GSK-3β inhibitor TDZD-8 prevents reduction of aquaporin-1 expression via activating autophagy under renal ischemia reperfusion injury

Qiaojuan Liu; Yonglun Kong; Xiangdong Guo; Baien Liang; Haixia Xie; Shan Hu; Mengke Han; Xiaoduo Zhao; Pinning Feng; Qianqian Lyu; Wei Dong; Xinling Liang; Weidong Wang; Chunling Li

doi:10.1096/fj.202100549R

GSK-3β inhibitor TDZD-8 prevents reduction of aquaporin-1 expression via activating autophagy under renal ischemia reperfusion injury

FASEB J. 2021 Aug;35(8):e21809. doi: 10.1096/fj.202100549R.

Authors

Qiaojuan Liu^{1

2}, Yonglun Kong¹, Xiangdong Guo¹, Baien Liang¹, Haixia Xie¹, Shan Hu¹, Mengke Han¹, Xiaoduo Zhao¹, Pinning Feng³, Qianqian Lyu⁴, Wei Dong⁵, Xinling Liang⁵, Weidong Wang^{1

6}, Chunling Li¹

Affiliations

¹ Institute of Hypertension, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China.
² Department of Immunology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China.
³ Department of Clinical Laboratory, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
⁴ MOE Key Laboratory of Marine Genetics and Breeding, College of Marine Life Sciences, Ocean University of China, Qingdao, China.
⁵ Division of Nephrology, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, China.
⁶ Department of Nephrology, The Seventh Affiliated Hospital, Sun Yat-sen University, Shenzhen, China.

PMID: 34314052
DOI: 10.1096/fj.202100549R

Abstract

Renal ischemia/reperfusion (I/R) injury is a main cause of acute kidney injury (AKI). Aquaporin (AQP)-1 water channel in the kidney is critical for the maintenance of water homeostasis and the urinary concentrating ability. Increasing evidence supports an important role of autophagy in the pathogenesis of AKI induced by renal I/R. The purpose of the present study is to investigate whether activation of autophagy prevents downregulation of AQP1 protein induced by renal I/R and potential molecular mechanisms. Renal I/R induced consistently reduced protein expression of AQP1, 2, and 3, as well as sodium cotransporters Na⁺ -K⁺ -2Cl^- cotransporter and α-Na,K-ATPase, which was associated with increased urine output and decreased creatinine clearance in rats. Renal I/R also suppressed autophagy and increased inflammatory responses in the kidney. 4-Benzyl-2-methyl-1,2,4-thiadiazolidine-3,5-dione (TDZD-8), the glycogen synthase kinase-3β inhibitor, ameliorated renal injury under I/R, activated autophagy and markedly increased expression of AQPs and sodium transporters in the kidney, which was associated with improved urine output and creatinine clearance in rats. Hypoxia/reoxygenation (H/R) induced suppression of autophagy and downregulation of AQP1 in murine inner medullary collecting duct 3 (IMCD3) cells, which was fully prevented by TDZD-8 treatment. Inhibition of autophagy by 3-methyladenine or Atg5 gene knockdown attenuated recovery of AQP1 protein expression induced by TDZD-8 in IMCD3 cells with H/R. Interleukin-1 beta (IL-1β) decreased the abundance of AQP1 protein in IMCD3 cells. H/R induced increases in protein expression of nod-like receptor pyrin domain-containing 3 and IL-1β, which was reversed by TDZD-8. In conclusion, TDZD-8 treatment prevented downregulation of AQP1 expression under renal I/R injury, likely via activating autophagy and decreasing IL-1β production.

Keywords: AQP1; GSK-3β; autophagy; ischemia/reperfusion.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Acute Kidney Injury / drug therapy*
Animals
Aquaporin 1 / metabolism*
Autophagy / drug effects
Cells, Cultured
Enzyme Inhibitors / pharmacology*
Glycogen Synthase Kinase 3 beta / antagonists & inhibitors*
Male
Mice
Rats
Rats, Sprague-Dawley
Reperfusion Injury / drug therapy*
Thiadiazoles / pharmacology*

Substances

4-benzyl-2-methyl-1,2,4-thiadiazolidine-3,5-dione
Aqp1 protein, mouse
Aqp1 protein, rat
Enzyme Inhibitors
Thiadiazoles
Aquaporin 1
Glycogen Synthase Kinase 3 beta
Gsk3b protein, rat