SIRT1 Activation Suppresses Corneal Endothelial–Mesenchymal Transition via the TGF-β/Smad2/3 Pathway

Endothelial–mesenchymal transition (EnMT) is the transversion of endothelial cells to mesenchymal cells under certain physiological or pathological conditions. When EnMT occurs in the corneal endothelium, corneal endothelial cells (CECs) lose their normal function and thus cannot maintain corneal cl...

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Main Authors: Yi Yu, Ruilin Guo, Jie Ling, Chenjia Xu, Minglu Ma, Xiaojuan Dong, Jing Wu, Ting Huang
Format: Article
Language:English
Published: MDPI AG 2024-12-01
Series:Current Issues in Molecular Biology
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Online Access:https://www.mdpi.com/1467-3045/46/12/827
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author Yi Yu
Ruilin Guo
Jie Ling
Chenjia Xu
Minglu Ma
Xiaojuan Dong
Jing Wu
Ting Huang
author_facet Yi Yu
Ruilin Guo
Jie Ling
Chenjia Xu
Minglu Ma
Xiaojuan Dong
Jing Wu
Ting Huang
author_sort Yi Yu
collection DOAJ
description Endothelial–mesenchymal transition (EnMT) is the transversion of endothelial cells to mesenchymal cells under certain physiological or pathological conditions. When EnMT occurs in the corneal endothelium, corneal endothelial cells (CECs) lose their normal function and thus cannot maintain corneal clarity. Studies have shown that the mechanism of EnMT in CECs involves the transforming growth factor-β (TGF-β) signaling pathway, and one of the important inhibitors of the TGF-β/Smad2/3 pathway is sirtuin-1 (SIRT1). In this study, we used a rat model of corneal endothelium injury and TGF-β1-treated human CECs to induce EnMT, aiming to explore whether SIRT1 activation inhibits corneal EnMT in vivo and in vitro. SIRT1 was activated and suppressed using resveratrol (RSV) and EX527, respectively. The endothelial markers and mesenchymal markers were measured by immunofluorescence and Western blot assays. Co-immunoprecipitation was used to detect the interaction between SIRT1 and Smad2/3. The results showed that after mechanical injury, the group treated with RSV-activated SIRT1 regained corneal transparency and recovered from edema faster than the control group. Moreover, RSV-activated SIRT1 downregulated the expression levels of alpha smooth muscle actin (α-SMA), vimentin, and Snail and upregulated the expression levels of E-cadherin and Na<sup>+</sup>/K<sup>+</sup>-ATPase both in vivo and in vitro, but these effects were reversed when SIRT1 was inhibited by EX527. SIRT1 also upregulated the expression levels of TGF-β receptor 1 and phosphorylated Smad2/3. The interaction between SIRT1 and Smad2/3 in vitro was confirmed by co-immunoprecipitation. Overall, our results indicate that SIRT1 activation inhibits corneal EnMT via the TGF-β/Smad2/3 pathway, which may be a potential therapeutic target for corneal endothelium dysfunction.
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spelling doaj-art-f8d2b44a1b4e414dbd60dd388e088d632025-08-20T02:53:19ZengMDPI AGCurrent Issues in Molecular Biology1467-30371467-30452024-12-014612138461385910.3390/cimb46120827SIRT1 Activation Suppresses Corneal Endothelial–Mesenchymal Transition via the TGF-β/Smad2/3 PathwayYi Yu0Ruilin Guo1Jie Ling2Chenjia Xu3Minglu Ma4Xiaojuan Dong5Jing Wu6Ting Huang7State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Guangzhou 510060, ChinaState Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Guangzhou 510060, ChinaState Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Guangzhou 510060, ChinaState Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Guangzhou 510060, ChinaState Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Guangzhou 510060, ChinaState Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Guangzhou 510060, ChinaState Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Guangzhou 510060, ChinaState Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Guangzhou 510060, ChinaEndothelial–mesenchymal transition (EnMT) is the transversion of endothelial cells to mesenchymal cells under certain physiological or pathological conditions. When EnMT occurs in the corneal endothelium, corneal endothelial cells (CECs) lose their normal function and thus cannot maintain corneal clarity. Studies have shown that the mechanism of EnMT in CECs involves the transforming growth factor-β (TGF-β) signaling pathway, and one of the important inhibitors of the TGF-β/Smad2/3 pathway is sirtuin-1 (SIRT1). In this study, we used a rat model of corneal endothelium injury and TGF-β1-treated human CECs to induce EnMT, aiming to explore whether SIRT1 activation inhibits corneal EnMT in vivo and in vitro. SIRT1 was activated and suppressed using resveratrol (RSV) and EX527, respectively. The endothelial markers and mesenchymal markers were measured by immunofluorescence and Western blot assays. Co-immunoprecipitation was used to detect the interaction between SIRT1 and Smad2/3. The results showed that after mechanical injury, the group treated with RSV-activated SIRT1 regained corneal transparency and recovered from edema faster than the control group. Moreover, RSV-activated SIRT1 downregulated the expression levels of alpha smooth muscle actin (α-SMA), vimentin, and Snail and upregulated the expression levels of E-cadherin and Na<sup>+</sup>/K<sup>+</sup>-ATPase both in vivo and in vitro, but these effects were reversed when SIRT1 was inhibited by EX527. SIRT1 also upregulated the expression levels of TGF-β receptor 1 and phosphorylated Smad2/3. The interaction between SIRT1 and Smad2/3 in vitro was confirmed by co-immunoprecipitation. Overall, our results indicate that SIRT1 activation inhibits corneal EnMT via the TGF-β/Smad2/3 pathway, which may be a potential therapeutic target for corneal endothelium dysfunction.https://www.mdpi.com/1467-3045/46/12/827SIRT1endothelial–mesenchymal transitioncorneal endothelial cellsTGF-β/Smad2/3
spellingShingle Yi Yu
Ruilin Guo
Jie Ling
Chenjia Xu
Minglu Ma
Xiaojuan Dong
Jing Wu
Ting Huang
SIRT1 Activation Suppresses Corneal Endothelial–Mesenchymal Transition via the TGF-β/Smad2/3 Pathway
Current Issues in Molecular Biology
SIRT1
endothelial–mesenchymal transition
corneal endothelial cells
TGF-β/Smad2/3
title SIRT1 Activation Suppresses Corneal Endothelial–Mesenchymal Transition via the TGF-β/Smad2/3 Pathway
title_full SIRT1 Activation Suppresses Corneal Endothelial–Mesenchymal Transition via the TGF-β/Smad2/3 Pathway
title_fullStr SIRT1 Activation Suppresses Corneal Endothelial–Mesenchymal Transition via the TGF-β/Smad2/3 Pathway
title_full_unstemmed SIRT1 Activation Suppresses Corneal Endothelial–Mesenchymal Transition via the TGF-β/Smad2/3 Pathway
title_short SIRT1 Activation Suppresses Corneal Endothelial–Mesenchymal Transition via the TGF-β/Smad2/3 Pathway
title_sort sirt1 activation suppresses corneal endothelial mesenchymal transition via the tgf β smad2 3 pathway
topic SIRT1
endothelial–mesenchymal transition
corneal endothelial cells
TGF-β/Smad2/3
url https://www.mdpi.com/1467-3045/46/12/827
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