三磷酸腺苷与神经生长因子对缺氧复氧后大鼠皮质神经元细胞活性及凋亡的影响

Effects of ATP and NGF on rat cortical neurons viability andapoptosis induced by anoxia-reoxygenation in vitro

目的探讨体外三磷酸腺苷(ATP)、神经生长因子(NGF)对缺氧复氧大鼠皮质神经元的保护作用。方法取体外培养8d的Wistar大鼠皮质神经元,随机分为正常对照组、缺氧组、ATP组和NGF组。用噻唑盐比色法(MTT)、乳酸脱氢酶(LDH)法测定细胞活性变化,流式细胞仪术、透射电镜检测细胞凋亡率和凋亡。结果与对照组比较,缺氧组神经元细胞活性明显降低,细胞凋亡率显著增加;ATP和NGF能明显改善缺氧对神经元细胞活性的影响,细胞凋亡率降低(P<0.05);与NGF组比较,ATP组神经元细胞活性增高,细胞凋亡率降低(P<0.05)。结论缺氧能够造成大鼠大脑皮质神经元的损伤并诱导其凋亡,ATP及NGF对缺氧复氧后大脑皮质神经元有保护作用。

Objective To investigate the effects of adenosine triphasphate （ATP） and nerve growth factor （NGF） on cell viability and apoptosis in cultured rat cortical neurons after anoxia-reoxygenation in vitro. Methods Cortical neurons cultured for 8 days were randomly divided into four groups ： control group, anoxic group（ anoxia for 6 hrs and reoxygenated for 24hrs） , NGF group（ anoxic group ＋ 40 ng/ml NGF in culture fluid） and ATP group（anoxic group ＋ 1μmol/L ATP in culture fluid）. The neurons viability were detected with the method of MTT and LDH. The apoptosis of neurons was examined by flow cytometry and electro-microscope. Results In comparison with control group, Anoxia inhibited the cell viability and increased the apoptosis rate and death of cortical neurons. ATP and NGF reversed upper progress. Compared with NGF, ATP improved the cell viability （P 〈 0. 05 ）, ameliorate the ratio of apoptosis （P 〈 0.05） greatly. Conclusions Anoxia damages cortical neurons and induces apoptosis Both ATP and NGF offer neuroprotective effects to anoxia-reoxygenation cultured rat cortical neurons.