Ochratoxin A (OTA) is universally known to induce nephrotoxicity via inducing oxidative stress and apoptosis, inhibiting protein synthesis and activating autophagy. Our previous studies have proved that OTA induces nephrotoxicity in vitro and in vivo by adjusting the NOD-like receptor protein 3 (NLRP3) inflammasome activation and caspase-1-dependent pyroptosis. Based on these findings, we further investigated the protective role of selenomethionine (SeMet) on OTA-caused nephrotoxicity using the Madin-Darby canine kidney (MDCK) epithelial cells as an in vitro model, proposing to offer a new way for remedying OTA-induced nephrotoxicity by nutritional manipulation. We measured the cell vitality, lactate dehydrogenase (LDH) activity and the expression of renal fibrotic genes, NLRP3 inflammasome and pyroptosis related genes. MTT and LDH results indicated that SeMet supplementation significantly mitigated 2.0 μg/ml OTA-induced cytotoxicity in MDCK cells (p < 0.05). Meanwhile, SeMet alleviated OTA induced increase of reactive oxygen species in MDCK cells. Then, the expressions of α-SMA, Vimentin, and TGF-β were detected both in mRNA and protein levels. The results indicated 8 μM SeMet supplementation could significantly downregulate the expression of OTA-induced renal fibrosis-related genes (p < 0.05). In addition, the upregulation of OTA-induced NLRP3 inflammasome and pyroptosis downstream genes was also significantly inhibited by 8 μM of SeMet (p < 0.05). In summary, SeMet could alleviate OTA-induced renal fibrotic genes expression and reduce NLRP3-caspase-1-dependent pyroptosis. Therefore, SeMet supplementation may become an effective approach for preserving animals from renal injury exposed to OTA.

中文翻译：

---

硒代蛋氨酸减轻赭曲霉毒素A诱导的MDCK细胞焦亡和肾纤维化因子的表达

众所周知，赭曲霉毒素 A (OTA) 通过诱导氧化应激和细胞凋亡、抑制蛋白质合成和激活自噬来诱导肾毒性。我们之前的研究证明，OTA 通过调节 NOD 样受体蛋白 3 (NLRP3) 炎性体激活和 caspase-1 依赖性细胞焦亡，在体外和体内诱导肾毒性。基于这些发现，我们以 Madin-Darby 犬肾 (MDCK) 上皮细胞为体外模型，进一步研究了硒代蛋氨酸 (SeMet) 对 OTA 引起的肾毒性的保护作用，提出了一种治疗 OTA 诱导的新方法。营养操作引起的肾毒性。我们测量了细胞活力、乳酸脱氢酶 (LDH) 活性以及肾纤维化基因、NLRP3 炎性体和细胞焦亡相关基因的表达。p  < 0.05)。同时，SeMet 减轻了 OTA 诱导的 MDCK 细胞中活性氧的增加。然后，在mRNA和蛋白质水平检测α-SMA、波形蛋白和TGF-β的表达。结果表明，补充 8 μM SeMet 可显着下调 OTA 诱导的肾纤维化相关基因的表达（p  < 0.05）。此外，8 μM SeMet 也显着抑制 OTA 诱导的 NLRP3 炎症小体和细胞焦亡下游基因的上调（p  < 0.05）。总之，SeMet 可以减轻 OTA 诱导的肾纤维化基因表达并减少 NLRP3-caspase-1 依赖性细胞焦亡。因此，SeMet 补充剂可能成为保护动物免受 OTA 肾损伤的有效方法。