Lim Minsu, Park Leejin, Shin Geewook, Hong Hekyung, Kang Incheol, Park Yongsoo
Department of Internal Medicine and Bioengineering, Hanyang University College of Medicine and Engineering, Seoul, Korea.
Ann N Y Acad Sci. 2008 Dec;1150:311-5. doi: 10.1196/annals.1447.011.
We herein report cytotoxicity of advanced glycation end-products (AGEs) on pancreatic beta cells. AGEs stimulated reactive oxygen species (ROS) generation but did not arrest proliferation of the INS-1 cell line. Pancreatic beta cell lines or primary cultured islets possess a receptor for AGE (RAGE), and its expression increased after AGE treatment. TUNEL staining and FACS analysis using annexin V/PI antibodies showed that apoptosis increased in INS-1 cells or primary cultured islets when incubated with BSA conjugated with glyceraldehyde (AGE2) or glucoaldehyde (AGE3), compared with those conjugated with glucose (AGE1). Reaction of INS-1 cells to Ki67, which is a cellular marker for proliferation, was also increased after AGE treatment. The ability of primary cultured islets to secrete insulin was retained even after AGE treatment under either low or high glucose conditions. The antiserum against RAGE partially prevented AGE-induced cellular events. Treatment of beta cells with the antioxidant metallothionein results in a significant reduction in pathologic changes. AGEs might be able to induce apoptosis as well as proliferation of pancreatic beta cell lines or primary cultured islets. Moreover, antibody array showed that RAD51 and RAD52 were significantly decreased in AGE2-treated INS-1 cells. AGEs might inhibit homologous DNA recombination for repairing DNA of INS-1 cells damaged by ROS generation. It might be suggested that treatment of AGEs resulted in ROS production and apoptosis through their receptor on pancreatic beta cells. AGEs might deteriorate function of pancreatic beta cells in patients with long-term hyperglycemia.
我们在此报告晚期糖基化终产物（AGEs）对胰腺β细胞的细胞毒性。AGEs刺激活性氧（ROS）生成，但未阻止INS-1细胞系的增殖。胰腺β细胞系或原代培养的胰岛具有AGE受体（RAGE），AGE处理后其表达增加。TUNEL染色以及使用膜联蛋白V/PI抗体的流式细胞术分析表明，与用葡萄糖偶联的（AGE1）相比，INS-1细胞或原代培养的胰岛在与甘油醛偶联的牛血清白蛋白（AGE2）或葡糖醛酸偶联的牛血清白蛋白（AGE3）孵育时，凋亡增加。AGE处理后，INS-1细胞对增殖细胞标志物Ki67的反应也增强。即使在低葡萄糖或高葡萄糖条件下进行AGE处理后，原代培养的胰岛分泌胰岛素的能力仍得以保留。抗RAGE抗血清部分阻止了AGE诱导的细胞事件。用抗氧化剂金属硫蛋白处理β细胞可导致病理变化显著减少。AGEs可能能够诱导胰腺β细胞系或原代培养的胰岛凋亡以及增殖。此外，抗体阵列显示在AGE2处理的INS-1细胞中RAD51和RAD52显著减少。AGEs可能抑制同源DNA重组以修复因ROS生成而受损的INS-1细胞的DNA。可能提示AGE处理通过其在胰腺β细胞上的受体导致ROS产生和凋亡。AGEs可能会使长期高血糖患者的胰腺β细胞功能恶化。
