Angiotensin (1-7) Inhibits Transforming Growth Factor-Β1-Induced Epithelial-Mesenchymal Transition of Human Keratinocyte Hacat Cells in vitro

Yueda Jihu; Ruobing Leng; Mengchang Liu; Hongjing Ren; Defu Xie; Chong Yao; Hong Yan

doi:10.2147/CCID.S441596

Angiotensin (1-7) Inhibits Transforming Growth Factor-Β1-Induced Epithelial-Mesenchymal Transition of Human Keratinocyte Hacat Cells in vitro

Clin Cosmet Investig Dermatol. 2024 May 8:17:1049-1058. doi: 10.2147/CCID.S441596. eCollection 2024.

Authors

Yueda Jihu^#^{1

2}, Ruobing Leng^#¹, Mengchang Liu^{1

2}, Hongjing Ren^{1

2}, Defu Xie^{1

2}, Chong Yao^{1

2}, Hong Yan^{1

2}

Affiliations

¹ Clinical College of Medicine, Southwest Medical University, Lu zhou, People's Republic of China.
² Department of Plastic and Burn Surgery, the Affiliated Hospital of Southwest Medical University, Lu zhou, People's Republic of China.

^# Contributed equally.

Abstract

Introduction: Angiotensin (1-7) (Ang-(1-7)) is an emerging component of the renin-angiotensin system (RAS) with effective anti-fibrosis properties and has been shown to interfere with epithelial-mesenchymal transition (EMT) by numerous studies. In recent years, EMT has been proposed as a new therapeutic target for skin fibrotic diseases such as keloids. However, the effect of Ang-(1-7) on EMT in skin is still unclear. Hence, the purpose of this study was to explore the effect of Ang-(1-7) on Transforming growth factor-β1(TGF-β1)-induced EMT of human immortalized keratinocytes HaCaT in vitro.

Methods: The study involved the use of the human immortalized keratinocyte cell line (HaCaT). The cells were cultured in high-glucose DMEM medium with 10% fetal bovine serum and 1% penicillin-streptomycin. Four groups were created for experimentation: control group (Group C), TGF-β1-treated group (Group T), Ang-(1-7)-treated group (Group A), and a group treated with both TGF-β1 and Ang-(1-7) (Group A + T). Various assays were conducted, including a cell proliferation assay using CCK-8 solution, a scratch wound healing assay to evaluate cell migration, and Western blotting to detect protein expressions related to cell characteristics. Additionally, quantitative real-time polymerase chain reaction (PCR) was performed to analyze epithelial-mesenchymal transition (EMT) related gene expression levels. The study aimed to investigate the effects of TGF-β1 and Ang-(1-7) on HaCaT cells.

Results: We found that Ang-(1-7) not only reduced the migration of HaCaT cells induced by TGF-β1 in vitro but also reduced the expression of α-SMA and vimentin, and restored the protein expression of E-cadherin and claudin-1. Mechanistically, Ang-(1-7) inhibits the phosphorylation levels of Smad2 and Smad3 in the TGF-β1 canonical pathway, and suppresses the expression of EMT-related transcription factors (EMT-TFs) such as SNAI2, TWIST1, and ZEB1.

Discussion: Taken together, our findings suggest that Ang-(1-7) inhibits TGF-β1-induced EMT in HaCaT cells in vitro by disrupting the TGF-β1-Smad canonical signaling pathway. These results may be helpful in the treatment of EMT in skin fibrotic diseases such as keloids.

Keywords: angiotensin (1-7); epithelial-mesenchymal transition; keratinocytes; transforming growth factor-β.

Grants and funding

This research was funded by Joint Project of Southwest Medical University and Luzhou city, the project number: 02×00150027. This research was also funded by the (Luzhou-Medical University) Cooperation Project (No. 2019LZXNYDZ08).