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《谷氨酰胺對(duì)大鼠視網(wǎng)膜興奮性損傷的保護(hù)作用》由會(huì)員上傳分享，免費(fèi)在線(xiàn)閱讀，更多相關(guān)內(nèi)容在學(xué)術(shù)論文-天天文庫(kù)。
1、摘要目的：研究腹腔注射谷氨酰胺對(duì)大鼠視網(wǎng)膜興奮性損傷中Bm3a表達(dá)的影響。方法：健康清潔型Wistar大鼠55只隨機(jī)分為三組，正常對(duì)照組5只，實(shí)驗(yàn)對(duì)照組25只、實(shí)驗(yàn)組25只。正常對(duì)照組不做任何處理，實(shí)驗(yàn)對(duì)照組和實(shí)驗(yàn)組中所有大鼠玻璃體腔注射2gl80mmol／1NMDA制作視網(wǎng)膜興奮性損傷模型，實(shí)驗(yàn)組在大鼠視網(wǎng)膜興奮性損傷模型制作7h后腹腔注射谷氨酰胺(0．75g／kg)。實(shí)驗(yàn)組及實(shí)驗(yàn)對(duì)照組分別在造模后8h、12h、16h、24h、48h各隨機(jī)處死5只大鼠，正常對(duì)照組大鼠在不同時(shí)間點(diǎn)隨機(jī)處死一只。HE染色光鏡下觀察各組視網(wǎng)膜組織的形態(tài)學(xué)變化，免疫組織化學(xué)染色及實(shí)時(shí)熒光定量RT-PCR方法檢測(cè)視網(wǎng)
2、膜Bm3a表達(dá)的變化。結(jié)果：HE染色顯示正常對(duì)照組大鼠視網(wǎng)膜組織結(jié)構(gòu)層次清楚，染色均勻，細(xì)胞形態(tài)規(guī)則。實(shí)驗(yàn)各組大鼠視網(wǎng)膜神經(jīng)節(jié)細(xì)胞排列紊亂，胞漿內(nèi)空泡樣變性，視網(wǎng)膜組織變薄，隨時(shí)間延長(zhǎng)神經(jīng)節(jié)細(xì)胞數(shù)目逐漸減少。實(shí)時(shí)熒光定量RT-PCR結(jié)果顯示實(shí)驗(yàn)各組Bm3amRNA均較正常組減少；造模后12h、16h、24h、48h，實(shí)驗(yàn)組Bm3amRNA均高于實(shí)驗(yàn)對(duì)照組，差異均有統(tǒng)計(jì)學(xué)意義；造模后8h，實(shí)驗(yàn)組與實(shí)驗(yàn)對(duì)照組Bm3amRNA差異無(wú)統(tǒng)計(jì)學(xué)意義。免疫組織化學(xué)染色結(jié)果顯示Bm3a只在視網(wǎng)膜神經(jīng)節(jié)細(xì)胞層表達(dá)，陽(yáng)性細(xì)胞計(jì)數(shù)及光密度值測(cè)定結(jié)果與實(shí)時(shí)熒光定量PCR結(jié)果相吻合。結(jié)論：1．成功建立大鼠視網(wǎng)膜興奮性損
3、傷模型；2．NMDA可致大鼠視網(wǎng)膜中Bm3a及其mRNA表達(dá)減少；3．腹腔注射谷氨酰胺可增加大鼠興奮性損傷視網(wǎng)膜中Bm3a的表達(dá)，提高神經(jīng)節(jié)細(xì)胞的存活率。碩士研究生：紀(jì)珍(眼科學(xué))指導(dǎo)老師：王大博教授關(guān)鍵詞：谷氨酰胺；視網(wǎng)膜；N一甲基天冬氨酸；Brn3aGlutamineProtectsRetinaFollowingExcitotocityInjuryinRatAbstractObjeetiveToobservetheeffectofglutamineonBm3aexpressioninretinaafterNMDA烈一methyl—D-aspartate)excitotocityinjury
4、inrat．MethodsFifty-fiveadultcleanWistarratswererandomlydividedintothreegroups：normalcontrolgroupincludingfiverats，experimentalcontrolgroupincludingtwenty—fiveratsthatweretreatedwithNMDA(2p1，80mmol／1)byintravitrealinjection，experimentalcontrolincludingtwenty-fiveratswhichweretreatedwithglutamine(0．75
5、9／kg)byintraperitonealinjectionsevenhoursafterNMDAinjected．Randomlychoosedfiveratsofexperimentalcontrolgroupandexperimentalgroupweresacrificedat8，12，16，24，48hourafterNMDAinjectedrespectively．Therats’retinawasHEstainedandobservedunderlightmicroscope．TheexpressionlevelofBrn3aweredeterminedbyimmunohist
6、ochemistryandReal—Timefluorescencequantitativereversetranscriptionpolymerasechainreaction(Real—TimeRT-PCR)．ResultsHEstainingshowedthatthestructureofretinaofnormalcontrolgroupwashistologicallywell-defined，uniforminstaining，andwithregularcellshape．Inexperimentalgroup，theretinalganglioncells(RGCs)werel
7、ooselyarrangedanditsnumberdecreasedwithtime．a(chǎn)ndtheretinabecamethinner．Real-TimeRT-PCRresultsshowedthatBrn3amRNAexpressiondecreasedinallexperimentaleyescomparedwithnormalcontroleyes(allP<0．05)．Therewas
