【摘要】： 目的:觀察氨、谷氨酸培養(yǎng)的SD乳鼠腦星形膠質(zhì)(astrocytes,AS)細(xì)胞水通道蛋白4(aquaporin-4,AQP4)的表達(dá),探討其和肝性腦病腦水腫發(fā)生的關(guān)系。以及分別應(yīng)用地塞米松、二甲基亞砜(Dimethyl sulfoxide,DMSO)和絲裂原活化蛋白激酶(mitogen activated protein kinases,MAPKs)信號(hào)轉(zhuǎn)導(dǎo)通路抑制劑U0126觀察對(duì)其表達(dá)的影響,尋求預(yù)防和治療肝性腦病腦水腫的藥物。方法:通過氨和谷氨酸培養(yǎng)SD乳鼠腦星形膠質(zhì)細(xì)胞,用倒置相差顯微鏡和電鏡觀察細(xì)胞形態(tài)及超微結(jié)構(gòu)、Fluo-3/Am熒光探針觀察細(xì)胞內(nèi)鈣離子變化、自動(dòng)生化分析儀測(cè)定LDH漏出量、采用免疫組織化學(xué)及逆轉(zhuǎn)錄聚合酶鏈?zhǔn)椒磻?yīng)檢測(cè)AQP4在蛋白和基因水平的表達(dá)及其變化,以及分別加用U0126、地塞米松、二甲基亞砜后觀察上述指標(biāo)的變化。結(jié)果:氨和谷氨酸可致培養(yǎng)的星形膠質(zhì)細(xì)胞死亡,星形膠質(zhì)細(xì)胞LDH漏出量增加,細(xì)胞內(nèi)鈣離子濃度增高,AQP4 mRNA和AQP4蛋白的表達(dá)增加。U0126、地塞米松、DMSO可使氨和谷氨酸培養(yǎng)星形膠質(zhì)細(xì)胞LDH漏出明顯減少,抑制細(xì)胞內(nèi)鈣離子的增加,同時(shí)使AQP4 mRNA和AQP4蛋白的表達(dá)減少。結(jié)論:1、氨和谷氨酸可致培養(yǎng)的星形膠質(zhì)細(xì)胞AQP4表達(dá)增加,且隨著氨濃度的升高AQP4表達(dá)也相應(yīng)增加。2、U0126、地塞米松、DMSO可以減少星形膠質(zhì)細(xì)胞AQP4的表達(dá),具有細(xì)胞保護(hù)作用。
[Abstract]:Aim: to investigate the expression of aquaporin 4 (aquaporin-4,AQP4) in astrocytes (astrocytes,AS) of SD rats cultured with ammonia and glutamate, and to explore the relationship between the expression of aquaporin 4 (aquaporin-4,AQP4) and the occurrence of cerebral edema in hepatic encephalopathy. The effects of Dexamethasone, dimethyl sulfoxide (Dimethyl sulfoxide,DMSO) and mitogen activated protein kinase (mitogen activated protein kinases,MAPKs) signal transduction pathway inhibitor U0126 on its expression were observed. To seek drugs for the prevention and treatment of cerebral edema in hepatic encephalopathy. Methods: the astrocytes of SD were cultured with ammonia and glutamate. The morphology and ultrastructure of the cells were observed by inverted phase contrast microscope and electron microscope. The changes of intracellular calcium were observed by Fluo-3/Am fluorescence probe. The leakage of LDH was measured by automatic biochemical analyzer, the expression and changes of AQP4 at protein and gene levels were detected by immunohistochemistry and reverse transcriptase polymerase chain reaction, and U0126 and dexamethasone were added respectively. The changes of above indexes were observed after dimethyl sulfoxide. Results: ammonia and glutamate could induce the death of cultured astrocytes, increase the amount of LDH leakage, increase the intracellular calcium concentration, and increase the expression of AQP4 mRNA and AQP4 protein, U0126, dexamethasone, U0126, dexamethasone, DMSO could significantly reduce the leakage of LDH from astrocytes cultured with ammonia and glutamate, inhibit the increase of intracellular calcium ion, and decrease the expression of AQP4 mRNA and AQP4 protein at the same time. Conclusion: 1. Ammonia and glutamate can induce the increase of AQP4 expression in cultured astrocytes, and the expression of AQP4 increases with the increase of ammonia concentration. 2Dexamethasone and DMSO can reduce the expression of AQP4 in astrocytes. It has cytoprotective effect.
【學(xué)位授予單位】：昆明醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R329

【參考文獻(xiàn)】

相關(guān)期刊論文 前3條

1 高維娟,錢濤;Ca~(2+)超載在缺血性神經(jīng)元損傷中的作用機(jī)制[J];河北醫(yī)學(xué);1999年02期

2 張緒清,聶青和;第十一講 糖皮質(zhì)激素在治療重型肝炎中的應(yīng)用及評(píng)價(jià)[J];實(shí)用肝臟病雜志;2004年02期

3 劉耳,王金兵,吳燕;細(xì)胞凍存保護(hù)劑二甲基亞砜的細(xì)胞毒作用[J];中國預(yù)防獸醫(yī)學(xué)報(bào);1994年04期

，

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