Wang Guangyu, Zhang Lijuan, Tan Jiaorong, Li Fei, Jin Yishan, He Limei, Yang Xin
Department of Endocrinology, Putuo People's Hospital of Tongji University, School of Medicine, Tongji University, Shanghai, 200060, China.
Department of Clinical Laboratory, Changning Maternity and Infant Health Hospital, East China Normal University, Shanghai, 200050, China.
Adv Biol (Weinh). 2025 Jul;9(7):e2400583. doi: 10.1002/adbi.202400583. Epub 2025 Apr 8.
Heart failure-induced renal tubular epithelial cell fibrosis is an important pathological process that leads to chronic kidney disease. This study is to investigate the regulatory mechanism of over-expression or knock-down SIRT1 gene, alleviating hypoxia-induced HK2 cell fibrosis in heart failure. The focus is on the SIRT1-FoxO1-FoxO3-Autophagy pathway. In vitro experiments are performed by HK2cell line to simulate the normal oxygen state (Normoxia) and the hypoxia state (Hypoxia) caused by heart failure, SIRT1 gene over-expression by transfected vectors, knock-down and Rapamycin (RAPA)-induced cellular autophagy, and the cell models are divided into four subgroups, named control group, oeSIRT1, siSIRT1 and siSIRT1+RAPA. Western blotting (WB), real-time qPCR, immunofluorescence (IF), ELISA, and transmission electron microscopy are used to quantitatively or semi-quantitatively analyze the expression of FoxO1, FoxO3, SIRT1, Beclin1, LC-3, α-SMA, E- Cadherin, and collagen-I in cells or supernatants. It is demonstrated that activation of SIRT1 regulates the expression and activity of FoxO1 and FoxO3, thereby affecting autophagy. This modulation leads to a reduction in HK2 fibrosis markers (α-SMA and E-cadherin) and extracellular matrix deposition (collagen I), which ultimately attenuates renal tubular epithelial cell fibrosis. These findings provide new insights into potential therapeutic strategies for treating heart failure-induced renal tubular epithelial cell fibrosis by targeting the SIRT1-FoxO1-FoxO3-Autophagy pathway.
心力衰竭诱导的肾小管上皮细胞纤维化是导致慢性肾脏病的一个重要病理过程。本研究旨在探讨过表达或敲低SIRT1基因对减轻心力衰竭中缺氧诱导的HK2细胞纤维化的调控机制。重点关注SIRT1-FoxO1-FoxO3-自噬通路。通过HK2细胞系进行体外实验，模拟正常氧状态（常氧）和心力衰竭引起的缺氧状态（缺氧），通过转染载体过表达SIRT1基因、敲低该基因以及用雷帕霉素（RAPA）诱导细胞自噬，细胞模型分为四个亚组，分别命名为对照组、oeSIRT1、siSIRT1和siSIRT1+RAPA。采用蛋白质免疫印迹法（WB）、实时定量聚合酶链反应、免疫荧光法（IF）、酶联免疫吸附测定法（ELISA）和透射电子显微镜对细胞或上清液中FoxO1、FoxO3、SIRT1、Beclin1、LC-3、α-平滑肌肌动蛋白（α-SMA）、E-钙黏蛋白和I型胶原的表达进行定量或半定量分析。结果表明，SIRT1的激活调节了FoxO1和FoxO3的表达及活性，从而影响自噬。这种调节导致HK2纤维化标志物（α-SMA和E-钙黏蛋白）减少以及细胞外基质沉积（I型胶原）减少，最终减轻肾小管上皮细胞纤维化。这些发现为通过靶向SIRT1-FoxO1-FoxO3-自噬通路治疗心力衰竭诱导的肾小管上皮细胞纤维化的潜在治疗策略提供了新的见解。
