大鼠視網(wǎng)膜神經(jīng)節(jié)細(xì)胞興奮性損傷過(guò)程中Mfn2作用的研究.pdf

1、分類(lèi)號(hào)：R775密級(jí)：不保密UDC：610學(xué)校代碼：11065碩士學(xué)位論文大鼠視網(wǎng)膜神經(jīng)節(jié)細(xì)胞興奮性損傷過(guò)程中大鼠視網(wǎng)膜神經(jīng)節(jié)細(xì)胞興奮性損傷過(guò)程中Mfn2Mfn2作用的研究作用的研究薛君發(fā)指導(dǎo)教師王大博教授學(xué)科專(zhuān)業(yè)名稱(chēng)眼科學(xué)論文答辯日期2016年5月20日AbstractOBJECTIVE:Toinvestigatethemechanismofmediationconductedbymitofusin2toNMDA(nmethyldas

2、partate)inducedretinalganglioncells(RGCs)excitotoxicityinrats.METHODS:FtyfiveadultSpragueDawleyratsweredividedintothreegroupsromly.3ul(40mM120nmoll)ofNMDAwasinjectedintothevitreouscavitytomakeexcitatymodel(with0.1MPBSinj

3、ectionascontrol).Eyesofeachgroupwereremovedoflensat1h12h24h48hafterintravitrealinjectionrespectively.Hematoxylineosinstainingofretinalcrosssectionswasconductedtoobservethemphologicalalteration.RealtimeFluescentQuantitati

4、vePCRwasappliedtoassessthelevelofBrn3amRNAtomeasurethedegreeofinjurytoRGCswithlevelofMfn2mRNAdetectedtoanalyzeditsexpressioninRGCs.ImmunohistochemistrywasalsousedtodetecttheconcentrationofMfn2expressedintheretina.RESULTS

5、:1.Hematoxylineosinstaining:Retinaofblankcontrolgroupwasinderunifmdyeingwithvividstructureofcellsineverylayer.Retinaganglioncellswereroundovalwhirangedinasinglelayer.IntheNMDAgroupsRGCswerehyicwhosespacebetweenthenucleus

6、cytomembranewaswidened.theyweredisderedalignedwitheachother.ThenumberofRGCsdecreasedthethicknessofinnerplexifmlayer(IPL)wasthinningastheprocessingtimewasprolongedwhileinPBSinjectiongroupsslightchangeswereobserved.2.Realt

7、imeFluescentQuantitativePCRofBrn3a:ComparedwiththeblankcontrolgrouptheexpressionofBrn3amRNAwaslowerineverygroupwithNMDAinjected.neverthelesstherewasnostatisticalsignificancebetweenPBSinjectiongroupsblankcontrolgroup(P0.0

8、5).3.RealtimeFluescentQuantitativePCRofMfn2:ComparedwithblankcontrolgrouptheexpressionlevelofretinalMfn2inNMDAgroupswashigherreachedapeakat24haftertheinjectionbegantodecreaseat48h.Exceptthelevelat1hafterinjection(P0.05)a

9、llofgroupswithNMDAinjectionweresignificantlydifferentcomparedwiththeblankcontrolgroup(P0.05).4.ImmunohistochemistryofMfn2:theexpressionofMfn2inblankcontrolgroupgroupswithPBSinjectionwasverylowinalloftheRGCslayertheinnerp

10、lexifmlayerouterplexifmlayerthephoteceptcelllayeratwhichthereweretracebrownishyellowparticlesdeposited.WhileMfn2expressioninRGCslayerwasincreasedinNMDAinjectiongroups.ThevalueofIODofimmunohistochemicalresultsareingoodagr

11、eementwiththeRTPCR’s.CONCLUSIONS:Mfn2mayparticipateinexcitotoxicityofRGCsunderglaucomaconditionmaybeoneofthekeynodesofoxidativestresscausedbymitochondrialdysfunction.PostgraduatePostgraduatestudent:student:JunfaJunfaXueX