神经细胞缺氧 /缺糖 5h后再给氧可诱导神经细胞凋亡和细胞坏死 ,并显著增加细胞内Ca²⁺浓度和LDH的释放 ,并随再给氧时间的延长而增加。三七总皂苷能降低神经细胞凋亡及坏死的百分率 ,降低细胞内Ca²⁺浓度 ,减少LDH的释放 ,三七总皂苷的作用随剂量增加而作用增强。结论 :三七总皂苷对缺血再灌注损伤后海马神经元的凋亡过程具有抑制作用 ,这种作用可能与其能降低细胞内Ca²⁺浓度有关。

Study on the Anti-apopotosis Induced by Hypoxia/hypoglycemia and Reoxygenation of Panax Notoginseng Saponins in Cultured Rat Hippocampal Neurons

Objective: To study the inhibitory effects of Panax Notoginseng Saponins(PNS) on apoptosis induced by hypoxia/hypoglycemia and reoxygenation in cultured rat hippocampal neurons. Method: Apoptosis were measured by flow cytometry, intracellular free calcium concentration([Ca²⁺]i)was measured with confocal laser scanning microscopy,morphological changes and neuronal necrosis were observed with fluorescence microscope, and meanwhile the leakage of lactic dehydrogenase(LDH) was measured. Result: Hypoxia/hypoglycemia cultures for 5 hours and reoxygenation induced neuronal apoptosis and necrosis,and significantly increased neuronal [Ca²⁺]i and the leakage of LDH. The effects were increased with the extending time of reoxygenation. PNS has could significantly decrease the percentage of neuronal apoptosis and necrosis,and reduce neuronal i and the leakage of LDH. Conclusion: PNS has inhibitory effect on neuronal apoptosis. This effect might be related to its effect of decreasing intracellular free calcium concentration.pocampalneurons.Method :Apoptosisweremeasuredbyflowcytometry ,intracel lularfreecalciumconcentration([Ca²⁺]i)wasmeasuredwithconfocallaserscanningmicroscopy ,morphologicalchangesandneuronalnecrosiswereobservedwithfluorescenc.