Shibayama R, Araki N, Nagai R, Horiuchi S
Department of Biochemistry, Kumamoto University School of Medicine, Japan.
Diabetes. 1999 Sep;48(9):1842-9. doi: 10.2337/diabetes.48.9.1842.
Prolonged incubation of proteins with reducing sugar produces advanced glycation end products (AGEs), which are implicated as factors for aging and diabetic complications. We previously demonstrated the presence of N(epsilon)-(carboxymethyl)lysine (CML), one of the main AGE structures, in human and animal tissues using a monoclonal anti-CML antibody (6D12). These findings suggest that CML structures present in vivo could serve as immunogens to generate autoantibodies. This suggestion was tested in the present study. First, plasma samples from diabetic rats reacted positively with AGE bovine serum albumin (BSA). These reactivities increased with the duration of diabetic states and were inhibited specifically by CML-BSA. Second, a fraction purified from plasma of diabetic patients, which bound to AGE-BSA, showed a positive reaction to CML-BSA and furthermore also to human lens proteins, which are known to undergo CML modification in vivo. Finally, patients with renal failure caused by diabetes or nondiabetic pathologies had a higher autoantibody activity against CML structure than that in normal subjects or diabetic patients without renal failure. These results indicate that CML accumulated in vivo serves as an immunological epitope to generate an autoantibody specific for CML that might be used as a potential marker for diabetic nephropathy or chronic renal failure.
蛋白质与还原糖长时间孵育会产生晚期糖基化终产物(AGEs),这些产物被认为是衰老和糖尿病并发症的因素。我们之前使用单克隆抗CML抗体(6D12)在人和动物组织中证实了主要AGE结构之一的Nε-(羧甲基)赖氨酸(CML)的存在。这些发现表明体内存在的CML结构可能作为免疫原产生自身抗体。本研究对这一推测进行了验证。首先,糖尿病大鼠的血浆样本与AGE牛血清白蛋白(BSA)发生阳性反应。这些反应性随着糖尿病状态的持续时间而增加,并被CML-BSA特异性抑制。其次,从糖尿病患者血浆中纯化的与AGE-BSA结合的部分,对CML-BSA呈阳性反应,并且对已知在体内会发生CML修饰的人晶状体蛋白也呈阳性反应。最后,由糖尿病或非糖尿病病理导致肾衰竭的患者针对CML结构的自身抗体活性高于正常受试者或无肾衰竭的糖尿病患者。这些结果表明体内积累的CML作为免疫表位产生针对CML的特异性自身抗体,这可能用作糖尿病肾病或慢性肾衰竭的潜在标志物。