Department of Nephropathy, The First Affiliated Hospital of Anhui Medical University, Hefei, China; Inflammation and Immune Mediated Diseases Laboratory of Anhui Province, Anhui Institute of Innovative Drugs, School of Pharmacy, Anhui Medical University, The Key Laboratory of Anti-inflammatory of Immune Medicines, Ministry of Education, Hefei 230032, China.
School of Basic Medical Sciences, Anhui Medical University, Anhui, China.
Biochem Pharmacol. 2020 Oct;180:114132. doi: 10.1016/j.bcp.2020.114132. Epub 2020 Jul 3.
Acute kidney injury (AKI), characterized by a rapid decline in renal function, is triggered by an acute inflammatory response that leads to kidney damage. An effective treatment for AKI is lacking. Using in vitro and in vivo AKI models, our laboratory has identified a series of anti-inflammatory molecules and their derivatives. In the current study, we identified the protective role of rutaecarpine (Ru) on renal tubules. We obtained a series of 3-aromatic sulphonamide-substituted Ru derivatives exhibiting enhanced renoprotective and anti-inflammatory function. We identified Compound-6c(Cpd-6c) as having the best activity and examined its protective effect against cisplatin nephropathy both in vivo and in vitro in cisplatin-stimulated tubular epithelial cells (TECs). Our results showed that Cpd-6c restored renal function more effectively than Ru, as evidenced by reduced blood urea nitrogen and serum creatinine levels in mice. Cpd-6c alleviated tubular injury, as shown by PAS staining and molecular analysis of kidney injury molecule-1 (KIM-1), with both prevention and treatment protocols in cisplatin-treated mice. Moreover, Cpd-6c decreased kidney inflammation, oxidative stress and programmed cell death. These results have also been confirmed in cisplatin-treated TECs. Using web-prediction algorithms, molecular docking, and cellular thermal shift assay (CETSA), we identified phosphodiesterase 4B (PDE4B) as a Cpd-6c target. In addition, we firstly found that PDE4B was up-regulated significantly in the serum of AKI patients. After identifying the function of PDE4B in cisplatin-treated tubular epithelial cells by siRNA transfection or PDE4 inhibitor rolipram, we showed that Cpd-6c treatment did not protect against cisplatin-induced injury in PDE4B knockdown TECs, thus indicating that Cpd-6c exerts its renoprotective and anti-oxidative effects via the PDE4B-dependent pathway. Collectively, Cpd-6c might serve as a potential therapeutic agent for AKI and PDE4B may be highly involved in the initiation and progression of AKI.
急性肾损伤（AKI）的特征是肾功能迅速下降，是由急性炎症反应引起的，导致肾脏损伤。目前缺乏有效的 AKI 治疗方法。我们实验室使用体外和体内 AKI 模型，已经确定了一系列抗炎分子及其衍生物。在本研究中，我们鉴定了 rutaecarpine（Ru）对肾小管的保护作用。我们获得了一系列 3-芳基磺酰胺取代的 Ru 衍生物，具有增强的肾保护和抗炎功能。我们确定了化合物 6c（Cpd-6c）具有最佳的活性，并在体内和体外研究了其对顺铂诱导的肾小管上皮细胞（TECs）损伤的保护作用。结果表明，Cpd-6c 比 Ru 更有效地恢复肾功能，表现为顺铂处理的小鼠血液中尿素氮和血清肌酐水平降低。Cpd-6c 通过 PAS 染色和肾损伤分子-1（KIM-1）的分子分析减轻肾小管损伤，在顺铂处理的小鼠中具有预防和治疗作用。此外，Cpd-6c 降低了肾脏炎症、氧化应激和程序性细胞死亡。这些结果在顺铂处理的 TECs 中也得到了证实。使用网络预测算法、分子对接和细胞热转移分析（CETSA），我们确定磷酸二酯酶 4B（PDE4B）是 Cpd-6c 的靶点。此外，我们首次发现 PDE4B 在 AKI 患者的血清中显著上调。通过 siRNA 转染或 PDE4 抑制剂 rolipram 鉴定了 PDE4B 在顺铂处理的肾小管上皮细胞中的功能后，我们表明 Cpd-6c 治疗不能保护 PDE4B 敲低的 TECs 免受顺铂诱导的损伤，这表明 Cpd-6c 通过 PDE4B 依赖性途径发挥其肾保护和抗氧化作用。总之，Cpd-6c 可能成为 AKI 的潜在治疗药物，而 PDE4B 可能高度参与 AKI 的发生和发展。
