七氟烷通过P~(70S6K)/Nrf2信号通路诱导神经元HO-1mRNA表达

目的探讨七氟烷(Sevoflurane)诱导神经元血红素氧合酶-1(HO-1)基因表达的信号转导通路。方法将培养7d的新生大鼠海马神经元随机分为4组:正常培养组(C组)、2%七氟烷组(S1组)、4%七氟烷组(S2组)和4%七氟烷+Rapamycin组(R组)。C组神经元按正常培养方法培养。S1组和S2组神经元分别给予2%或4%七氟烷处理60min后继续培养24h。R组在神经元给予4%七氟烷处理同时在培养液中加入Rapamycin使其终浓度为10nmol.L-1后同S2组处理。收集神经元进行HO-1mRNA和P70S6K、Nrf2、AP-1和HO-1蛋白表达的检测。结果S1组P70S6K和Nrf2蛋白表达增加(vsC组,P<0.01),HO-1mRNA和HO-1蛋白表达增加(vsC组,P<0.01),AP-1蛋白表达变化不明显(vsC组,P>0.05)。S2组P70S6K和Nrf2蛋白表达增加(vsS1组,P<0.05),HO-1mRNA和HO-1蛋白表达增加(vsS1组,P<0.05),AP-1蛋白表达变化不明显(vsS1组,P>0.05)。R组P70S6K和Nrf2蛋白表达减少(vsS2组,P<0.01),HO-1mRNA和HO-1蛋白表达减少(vsS2组,P<0.01),AP-1蛋白表达变化不明显(vsS2组,P>0.05)。结论Sevoflurane通过P70S6K/Nrf2信号通路诱导神经元HO-1mRNA表达。

Sevoflurane induces neuron HO-1 mRNA expression via P'70S6K/Nrf2 kinase signal pathways

Aim To investigate signal pathway associated with sevoflurane-induced neuron hemeoxygenase-1(HO-1) expression.Methods Newborn(24～48 h) Wistar rats were decapitated and hippocampus tissue was digested with 0.125% trypsin,and suspended in a medium containing DMEM supplemented to 25 mmol·L-1glucose,10% fetal bovine serum,10% horse serum,and 2 mmol·L-1glutamine.Cells were plated at 1.0×106·L-1 on poly-Dlysine-treated 6-well(2 ml/well) plates and were treated with 10 μmol·L-1 cytosine arabinoside on day 4 to minimize glial growth.One-half of the medium was replaced twice a week with medium consisting of DMEM(4.5 g·L-1 glucose)/F12(1 :1),5% fetal bovine serum and 5% horse serum.Cells were used after growing for 7 days.Cells were randomly divided into 4 groups:control group(C group),2% Sevoflurane group(S1 group),4% Sevoflurane group(S2 group)and 4% Sevoflurane+Rapamycin group(R group).Control cells were cultured normally.Group S1 and S2 cells were preconditioned with 2% or 4% Sevoflurane,then cultured normally for 24 hours.Group R cells culture medium was added with 10 nmol·L-1 Rapamycin and preconditioned with 4% Sevoflurane,then cultured normally for 24 hours.The expression of HO-1 mRNA,P70S6K,Nrf2,AP-1 and HO-1 protein was detected.Results 2% Sevoflurane enhanced neuron expression of P70S6K and Nrf2(vs group C,P<0.01),and increased neuron expression of HO-1 mRNA and HO-1(vs group C,P<0.01).4% Sevoflurane enhanced neuron expression of P70S6K and Nrf2(vs group S1,P<0.05),and increased neuron expression of HO-1 mRNA and HO-1(vs group S1,P<0.05).Rapamycin inhibited neuron expression of P70S6K and Nrf2(vs group S2,P<0.01),and inhibited neuron expression of HO-1 mRNA and HO-1(vs group S2,P<0.01).There was no difference of AP-1 protein expression had in each group.Conclusion Sevoflurane induces neuron HO-1 mRNA expression via P70S6K/Nrf2 kinase signal pathways.