知识与财富的链接
目的: 研究H2S对缺氧/复氧后神经元存活信号转导通路PI3K/Akt/P70S6K的影响。方法: 取96孔和6孔培养板上培养7 d的海马神经元,正常培养组（C组）神经元按正常培养方法培养。NaHS组在进行缺氧/复氧时加入NaHS使其终浓度为150 μmol/L。Tri组、Rap组和Tri+Rap组在加入150 μmol/L NaHS的同时分别加入triciribin 10 μmol/L、rapamycin 10 nmol/L或triciribin 10 μmol/L+rapamycin 10 nmol/L。96孔培养板的神经元进行细胞存活力的检测。6孔培养板的神经元进行神经元纯度鉴定、神经元凋亡、cAMP和PI3-K、Akt和P70S6K蛋白表达的检测。结果: NaHS显著增加了cAMP的浓度和PI3、Akt、P70S6K蛋白的表达,同时增加了神经元存活率、降低了神经元凋亡率（P<0.01 vs C组和A/R组）。Triciribin抑制了Akt和P70S6K表达同时降低了神经元存活率、升高了神经元凋亡率（P<0.05,P<0.01 vs NaHS组）。Rapamycin抑制了P70S6K表达同时降低了神经元存活率、升高了神经元凋亡率（P<0.05,P<0.01 vs NaHS组）。结论： H2S通过cAMP激活了PI3K/Akt/P70S6K信号通路,抑制缺氧/复氧后海马神经元的凋亡。
AIM: To investigate the effect of hydrogen sulfide on neuron apoptosis through PI3-K/Akt/P70S6K cell-survival signal transduction pathways after neuron anoxia-reoxygenation.METHODS: Newborn (24-48 h) Wistar rats were decapitated.The hippocampus tissue was dissected and cells were suspended.Cells were plated at 1.0×108 cells/L on poly-dlysine-treated 96-well (100 μL/well) plates and 6-well (2 mL/well) plates.Cells were used after 7 days.For anoxia-reoxygenation (oxygen glucose deprivation,OGD) experiments,cells were washed three times in a glucose-free balanced salt solution (BSS).They were then placed in deoxygenated glucose-free medium and cultured under 95% N2,5% CO2 in an anaerobic chamber equilibrated to 37 ℃ and 100% humidity for 45 min.OGD was terminated by replacement of stored medium and by returning the cultures to a standard incubator maintained at 37 ℃ in 95% air,5% CO2.In experimental group,cells were respectively carried out OGD,OGD+150 μmol/L NaHS,OGD+150 μmol/L NaHS+10 μmol/L triciribin,OGD+150 μmol/L NaHS+10 nmol/L rapamycin and OGD+150 μmol/L NaHS+10 μmol/L triciribin+10 nmol/L rapamycin.Control cells were cultured normally.24 h later,neuron viability and apoptosis were measured.The level of cAMP and protein expression of PI3-K,Akt and P70S6K were detected.RESULTS: NaHS enhanced concentration of cAMP and expression of PI3-K,Akt and P70S6K.Meanwhile,increased neuron viability and decreased neuron apoptosis (P<0.01 vs group C or group I/R) were observed.Triciribin inhibited Akt and P70S6K,as well as increased neuron apoptosis and decreased neuron viability (P<0.05,P<0.01 vs group NaHS).Rapamycin inhibited P70S6K,as well as increased neuron apoptosis and decreased neuron viability (P<0.05,P<0.01 vs group NaHS).CONCLUSION: H2S inhibits hippocampus neuron apoptosis and protects neuron from anoxia-reoxygenation injury through cAMP-mediated PI3-K/Akt/P70S6K kinase cell-survival signaling pathways.
