延胡索乙素对乳鼠海马神经元缺氧缺糖损伤保护作用的研究

Study of the Protective Effect of Tetrahydropalmatine on the Injury of Suckling Rat Hippocampal Neurons due to Hypoxia and Sugar Deficiency

目的:探讨延胡索乙素对乳鼠海马神经元细胞因缺糖缺氧所致损伤的保护作用。方法:建立乳鼠海马神经元细胞缺糖缺氧损伤模型。利用CCK法和预实验确定加药浓度后,将培养的细胞随机分成6组:正常组、模型组、延胡索乙素低、中、高治疗组(2、4、8μg/mL)和阳性药物对照组尼莫地平(8μg/mL)。观察延胡索乙素对乳鼠海马神经元乳酸脱氢酶(LDH)的释放量,细胞一氧化氮(NO)和丙二醛(MDA)含量,一氧化氮合酶(NOS)、超氧化歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-PX)活性的影响;采用流式细胞仪检测各组细胞的凋亡率。结果:与模型组相比,延胡索乙素中、高剂量组均能显著增加细胞内SOD以及GSH-PX的活性(P<0.01);能够显著减少细胞内LDH外漏以及抑制NOS的活性(P<0.01);能够显著减少细胞内NO以及MDA的生成量(P<0.01)。结论:延胡索乙素对乳鼠海马神经元细胞的缺糖缺氧损伤显示出良好的保护作用,其机制可能与抗氧化、抑制细胞凋亡有关。

Objective To investigate the protective effect of tetrahydropalmatine on the injury of suckling rat hippocampal neurons due to hypoxia and sugar deficiency. Methods To establish the model of hypoglycemic anoxic injury in suckling rat hippocampal neurons. CCK method and preliminary experiment were used to determine the drug concentration, the cultured cells were randomly divided into 6 groups: normal group, model group, low, medium and high tetrahydropalmatine treatment group(2, 4, 8 μg/mL) and positive control group nimodipine(8 μg/mL). To study the effect of tetrahydropalmatine on the release quantities of lactate dehydrogenase(LDH), nitric oxide(NO) and malondialdehyde(MDA) content, nitric oxide synthase(NOS), super oxide dismutase(SOD) and glutathione peroxidase(GSH-PX) activities in suckling rat hippocampal neurons;Cell apoptosis rate was detected by flow cytometry. Results Compared with the model group, the activity of SOD and GSH-PX in the cells were significantly increased in both middle and high dose groups(P<0.01). It could significantly reduce the intracellular LDH leakage and inhibit the activity of NOS(P<0.01). The productions of NO and MDA in cells(P<0.01) were reduced by tetrahydropalmatine;It could obviously inhibit early apoptosis of cells;the protein expression levels of caspase-3 and caspase-8 were suppressed by tetrahydropalmatine. Conclusion Tetrahydropalmatine showed a good protective effect on the hypoglycemic anoxia damage in suckling rat hippocampal neurons, and its mechanism may be related to anti-oxidation and inhibition of apoptosis.