Estrogen inhibits TGF‑&beta;1‑stimulated cardiac fibroblast differentiation and collagen synthesis by promoting Cdc42

Xu,Jingyi; Wang,Feng; Li,Yuan; Huang,Ying; Li,Ping; Zhang,Yiqing; Xu,Guidong; Sun,Kangyun

doi:10.3892/mmr.2024.13246

Estrogen inhibits TGF‑β1‑stimulated cardiac fibroblast differentiation and collagen synthesis by promoting Cdc42

Authors:
- Jingyi Xu
- Feng Wang
- Yuan Li
- Ying Huang
- Ping Li
- Yiqing Zhang
- Guidong Xu
- Kangyun Sun
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Affiliations: Department of Central Laboratory, The Affiliated Suzhou Hospital of Nanjing Medical University, Suzhou Municipal Hospital, Gusu School, Nanjing Medical University, Suzhou, Jiangsu 215008, P.R. China, Department of Pharmacy, The Affiliated Suzhou Hospital of Nanjing Medical University, Suzhou Municipal Hospital, Gusu School, Nanjing Medical University, Suzhou, Jiangsu 215008, P.R. China, Department of Cardiology, The Affiliated Suzhou Hospital of Nanjing Medical University, Suzhou Municipal Hospital, Gusu School, Nanjing Medical University, Suzhou, Jiangsu 215008, P.R. China
Published online on: May 17, 2024 https://doi.org/10.3892/mmr.2024.13246
Article Number: 123
Copyright: © Xu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

17β‑estradiol (E2) can inhibit cardiac fibrosis in female patients with heart failure (HF) and activate cell division cycle 42 (Cdc42), however it is unknown whether 17β‑estradiol (E2) can ameliorate differentiation and collagen synthesis in TGF‑β1‑stimulated mouse cardiac fibroblasts (MCFs) by regulating cell division cycle 42 (Cdc42). The present study aimed to investigate the roles of estrogen and Cdc42 in preventing myocardial fibrosis and the underlying molecular mechanisms. An ELISA was used to measure the levels of E2 and Cdc42 in the serum of patients with heart failure (HF), and western blotting was used to measure the expression levels of Cdc42 in TGF‑β1‑stimulated immortalized MCFs. MCFs were transfected with a Cdc42 overexpression (OE) lentivirus or small interfering RNA (siRNA), or treated with a Cdc42 inhibitor (MLS‑573151), and the function of Cdc42 was assessed by western blotting, immunofluorescence staining, reverse transcription‑quantitative PCR and dual‑luciferase reporter assays. Western blotting and immunofluorescence staining were performed to verify the protective effect of E2 on TGF‑β1‑stimulated MCFs, and the association between the protective effect and Cdc42. The results demonstrated that Cdc42 levels were increased in the serum of patients with HF and were positively correlated with the levels of E2; however, Cdc42 levels were decreased in TGF‑β1‑stimulated MCFs. Cdc42 inhibited MCF differentiation and collagen synthesis, as indicated by the protein expression of α‑smooth muscle actin, collagen I and collagen III. Mechanistically, Cdc42 inhibited the transcription of TGF‑β1 by promoting the expression of p21 (RAC1)‑activated kinase 1 (Pak1)/JNK/c‑Jun signaling pathway proteins and inhibiting the activity of the Tgfb1 gene promoter. In addition, E2 inhibited the differentiation and collagen synthesis of TGF‑β1‑stimulated MCFs, and promoted the protein expression of Pak1, JNK and c‑Jun, consistent with the effects of Cdc42, whereas the effects of E2 were abolished when Cdc42 was knocked down. The aforementioned findings suggested that E2 could inhibit differentiation and collagen synthesis in TGF‑β1‑stimulated MCFs by regulating Cdc42 and the downstream Pak1/JNK/c‑Jun signaling pathway.

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