Damaging effects of advanced glycation end-products in the murine macrophage cell line J774A.1
Damaging effects of advanced glycation end-products in the murine macrophage cell line J774A.1
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DOI：
10.1016/s0887-2333(02)00016-4
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发表时间：
2002-08-01
影响因子：
3.2
通讯作者：
Cottalasso, D
中科院分区：
文献类型：
--
作者：
Bassi, AM;Ledda, S;Cottalasso, D
还原糖（如醛糖）与蛋白质的相互作用以及随后的分子重排产生不可逆的晚期糖基化终产物（AGEs），这是一类非酶糖化蛋白或脂质。AGEs形成交联，捕获大分子并释放活性氧中间体。AGEs与衰老有关，并增加了几种相关疾病。本研究的目的是在鼠巨噬细胞系J774A.1中评估48 It暴露于含有已知量戊糖苷的糖化血清的影响，戊糖苷是一种在糖尿病和尿毒症受试者的血浆和组织中发现的众所周知的AGE。将胎牛血清与核糖（50 mM）在37 ℃下孵育7天以获得约10 nmol/ml的戊糖苷。研究的细胞毒性参数是通过中性红摄取、乳酸脱氢酶释放和四唑盐试验的细胞形态和活力。在培养基中，并在细胞内室，结合和游离戊糖苷进行了评价，通过HPLC，作为敏感和特异性标记物，和硫代巴比妥酸反应物质（TBARS），作为脂质过氧化程度的指标。我们的研究结果证实，巨噬细胞能够采取戊糖苷。可以想象，结合的戊糖苷被降解，游离的戊糖苷在细胞内释放，然后释放到培养基中。培养基中AGE的增加与TBAR的增加相结合，这意味着发生了氧化应激;观察到了显著的细胞毒性作用，随后游离戊糖苷和TBAR释放到培养基中。(C)2002爱思唯尔科技有限公司。保留所有权利。
The interaction of reducing sugars, such as aldose, with proteins and the subsequent molecular rearrangements, produces irreversible advanced glycation end-products (AGEs), a heterogeneous class of non-enzymatic glycated proteins or lipids. AGEs form cross-links, trap macromolecules and release reactive oxygen intermediates. AGEs are linked to aging, and increase in several related diseases. The aim of this study was to assess, in a murine macrophage cell line, J774A.1, the effects of 48 It of exposure to glycated serum containing a known amount of pentosidine, a well-known AGE found in the plasma and tissues of diabetic and uremic subjects. Fetal bovine serum was incubated with ribose (50 mm) for 7 days at 37 degreesC to obtain about 10 nmol/ml of pentosidine. The cytotoxic parameters studied were cell morphology and viability by neutral red uptake, lactate dehydrogenase release and tetrazolium salt test. In the medium and in the intracellular compartment, bound and free pentosidine were evaluated by HPLC, as sensitive and specific glycative markers, and thiobarbituric acid reactive substances (TBARs), as index of the extent of lipid peroxidation. Our results confirm that macrophages are able to take up pentosidine. It is conceivable that bound pentosidine is degraded and free pentosidine is released inside the cell and then into the medium. The AGE increase in the medium was combined with an increase in TBARs, meaning that an oxidative stress occurred; marked cytotoxic effects were observed, and were followed by the release of free pentosidine and TBARs into the culture medium. (C) 2002 Elsevier Science Ltd. All rights reserved.