氧化槐定碱对新生大鼠海马神经元缺氧损伤的保护作用及其作用机制

Protective effects and mechanisms of OSR on primary cultured hippocampus neurons subjected to anoxic injury in neonatal rat

目的:研究氧化槐定碱(oxysophoridine,OSR)对原代培养的新生大鼠海马神经元缺氧损伤的保护作用及其机制。方法:以原代培养的新生大鼠海马神经元为研究对象,用无糖培养液结合物理性缺氧建立缺氧损伤模型,测定神经细胞的存活率、乳酸脱氢酶(LDH)漏出率以及细胞中丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-PX)、一氧化氮合酶(NOS)和一氧化氮(NO)水平的变化。采用荧光双波长分光光度计测定神经细胞内游离钙离子浓度([Ca2+]i)的变化。结果:缺氧损伤模型组能在短时间内造成神经元的死亡,LDH漏出率的增多,MDA,NO含量增多,NOS活性升高,SOD,GSH-PX活性降低,细胞内[Ca2+]i增加。OSR组(0.625,5,10μg.L-1)能不同程度地降低缺氧模型对神经元的损伤。结论:氧化槐定碱对新生大鼠海马神经元缺氧损伤具有明显的保护作用,其机制可能与减轻细胞内钙离子超载以及抗氧化损伤有关。

Objective: To investigate the protective effects of oxysophoridine(OSR) on primary cultured hippocampus neurons subjected to anoxia injury in neonatal rats and its mechanism.Method: The model of anoxia injury of hippocampus neurons in neonatal rats were primarily cultured in vitro by physical oxygen deficiency using glucose-free culture fluid.The survival rate of neurons,the leaking rate of lactate dehydrogenase(LDH),the intracellular contents of malondialdehyde(MDA) and nitric oxide(NO),the activities of superoxide dismutase(SOD),glutathione peroxidase(GSH-PX) and nitric oxide synthase(NOS) were measured.The intracellular free calcium concentration(i) in hippocampus neurons were detected with Ca2+-sensitive dual wavelength fluorescence spectrophotometer.Result: Neuron death occurred in the anoxia injury model group with increase of LDH leaking rate,the contents of NO,MDA,intracellular i and the elevated activity of NOS while decreased activities of SOD and GSH-PX.The hippocampus neurons subjected to anoxia injury were alleviated in OSR(0.625,5,10 μg·L-1) group.Conclusion: OSR has significant protective effects on hippocampus neurons subjected to anoxic injury.The mechanism of its protective effect may relate to its reduction of calcium overload and against oxidation injury.