Gasiorowski K, Brokos B, Kulma A, Ogorzałek A, Skórkowska K
Wroclaw Medical University, Department of Basic Medical Sciences, Kochanowskiego 14, 51- 601 Wrocław, Poland.
Cell Mol Biol Lett. 2001;6(3):649-75.
An antimutagenic activity of fluphenazine, todralazine, anthocyanins and alkylresorcinols was established in a battery of short-term cytogenetic tests. One of the possible mechanisms of their antimutagenic action could be an increase in apoptotic elimination of heavily-damaged cells from a culture. In this paper we provide data on quantitative estimation of the antimutagens' impact on apoptosis in lymphocyte cultures exposed in the G(0)-phase to genotoxic agents: hydrogen peroxide (0.2mM, 20 min.) or benzo[a]pyrene (40 microM, 90 min.), and then cultured for 36 hrs in the presence of a lectin (PHA-M, 1% v/v) and each of the tested antimutagens. Apoptosis was estimated by means of microscopic examination of cell smears stained with a mixture of fluorochromes (ethidium bromide/acridine orange) as well as of the results of DNA separation with the field inversion gel electrophoresis. By microscopic examination we assessed that the frequencies of cells exhibiting morphological features of apoptosis considerably increased in the cultures containing the antimutagens. The FIGE separation of DNA from those cultures proved that the DNA content in the 30-50 kb domain was markedly elevated, as compared with the control cultures that did not contain antimutagens. It was established in the regression analysis that the apoptosis-enhancing effect significantly depended on the concentration of each tested antimutagen in a culture medium. However, marked differences of apoptosis-enhancing potency were noticed among the four antimutagens. The multicriterial analysis proved that the apoptosis-enhancing effects of fluphenazine and also, to a smaller extent, by alkylresorcinols, were many times stronger than those of anthocyanins and of todralazine. The results suggest that the enhancement of apoptosis by fluphenazine and by alkylresorcinols can explain a major part of their antimutagenic activity, whereas in the case of anthocyanins and of todralazine other mechanisms of antimutagenic action should be sought for.
在一系列短期细胞遗传学试验中确定了氟奋乃静、托屈嗪、花色苷和烷基间苯二酚的抗诱变活性。它们抗诱变作用的一种可能机制可能是增加从培养物中凋亡清除严重受损细胞的能力。在本文中，我们提供了关于抗诱变剂对处于G(0)期的淋巴细胞培养物中凋亡影响的定量估计数据，这些淋巴细胞培养物暴露于遗传毒性剂：过氧化氢（0.2mM，20分钟）或苯并[a]芘（40μM，90分钟），然后在存在凝集素（PHA-M，1% v/v）和每种测试抗诱变剂的情况下培养36小时。通过用荧光染料混合物（溴化乙锭/吖啶橙）染色的细胞涂片的显微镜检查以及场反转凝胶电泳的DNA分离结果来估计凋亡。通过显微镜检查，我们评估了在含有抗诱变剂的培养物中表现出凋亡形态特征的细胞频率显著增加。从那些培养物中进行的场反转凝胶电泳DNA分离证明，与不含抗诱变剂的对照培养物相比，30-50kb区域的DNA含量明显升高。回归分析表明，凋亡增强作用显著取决于培养基中每种测试抗诱变剂的浓度。然而，在这四种抗诱变剂之间注意到凋亡增强效力存在显著差异。多标准分析证明，氟奋乃静以及在较小程度上烷基间苯二酚的凋亡增强作用比花色苷和托屈嗪强许多倍。结果表明，氟奋乃静和烷基间苯二酚诱导的凋亡增强可以解释它们抗诱变活性的主要部分，而对于花色苷和托屈嗪，则应寻找其他抗诱变作用机制。
