Sugimoto N, Ozutsumi K, Matsuda M
Eur J Epidemiol. 1985 Dec;1(4):264-73. doi: 10.1007/BF00237101.
The morphological alterations (bleb-balloon formation) induced by Clostridium perfringens type A enterotoxin in HeLa and Vero cells were studied under defined extracellular conditions. The action of enterotoxin was found to depend on the temperature but not on energy metabolism. The morphological alterations by the enterotoxin occurred in phosphate buffered saline containing Ca2+ and Mg2+. Of the constituents of the buffered saline, Ca2+ was essential for the morphological alterations and other ions were interchangeable. The morphological alterations by the enterotoxin occurred also in 10 mM Hepes-Na buffer, pH 7.2 containing NaCl, KCl or choline chloride at a concentration of over ca. 50 mM and in 10 mM Hepes-Ca buffer, pH 7.2 containing CaCl2 at a concentration of over ca. 50 mM. Addition of sucrose to the medium prevented induction of the morphological alterations. The amount of sucrose necessary to protect the cells increased with increase in NaCl, KCl or CaCl2 concentration in the medium. A calcium ionophore A23187 mimicked the action of enterotoxin. Examination of the cation contents of the cells by atomic absorption spectrophotometry showed early and rapid increase of Ca2+ during intoxication with concomitant changes in Na+, K+ and Mg2+ that reduced the ion concentration gradients between inside and outside of the cell present before toxin treatment. The mechanism of action of C. perfringens type A enterotoxin is discussed on the basis of these findings.
在特定的细胞外条件下，研究了A型产气荚膜梭菌肠毒素在HeLa细胞和Vero细胞中诱导的形态学改变（泡状-气球样形成）。发现肠毒素的作用取决于温度，而不取决于能量代谢。肠毒素引起的形态学改变发生在含有Ca2+和Mg2+的磷酸盐缓冲盐水中。在缓冲盐的成分中，Ca2+对于形态学改变是必需的，其他离子可以相互替换。肠毒素引起的形态学改变也发生在含有浓度超过约50 mM的NaCl、KCl或氯化胆碱的10 mM Hepes-Na缓冲液（pH 7.2）中，以及含有浓度超过约50 mM的CaCl2的10 mM Hepes-Ca缓冲液（pH 7.2）中。向培养基中添加蔗糖可防止形态学改变的诱导。保护细胞所需的蔗糖量随着培养基中NaCl、KCl或CaCl2浓度的增加而增加。钙离子载体A23187模拟了肠毒素的作用。通过原子吸收分光光度法检测细胞的阳离子含量表明，中毒期间Ca2+早期快速增加，同时Na+、K+和Mg2+发生变化，降低了毒素处理前细胞内外的离子浓度梯度。基于这些发现，讨论了A型产气荚膜梭菌肠毒素的作用机制。
