PHLDA3 inhibition attenuates endoplasmic reticulum stress‑induced apoptosis in myocardial hypoxia/reoxygenation injury by activating the PI3K/AKT signaling pathway

Liu,Kai; Chen,Ying; Ai,Fen; Li,Yun-Qian; Zhang,Kun; Zhang,Wei-Tong

doi:10.3892/etm.2021.10045

PHLDA3 inhibition attenuates endoplasmic reticulum stress‑induced apoptosis in myocardial hypoxia/reoxygenation injury by activating the PI3K/AKT signaling pathway

Authors:
- Kai Liu
- Ying Chen
- Fen Ai
- Yun-Qian Li
- Kun Zhang
- Wei-Tong Zhang
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Affiliations: Department of Geriatric Center, Hainan General Hospital, Hainan Affiliated Hospital of Hainan Medical University, Haikou, Hainan 570311, P.R. China, Department of Emergency, The Central Hospital of Wuhan, Tongji Medical College, Hua Zhong University of Science and Technology, Wuhan, Hubei 430000, P.R. China, Department of General Surgery, Hainan General Hospital, Hainan Affiliated Hospital of Hainan Medical University, Haikou, Hainan 570311, P.R. China
Published online on: April 14, 2021 https://doi.org/10.3892/etm.2021.10045
Article Number: 613
Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Endoplasmic reticulum stress (ERS)‑induced apoptosis serves a crucial role in the pathogenesis of myocardial ischemia/reperfusion injury (MIRI). Previous studies have confirmed that pleckstrin homology‑like domain family A member 3 (PHLDA3) is an important mediator in ERS‑associated apoptosis. The aim of the current study focused on whether PHLDA3 served protective effects on hypoxia/reoxygenation (H/R)‑injured cardiomyocytes by inhibiting ERS‑induced apoptosis. Furthermore, the molecular mechanisms associated with the PI3K/AKT signaling pathway were investigated. Primary neonatal rat cardiomyocytes were isolated and randomized into four groups: i) Control + adenovirus encoding scrambled short hairpin RNA (AdshRNA); ii) control + adenoviral vectors encoding PHLDA3 shRNA (AdshPHLDA3); iii) H/R+ AdshRNA and iv) H/R+AdshPHLDA3. AdshPHLDA3 was used to knock down PHLDA3. An H/R injury model was constructed by treatment with hypoxia for 4 h followed by reoxygenation for 6 h. A PI3K/AKT inhibitor, LY294002, was supplemented in mechanistic studies. Cell viability and LDH/CK releases were detected to evaluate myocardial damage. Flow cytometry assays were used to assess apoptotic response. Western blotting assays were used to detect protein expression. The results demonstrated that H/R induced myocardial damage and increased PHLDA3 expression. ERS‑induced apoptosis was significantly increased following H/R injury, as indicated by increased apoptotic rates and ERS‑associated protein expression, including those of CHOP, 78 kDa glucose‑regulated protein and caspase‑12. However, PHLDA3 inhibition following AdshPHLDA3 transfection reversed cell damage and ERS‑associated apoptosis on H/R injury. Studies for molecular mechanisms concluded that the apoptosis‑inhibition effects and cardioprotective roles of PHLDA3 inhibition were induced partly by the activation of the PI3K/AKT pathway, which was verified by LY294002 treatment. In conclusion, in the process of H/R injury, PHLDA3 inhibition reduced ERS‑induced apoptosis and H/R injury by activating the PI3K/AKT pathway. PHLDA3 may be a therapeutic target for the treatment of MIRI.

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