Königs Maika, Schwerdt Gerald, Gekle Michael, Humpf Hans-Ulrich
Institut für Lebensmittelchemie, Westfälische Wilhelms-Universität Münster, Germany.
Mol Nutr Food Res. 2008 Jul;52(7):830-9. doi: 10.1002/mnfr.200700439.
Toxic effects of the mycotoxin deoxynivalenol (DON) observed in animals range from diarrhea, vomiting, gastro-intestinal inflammation to necrosis of several tissues. In the last years, DON has been tested in hepatocytes of several animal species for its cytotoxicity. However, these tests are limited to the use of animal cells. No studies using human hepatocytes are available. Further investigations with the human hepatocellular liver carcinoma cell line HepG2 might be limited due to the disadvantages of cell lines (e. g. immortalization, tumor derivation, longtime cultivation) and do not necessarily reflect the response of normal human cells. In order to overcome this problem and to be closer to the human situation, we studied the effect of DON in human primary hepatocytes and compared these data to the effects in the HepG2 cell line. Cell viability, apoptotic and necrotic cell death, albumin secretion and metabolic activity were determined. It could be demonstrated that DON has a distinct cytotoxic effect on human primary hepatocytes. Viability, protein content and albumin secretion were reduced in a dose-dependent manner. The apoptotic key enzyme caspase-3 was activated, while LDH release occurred only after long incubation time due to a secondary necrosis. Furthermore, we studied the metabolism of DON using LC-MS/MS. DON was neither metabolized by primary hepatocytes cells nor by the HepG2 cell line.
在动物身上观察到的霉菌毒素脱氧雪腐镰刀菌烯醇（DON）的毒性作用范围从腹泻、呕吐、胃肠道炎症到多个组织的坏死。在过去几年中，DON已在几种动物的肝细胞中进行了细胞毒性测试。然而，这些测试仅限于使用动物细胞。尚无使用人类肝细胞的研究。由于细胞系存在的缺点（如永生化、肿瘤来源、长期培养），对人肝细胞肝癌细胞系HepG2的进一步研究可能受到限制，且不一定能反映正常人类细胞的反应。为了克服这一问题并更接近人类实际情况，我们研究了DON对原代人肝细胞的影响，并将这些数据与在HepG2细胞系中的影响进行了比较。测定了细胞活力、凋亡和坏死性细胞死亡、白蛋白分泌和代谢活性。结果表明，DON对原代人肝细胞具有明显的细胞毒性作用。活力、蛋白质含量和白蛋白分泌呈剂量依赖性降低。凋亡关键酶半胱天冬酶-3被激活，而乳酸脱氢酶（LDH）释放仅在长时间孵育后由于继发性坏死才发生。此外，我们使用液相色谱-串联质谱法（LC-MS/MS）研究了DON的代谢情况。DON既不被原代肝细胞代谢，也不被HepG2细胞系代谢。
