Ying Shao1, Chuan Lv1, Can Wu1, Yuehong Zhou2, Qiuyue Wang1. 1. Department of Endocrinology, the First Hospital Affiliated to China Medical University, Shenyang, Liaoning, China. 2. Division of Endocrinology, Shenyang NO.8 hospital, Shenyang, Liaoning, China.
Abstract
BACKGROUND: Silent information regulator 1 (Sirt1) plays a protective role in kidney. Sirt1 suppresses activation of hypoxia-inducible factor-1 alpha (HIF-1α), with MircroRNA-217 (Mir-217) being closely related to Sirt1. The relationship of Sirt1, HIF-1α and Mir-217, however, has never been reported in high glucose cultured rat glomerular mesangial cells (RMCs). Thus, we explored the role of Mir-217 on inflammation and fibrosis in RMCs cultured with high glucose in vitro through Sirt1/HIF-1α signaling pathway. METHODS: Rat glomerular mesangial cells were pre-incubated with Sirt1 activator Resveratrol prior to high glucose treatment. Furthermore the cells were transiently transfected with Sirt1 small interfering RNA (siRNA), HIF-1α siRNA and Mir-217 inhibitor using Lipofectamine 2000. Real-time PCR was used to analyse the expression of Mir-217, Sirt1 mRNA and HIF-1α mRNA; Western Blot was used to observe protein expression of Sirt1, HIF-1α, connective tissue growth factor, endothelin-1 and fibronectin; enzyme-linked immunosorbent assay was used to detect protein expression of transforming growth factor-β1 and vascular endothelial growth factor. RESULTS: High glucose increased Mir-217 expression. High glucose decreased Sirt1 expression, accompanied by the increased HIF-1α expression and then promoted inflammation and fibrosis. In addition, Mir-217 gene silencing or Resveratrol could suppress the expression of HIF-1α, which in turn restrained inflammation and fibrosis in rat glomerular mesangial cells cultured with high glucose. CONCLUSION: This study clarified the role of Mir-217 in high glucose cultured rat glomerular mesangial cells through Sirt1/HIF-1α signaling pathway and provided new therapeutic targets for diabetic nephropathy.
BACKGROUND:Silent information regulator 1 (Sirt1) plays a protective role in kidney. Sirt1 suppresses activation of hypoxia-inducible factor-1 alpha (HIF-1α), with MircroRNA-217 (Mir-217) being closely related to Sirt1. The relationship of Sirt1, HIF-1α and Mir-217, however, has never been reported in high glucose cultured rat glomerular mesangial cells (RMCs). Thus, we explored the role of Mir-217 on inflammation and fibrosis in RMCs cultured with high glucose in vitro through Sirt1/HIF-1α signaling pathway. METHODS:Rat glomerular mesangial cells were pre-incubated with Sirt1 activator Resveratrol prior to high glucose treatment. Furthermore the cells were transiently transfected with Sirt1 small interfering RNA (siRNA), HIF-1α siRNA and Mir-217 inhibitor using Lipofectamine 2000. Real-time PCR was used to analyse the expression of Mir-217, Sirt1 mRNA and HIF-1α mRNA; Western Blot was used to observe protein expression of Sirt1, HIF-1α, connective tissue growth factor, endothelin-1 and fibronectin; enzyme-linked immunosorbent assay was used to detect protein expression of transforming growth factor-β1 and vascular endothelial growth factor. RESULTS: High glucose increased Mir-217 expression. High glucose decreased Sirt1 expression, accompanied by the increased HIF-1α expression and then promoted inflammation and fibrosis. In addition, Mir-217 gene silencing or Resveratrol could suppress the expression of HIF-1α, which in turn restrained inflammation and fibrosis in rat glomerular mesangial cells cultured with high glucose. CONCLUSION: This study clarified the role of Mir-217 in high glucose cultured rat glomerular mesangial cells through Sirt1/HIF-1α signaling pathway and provided new therapeutic targets for diabetic nephropathy.
Authors: Lei Bao; Cecilia S Chau; Zhengdeng Lei; Hong Hu; Angelina G Chan; Kyle T Amber; Mark Maienschein-Cline; Maria M Tsoukas Journal: Arch Dermatol Res Date: 2021-01-12 Impact factor: 3.017