Labieniec Magdalena, Biernat Monika, Gabryelak Teresa
Department of General Biophysics, Institute of Biophysics, University of Lodz, 12/16 Banacha Street, 90-237 Lodz, Poland.
Cell Biol Int. 2007 Jul;31(7):683-90. doi: 10.1016/j.cellbi.2006.12.005. Epub 2006 Dec 30.
The exposure of freshwater mussels Unio tumidus to phenolic compounds (tannic, ellagic and gallic acid) in vivo caused changes in proteins and DNA function of digestive gland cells. The mussels were exposed to various concentrations of tested polyphenols (60, 200 and 500 microM) for 24 and 48 h and their antioxidant and pro-oxidant effects were determined. The number of SH-groups was quantified spectrophotometrically using Ellman's reagent. Oxidative modification of proteins increased in the digestive gland cells in a dose- and time-dependent manner. The level of nuclear DNA damage was investigated using the comet assay. The results revealed that polyphenolic acids induce single and double-strand breaks in DNA. The highest changes were observed for tannic and gallic acids and the smallest ones for ellagic acid. 1h of DNA repair process was also studied using the same method. The data obtained in this experiment demonstrate that the most effective DNA repair occurs in the cells exposed to phenolic compounds for 24h. A longer incubation (up to 48 h) does not decrease the capacity of the repair mechanism. The antioxidant activity of the tested phenols was analyzed spectrofluorimetrically using a fluorescence probe DCFH-DA (dichlorofluorescein-diacetate). The experimental data showed that the tested acids can act as antioxidants when used at higher doses (200 and 500 microM) against the reactive oxygen species present in the digestive gland cells. The most effective was ellagic acid, also applied at the smallest dose of 60 microM, in comparison with tannic and gallic acids. In conclusion, our results demonstrate that chosen water-soluble polyphenols, which are located in various plant tissues and are also found in the aquatic environment, can influence organisms living in the water. They can be exposed to these chemicals that cause morphological alterations and changes in certain physiological processes in their organs (i.e. digestive gland cells of bivalve molluscs).
将淡水贻贝圆顶珠蚌暴露于酚类化合物（单宁酸、鞣花酸和没食子酸）中，会导致其消化腺细胞的蛋白质和DNA功能发生变化。将贻贝暴露于不同浓度的受试多酚（60、200和500微摩尔）中24小时和48小时，并测定其抗氧化和促氧化作用。使用埃尔曼试剂通过分光光度法定量SH基团的数量。消化腺细胞中蛋白质的氧化修饰呈剂量和时间依赖性增加。使用彗星试验研究核DNA损伤水平。结果表明，多酚酸可诱导DNA单链和双链断裂。单宁酸和没食子酸引起的变化最大，而鞣花酸引起的变化最小。还使用相同方法研究了1小时的DNA修复过程。本实验获得的数据表明，在暴露于酚类化合物24小时的细胞中发生了最有效的DNA修复。更长时间的孵育（长达48小时）不会降低修复机制的能力。使用荧光探针DCFH-DA（二氯荧光素二乙酸酯）通过荧光分光光度法分析受试酚类的抗氧化活性。实验数据表明，受试酸在较高剂量（200和500微摩尔）下可作为抗氧化剂，对抗消化腺细胞中存在的活性氧。与单宁酸和没食子酸相比，最有效的是鞣花酸，其最小剂量为60微摩尔时也有效果。总之，我们的结果表明，存在于各种植物组织中且在水生环境中也能发现的特定水溶性多酚，可影响生活在水中的生物。它们可能会接触到这些化学物质，从而导致其器官（即双壳贝类的消化腺细胞）出现形态改变和某些生理过程的变化。
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