Ghrelin protects the myocardium with hypoxia/reoxygenation treatment through upregulating the expression of growth hormone, growth hormone secretagogue receptor and insulin-like growth factor-1, and promoting the phosphorylation of protein kinase B

Liu,Yang; Liu,Yanling; Li,Guolin; Chen,Zhengrong; Gu,Guixiong

doi:10.3892/ijmm.2018.3886

Ghrelin protects the myocardium with hypoxia/reoxygenation treatment through upregulating the expression of growth hormone, growth hormone secretagogue receptor and insulin-like growth factor-1, and promoting the phosphorylation of protein kinase B

Authors:
- Yang Liu
- Yanling Liu
- Guolin Li
- Zhengrong Chen
- Guixiong Gu
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Affiliations: Department of Child Hygiene, Children's Hospital of Soochow University, Suzhou, Jiangsu 215000, P.R. China, Department of Pediatrics, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China, Department of Pediatrics, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China, Department of Respiratory Disease, Children's Hospital of Soochow University, Suzhou, Jiangsu 215000, P.R. China, Department of Child Hygiene, Children's Hospital of Soochow University, Suzhou, Jiangsu 215000, P.R. China
Published online on: September 18, 2018 https://doi.org/10.3892/ijmm.2018.3886
Pages: 3037-3046
Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Ghrelin is an endogenous ligand of growth hormone (GH) secretagogue receptor (GHSR) and has a number of biological effects, including heart protection. The present study aimed to reveal the positive effect of ghrelin on myocardium with hypoxia/reoxygenation (H/R) treatment and the involved molecular mechanisms. Successful construction of lentiviral expression vector (ghrelin‑pLVX‑Puro) was confirmed by colony polymerase chain reaction (PCR) verification. Primary rat cardiac myocytes were isolated and identified by immunofluorescence staining. Existence of red fluorescence of α‑sarcomeric actinin indicated the successful isolation. Following ghrelin transfection and H/R treatment, primary cells were divided into four groups: Control, H/R, empty (empty pLVX‑Puro + H/R) and ghrelin (ghrelin‑pLVX‑Puro + H/R). Cell viability and apoptosis were evaluated by Cell Counting Kit‑8 (CCK‑8) and Hoechst staining, respectively. The cell viability in the ghrelin group was significantly higher than that in the empty control group (P<0.05). The apoptosis rate in the ghrelin group was significantly lower than that in the empty control group (P<0.05). An ex vivo rat cardiac perfusion model was established. Following ghrelin incubation and H/R treatment, ex vivo myocardium was divided into four groups: Control, sham, H/R and ghrelin (ghrelin + H/R). Immunohistochemical analysis demonstrated that ghrelin increased the integrity of cardiac myocytes, and decreased shrinkage and apoptosis. mRNA and protein expression levels of GH, GHSR, insulin‑like growth factor‑1 (IGF‑1), protein kinase B (Akt), phosphorylated Akt (p‑Akt) were determined by reverse transcription (RT)‑PCR, western blot analysis and immunohistochemical analysis. Ghrelin upregulated the mRNA and protein expression levels of GH, GHSR and IGF‑1, and increased the ratio of p‑Akt to Akt protein level (p‑Akt/Akt) in cardiac myocytes and myocardial tissues with H/R treatment. In conclusion, ghrelin protected the myocardium with H/R treatment through upregulating the expression of GH, GHSR and IGF‑1, and promoting the phosphorylation of Akt. This would provide promising insights into the treatment of hypoxic myocardial injury by ghrelin.

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