Department of Pharmacology and Toxicology, Faculty of Pharmacy, Comenius University in Bratislava, Slovak Republic.
Arch Razi Inst. 2024 Apr 30;79(2):307-314. doi: 10.32592/ARI.2024.79.2.307. eCollection 2024 Apr.
The compound 1-methyltryptophan (1-MT) has been shown to act protectively in renal ischemia-reperfusion injury. Toll-like receptor 4 signaling is also a regular process of epithelial-mesenchymal transition (EMT) that can after ischemia-reperfusion injury (IRI) result in an increase in renal fibrosis. EMT is associated with specific transcription factors: Snai1, Snai2, Zeb1, and Twist. 1-MT could regulate EMT and act as an antifibrotic agent. This study aimed to investigate the effect of 1-MT on EMT transcription factors in tubular epithelial cells that underwent 30 min. Renal tubular epithelial cells (TECs) were isolated from Lewis rats using a standard protocol with FeO magnetic separation and selective media as previously mentioned. Cells were cultivated and divided into 4 groups, namely C-TECs: control cells, IRI-TECs: IRI-induced TECs, D-IRI-TECs: IRI-induced TECs treated with 1-methyl-D-tryptophan, and L-IRI-TECs: IRI-induced TECs treated with 1-methyl-L-tryptophan. IRI was induced in all groups for 30 min by mineral oil (except for C-TECs) followed by 48-hour reperfusion. RNA and proteins were isolated from harvested cells. Using a semi-quantitative polymerase chain reaction, we assessed the relative mRNA expression of EMT transcription factors Snai1, Snai2, Zeb1, and Twist. Hereby, we showed that the treatment of ischemia-induced TECs with both 1-MT isomers lowered the expression of EMT transcription factors Snai1 and Zeb1 which were increased by ischemia and reperfusion of TECs. This could act favorably in renal IRI decreasing EMT and renal fibrosis, therefore showing the potential of 1-MT as a part of therapy in renal transplantation aimed at renal ischemia-reperfusion injury.
1-甲基色氨酸（1-MT）已被证明在肾缺血再灌注损伤中具有保护作用。Toll 样受体 4 信号也是上皮-间充质转化（EMT）的一个常规过程，它可以在缺血再灌注损伤（IRI）后导致肾纤维化增加。EMT 与特定的转录因子有关：Snai1、Snai2、Zeb1 和 Twist。1-MT 可以调节 EMT 并发挥抗纤维化作用。本研究旨在探讨 1-MT 对经历 30 分钟的肾小管上皮细胞 EMT 转录因子的影响。使用 FeO 磁分离和前面提到的选择性培养基的标准方案从 Lewis 大鼠中分离出肾小管上皮细胞（TEC）。将细胞培养并分为 4 组，即 C-TECs：对照细胞、IRI-TECs：IRI 诱导的 TECs、D-IRI-TECs：用 1-甲基-D-色氨酸处理的 IRI 诱导的 TECs 和 L-IRI-TECs：用 1-甲基-L-色氨酸处理的 IRI 诱导的 TECs。所有组的 IRI 诱导时间均为 30 分钟，用矿物油（除 C-TECs 外）诱导，然后再进行 48 小时再灌注。从收获的细胞中分离 RNA 和蛋白质。使用半定量聚合酶链反应，我们评估了 EMT 转录因子 Snai1、Snai2、Zeb1 和 Twist 的相对 mRNA 表达。结果表明，两种 1-MT 异构体处理缺血诱导的 TECs 可降低 EMT 转录因子 Snai1 和 Zeb1 的表达，而 Snai1 和 Zeb1 的表达在 TECs 的缺血和再灌注中增加。这可能有利于肾 IRI 减少 EMT 和肾纤维化，因此显示了 1-MT 作为旨在减少肾缺血再灌注损伤的肾移植治疗的一部分的潜力。
