Abstract: TH-PO543
High Phosphate-Inducing Valvular Interstitial-Endothelium Cross-Talk Through mir-382/SOD2 Axis in CKD with Valve Injury
Session Information
- Bone and Mineral Metabolism: Basic
November 07, 2019 | Location: Exhibit Hall, Walter E. Washington Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Bone and Mineral Metabolism
- 401 Bone and Mineral Metabolism: Basic
Authors
- Wang, Li-ting, Institute of Nephrology, ZhongDa Hospital, school of medicine, Southeast University, Nanjing, China
- Zhang, Yu-xia, Institute of Nephrology, ZhongDa Hospital, school of medicine, Southeast University, Nanjing, China
- Chen, Si-Jie, Institute of Nephrology, ZhongDa Hospital, school of medicine, Southeast University, Nanjing, China
- Guo, Yu, Institute of Nephrology, ZhongDa Hospital, school of medicine, Southeast University, Nanjing, China
- Wang, Xiao-chen, Institute of Nephrology, ZhongDa Hospital, school of medicine, Southeast University, Nanjing, China
- Ni, Li-Hua, Institute of Nephrology, ZhongDa Hospital, school of medicine, Southeast University, Nanjing, China
- Song, Kaiyun, Institute of Nephrology, ZhongDa Hospital, school of medicine, Southeast University, Nanjing, China
- Zhang, Xiaoliang, Institute of Nephrology, ZhongDa Hospital, school of medicine, Southeast University, Nanjing, China
- Liu, Bi-Cheng, Institute of Nephrology, ZhongDa Hospital, school of medicine, Southeast University, Nanjing, China
- Tang, Ri-ning, Institute of Nephrology, ZhongDa Hospital, school of medicine, Southeast University, Nanjing, China
Background
CKD valve injury is the main cause of CVD among CKD patients, but it underlying mechanisms are still unknown. Previous studies demonstrated valve interstitial cells (VICs) participate in valve injury to produce excessive quantities of the valvular ECM. Recent findings suggested valve interstitial-endothelium crosstalk are closely related to valve homeostasis and injury. Hence, we want to investigate the mechanism between VECs and VICs upon high phosphate (HP) stimulation.
Methods
We used c57/b mouse and HP-stimulated VIC as in vivo and in vitro model of CKD, respectively. Transwell migration were performed to determine VIC could aggravate valve endothelial cell（VEC） EndMT upon HP stimulation by qPCR, WB and immunofluorescence. ELISA were performed to detect the expression of TGFβ-1 in VIC cell supernatant. The expression of key factors involved in EndMT process, such as CD31, VE-cadherin, α-SMA, FSP1 were evaluated by western blotting and immunofluorescence. qRT-PCR was used to measure levels of miR-382 in VEC.
Results
TGFβ-1were significantly increased in VIC upon HP stimulation. Transwell migration were proved VIC could aggravate VEC endothelial-to-mesenchymal transition (EndMT) upon HP stimulation, with the up-regulation of mesenchymal markers (FSP1 and α-SMA) and stem cell markers (CD44 and CD10) and down-regulation of the endothelial marker (CD31, VE-cadherin), consistent with CKD aortic valve samples. Knockdown of EC miR-382, which was up-regulated by TGFβ1, could attenuate TGFβ1-induced loss of the endothelial marker VE-cadherin and CD31. miR-382 was confirmed by 3'-untranslated region reporter assay to target superoxide dismutase 2 (SOD2) that were downregulated at the protein level by TGFβ1. Knockdown of miR-382 attenuated TGFβ1-induced downregulation of SOD2. Overexpression of SOD2 ameliorated loss of the endothelial marker and CKD aortic leaflets thickening.
Conclusion
VIC could secret TGFβ-1 upon HP stimulation, which lead to EndMT through mir-382/SOD2 axis. This could cause CKD aortic leaflets thickening and aggravate valve injury.
Pattern diagram