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Kindlin-2 regulates the differentiation of 3T3-L1 preadipocytes: implications for wound healing

BACKGROUND: Adipose tissue has been proven to play a crucial role in wound healing, while kindlin-2, an integrin-associated protein, has been shown to regulate cell adhesion, migration, and differentiation. This study aimed to explore its involvement in the cell differentiation of 3T3-L1 preadipocyt...

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Autores principales: Pan, Yuyan, Wang, Qiang, Luan, Wenjie, Shi, Yuedong, Liu, Jiaqi, Qi, Fazhi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: AME Publishing Company 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7944273/
https://www.ncbi.nlm.nih.gov/pubmed/33708975
http://dx.doi.org/10.21037/atm-21-176
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author Pan, Yuyan
Wang, Qiang
Luan, Wenjie
Shi, Yuedong
Liu, Jiaqi
Qi, Fazhi
author_facet Pan, Yuyan
Wang, Qiang
Luan, Wenjie
Shi, Yuedong
Liu, Jiaqi
Qi, Fazhi
author_sort Pan, Yuyan
collection PubMed
description BACKGROUND: Adipose tissue has been proven to play a crucial role in wound healing, while kindlin-2, an integrin-associated protein, has been shown to regulate cell adhesion, migration, and differentiation. This study aimed to explore its involvement in the cell differentiation of 3T3-L1 preadipocytes and its role in wound healing. METHODS: Cell adhesion, Cell Counting Kit-8 (CCK-8), Transwell, and in vitro wound healing assays, along with adipogenic and osteogenic differentiation induction were performed in 3T3-L1 preadipocytes in which kindlin-2 was knocked down or overexpressed. In vivo, kindlin-2 (+/−) transgenic mice were constructed, and wound healing was analyzed by immunohistochemistry (IHC) in a mouse dorsal wound model. Real-time polymerase chain reaction (RT-PCR) and western blotting were performed to analyze the expression of adipokines and adipogenic markers in mouse wound tissues. Adipogenic differentiation induction of adipose tissue stromal vascular fraction (SVF) were performed, and the expression of adipogenic markers in SVF was detected by western blotting. The target signaling pathway highly related to adipogenic differentiation was explored by computational biology and verified by western blotting. RESULTS: Knockdown of kindlin-2 was found to inhibit the adhesion, migration, and adipogenic differentiation of 3T3-L1 preadipocytes while promoting their osteogenic differentiation. In contrast, kindlin-2 overexpression resulted in increased adhesion, migration, and adipogenic differentiation of 3T3-L1 preadipocytes while reducing osteogenic differentiation. In vivo, downregulation of kindlin-2 inhibited adipogenesis in kindlin-2 transgenic mice, resulting in delayed wound healing by inhibiting inflammation, angiogenesis, collagen remodeling, and wound contraction. Mechanistically, we found that kindlin-2 could regulate adipogenic differentiation through PI3K/AKT/mTOR signaling pathway. CONCLUSIONS: Our study revealed the essential role that kindlin-2 has in the differentiation and wound healing of 3T3-L1 preadipocytes, which offers a theoretical basis for further research and a novel strategy for wound healing.
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spelling pubmed-79442732021-03-10 Kindlin-2 regulates the differentiation of 3T3-L1 preadipocytes: implications for wound healing Pan, Yuyan Wang, Qiang Luan, Wenjie Shi, Yuedong Liu, Jiaqi Qi, Fazhi Ann Transl Med Original Article BACKGROUND: Adipose tissue has been proven to play a crucial role in wound healing, while kindlin-2, an integrin-associated protein, has been shown to regulate cell adhesion, migration, and differentiation. This study aimed to explore its involvement in the cell differentiation of 3T3-L1 preadipocytes and its role in wound healing. METHODS: Cell adhesion, Cell Counting Kit-8 (CCK-8), Transwell, and in vitro wound healing assays, along with adipogenic and osteogenic differentiation induction were performed in 3T3-L1 preadipocytes in which kindlin-2 was knocked down or overexpressed. In vivo, kindlin-2 (+/−) transgenic mice were constructed, and wound healing was analyzed by immunohistochemistry (IHC) in a mouse dorsal wound model. Real-time polymerase chain reaction (RT-PCR) and western blotting were performed to analyze the expression of adipokines and adipogenic markers in mouse wound tissues. Adipogenic differentiation induction of adipose tissue stromal vascular fraction (SVF) were performed, and the expression of adipogenic markers in SVF was detected by western blotting. The target signaling pathway highly related to adipogenic differentiation was explored by computational biology and verified by western blotting. RESULTS: Knockdown of kindlin-2 was found to inhibit the adhesion, migration, and adipogenic differentiation of 3T3-L1 preadipocytes while promoting their osteogenic differentiation. In contrast, kindlin-2 overexpression resulted in increased adhesion, migration, and adipogenic differentiation of 3T3-L1 preadipocytes while reducing osteogenic differentiation. In vivo, downregulation of kindlin-2 inhibited adipogenesis in kindlin-2 transgenic mice, resulting in delayed wound healing by inhibiting inflammation, angiogenesis, collagen remodeling, and wound contraction. Mechanistically, we found that kindlin-2 could regulate adipogenic differentiation through PI3K/AKT/mTOR signaling pathway. CONCLUSIONS: Our study revealed the essential role that kindlin-2 has in the differentiation and wound healing of 3T3-L1 preadipocytes, which offers a theoretical basis for further research and a novel strategy for wound healing. AME Publishing Company 2021-02 /pmc/articles/PMC7944273/ /pubmed/33708975 http://dx.doi.org/10.21037/atm-21-176 Text en 2021 Annals of Translational Medicine. All rights reserved. https://creativecommons.org/licenses/by-nc-nd/4.0/Open Access Statement: This is an Open Access article distributed in accordance with the Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License (CC BY-NC-ND 4.0), which permits the non-commercial replication and distribution of the article with the strict proviso that no changes or edits are made and the original work is properly cited (including links to both the formal publication through the relevant DOI and the license). See: https://creativecommons.org/licenses/by-nc-nd/4.0 (https://creativecommons.org/licenses/by-nc-nd/4.0/) .
spellingShingle Original Article
Pan, Yuyan
Wang, Qiang
Luan, Wenjie
Shi, Yuedong
Liu, Jiaqi
Qi, Fazhi
Kindlin-2 regulates the differentiation of 3T3-L1 preadipocytes: implications for wound healing
title Kindlin-2 regulates the differentiation of 3T3-L1 preadipocytes: implications for wound healing
title_full Kindlin-2 regulates the differentiation of 3T3-L1 preadipocytes: implications for wound healing
title_fullStr Kindlin-2 regulates the differentiation of 3T3-L1 preadipocytes: implications for wound healing
title_full_unstemmed Kindlin-2 regulates the differentiation of 3T3-L1 preadipocytes: implications for wound healing
title_short Kindlin-2 regulates the differentiation of 3T3-L1 preadipocytes: implications for wound healing
title_sort kindlin-2 regulates the differentiation of 3t3-l1 preadipocytes: implications for wound healing
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7944273/
https://www.ncbi.nlm.nih.gov/pubmed/33708975
http://dx.doi.org/10.21037/atm-21-176
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