增生性玻璃体视网膜病变中组织型转谷氨酰胺酶的表达

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1、增生性玻璃体视网膜病变中组织型转谷氨酰胺酶的表达作者：王辉　刘哲丽　郝旭红　徐丽【摘要】　　目的:检测增生性玻璃体视网膜病变（PVR）视网膜前膜和视网膜色素上皮（RPE）细胞中组织型转谷氨酰胺酶(tTG)的表达情况，探讨tTG是否参与PVR的发病过程。方法:PVR患者21例,收集视网膜前膜，C级8例，D级13例,行tTG免疫组织化学染色。另对培养3～5代的人RPE细胞分别用含有100mL/LPVR玻璃体液、正常玻璃体液和PBS的DMEM培养液进行孵育24h后行tTG免疫细胞化学染色。光镜下观察，比较C级和D级膜的表达阳性率，显微图像分析测量3

2、组RPE细胞染色的平均灰度值。结果：在PVR视网膜前膜中tTG呈阳性表达（81％），C级和D级膜表达阳性率无差异（C级88％，D级77％，P>0.05）。培养的人RPE细胞表达tTG，在PVR患者的玻璃体液作用下，tTG表达的平均灰度值为137.0±2.6，与正常玻璃体液组（143.5±2.9）及PBS对照组（143.6±3.0）相比，表达水平升高（P<0.05）。结论：PVR视网膜前膜和培养的人RPE细胞tTG表达阳性，在PVR患者的玻璃体液作用下，RPE细胞的tTG表达水平升高，这表明tTG参与了PVR的发病过程，在PVR视网

3、膜前膜的形成过程中可能有重要作用。12【关键词】增生性玻璃体视网膜病变　组织型转谷氨酰胺酶　视网膜色素上皮细胞　　TissuetransglutaminaseinproliferativevitreoretinopathyAbstractAIM:Toinvestigatetheexpressionoftissuetransglutaminase(tTG)inproliferativevitreoretinopathy(PVR)epiretinalmembranesandretinalpigmentepithelium(RPE)cellswhi

4、chisthoughttobeakeyelementinPVRformation.METHODS:Twenty-onesamplesofepiretinalmembraneswereobtainedfrompatientswithPVRinvolvedpreviousrhegmatogenousretinaldetachment.EightsampleswereatCstageofPVR,13sampleswereatDstage.Thesampleswereselectedtoperformimmunohistochemicalstaini

5、ng.Third-tofifth-passagehumanRPEcellswereincubatedbyserum-freeDMEMcontaining100mL/LPVRvitreoussamples,normalvitreoussamplesorPBSandstainedimmunocytochemically.PositiveratesofexpressionwerecalculatedinbothCandDstagemembranesunderlightmicroscopy.Thestainingaveragegrayvaluesof

6、thethreegroupRPEcellsweredetectedwithimageanalyzingsystem.RESULTS:TheexpressionoftTGwaspresentinPVRepiretinalmembranes(80%).12PositivestainingwasmostprominentinECM,fibroblast-likecellsandmacrophages.TherewasnosignificantdifferenceinpositiveratesbetweenCandDstagemembranes(P&

7、gt;0.05).CulturedhumanRPEcellsexpressedtTG,andtheexpressionlevelofRPEcellsincubatedwithPVRvitreoussampleswashigherthanthoseincubatedwithnormalvitreoussamplesandPBS(P<0.05).CONCLUSION:ThefindingsdemonstratedthattTGispresentinPVRepiretinalmembranesandculturedhumanRPEcells.

8、ItsamountisstimulatedtoincreasebythePVRvitreoushumor.tTGtakespartintheprocessofPVR