Gadelha F R, Moreno S N, De Souza W, Cruz F S, Docampo R
Instituto de Microbiologia, Universidade Federal do Rio de Janeiro, Brazil.
Mol Biochem Parasitol. 1989 May 1;34(2):117-26. doi: 10.1016/0166-6851(89)90003-0.
The first morphological alteration observed in Trypanosoma cruzi different stages upon incubation with crystal violet was mitochondrial swelling. The use of digitonin to solubilize T. cruzi plasma membrane allowed the demonstration of an uncoupling action of crystal violet on epimastigote mitochondria in situ. Low concentrations of crystal violet (20-50 microM) or carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP; 0.5 microM) uncoupled the respiratory control mechanism. The inhibition of State 3 respiration by oligomycin was released by crystal violet or FFCCP. Crystal violet released respiratory control, and enhanced ATPase activity of digitonin-permeabilized epimastigotes. Higher concentrations of crystal violet inhibited mitochondrial respiration. The uncoupled effect of crystal violet was stimulated by inorganic phosphate. In addition, crystal violet inhibited endongenous and glucose-stimulated respiration of the intact epimastigotes, and inhibited the Mg2+-ATPase in the epimastigote mitochondrial fractions. The inhibition of this Mg2+-ATPase increased up to pH 9.0 and decreased with increasing protein concentration. These data indicate that the T. cruzi mitochondrion is apparently the main target of crystal violet toxicity.
在用结晶紫孵育后，在克氏锥虫不同阶段观察到的第一个形态学改变是线粒体肿胀。使用洋地黄皂苷溶解克氏锥虫细胞膜，证实了结晶紫对体内前鞭毛体线粒体的解偶联作用。低浓度的结晶紫（20 - 50微摩尔）或羰基氰对三氟甲氧基苯腙（FCCP；0.5微摩尔）使呼吸控制机制解偶联。寡霉素对状态3呼吸的抑制作用被结晶紫或FFCCP解除。结晶紫解除了呼吸控制，并增强了经洋地黄皂苷通透处理的前鞭毛体的ATP酶活性。较高浓度的结晶紫抑制线粒体呼吸。结晶紫的解偶联作用受到无机磷酸盐的刺激。此外，结晶紫抑制完整前鞭毛体的内源性呼吸和葡萄糖刺激的呼吸，并抑制前鞭毛体线粒体部分中的Mg2 + -ATP酶。这种Mg2 + -ATP酶的抑制作用在pH 9.0时增强，并随着蛋白质浓度的增加而降低。这些数据表明，克氏锥虫线粒体显然是结晶紫毒性的主要靶点。
