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目的:观察川芎嗪预给药对胎鼠缺氧/复氧海马神经细胞凋亡的影响。方法:胎鼠海马神经细胞培养鉴定后,随机分为5组:正常组(C组)、缺氧/复氧损伤组(A/R组)、不同剂量川芎嗪组(L组、M组和H组)。C组不制备缺氧/复氧模型;A/R组制备缺氧/复氧模型;L组、M组和H组加入川芎嗪,终浓度分别为60、200和800μg/mL,孵育1h后制备缺氧/复氧模型。缺氧/复氧模型制备方法:海马神经细胞置入90%N2加10%CO2培养箱中孵育2h诱导缺氧,然后放入37℃、5%CO2培养箱中复氧24h。处理结束后在倒置显微镜下观察海马神经细胞形态,用透射电镜观察海马神经细胞超微结构的变化,测定海马神经细胞凋亡率、细胞活力。结果:与C组比较,A/R组海马神经细胞活力明显下降(P<0.01);与A/R组比较,L组、M组和H组预给药均可升高神经细胞活力(P<0.05);其中M组效果最明显,与L组、H组比较,差异均有显著性意义(P<0.05)。与C组比较,A/R组后海马神经元凋亡比例明显增加(P<0.01)。与A/R组比较,L组、M组和H组预给药均可不同程度降低海马神经细胞的凋亡率(P<0.01)。其中M组效果最为显著,与L组、H组比较,差异均有非常显著性意义(P<0.01)。结论:川芎嗪有明显的神经保护作用,该作用具有剂量相关性,适中浓度的川芎嗪保护作用最显著。
Objective: To observe the effect of ligustrazine on apoptosis of fetal rat hypoxia / reoxygenation hippocampus neurons. Methods: Fetal rat hippocampal neurons were cultured and identified and randomly divided into 5 groups: normal group (C group), hypoxia / reoxygenation injury group (A / R group), different doses of Ligustrazine group (L group, M group and H group). Hypoxia / reoxygenation model was prepared in A / R group; Ligustrazine was added to L group, M group and H group, the final concentrations were 60, 200 and 800μg / mL, respectively. After incubation for 1h Preparation of hypoxia / reoxygenation model. Hypoxia / reoxygenation model preparation method: Hippocampal neurons were placed in 90% N2 plus 10% CO2 incubator incubated for 2h to induce hypoxia, and then placed in 37 ℃, 5% CO2 incubator reoxygenation 24h. After the treatment, the morphological changes of hippocampal neurons were observed under an inverted microscope. The changes of the ultrastructure of hippocampal neurons were observed by transmission electron microscopy. The apoptosis rate and cell viability of hippocampal neurons were measured. RESULTS: Compared with group C, the viability of neurons in hippocampus of A / R group was significantly decreased (P <0.01). Compared with A / R group, pretreatment with L, M and H groups increased neuronal activity <0.05). The effect of M group was the most obvious, and there was significant difference between L group and H group (P <0.05). Compared with group C, the percentage of apoptotic hippocampal neurons in A / R group was significantly increased (P <0.01). Compared with A / R group, L group, M group and H group pretreatment can reduce the apoptosis rate of hippocampus neurons to some extent (P <0.01). Among them, M group had the most significant effect, and there was significant difference between L group and H group (P <0.01). Conclusion: Tetramethylpyrazine has obvious neuroprotective effect. The effect is dose-dependent, and the protective effect of ligustrazine with moderate concentration is the most significant.