本文選題：硫化氫　切入點(diǎn)：氧糖缺失/恢復(fù)　出處：《華中科技大學(xué)》2013年碩士論文　論文類型：學(xué)位論文

【摘要】：目的研究外源性硫化氫（hydrogensulfide，H_2S）對大鼠海馬神經(jīng)元氧糖缺失/恢復(fù)（oxygen-glucosedeprivationandrecovery,OGD/R)損傷后的作用，并探討海馬神經(jīng)元OGD/R損傷后缺氧誘導(dǎo)因子-1α（hypoxia-induciblefactor-1α,HIF-la）的變化及H_2S的干預(yù)作用。 方法孕16-18d胎鼠海馬神經(jīng)元原代培養(yǎng)，培養(yǎng)7d后神經(jīng)元特異性烯醇化酶（neuron-specificenolase，NSE）免疫組化法作神經(jīng)元鑒定，培養(yǎng)第8天時OGD1h/R24h造成缺氧/復(fù)氧損傷，缺氧同時給予硫氫化鈉（sodiumhydrosulfide,NaHS）干預(yù)。將大鼠海馬神經(jīng)元隨機(jī)分為3組：正常培養(yǎng)組（Ⅰ組）、OGD/R組（Ⅱ組）、OGD/R+NaHS組（Ⅲ組），Ⅲ組又根據(jù)NaHS濃度分為Ⅲ1-5亞組；NaHS濃度分別為25、50、100、200、400μmol/L。MTT法測定各組細(xì)胞活力，比色法檢測各組培養(yǎng)液中乳酸脫氫酶（lactatedehydrogenase,LDH）活性，流式法檢測各組細(xì)胞凋亡，RT-PCR法檢測各組細(xì)胞Caspase-3mRNA,HIF-1mRNA表達(dá)。 結(jié)果（1）與正常組相比較，模型組細(xì)胞活力明顯降低，培養(yǎng)液中LDH漏出增多，凋亡加重，Caspase-3mRNA、HIF-1αmRNA表達(dá)增高（P＜0.01）；（2）與模型組相比較，Ⅲ1組細(xì)胞活力、培養(yǎng)液中LDH漏出、凋亡、Caspase-3mRNA、HIF-1αmRNA表達(dá)的差異無統(tǒng)計(jì)學(xué)意義（P＞0.05）；（3）與模型組相比較，Ⅲ2-4組細(xì)胞活力明顯增高，培養(yǎng)液中LDH漏出減輕，凋亡減輕，Caspase-3mRNA表達(dá)降低，HIF-1αmRNA表達(dá)明顯增高（P＜0.01）；(4）與模型組相比較，Ⅲ5組細(xì)胞活力明顯降低，，培養(yǎng)液中LDH漏出增多，凋亡加重，Caspase-3mRNA表達(dá)增高，HIF-1αmRNA表達(dá)明顯降低（P＜0.01）。 結(jié)論低濃度H_2S（25μmol/L）對海馬神經(jīng)元OGD/R損傷無明顯影響，中濃度H_2S（50-200μmol/L）可以減輕海馬神經(jīng)元OGD/R損傷，增加細(xì)胞活力，減輕細(xì)胞凋亡，但高濃度H_2S（400μmol/L）則可加重其損傷；海馬神經(jīng)元OGD/R損傷后HIF-1αmRNA表達(dá)增強(qiáng)，適當(dāng)濃度H_2S（50-200μmol/L）可能通過上調(diào)HIF-lα的表達(dá)發(fā)揮保護(hù)作用。
[Abstract]:Objective to study the effect of exogenous hydrogen sulfide (H2S) on oxygen-glucose privacy and recovery of oxygen-glucose privacy and recovery after oxygen-glucose deprivation and recovery of hypoxic-inducible factor-1 偽 hypoxia-inducible factor-1 偽 (HIF-laA) in hippocampal neurons after OGD/R injury in rats and to explore the effect of H2S on HIF-la-la-induced by hypoxia inducible factor 1 偽 hypoxia-inducible factor-1 偽 (HIF-laa) in hippocampal neurons after injury of hypoxia inducible factor 1 偽 hypoxia-inducible factor-1 偽 (HIF-laa). Methods Neuron specific enolase neuron-specific enolase (NSE) immunohistochemical method was used to identify the neurons in hippocampal neurons of fetal rats on the 16th to 18th day of gestation. Hypoxia / reoxygenation injury was induced by OGD1h/R24h on the 8th day of culture. The hippocampal neurons of rats were randomly divided into three groups: normal culture group (group 鈪

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