摘要
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.
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.
出处
《中华眼科杂志》
CAS
CSCD
北大核心
2010年第7期580-584,共5页
Chinese Journal of Ophthalmology
基金
基金项目：上海市自然科学基金（08za1412000）
关键词
糖尿病视网膜病变
糖尿病
实验性
糖基转移酶类
糖基化
Diabetic retinopathy
Diabetes mellitus,experimental
Glycosyltransferases
Glycosylation
