Pirfenidone attenuates homocysteine‑induced apoptosis by regulating the connexin 43 pathway in H9C2 cells

Chen,Kai; Chen,Ling; Ouyang,Yuanshuo; Zhang,Liang; Li,Xinzhi; Li,Li; Si,Junqiang; Wang,Li; Ma,Ketao

doi:10.3892/ijmm.2020.4497

Pirfenidone attenuates homocysteine‑induced apoptosis by regulating the connexin 43 pathway in H9C2 cells

Authors:
- Kai Chen
- Ling Chen
- Yuanshuo Ouyang
- Liang Zhang
- Xinzhi Li
- Li Li
- Junqiang Si
- Li Wang
- Ketao Ma
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Affiliations: Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Medicine School of Shihezi University, Shihezi, Xinjiang 832008, P.R. China, The Third Department of Cardiology, The First Affiliated Hospital of The Medical College, Shihezi University, Shihezi, Xinjiang 832008, P.R. China
Published online on: February 13, 2020 https://doi.org/10.3892/ijmm.2020.4497
Pages: 1081-1090
Copyright: © Chen et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Pirfenidone (PFD) is an anti‑fibrotic agent that is clinically used in the treatment of idiopathic pulmonary fibrosis. PFD has been shown to exert protective effects against damage to orbital fibroblasts, endothelial cells, liver cells and renal proximal tubular cells; however, its effect on myocardial cell apoptosis remains unclear. The present study aimed to characterize the effects of PFD on homocysteine (Hcy)‑induced cardiomyocyte apoptosis and investigated the underlying mechanisms. H9C2 rat cardiomyocytes were pre‑treated with PFD for 30 min followed by Hcy exposure for 24 h. The effects of PFD on cell cytotoxicity were evaluated by CCK‑8 assay. The apoptosis rate of each group was determined by flow cytometry. The protein and mRNA levels of connexin 43 (Cx43), Bax, B‑cell lymphoma‑2 (Bcl‑2) and caspase‑3 were measured by western blot analysis and reverse transcription‑quantitative PCR, respectively. The present results demonstrated that the apoptotic rate increased following Hcy exposure, whereas the apoptotic rate significantly decreased following PFD pre‑treatment. Furthermore, the ratio of Bax/Bcl2 was upregulated following Hcy exposure, and Hcy upregulated the expression levels of cleaved caspase‑3 and Cx43. Notably, these effects were prevented by PFD. Additionally, the effects of PFD were inhibited by the Cx43 agonist, AAP10. In summary, the findings of the present study demonstrate that PFD protects H9C2 rat cardiomyocytes against Hcy‑induced apoptosis by modulating the Cx43 signaling pathway.

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