中华眼科杂志
中華眼科雜誌
중화안과잡지
Chinese Journal of Ophthalmology
2010年
7期
580-584
,共5页
刘堃%刘海芸%叶纹%江建海%许迅
劉堃%劉海蕓%葉紋%江建海%許迅
류곤%류해예%협문%강건해%허신
糖尿病视网膜病变%糖尿病,实验性%糖基转移酶类%糖基化
糖尿病視網膜病變%糖尿病,實驗性%糖基轉移酶類%糖基化
당뇨병시망막병변%당뇨병,실험성%당기전이매류%당기화
Diabetic retinopathy%Diabetes mellitus,experimental%Glycosyltransferases%Glycosylation
目的 观察糖尿病大鼠视网膜组织糖基转移酶表达改变情况,为进一步研究糖基转移酶及其介导的酶促糖基化反应参与糖尿病性视网膜病变发病机制建立研究基础.方法 实验研究.STZ诱导建立糖尿病大鼠模型,应用RT-PCR法观察已明确基因序列的部分糖基转移酶在糖尿病大鼠视网膜组织的表达情况.提取视网膜组织蛋白,SDS-PAGE电泳,RCA-I凝集素染色以及大鼠眼球组织切片RCA-I凝集素染色,观察视网膜组织蛋白糖基化修饰变化情况.对实时荧光定量PCR及组织切片凝集素分析的相关结果采用独立样本t检验进行统计学分析.结果 实时PCR检测:6个受检糖基转移酶在糖尿病大鼠组及正常大鼠组相对基因表达量分别为:O-linked N-acetylglucosamine transferase(0.16.50±0.1160, 0.1160±0.0363);UDP-Gal:betaGlcNAc beta1,3-galactosyltransferase (0.0186±0.0122,0.0152±0.0047);alpha 1,4-galactosyltransferase(0.0040±0.0040,0.00.54±0.0022):mannoside acetylghcosaminyltransferase 1(0.0228±0.0166,0.0187±0.0050);UDP-glucose ceramide glucosyltransferase(0.0129±0.0096,0.0116±0.0040);UDP-Gal:betaGlcNAc beta 1,4-galactosyltransferase 1(0.0157±0.0010,0.0081±0.0016).糖尿病大鼠视网膜组织中β1,4半乳糖基转移酶-1(UDP-Gal:betaGlcNAc beta 1,4-galactosyltransferase 1,GalT-1)的表达上调(t=6.847,P=0.002);蛋白凝集素及组织凝集素染色分析发现:糖尿病大鼠视网膜组织中Galβ 1→4GlcNAc糖基化基团修饰表达上调.结论 糖尿病状态下,大鼠视网膜组织中β1,4半乳糖基转移酶-1的表达及其介导的酶促糖基化反应上调.初步的实验结果为进一步研究糖基转移酶及其介导的酶促糖基化反应参与糖尿病视网膜病变的发病机制建立了相关实验基础.
目的 觀察糖尿病大鼠視網膜組織糖基轉移酶錶達改變情況,為進一步研究糖基轉移酶及其介導的酶促糖基化反應參與糖尿病性視網膜病變髮病機製建立研究基礎.方法 實驗研究.STZ誘導建立糖尿病大鼠模型,應用RT-PCR法觀察已明確基因序列的部分糖基轉移酶在糖尿病大鼠視網膜組織的錶達情況.提取視網膜組織蛋白,SDS-PAGE電泳,RCA-I凝集素染色以及大鼠眼毬組織切片RCA-I凝集素染色,觀察視網膜組織蛋白糖基化脩飾變化情況.對實時熒光定量PCR及組織切片凝集素分析的相關結果採用獨立樣本t檢驗進行統計學分析.結果 實時PCR檢測:6箇受檢糖基轉移酶在糖尿病大鼠組及正常大鼠組相對基因錶達量分彆為:O-linked N-acetylglucosamine transferase(0.16.50±0.1160, 0.1160±0.0363);UDP-Gal:betaGlcNAc beta1,3-galactosyltransferase (0.0186±0.0122,0.0152±0.0047);alpha 1,4-galactosyltransferase(0.0040±0.0040,0.00.54±0.0022):mannoside acetylghcosaminyltransferase 1(0.0228±0.0166,0.0187±0.0050);UDP-glucose ceramide glucosyltransferase(0.0129±0.0096,0.0116±0.0040);UDP-Gal:betaGlcNAc beta 1,4-galactosyltransferase 1(0.0157±0.0010,0.0081±0.0016).糖尿病大鼠視網膜組織中β1,4半乳糖基轉移酶-1(UDP-Gal:betaGlcNAc beta 1,4-galactosyltransferase 1,GalT-1)的錶達上調(t=6.847,P=0.002);蛋白凝集素及組織凝集素染色分析髮現:糖尿病大鼠視網膜組織中Galβ 1→4GlcNAc糖基化基糰脩飾錶達上調.結論 糖尿病狀態下,大鼠視網膜組織中β1,4半乳糖基轉移酶-1的錶達及其介導的酶促糖基化反應上調.初步的實驗結果為進一步研究糖基轉移酶及其介導的酶促糖基化反應參與糖尿病視網膜病變的髮病機製建立瞭相關實驗基礎.
목적 관찰당뇨병대서시망막조직당기전이매표체개변정황,위진일보연구당기전이매급기개도적매촉당기화반응삼여당뇨병성시망막병변발병궤제건립연구기출.방법 실험연구.STZ유도건립당뇨병대서모형,응용RT-PCR법관찰이명학기인서렬적부분당기전이매재당뇨병대서시망막조직적표체정황.제취시망막조직단백,SDS-PAGE전영,RCA-I응집소염색이급대서안구조직절편RCA-I응집소염색,관찰시망막조직단백당기화수식변화정황.대실시형광정량PCR급조직절편응집소분석적상관결과채용독립양본t검험진행통계학분석.결과 실시PCR검측:6개수검당기전이매재당뇨병대서조급정상대서조상대기인표체량분별위:O-linked N-acetylglucosamine transferase(0.16.50±0.1160, 0.1160±0.0363);UDP-Gal:betaGlcNAc beta1,3-galactosyltransferase (0.0186±0.0122,0.0152±0.0047);alpha 1,4-galactosyltransferase(0.0040±0.0040,0.00.54±0.0022):mannoside acetylghcosaminyltransferase 1(0.0228±0.0166,0.0187±0.0050);UDP-glucose ceramide glucosyltransferase(0.0129±0.0096,0.0116±0.0040);UDP-Gal:betaGlcNAc beta 1,4-galactosyltransferase 1(0.0157±0.0010,0.0081±0.0016).당뇨병대서시망막조직중β1,4반유당기전이매-1(UDP-Gal:betaGlcNAc beta 1,4-galactosyltransferase 1,GalT-1)적표체상조(t=6.847,P=0.002);단백응집소급조직응집소염색분석발현:당뇨병대서시망막조직중Galβ 1→4GlcNAc당기화기단수식표체상조.결론 당뇨병상태하,대서시망막조직중β1,4반유당기전이매-1적표체급기개도적매촉당기화반응상조.초보적실험결과위진일보연구당기전이매급기개도적매촉당기화반응삼여당뇨병시망막병변적발병궤제건립료상관실험기출.
Objective To investigate whether hyperglycemia affect the expression of glycosyltransferases and Enzyme-catalyzed glycosylation in the retina of diabetic rat.Method It was an experimental study.RT-PcR was used to analyze the mRNA level of six glycosyltransferases in the retina of steptozocin diabetic rats:Lectin blot assay with RCA-I was performed to investigate the level of Galβ 1→4GlcNAc or N-glycans on total retinal glycoproteins.Results mRNA level of six glycosyltransferases in the retina of streptocin diabetic rats and of normal rat is:O-linked N-acetylglucosamine transferase(0.1650±0.1160 versus 0.1160±0.036);UDP-Gal:betaGlcNAc beta1,3-galactosyltransferase(0.0186±0.0122 versus 0.0152±0.0047);alpha 1,4-galactosyltransferase(0.0040±0.0040 versus 0.0054±0.0022);mannoside acetylglucosaminyltransferase 1(0.0228±0.0166 versus 0.0187±0.0050);UDP-glucose ceramide glucosyltransferase(0.0129±0.0096 versus 0.0116±0.0040);UDP-Gal:betaGlcNAc beta 1,4-galactosyltransferase 1(0.0157±0.0010 versus 0.0081±0.0016).The mRNA level of UDP-Gal:betaGlcNAc beta 1,4-galactosyltransferase 2 was up-regulated in the retina of streptocin diabetic rats(t=6.847,P=0.002).Consistent with this,the level of Galβ 1→4GlcNAc of glycoproteins in streptocin diabetic rat's retina was strengthened compared with that in control rats.Conclusions The results from this study showed that Hyperglycemia could up-regulate the expression of UDP-Gal:betaGlcNAc beta 1,4-galactosyltransferase 1 and strengthed the level of Galβ 1→4GlcNAc of glycoproteins in the retina of streptocin diabetic rats.Our initial results will contribute to the research for the relation between the Enzymecatalyzed Glycosylation and the etiopathogenesis of diabetic retinopathy.
