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铝盐和铅盐对人皮肤成纤维细胞脂质过氧化及细胞存活的影响。

Effect of aluminum and lead salts on lipid peroxidation and cell survival in human skin fibroblasts.

作者信息

Dominguez M C, Sole E, Goñi C, Ballabriga A

机构信息

Unitat de Recerca Biomèdica, Hospital Universitari Materno-infantil Vall d'Hebron, Barcelona, Spain.

出版信息

Biol Trace Elem Res. 1995 Jan-Mar;47(1-3):57-67. doi: 10.1007/BF02790101.

Abstract

The aim of this study was to see whether aluminum (Al) and lead (Pb) salts are toxic for cultured human fibroblasts under different experimental conditions, in the controllable situation offered by cell cultures. Cell survival and membrane lipid peroxidation served as markers of Al and Pb toxicity. Evaluation of the living cells was carried out using a colorimetric method, the mitochondrial reduction of 1-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT). Lipoperoxidation assay was performed on whole cell homogenates by measuring thiobarbituric acid-reactive substances (TBARS) produced after incubation with ascorbic acid-ferrous sulfate. Al(III) and Pb(II) salts (300 microM) produce a considerable decrease in cell survival after an exposure period of 4d, evident with the three fetal calf serum concentrations in the culture media: 2, 5, and 10%. Taking into account in vitro cell aging, the cytotoxic effects of Al(III) and Pb(II) are greater in senescent fibroblasts than in young cells. Lead-induced cytotoxicity is higher than Al-induced cytotoxicity. A mechanism that contributes to cellular toxicity is membrane lipid peroxidation; our results demonstrate that Al(III) and Pb(II) ions, 400 microM, exert an antioxidant-like effect or a pro-oxidant action on cell membranes depending on exposure time. We describe significant increases in TBARS formation associated with the presence of 400 microM Al(III) or Pb(II) salts in the culture media. Our study also revealed that these heavy metals induce a cell age-dependent action on membrane lipoperoxidation that is greater in senescent fibroblasts and this could have severe consequences for maintenance of cellular integrity.

摘要

本研究的目的是在细胞培养所提供的可控条件下,观察铝(Al)盐和铅(Pb)盐在不同实验条件下对培养的人成纤维细胞是否有毒性。细胞存活率和膜脂质过氧化作为铝和铅毒性的标志物。使用比色法,即1-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)的线粒体还原法对活细胞进行评估。通过测量与抗坏血酸 - 硫酸亚铁孵育后产生的硫代巴比妥酸反应性物质(TBARS),对全细胞匀浆进行脂质过氧化测定。在4天的暴露期后,Al(III)和Pb(II)盐(300 microM)会使细胞存活率显著降低,在培养基中三种胎牛血清浓度(2%、5%和10%)下均很明显。考虑到体外细胞衰老,Al(III)和Pb(II)对衰老成纤维细胞的细胞毒性作用比对年轻细胞更大。铅诱导的细胞毒性高于铝诱导的细胞毒性。导致细胞毒性的一种机制是膜脂质过氧化;我们的结果表明,400 microM的Al(III)和Pb(II)离子根据暴露时间对细胞膜发挥抗氧化样作用或促氧化作用。我们描述了与培养基中存在400 microM Al(III)或Pb(II)盐相关的TBARS形成显著增加。我们的研究还表明,这些重金属对膜脂质过氧化具有细胞年龄依赖性作用,在衰老成纤维细胞中更大,这可能对维持细胞完整性产生严重后果。

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