cAMP-response element binding protein mediates podocyte injury in diabetic nephropathy by targeting lncRNA DLX6-AS1
cAMP-response element binding protein mediates podocyte injury in diabetic nephropathy by targeting lncRNA DLX6-AS1
复制标题
cAMP反应元件结合蛋白通过靶向lncRNA DLX6-AS1介导糖尿病肾病足细胞损伤
DOI：
10.1016/j.metabol.2022.155155
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发表时间：
2022
影响因子：
9.8
通讯作者：
Zhangsuo Liu
中科院分区：
文献类型：
--
作者：
Wen Zheng;Jia Guo;Xiaoqing Lu;Yongxia Qiao;Dong Liu;Shaokang Pan;Lulu Liang;Chang Liu;Hongchao Zhu;Zhangsuo Liu;Zhangsuo Liu
进行性蛋白尿是糖尿病肾病（DN）最早的临床特征之一。在我们先前的研究中，发现lncRNA DLX 6-AS 1（DLX 6-AS 1，小鼠中为Dlx 6 os 1）与DN患者的蛋白尿程度相关。此外，Dlx 6 os 1的缺乏在db/db小鼠模型中关闭炎症反应中是关键的。本研究采用JASPAR数据库和DNA pull down结合液相色谱-串联质谱技术，对DN患者DLX 6-AS 1的表达进行了研究，并对DLX 6-AS 1的表达进行了分析。然后进行双荧光素酶报告基因测定和染色质免疫沉淀以确认结合位点。我们还在db/db小鼠模型和体外培养的人足细胞中研究了DN进展的调控因素，结果表明cAMP反应元件结合蛋白（CREB）在DN中高表达，并与DLX 6-AS 1密切相关。在db/db小鼠和培养的足细胞中，CREB沉默显著降低DLX 6-AS 1或Dlx 6 os 1的水平，并减轻肾损伤。机制上，CREB过表达通过靶向DLX 6-AS 1加重肾脏炎症并破坏足细胞结构。CREB在足细胞损伤中的损伤作用也被选择性抑制剂666-15以剂量依赖的方式抑制。在体内，666-15抑制CREB可显著减轻蛋白尿，并改善足细胞中的炎症浸润。我们的研究结果表明，CREB是足细胞损伤的关键介质，并通过调节DLX 6-AS 1起作用。因此，CREB可能是治疗DN的一个有效和潜在的治疗靶点。
Progressive proteinuria is one of the earliest clinical features of diabetic nephropathy (DN). In our previous study, lncRNA DLX6-AS1 (DLX6-AS1, Dlx6os1 in the mouse) was found to be associated with the extent of albuminuria in DN patients. Furthermore, the lack of Dlx6os1 was pivotal in switching off the inflammatory response in db/db mouse model. However, the regulatory factors responsible for elevated DLX6-AS1 in DN remains unknown.To identify potential regulatory factors for DLX6-AS1, JASPAR database and DNA pull down combined subsequent liquid chromatography-tandem mass spectrometry were used. Dual-luciferase reporter assay and chromatin immunoprecipitation were then performed to confirm binding sites. We also investigated the effects of the regulatory factors on DN progression in db/db mouse model and cultured human podocytes.Our analyses demonstrated that cAMP-response element binding protein (CREB) was highly expressed and closely associated with DLX6-AS1 in DN. In db/db mouse and in cultured podocytes, CREB silencing significantly reduced the level of DLX6-AS1 or Dlx6os1 and attenuated renal damage. Mechanistically, CREB overexpression aggravated renal inflammation and destroyed the structure of podocytes by targeting DLX6-AS1. The damaging role of CREB in podocyte injury was also inhibited by 666-15, a selective inhibitor, in a dose-dependent manner. In vivo, the inhibition of CREB by 666-15 significantly attenuated albuminuria and ameliorated inflammatory infiltration in podocytes.Our findings indicated that CREB is a key mediator of podocyte injury and acts by regulating DLX6-AS1. Thus, CREB may be an effective and potential therapeutic target for the treatment of DN.