Majeed S Abdul, Nambi K S N, Taju G, Vimal S, Venkatesan C, Hameed A S Sahul
OIE Reference Laboratory for WTD, PG and Research Department of Zoology, C. Abdul Hakeem College, Melvisharam, 632 509, Vellore District, Tamil Nadu, India.
Environ Sci Pollut Res Int. 2014 Dec;21(23):13539-50. doi: 10.1007/s11356-014-3279-8. Epub 2014 Jul 16.
The cytotoxicity, genotoxicity and oxidative stress of malachite green (MG) was investigated using the fish Channa striata kidney (CSK) and Channa striata gill (CSG) cell lines. Five concentrations ranging from 0.001 to 10 μg mL(-1) were tested in three independent experiments. Cytotoxicity was assessed by 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, Rhodamine 123 and Alamar Blue. The mitochondrial changes and apoptosis of MG-exposed cells were observed by Rhodamine 123 and acridine orange/ethidium bromide (AO/EB) staining, respectively. In vitro potential DNA damaging effect of MG was tested using comet assay. Mitochondrial damage, apoptosis and DNA fragmentation increased in a concentration-dependent manner. Additionally, DNA electrophoretic mobility experiments were carried out to study the binding effect of MG to double-stranded DNA (dsDNA) of cells. DNA shift mobility experiments showed that MG is capable of strongly binding to linear dsDNA causing its degradation. Biochemical parameters such as lipid peroxidation (MDA), catalase (CAT) activity and reduced glutathione (GSH) levels were evaluated after exposure to MG. In CSK and CSG cell lines exposed to MG for 48 h, a significant increase in lipid peroxidation, which might be associated with decreased levels of reduced glutathione and catalase activity in these cell lines (p < 0.001), was observed.
利用条纹鳢肾脏(CSK)和条纹鳢鳃(CSG)细胞系研究了孔雀石绿(MG)的细胞毒性、遗传毒性和氧化应激。在三个独立实验中测试了0.001至10μg mL(-1)的五个浓度。通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)法、罗丹明123和alamar蓝评估细胞毒性。分别通过罗丹明123和吖啶橙/溴化乙锭(AO/EB)染色观察MG暴露细胞的线粒体变化和凋亡。使用彗星试验测试MG在体外潜在的DNA损伤作用。线粒体损伤、凋亡和DNA片段化呈浓度依赖性增加。此外,进行了DNA电泳迁移实验以研究MG与细胞双链DNA(dsDNA)的结合作用。DNA迁移率实验表明MG能够与线性dsDNA强烈结合并导致其降解。在暴露于MG后评估了脂质过氧化(MDA)、过氧化氢酶(CAT)活性和还原型谷胱甘肽(GSH)水平等生化参数。在暴露于MG 48小时的CSK和CSG细胞系中,观察到脂质过氧化显著增加,这可能与这些细胞系中还原型谷胱甘肽水平降低和过氧化氢酶活性降低有关(p < 0.001)。