糖尿病视网膜病变与非酶糖化终产物的关系

Pathological impairment of microcirculation and abnormal deposition of AGEs on retinae in diabetic ratinopathy of rats

目的建立非酶糖化终产物(AGEs)检测方法,研究糖尿病视网膜病变与AGEs的关系。方法以体外孵育的AGEs免疫新西兰大白兔获得抗体;ELISA法标记糖尿病大鼠视网膜上AGEs的沉积;HE染色观察其病理改变,spearman检验统计两者与糖尿病病程的相关性。结果多克隆抗体经免疫双扩散与竞争抑制性ELISA实验被证实与AGEs特异结合。糖尿病大鼠视网膜抗AGEs染色阳性,对照组为阴性(P<0.01);其微血管周细胞密度为4.68±0.528个/油镜视野,对照组为6.74±0.532(P<0.01);其AGEs沉积程度与周细胞密度负相关,两者异常度随糖尿病病程延长而逐步加重,P<0.01。结论糖尿病代谢紊乱导致AGEs在视网膜上沉积,后者和以周细胞丢失为代表的早期视网膜微血管病有密切关系。

Objective To set up an immunoassay of advanced glycosylation end products(AGEs )in vivo,and further study the correlation between microcirculation changes a nd the accumulation of AGEs on retinae in diabetic rats.Methods Polyclonal an ti-AGE antibody was prepared in New Zealand rabbits using the AGE-BSA incubati ng in vitro as an antigen.Then this antibody was used to assess the deposition of AGEs in retina of streptozotocin-induced diabetes rats by ELISA,and the pa thological impairments of vascular tissues were observed by HE staining.The cor relations between the course of diabetes and the loss of pericytes or AGE-speci fic staining were analyzed using SPSS statistic software.Results The AGE-stain ing in retina of diabetic rats(8weeks course of disease)was positive scored a s“++”,while that of normal rats was negtive(P<0.01).The average amo unt of pericytes of capillary vessel per10×100microscope visual field was 4.68±0.528in diabetes group and6.74±0.532in control group ( P<0.01).The data from diabetic rats were analyzed with spearman rank-cor relation test with the correlative coefficient -0.900,0.896and-0. 867,respectively (P<0.01).Conclusion The metabolic abnormality in di abetes result in AGE deposition in vascular tissues of retina.It has a close re lation with the loss of pericytes of capillary vessels that resemble the early p athological impairment of microcirculation in diabetic retinopathy.