Teng J, Sun F, Wang X, Fang Z, Sun Y, Li J
Department of Nephrology, Yantaishan Hospital, Yantai City, Shandong Province, 264003, China.
Department of Clinical Laboratory, Yantaishan Hospital, Yantai City, Shandong Province, 264003, China.
J Physiol Pharmacol. 2024 Oct;75(5). doi: 10.26402/jpp.2024.5.02. Epub 2024 Dec 4.
Melatonin (Mel) has been documented to modulate epithelial-mesenchymal transition (EMT) in cellular systems. The interstitial transformation of renal tubular epithelial cells constitutes a key pathogenic mechanism underlying renal fibrosis. This study aims to elucidate the role of Mel in the EMT process of renal tubular epithelial cells. A rat model of unilateral ureteral obstruction (UUO) was developed through unilateral ureteral ligation, followed by treatment with Mel (5, 10, and 20 mg/kg). Subsequent analyses included histopathological examination, measurement of creatinine and blood urea nitrogen levels, immunofluorescence analysis of fibronectin (FN), and immunohistochemical analysis of alpha-smooth muscle actin (α-SMA). Transforming growth factor-beta 1 (TGF-β1) initiates a fibrotic response in NRK-52E cells, which is subsequently treated with Mel (0. 1, 1, and 10 μmol/L). Evaluates cell viability, migration, and the expression of EMT related proteins, including FN, α-SMA, collagenase I, and E-cadherin. To validate the regulatory interaction between miR-153-3p and Forkhead transcription factor o subfamily member 3A (FOXO3A), cells are transfected with miR-153-3p mimics or siRNA targeting FOXO3A (si-FOXO3A). In results Mel exhibits a dose-dependent capacity to ameliorate renal injury and rectify glomerular structural abnormalities in UUO rat models. In comparison to UUO model rats, melatonin significantly reduced the expression levels of FN and α-SMA. In vitro studies demonstrated that Mel inhibited the activity and migratory behavior of cells, as well as the protein expression levels of FN, α-SMA, and collagenase I in NRK-52E cells. Relative to the control group, UUO rats and TGF-β1-induced NRK-52E cells exhibited elevated expression of miR-153-3p and reduced FOXO3A. In vitro models further revealed that upregulation of miR-153-3p or downregulation of FOXO3A can negate the protective effects of Melatonin on TGF-β1-induced EMT. Concluding, Mel inhibits miR-153-3p, thereby promoting the transcription of FOXO3A in UUO rats, which alleviates renal injury and attenuates TGF-1-induced EMT in cells.
褪黑素（Mel）已被证明可在细胞系统中调节上皮-间质转化（EMT）。肾小管上皮细胞的间质转化是肾纤维化的关键致病机制。本研究旨在阐明Mel在肾小管上皮细胞EMT过程中的作用。通过单侧输尿管结扎建立单侧输尿管梗阻（UUO）大鼠模型，随后用Mel（5、10和20mg/kg）进行治疗。后续分析包括组织病理学检查、肌酐和血尿素氮水平的测定、纤连蛋白（FN）的免疫荧光分析以及α-平滑肌肌动蛋白（α-SMA）的免疫组织化学分析。转化生长因子-β1（TGF-β1）在NRK-52E细胞中引发纤维化反应，随后用Mel（0.1、1和10μmol/L）进行处理。评估细胞活力、迁移以及EMT相关蛋白的表达，包括FN、α-SMA、胶原酶I和E-钙黏蛋白。为了验证miR-153-3p与叉头转录因子o亚家族成员3A（FOXO3A）之间的调控相互作用，用miR-153-3p模拟物或靶向FOXO3A的小干扰RNA（si-FOXO3A）转染细胞。结果显示，Mel在UUO大鼠模型中具有剂量依赖性改善肾损伤和纠正肾小球结构异常的能力。与UUO模型大鼠相比，褪黑素显著降低了FN和α-SMA的表达水平。体外研究表明，Mel抑制细胞活性和迁移行为，以及NRK-52E细胞中FN、α-SMA和胶原酶I的蛋白表达水平。相对于对照组，UUO大鼠和TGF-β1诱导的NRK-52E细胞中miR-153-3p表达升高，FOXO3A表达降低。体外模型进一步表明，miR-153-3p的上调或FOXO3A的下调可抵消褪黑素对TGF-β1诱导的EMT的保护作用。结论是，Mel抑制miR-153-3p，从而促进UUO大鼠中FOXO3A的转录，减轻肾损伤并减弱TGF-1诱导的细胞EMT。
