不同浓度七氟醚对大鼠原代皮质神经元的影响

Effect of different concentrations of sevoflurane on neurotoxicity of primary cultured rat cortical neurons

目的评价不同浓度七氟醚对大鼠原代皮质神经元的毒性作用。方法出生24h内的SD大鼠48只,体外培养大鼠皮质神经元并接种于6孔板(2cm)或12孔板(4.5 mm)培养皿,采用随机数字表法分为四组:1%七氟醚处理组(A组)、2%七氟醚处理组(B组)、4%七氟醚处理组(C组)和对照组(D组),每组12只。A、B、C组神经元分别给予1%、2%和4%浓度的七氟醚处理6h,D组神经元正常培养,采用CCK-8法测定细胞存活率,采用乳酸脱氢酶(LDH)法测定细胞死亡率,采用免疫蛋白印迹(Western blot)法测定凋亡蛋白caspase-3 17kDa片段评价细胞凋亡和促凋亡蛋白Bid、Bim和Puma的表达。结果 B、C组神经元的存活率明显低于D组,死亡率明显高于D组,caspase-3的17kDa片段、Bid、Bim和Puma表达明显高于D组(P<0.05);C组神经元的存活率明显低于B组,死亡率明显高于B组,caspase-3的17kDa片段、Bid、Bim和Puma表达明显高于B组(P<0.05)。A、D组各指标差异均无统计学意义。结论随着吸入七氟醚浓度的升高,七氟醚可能通过促进促凋亡蛋白Bid、Bim和Puma的表达增加对神经元的毒性作用。

Objective To explore the relationship of sevoflurane neurotoxicity with the expression of Bid,Bim,Puma.Methods The cortical neuron from newborn SD rat（within 24h）were seeded in 6or 12 well plate,and then randomly divided into 4groups.Rat culture cortical neurons in vitro exposed in 1%,2%,4% and 0% sevoflurane for 6hwere divided into A,B,C and D group.The effect of neuron viability,death and apoptosis were assessed using CCK-8,LDH and caspase-3cleavage 17 kDa expression assay.The expressions of Bid,Bim and Puma were assessed by western blot.Results Compared with group D,there were significant increases of neuron death and apoptosis,but a decrease of neuron viability,and upregulated expressions of Bid,Bim and Puma in group B（P〈0.05）;Compared with group B,Group C had increased death and apoptosis and decreased viability of neurons,as well as upregulated expressions of Bid,Bim and Puma（P〈0.05）.Conclusion Along with the increase of the concentration,sevoflurane neurotoxicity was increased by upregulation of Bid,Bim,Puma expression.