Garza-Ocañas L, Hsieh G C, Acosta D, Torres-Alanís O, Piñeyro-López A
Departamento de Farmacología y Toxicología, Facultad de Medicina, Universidad Autónoma de Nuevo, León, México.
Toxicology. 1992;73(2):191-201. doi: 10.1016/0300-483x(92)90102-k.
The present study was undertaken to assess and compare the in vitro cytotoxicity of toxins T-514 and T-544 of buckthorn (Karwinskia humboldtiana) using primary cultures of rat hepatocytes and keratinocytes. Cell cultures were exposed to 6, 12, 25 and 50 microM toxins for 2-, 4-, 6- and 24-h periods. Cytotoxicity was determined by release of the cytoplasmic enzyme, lactate dehydrogenase (LDH), in culture media, methylthiazoltetrazolium (MTT) reduction and neutral red (NR) uptake. An increase in LDH leakage was observed in liver cell cultures as early as 2 h with 50 microM T-544 and with 6 microM T-514 and T-544 at 6 h and 24 h, respectively. In the NR assay the toxicity was evident at 2 h with 12 microM T-514 and T-544 and with 6 microM concentrations of both toxins at 6 h. On the other hand, a decrease in MTT reduction was detected at 4 h with 50 microM concentrations of both toxins and with 25 microM T-544 and 12 microM T-514 at 6 h and 6 microM T-514 and T-544 at 24 h. Both toxins were shown to be highly hepatotoxic; T-514 was more toxic than T-544. In the skin cell cultures, the toxicity of the toxins was not as severe and was not expressed until 12 h of exposure.
本研究旨在利用大鼠肝细胞和角质形成细胞的原代培养物，评估并比较鼠李（卡氏鼠李）毒素T - 514和T - 544的体外细胞毒性。将细胞培养物暴露于6、12、25和50微摩尔的毒素中2、4、6和24小时。通过测定培养基中细胞质酶乳酸脱氢酶（LDH）的释放、噻唑蓝（MTT）还原和中性红（NR）摄取来确定细胞毒性。在肝细胞培养物中，早在2小时时，50微摩尔的T - 544以及在6小时和24小时时6微摩尔的T - 514和T - 544就观察到LDH泄漏增加。在NR测定中，12微摩尔的T - 514和T - 544在2小时时以及两种毒素6微摩尔浓度在6小时时毒性明显。另一方面，在4小时时，两种毒素50微摩尔浓度以及在6小时时25微摩尔的T - 544和12微摩尔的T - 514，在24小时时6微摩尔的T - 514和T - 544均检测到MTT还原减少。两种毒素均显示出高度肝毒性；T - 514比T - 544毒性更强。在皮肤细胞培养物中，毒素的毒性没有那么严重，直到暴露12小时才表现出来。
