【作者】 谭少华；

【导师】 徐安定；

【作者基本信息】 暨南大学 ， 神经病学， 2004， 硕士

【摘要】 目的：确定内皮素(endothelin,ET)-1能否直接诱导皮层神经元释放一氧化氮(Nitric oxide,NO)，以及所经过的ET受体亚型。 方法：神经元培养取自SD乳鼠(出生24h内)大脑皮层细胞，体外原代培养7d用于实验。首先以不同浓度的ET-1(10 nM，100 nM)刺激神经元，在加药前，加药后30 min和加药后6h收集培养基，以硝酸还原酶法检测培养基中NO的浓度。接着，在10 nM ET-1处理的同时，分别加入BQ123(选择性ET_A受体拮抗剂)、BQ788(选择性ET_B受体拮抗剂)、PD145065(非选择性ET受体拮抗剂)，作用6h后检测培养基中NO浓度。 结果：10nM及100 nM ET-1作用6 h后，神经元培养液中NO的浓度明显高于对照组(P值分别为0.029，0.015)，而二者之间则没有显著性差异(P=1.000)。BQ788和PD145065可分别完全阻断ET-1刺激神经元释放NO的作用，使其培养液中NO浓度保持与对照组一致水平(各P＞0.05)，而明显低于单独ET-1组(各P＜0.05)。BQ123对ET-1刺激神经元释放NO无影响，其培养液中NO浓度与单独ET-1组相近(P=0.987)，均高于对照组(各P＜0.05)。 结论：外源性ET-1可通过ET_B受体直接诱导培养SD大鼠大脑皮层神经元释放NO。更多还原

【Abstract】 Objective: To observe the effect of endothelin (ET)-l on nitric oxide (NO) release in cultured rat cortical neurons and the ET receptor subtype(s) involved.Methods: Primary cortical neuron cultures were prepared from neonate SD rat brain and incubated for 7 days into confluence. Firstly, the cultures were stimulated with ET-1 (10 nM, 100 nM, respectively), the medium was collected at 0 min, 30 min, 6 h interval for detection of NO concentration using nitrate reductase method. Thereafter, in addition to 10 nM ET-1, BQ123 (a selective ETA receptor antagonist), BQ788 (a selective ETa receptor antagonist) or PD145065 (a non-selective ET receptor antagonist) was simultaneously added into the cultures, respectively, and the medium was collected 6 h after the administration for NO concentration measurement.Results: NO concentration in Culture medium significant increased and washigher than that in control group after treated with 10 nM ET-1 and 100 nM ET-1 for 6 h (P=0.029, 0.015, respectively), and no significant difference of NO levels in 10 nM ET-1 and 100 nM ET-1 was observed. BQ788 or PD145065, but not BQ123, completely blocked the effect of ET-1 on NO release from cultured neurons, making the NO levels in medium unchanged as compared to control (P>0.05, respectively) and obviously lower than that in ET-1 group (P<0.05, respectively).Conclusions: ET-1 can directly stimulate cultured cortex neurons to release NO via ETB receptors.更多还原