Yamaguchi M, Nakamura N, Nakano K, Kitagawa Y, Shigeta H, Hasegawa G, Ienaga K, Nakamura K, Nakazawa Y, Fukui I, Obayashi H, Kondo M
The First Department of Internal Medicine, Kyoto Prefectural University of Medicine, Japan.
Diabet Med. 1998 Jun;15(6):458-62. doi: 10.1002/(SICI)1096-9136(199806)15:6<458::AID-DIA601>3.0.CO;2-Q.
Crossline is a novel advanced glycation endproduct (AGE) which has both a crosslink and fluorescence similar to AGE-protein in vivo. To assess the association of AGEs to the development of diabetic retinopathy we developed a sensitive and specific enzyme-linked immunosorbent assay (ELISA) for crossline in blood samples and investigated the association of the development of retinopathy and erythrocyte membrane protein (EMP)-crossline concentrations in patients with Type 2 diabetes mellitus (Type 2 DM). Crossline formation in EMP exceeded that in haemoglobin and was detectable in normal EMP samples without pretreatment by this ELISA system. Mean (+/-SE) EMP crossline levels were elevated 1.6-fold in diabetic patients without retinopathy (7.6 +/- 0.5 pmol mg(-1), p < 0.005), 2.2-fold in diabetic patients with non-proliferative retinopathy (10.5 +/- 0.6 pmol mg(-1), p < 0.001) and 2.6-fold in diabetic patients with proliferative retinopathy (12.0 +/- 0.6 pmol mg(-1), p < 0.001) compared with healthy control subjects (4.7 +/- 0.5 pmol mg(-1)). Type 2 DM patients with retinopathy had significantly higher EMP-crossline levels than those without retinopathy (p < 0.005). Our data suggest that elevated EMP-crossline concentrations are associated with the presence of retinopathy in patients with Type 2 DM and EMP-crossline measured by our ELISA may provide a useful marker for assessing the role of glycation in the development of diabetic retinopathy.
交联物是一种新型的晚期糖基化终产物（AGE），其在体内具有与AGE-蛋白质相似的交联和荧光特性。为了评估AGEs与糖尿病视网膜病变发生发展的关联，我们开发了一种灵敏且特异的酶联免疫吸附测定法（ELISA），用于检测血样中的交联物，并研究2型糖尿病（2型DM）患者视网膜病变的发生发展与红细胞膜蛋白（EMP）-交联物浓度之间的关联。EMP中的交联物形成超过了血红蛋白中的交联物形成，并且在未经预处理的正常EMP样品中，通过该ELISA系统即可检测到。与健康对照受试者（4.7±0.5 pmol mg⁻¹）相比，无视网膜病变的糖尿病患者的平均（±SE）EMP交联物水平升高了1.6倍（7.6±0.5 pmol mg⁻¹，p<0.005），非增殖性视网膜病变的糖尿病患者升高了2.2倍（10.5±0.6 pmol mg⁻¹，p<0.001），增殖性视网膜病变的糖尿病患者升高了2.6倍（12.0±0.6 pmol mg⁻¹，p<0.001）。患有视网膜病变的2型DM患者的EMP-交联物水平显著高于无视网膜病变的患者（p<0.005）。我们的数据表明，升高的EMP-交联物浓度与2型DM患者视网膜病变的存在相关，并且我们通过ELISA测定的EMP-交联物可能为评估糖基化在糖尿病视网膜病变发生发展中的作用提供一个有用的标志物。
