Abundant Lipid and Protein Components of Drusen 英文参考文献.docVIP

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Abundant Lipid and Protein Components of Drusen 英文参考文献.doc

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Abundant Lipid and Protein Components of Drusen 英文参考文献

AbundantLipidandProteinComponentsofDrusen LanWang1,MarkE.Clark1,DavidK.Crossman2,KyokoKojima3,JeffreyD.Messinger1,JamesA. Mobley4,ChristineA.Curcio1* 1DepartmentofOphthalmology,UniversityofAlabamaatBirmingham,Birmingham,Alabama,UnitedStatesofAmerica,2DepartmentofMicrobiology,Universityof AlabamaatBirmingham,Birmingham,Alabama,UnitedStatesofAmerica,3DepartmentofEpidemiology,UniversityofAlabamaatBirmingham,Birmingham,Alabama, UnitedStatesofAmerica,4DepartmentofSurgery,UniversityofAlabamaatBirmingham,Birmingham,Alabama,UnitedStatesofAmerica Abstract Background:Drusenareextracellularlesionscharacteristicofagingandage-relatedmaculopathy,amajorretinaldiseaseof theelderly.Wedeterminedtherelativeproportionsoflipidsandproteinsindrusencappedwithretinalpigmentepithelium (RPE)andinRPEisolatedfromnon-macularregionsof36humanretinaswithgrosslynormalmaculasobtained,6hrafter death. Methodology/Principal Findings: Druse pellets were examined by light and electron microscopy. Component proteins were extracted using novel methods for preserved tissues, separated, subjected to tryptic digestion and LC-MS(MS)2 analysisusinganiontrapmassspectrometer,andidentifiedwithreferencetodatabases.Lipidclasseswereseparatedusing thin layer chromatography and quantified by densitometry. Major druse components were esterified cholesterol (EC), phosphatidylcholine (PC), and protein (37.5613.7, 36.9612.9, and 43.0611.5ng/druse, respectively). Lipid-containing particles (median diameter, 77nm) occupied 37–44% of druse volume. Major proteins include vitronectin, complement component 9, apoE, and clusterin, previously seen in drusen, and ATP synthase subunit b, scavenger receptor B2, and retinoldehydrogenase5,previouslyseeninRPE.DrusenandRPEhadsimilarproteinprofiles,withhigherintensitiesand greater variability in drusen. C8, part of the complement membrane attack complex, was localized in drusen by immunofluorescence. Conclusions/Significance: Atleast40%ofdrusecontentiscomprisedbylipidsdominatedbyECandPC,