Czerucka D, Nano J L, Bernasconi P, Rampal P
Laboratoire de Gastroentérologie et Nutrition, Faculté de Médecine, Nice.
Gastroenterol Clin Biol. 1991;15(1):22-7.
In vivo, Clostridium difficile acts by releasing 2 toxins: toxin A, an enterotoxin, and toxin B, a cytotoxin. This study was performed to determine: a) whether the rat epithelial intestinal cell line IRD 98 responds to Clostridium difficile toxin A and B; b) whether the yeast Saccharomyces boulardii has an effect on this model. Evaluation of 3H-thymidine incorporation into IRD 98 cells exposed to toxin B revealed that DNA synthesis was inhibited for low concentrations (10 ng/ml). For higher concentrations, DNA synthesis was not modified. Evaluation of 14C-leucine incorporation into IRD 98 cells exposed to toxin B revealed that this toxin affected protein synthesis. Whereas cholera toxin stimulates adenylate cyclase in IRD 98 cells, cAMP levels in cells exposed to various quantities of toxin A was similar to that in control cells. As opposed to cholera toxin, which does not affect the cytoskeletal structure of IRD 98 cells, 7 micrograms/ml of toxin A and even smaller amounts of toxin B (1 ng/ml) were found to cause structural alterations by immunofluorescence studies. Prior exposition of cells to Saccharomyces boulardii reduced or prevented the rounding of cells in the presence of these toxins. IRD 98 cells can thus be considered a good model for in vitro investigation of the effects of Clostridium difficile toxins A and B, and findings suggest that Saccharomyces boulardii has a protective effect against the action of these toxins.
在体内,艰难梭菌通过释放两种毒素发挥作用:毒素A,一种肠毒素;毒素B,一种细胞毒素。进行这项研究的目的是确定:a)大鼠上皮肠细胞系IRD 98是否对艰难梭菌毒素A和B有反应;b)布拉酵母菌对该模型是否有影响。对暴露于毒素B的IRD 98细胞中3H-胸腺嘧啶核苷掺入情况的评估显示,低浓度(10 ng/ml)时DNA合成受到抑制。对于更高浓度,DNA合成未发生改变。对暴露于毒素B的IRD 98细胞中14C-亮氨酸掺入情况的评估显示,该毒素影响蛋白质合成。霍乱毒素可刺激IRD 98细胞中的腺苷酸环化酶,而暴露于不同量毒素A的细胞中的cAMP水平与对照细胞中的相似。与不影响IRD 98细胞细胞骨架结构的霍乱毒素不同,免疫荧光研究发现,7微克/毫升的毒素A甚至更少量的毒素B(1 ng/ml)会导致结构改变。在这些毒素存在的情况下,预先将细胞暴露于布拉酵母菌可减少或防止细胞变圆。因此,IRD 98细胞可被视为体外研究艰难梭菌毒素A和B作用的良好模型,研究结果表明布拉酵母菌对这些毒素的作用具有保护作用。