Antiglycation effect of gliclazide on in vitro AGE formation from glucose and methylglyoxal
Antiglycation effect of gliclazide on in vitro AGE formation from glucose and methylglyoxal
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DOI：
10.3181/0705-bc-131
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发表时间：
2008-02-01
影响因子：
3.2
通讯作者：
Hamada, Yoji
中科院分区：
文献类型：
--
作者：
Li, Weiguo;Ota, Kimiko;Hamada, Yoji
格列齐特是一种广泛用于治疗糖尿病的磺酰脲类药物，已知其可抑制活性氧。为了阐明其抗氧化能力是否干扰糖化过程，我们孵育牛血清白蛋白（BSA）与1 M葡萄糖或1 mM甲基乙二醛，在存在或不存在格列齐特，并观察形成的晚期糖化终产物（AGES）。通过AGE特异性荧光、酶联免疫吸附试验（ELISA）和Western印迹法评估AGE的产生。通过将BSA与1 M葡萄糖或1 mM甲基乙二醛孵育，在320/383 nm和335/385 nm的激发/发射波长处的荧光明显增加，并且1 mM格列齐特以剂量依赖性方式在两个波长处显著减弱荧光增强。格列齐特对葡萄糖诱导的荧光的抑制作用几乎与氨基胍（一种公认的抗糖化剂）相当，而格列齐特对甲基甘氨酸衍生的荧光形成的抑制作用低于氨基胍。用ELISA法测定的AGE浓度显示相似的结果。BSA与1 M葡萄糖或1 mM甲基乙二醛的孵育产生了羧甲基赖氨酸或精氨酸嘧啶的明显增加。这两个AGES显着降低1 mM格列齐特和葡萄糖衍生的羧甲基赖氨酸的减少是由氨基胍引起的。Western blotting结果支持ELISA结果。据我们所知，本研究首次提供了格列齐特对葡萄糖和丙酮醛体外形成AGE的抗糖化作用的证据。
Gliclazide, a sulfonylurea widely used for treatment of diabetes mellitus, is known to scavenge reactive oxygen species. To clarify whether its antioxidative ability interferes with the glycation processes, we incubated bovine serum albumin (BSA) with 1 M glucose or 1 mM methylglyoxal, in the presence or absence of gliclazide, and observed the formation of advanced glycation end products (AGES). AGE production was assessed by AGE-specific fluorescence, an enzyme-linked immunosorbent assay (ELISA), and Western blotting. The fluorescence at excitation/emission wavelengths of 320/383 nm and 335/385 nm was definitely increased by incubating BSA with 1 M glucose or 1 mM methylglyoxal, and 1 mM gliclazide significantly blunted the fluorescent augmentation, in both wavelengths, in a dose-dependent fashion. Gliclazide almost equaled to aminoguanidine, a putative antiglycation agent, in the inhibitory effect on the glucose-induced fluorescence, while the methylglyoxal-derived fluorescent formation was less suppressed by gliclazide than by aminoguanidine. The AGE concentrations determined by ELISA showed similar results. Incubation of BSA with 1 M glucose or 1 mM methylglyoxal yielded an apparent increase in carboxymethyllysine or argpyrimidine. Both AGES were significantly lowered by 1 mM gliclazide and a reduction of glucose-derived carboxymethyllysine was comparable to that caused by aminoguanidine. The results of Western blotting supported the findings in ELISA. To our knowledge, the present study provides the first evidence of the antiglycation effect of gliclazide on in vitro AGE formation from glucose and methylglyoxal.