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1、地塞米松诱导大鼠白内障形态学及酶活性变化论文王建伟,严宏,王永强,哈文静,丁正华,郭勇【关键词】白内障Alterationofmorphologyandenzymeactivityindexamethasoninducedcataractinratlens【Abstract】AIM:Toobservethemorphologicalfeaturesandenzymeactivitiesindexamethasoninducedorganculturedratlensandtostudythemechanisms
2、ofsteroidcataract.METHODS:Onehundredlensesculturedinvitroethasoninducedcataractgroup(DMEM+dexamethason10μmol/L).Folloicroscope.Ond1,3,5and7,12lensesofeachgroupogenizedandtheactivitiesofenzymes(catalase,superoxidedismutase,lactatedehydrogenase)easured,respect
3、ively.Tadeintospecimenoftransmissionelectronmicroscopyfortheobservationoftheultrastructureond7.RESULTS:Thelensesofcontrolgroupexhibitedmistlikeopacityandthoseofcataractgrouphaddensenuclearopacityond7.Ond3,theactivitiesofcatalase,superoxidedismutaseandlactate
4、dehydrogenasedecreasedby34.51%,7.78%,15.73%respectivelyparedicroscope,thearrangementoffibercellsincontrolgroupembrane,celljunctionandorganelleal,entoffibercellsalappearingcellsitochondria,alargeamountofvacuolesandexpandedextracellularlacunae.CONCLUSION:Lense
5、shavenuclearcataractindexamethasoninducedorganculturedratlens.Oxidativestressmaybeinvolvedinthesteroidinducedcataractformation.【Keyutase;lactatedehydrogenase;transmissionelectronmicroscope【摘要】目的:观察地塞米松诱导离体大鼠激素性白内障的形态学和酶活性变化,探讨激素性白内障发病机制.方法:大鼠的100只透明晶状体随机分为对照
6、组(DMEM)、地塞米松诱导的白内障组(DMEM+地塞米松10μmol/L),体外培养7d.freela公司产品,CAT,SOD,LDH测试盒购自南京建成生物工程研究所.1.2方法1.2.1离体晶状体培养以颈椎脱臼法处死SD大鼠,取其眼球,剔除肌肉、筋膜等组织,安尔碘浸泡5min,生理盐水冲洗3次,将眼球浸于DMEM液中,由后极部剪开巩膜,取出晶状体.尽量去除晶状体表面的玻璃体.将晶状体放入含5×104u/L青霉素和5×104u/L链霉素的DMEM培养基中,于37℃下,置50mL/LCO2细胞培养箱中培养8h后
7、,弃去混浊晶状体,留取100只透明晶状体备用.1.2.2激素性白内障模型的建立[5]将100只备用晶状体按随机数字大小等分为两组:正常对照组(A:DMEM),地塞米松诱导的激素性白内障组(B:DMEM+地塞米松10μmol/L).分别放入含培养基的无菌24孔板中,于37℃下,置50mL/LCO2细胞培养箱中培养,隔日换液.培养中每日用解剖显微镜观察晶状体,记录晶状体混浊程度.各组分别于第1,3,5,7日取出12只晶状体,称质量,-70℃冰箱冷冻保存.1.2.3晶状体混浊程度的动态观察利用解剖显微镜,每日观察、记
8、录晶状体混浊程度.参考Mathur等[6]离体晶状体混浊的分级方法,将晶状体混浊程度分为0~5级,0级=透明晶状体,1级=晶状体呈雾状混浊,2级=晶状体皮质和核之间出现可见分界,3级=轻度晶状体核混浊,4级=重度晶状体核混浊,5级=晶状体完全混浊(Fig1).1.2.4酶活性测定分别于第1,3,5,7日,从各组中取12只晶状体,按50g/L(100SX下观察超微结构.统计学处理:使用S