Zhang Xianjing, Zhang Jiarong, Xu Xiaojun, Chen Suzhen, Gao Fei
Department of Endocrinology and Metabolism, Shanghai Clinical Center for Diabetes, Shanghai Diabetes Institute, Shanghai Key Laboratory of Diabetes Mellitus, Shanghai Jiao Tong University School of Medicine Affiliated Sixth People's Hospital, Shanghai, China.
Department of Pharmacy, The Fourth Affiliated Hospital, Zhejiang University School of Medicine, Yiwu, Zhejiang, China.
Metabol Open. 2024 Aug 29;23:100316. doi: 10.1016/j.metop.2024.100316. eCollection 2024 Sep.
Diabetic Nephropathy (DN) has become the leading cause of end-stage renal disease worldwide. Studies have indicated that Transforming Growth Factor beta1 (TGFβ1) is the most potent factor contributing to renal fibrosis, and understanding the exact pathogenic mechanism of renal fibrosis is crucial for alleviating the condition. Previous research has identified Yin Yang 1 (YY1) as an effective inhibitor of TGF-β1. Our study, through dual-luciferase reporter gene assays and Western blot experiments, screened and obtained the small molecule compound PdⅡ. Subsequently, validation in a high-glucose-induced renal mesangial cell injury model showed that PdⅡ treatment significantly increased the expression of YY1 protein and mRNA, while correspondingly reducing the expression of TGFβ1 protein and mRNA. Dual-luciferase reporter gene assay results revealed that, compared to the control group, the luciferase transcription activity of YY1 molecules increased in the PdⅡ treatment group, and the luciferase transcription activity of TGFβ1 decreased. By further designing mutations in the binding sites between TGFβ1 and YY1 on the promoter, transfecting fluorescent enzyme reporter gene plasmids with TGFβ1 mutant promoter into mesangial cells damaged by high glucose, and then treating the cells with PdⅡ, it was observed that the luciferase transcription activity of TGFβ1 did not decrease. Therefore, these results suggest that PdⅡ may inhibit TGFβ1 transcriptional activity by activating YY1, thereby slowing down the progression of diabetic nephropathy.
糖尿病肾病（DN）已成为全球终末期肾病的主要原因。研究表明，转化生长因子β1（TGFβ1）是导致肾纤维化的最主要因素，了解肾纤维化的确切致病机制对于缓解病情至关重要。先前的研究已确定阴阳1（YY1）是TGF-β1的有效抑制剂。我们的研究通过双荧光素酶报告基因检测和蛋白质印迹实验，筛选并获得了小分子化合物PdⅡ。随后，在高糖诱导的肾系膜细胞损伤模型中的验证表明，PdⅡ处理显著增加了YY1蛋白和mRNA的表达，同时相应降低了TGFβ1蛋白和mRNA的表达。双荧光素酶报告基因检测结果显示，与对照组相比，PdⅡ处理组中YY1分子的荧光素酶转录活性增加，而TGFβ1的荧光素酶转录活性降低。通过进一步设计TGFβ1与YY1在启动子上结合位点的突变，将带有TGFβ1突变启动子的荧光酶报告基因质粒转染至高糖损伤的系膜细胞中，然后用PdⅡ处理细胞，观察到TGFβ1的荧光素酶转录活性并未降低。因此，这些结果表明，PdⅡ可能通过激活YY1来抑制TGFβ1的转录活性，从而减缓糖尿病肾病的进展。
