中华眼底病杂志
中華眼底病雜誌
중화안저병잡지
CHINESE JOURNAL OF OCULAR FUNDUS DISEASES
2013年
1期
30-34
,共5页
谢琳%唐仕波%史煜%朱晓波
謝琳%唐仕波%史煜%硃曉波
사림%당사파%사욱%주효파
糖尿病视网膜病变/病理生理学%DNA,线粒体/病理生理学%动物实验
糖尿病視網膜病變/病理生理學%DNA,線粒體/病理生理學%動物實驗
당뇨병시망막병변/병리생이학%DNA,선립체/병리생이학%동물실험
Diabetic retinopathy/pathophysiology%DNA,mitochondrial/pathophysiology%Animal experimentation
目的 观察不同病程糖尿病大鼠视网膜血管细胞线粒体DNA (mtDNA)损伤及黏附分子表达、细胞凋亡情况.方法 Sprague-Dawley大鼠100只,分为正常对照组和实验组.实验组大鼠腹腔注射链尿佐菌素诱导糖尿病模型,造模成功后依照病程分为DR1、DR2、DR3月组,正常对照组分为NR1、NR2、NR3月组.提取各组大鼠视网膜血管DNA,联合Fpg酶切Southern印迹杂交检测未损伤mtDNA含量；实时荧光逆转录聚合酶链反应(RT-PCR)观察mtDNA编码基因环氧酶-1(COX-1)及转录因子A(mtTFA)mRNA的表达；消化铺片TdT介导的dUTP缺口末端标记(TUNEL)免疫荧光、细胞间黏附因子(ICAM-1)免疫组织化学染色,分别观察细胞凋亡及黏附分子的表达.结果 不同病程糖尿病大鼠视网膜未损伤mtDNA含量与不同鼠龄正常对照组比较,糖尿病大鼠未损伤mtDNA含量明显减少,且随着病程延长减少更明显；COX-1及mtTFA mRNA表达相应降低；糖尿病大鼠随病程延长,TUNEL、ICAM-1阳性细胞数增多.结论 糖尿病大鼠随着病程延长,视网膜血管细胞mtDNA损伤加重,其编码基因及转录调控基因mRNA的表达均相应下降,黏附分子表达上调,细胞凋亡增加.
目的 觀察不同病程糖尿病大鼠視網膜血管細胞線粒體DNA (mtDNA)損傷及黏附分子錶達、細胞凋亡情況.方法 Sprague-Dawley大鼠100隻,分為正常對照組和實驗組.實驗組大鼠腹腔註射鏈尿佐菌素誘導糖尿病模型,造模成功後依照病程分為DR1、DR2、DR3月組,正常對照組分為NR1、NR2、NR3月組.提取各組大鼠視網膜血管DNA,聯閤Fpg酶切Southern印跡雜交檢測未損傷mtDNA含量；實時熒光逆轉錄聚閤酶鏈反應(RT-PCR)觀察mtDNA編碼基因環氧酶-1(COX-1)及轉錄因子A(mtTFA)mRNA的錶達；消化鋪片TdT介導的dUTP缺口末耑標記(TUNEL)免疫熒光、細胞間黏附因子(ICAM-1)免疫組織化學染色,分彆觀察細胞凋亡及黏附分子的錶達.結果 不同病程糖尿病大鼠視網膜未損傷mtDNA含量與不同鼠齡正常對照組比較,糖尿病大鼠未損傷mtDNA含量明顯減少,且隨著病程延長減少更明顯；COX-1及mtTFA mRNA錶達相應降低；糖尿病大鼠隨病程延長,TUNEL、ICAM-1暘性細胞數增多.結論 糖尿病大鼠隨著病程延長,視網膜血管細胞mtDNA損傷加重,其編碼基因及轉錄調控基因mRNA的錶達均相應下降,黏附分子錶達上調,細胞凋亡增加.
목적 관찰불동병정당뇨병대서시망막혈관세포선립체DNA (mtDNA)손상급점부분자표체、세포조망정황.방법 Sprague-Dawley대서100지,분위정상대조조화실험조.실험조대서복강주사련뇨좌균소유도당뇨병모형,조모성공후의조병정분위DR1、DR2、DR3월조,정상대조조분위NR1、NR2、NR3월조.제취각조대서시망막혈관DNA,연합Fpg매절Southern인적잡교검측미손상mtDNA함량；실시형광역전록취합매련반응(RT-PCR)관찰mtDNA편마기인배양매-1(COX-1)급전록인자A(mtTFA)mRNA적표체；소화포편TdT개도적dUTP결구말단표기(TUNEL)면역형광、세포간점부인자(ICAM-1)면역조직화학염색,분별관찰세포조망급점부분자적표체.결과 불동병정당뇨병대서시망막미손상mtDNA함량여불동서령정상대조조비교,당뇨병대서미손상mtDNA함량명현감소,차수착병정연장감소경명현；COX-1급mtTFA mRNA표체상응강저；당뇨병대서수병정연장,TUNEL、ICAM-1양성세포수증다.결론 당뇨병대서수착병정연장,시망막혈관세포mtDNA손상가중,기편마기인급전록조공기인mRNA적표체균상응하강,점부분자표체상조,세포조망증가.
Objective To observe the oxidative damage of mtDNA,apoptosis and expression of adhesion molecules in retinal capillary cells of diabetic rat with different disease courses.Methods One hundred Sprague-Dawley rats were randomly divided into the control group and the experimental group.The rats of experimental group were induced with streptozotocin (STZ) injection creating a diabetic model.Then they were divided into DR1m,DR2m DR3m group according to disease courses.The rats of control group were divided into NR1m,NR2m,NR3m group.Rat retinal capillaries were prepared,and then the contents of undamaged mtDNA were examined by Southern blot combined with Fpg.The expression of cyclooxygenase (COX)-1 encoded by mtDNA and transcription factors A (mtTFA) mRNA were detected by real-time quantitative polymerase chain reaction (RT-PCR).Apoptosis and expression of intercellular adhesion molecule-1 (ICAM-1) were detected by terminal dUT nick end-labeling (TUNEL) immunofluorescence and immunohistochemistry respectively.Results The contents of undamaged mtDNA in rats of DR1m,DR2m,DR3m were less than those of NR1m、NR2m、NR3m.The contents of undamaged mtDNA in diabetic rats decreased with the increase of disease courses.In addition,the mRNA levels of COX-1 and mtTFA were down-regulated in diabetic rats.The positive cells of TUNEL and ICAM-1TUNEL and ICAM-1 in diabetic rats increased with the increase of disease courses.Conclusion With the increase of disease courses,mtDNA damage and apoptotic cells are increased,while the expression of mRNA encoded by mtDNA and ICAM-1 decreased in retinal capillarv cells in diabetic rats.
