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1、中华糖尿病杂志2004年第12卷第4期ChinJDiabetes,August2004,Vol12,No4·273··基础论著·糖尿病大鼠视网膜病变细胞凋亡的观察潘琳周水平郭艳茹李光伟【摘要】目的探讨早、中、晚期不同病程糖尿病(DM)大鼠视网膜病变(DR)毛细血管周细胞、内皮细胞凋亡的发生,为研究DR发病机制提供形态学依据。方法90只雄性SD大鼠随机分成正常对照组(n=30)及链脲佐菌素(STZ)糖尿病大鼠6、9、12个月模型组(n=20)。各实验组分别按期处死10只大鼠,取眼球作视网膜消化铺片,PAS染色及核苷酸末
2、端转移酶介导dUTP缺口翻译法(TUNEL法)特异性标记凋亡细胞,光镜下观察。结果6个月模型组,TUNEL标记法显示周细胞凋亡,但未见内皮细胞凋亡。9个月模型组,PAS染色、TUNEL标记法显示周细胞、内皮细胞均有凋亡,周细胞及内皮细胞出现核固缩、核破碎、凋亡小体形成等细胞凋亡形态学改变。12个月模型组,内皮细胞凋亡多见,部分区域每个高倍视野下可见2~3个不同时期的内皮细胞凋亡。结论DR期间内皮细胞及细胞周细胞均可发生凋亡,DR早期以周细胞凋亡为主,中晚期内皮细胞发生凋亡,并逐渐增多。【关键词】糖尿病视网膜病变;内皮
3、细胞;周细胞;细胞凋亡ExperimentalstudyonprogrammedretinalcelldeathindiabeticratsPANLin,ZHOUShui2ping,GUOYan2ru,etal.Departmentofcellbiology,China2JapanFriendshipInstitutesofClinicalMedSci2ence,China2JapanFriendshipHospital,Beijing100029,China【Abstract】ObjectiveTodetermin
4、etheoccurrenceofcelldeathinsituindiabetic2likeretinopathyfromthestreptozotocin(STZ)2induceddiabeticratsafter6,9,12months1Methods90maleSprague2Dawleyratswererandomlydividedintonormalcontrolgroup(n=30)anddiabeticmodelgroup1STZwereusedtoinducediabetesinrats1Ratsin
5、eachgroupwerefrequentlyexaminedwithbloodglucoseandweight,andkilledafter6,9,12months(n=20)1Thesectionspreparedfromthetrypsin2digestedretinawerestudieswithperiodicacid2schiff2hematoxylinstain.Theterminaldeoxynucleotidyltransferase2mediat2eddUTPnickendlabeling(TUN
6、EL)reactionwasusedtodetectthepreferentiallyapoptoticDNAfrag2mentation1ResultsRetinalmicrovesselsof6monthdiabeticratsshowedTUNEL2positivepericytes,whichwereabsentincontrolrats1Retinalmicrographsof9,12monthdiabeticratsshowedpericytesandendothelialcellapoptosisres
7、pectivelywithTUNELreactionandPASstain1At12months,thenumbersofapoptoticendothelialcellsweremore,andeachofthepicturesshowedTUNRL2positiveendothelialcellorapoptoticbodiesunderthelightmicroscope1ConclusionsThesefindingsindicatethatdiabetesleadstotheaccelerateddeath
8、insituofpericyteintheearlystagesofretinopathy,andwiththeadvanceofdiseasetheendothelialcellapoptosisoccurs1【Keywords】Diabeticretinopathy;Endothelialcell;Pericyte;Apoptosis糖尿病