七氟醚对神经元凋亡保护作用的剂量-效应关系

Dose-efficiency relation of the protective effect of sevoflurane on neuronal apoptosis

目的研究七氟醚在原代神经元培养细胞缺氧/缺糖(oxygen and glucose deprivation,OGD)模型中的脑保护作用的剂量-效应关系。方法共使用103孔培养细胞。对照组(n=8)仅行OGD而不加任何试剂;七氟醚(Sev)1、2、3组(每浓度组,n=8)分别加入与1、2、3MAC相对应浓度的溶解于PBS液的七氟醚,分别在OGD前、中、后给予七氟醚;OGD+PBS液组(n=2)只给予不含溶解七氟醚的PBS液。另取1孔神经元,不改变其培养液的成分,只在37℃孵育3h以作为阴性对照。OGD时间3h。OGD结束4h后采用双参数流式细胞仪法测定神经元凋亡情况。结果与对照组及PBS组相比,七氟醚各剂量组FITC值降低,并呈剂量依赖性(P<0.01)。结论在OGD前、中、后给予1、2、3MAC七氟醚都能呈剂量依赖性减少神经元的凋亡。

Objective To determine the effects of sevoflurane in vitro on neuronal apoptosis in the presence of oxygen and glucose deprivation（OGD）. Methods Neuronal cell cultures were prepared from neonatal rat cortex and were used between 10 and 14 days in vitro. Ninety -six culture dishes （n =8 for each concentration group） were exposed to 0, 1,2 or 3 MAC sevoflurane for 30 minutes before,during,and after OGD. The remaining 7 dishes were treated as follows： one had no media change （control）,six dishes were incubated in PBS only before, during, and after OGD. The cultures were then deprived of oxygen and glucose for 3 hours. Four hours after exposure, flow eytometric （FACS） analysis was applied to Annexin-V and Propidium Iodide （PI） stained cells to evaluate the degree of the apoptosis. Results Exposure of cultured neurons to sevoflurane elicited a significant inhibition of OGD-induced neuronal apoptosis （P〈0.01）. The effect was demonstrated in a concentration-dependent manner at the concentrations as low as 1.0 MAC （P〈0.01）. Conclusion Sevoflurane at 1.0,2.0,or 3.0 MAC can attenuate in vitro the neuronal apoptosis in a concentrationdependent manner whether it is given before,during or after OGD.