Abstract：
BACKGROUND: Senolytic combination of dasatinib and quercetin (DQ) is the most studied senolytics drugs used to treat various age-related diseases. However, its protective activity against diabetic kidney disease (DKD) and underlying mechanisms are uncertain.
OBJECTIVE: To investigate the functions and potential mechanisms of the senolytics DQ on DKD.
METHODS: Diabetic db/db mice were administrated DQ or transfected with over-expressed PPARα or shPPARα vector. The positive control group was administered irbesartan. Renal function and fibrotic changes in kidney tissue were tested. Single-cell RNA-seq (scRNA-seq) was conducted to analyze the differential transcriptome between the diabetic and control mice. Molecular docking simulation was used to assess the combination of DQ and potential factors. Moreover, tubular epithelial cells under high-glucose (HG) conditions were incubated with DQ and transfected with or without over-expressed PPARα/siPPARα vector.
RESULTS: DQ significantly improved renal function, histopathological and fibrotic changes, alleviated lipid deposition, and increased ATP levels in mice with DKD. DQ reduced multiple fatty acid oxidation (FAO) pathway-related proteins and up-regulated PPARα in db/db mice. Overexpression of PPARα upregulated the expression of PPARα-targeting downstream FAO pathway-related proteins, restored renal function, and inhibited renal fibrosis in vitro and in vivo. Moreover, molecular docking and dynamics simulation analyses indicated the nephroprotective effect of DQ via binding to PPARα. Knockdown of PPARα reversed the effect of DQ on the FAO pathway and impaired the protective effect of DQ during DKD.
CONCLUSIONS: For the first time, DQ was found to exert a renal protective effect by binding to PPARα and attenuating renal damage through the promotion of FAO in DKD.
OBJECTIVE: To investigate the functions and potential mechanisms of the senolytics DQ on DKD.
METHODS: Diabetic db/db mice were administrated DQ or transfected with over-expressed PPARα or shPPARα vector. The positive control group was administered irbesartan. Renal function and fibrotic changes in kidney tissue were tested. Single-cell RNA-seq (scRNA-seq) was conducted to analyze the differential transcriptome between the diabetic and control mice. Molecular docking simulation was used to assess the combination of DQ and potential factors. Moreover, tubular epithelial cells under high-glucose (HG) conditions were incubated with DQ and transfected with or without over-expressed PPARα/siPPARα vector.
RESULTS: DQ significantly improved renal function, histopathological and fibrotic changes, alleviated lipid deposition, and increased ATP levels in mice with DKD. DQ reduced multiple fatty acid oxidation (FAO) pathway-related proteins and up-regulated PPARα in db/db mice. Overexpression of PPARα upregulated the expression of PPARα-targeting downstream FAO pathway-related proteins, restored renal function, and inhibited renal fibrosis in vitro and in vivo. Moreover, molecular docking and dynamics simulation analyses indicated the nephroprotective effect of DQ via binding to PPARα. Knockdown of PPARα reversed the effect of DQ on the FAO pathway and impaired the protective effect of DQ during DKD.
CONCLUSIONS: For the first time, DQ was found to exert a renal protective effect by binding to PPARα and attenuating renal damage through the promotion of FAO in DKD.
摘要：
背景：达沙替尼和槲皮素（DQ）的抗衰老组合是研究最多的用于治疗各种与年龄相关的疾病的抗衰老药物。然而,其对糖尿病肾病（DKD）的保护活性和潜在机制尚不确定。
目的：探讨解毒剂DQ对DKD的作用及可能机制。
方法:糖尿病db/db小鼠给予DQ或用过表达的PPARα或shPPARα载体转染。阳性对照组给予厄贝沙坦。检测肾功能和肾组织纤维化变化。进行单细胞RNA-seq(scRNA-seq)以分析糖尿病小鼠和对照小鼠之间的差异转录组。使用分子对接模拟来评估DQ和潜在因素的组合。此外，在高葡萄糖（HG）条件下，将肾小管上皮细胞与DQ一起孵育，并在有或没有过表达的PPARα/siPPARα载体的情况下转染。
结果：DQ显著改善肾功能，组织病理学和纤维化变化，减轻脂质沉积，DKD小鼠的ATP水平升高。DQ减少db/db小鼠中多种脂肪酸氧化（FAO）途径相关蛋白并上调PPARα。PPARα的过表达上调PPARα靶向下游FAO途径相关蛋白的表达,肾功能恢复,并在体外和体内抑制肾脏纤维化。此外，分子对接和动力学模拟分析表明，DQ通过与PPARα结合具有肾保护作用。PPARα的敲除逆转了DQ对FAO途径的影响，并损害了DQ在DKD期间的保护作用。
结论：第一次，发现DQ通过与PPARα结合而发挥肾脏保护作用，并通过在DKD中促进FAO减轻肾脏损害。
目的：探讨解毒剂DQ对DKD的作用及可能机制。
方法:糖尿病db/db小鼠给予DQ或用过表达的PPARα或shPPARα载体转染。阳性对照组给予厄贝沙坦。检测肾功能和肾组织纤维化变化。进行单细胞RNA-seq(scRNA-seq)以分析糖尿病小鼠和对照小鼠之间的差异转录组。使用分子对接模拟来评估DQ和潜在因素的组合。此外，在高葡萄糖（HG）条件下，将肾小管上皮细胞与DQ一起孵育，并在有或没有过表达的PPARα/siPPARα载体的情况下转染。
结果：DQ显著改善肾功能，组织病理学和纤维化变化，减轻脂质沉积，DKD小鼠的ATP水平升高。DQ减少db/db小鼠中多种脂肪酸氧化（FAO）途径相关蛋白并上调PPARα。PPARα的过表达上调PPARα靶向下游FAO途径相关蛋白的表达,肾功能恢复,并在体外和体内抑制肾脏纤维化。此外，分子对接和动力学模拟分析表明，DQ通过与PPARα结合具有肾保护作用。PPARα的敲除逆转了DQ对FAO途径的影响，并损害了DQ在DKD期间的保护作用。
结论：第一次，发现DQ通过与PPARα结合而发挥肾脏保护作用，并通过在DKD中促进FAO减轻肾脏损害。
