Humanin inhibits lymphatic endothelial cells dysfunction to alleviate myocardial infarction-reperfusion injury via BNIP3-mediated mitophagy

Abstract

Objective

Acute myocardial infarction (AMI) ranks among the top contributors to sudden death and disability worldwide. It should be noted that current therapies always cause increased reperfusion damage. Evidence suggests that humanin (HN) reduces mitochondrial dysfunction to have cardio-protective effects against MI-reperfusion injury. In this context, we hypothesized that HN may attenuate MI-reperfusion injury by alleviating lymphatic endothelial cells dysfunction through the regulation of mitophagy.

Materials and methods

In this study, primary lymphatic endothelial cells were selected as the experimental model. Cells were maintained under 1% O₂ to induce a hypoxic phenotype. For in vivo experiments, the left coronary arteries of C57/BL6 mice were clamped for 45 min followed by 24 h reperfusion to develop MI-reperfusion injury. The volume of infarcted myocardium in MI-reperfusion injury mouse models were TTC staining. PCR and western blot were used to quantify the expression of autophagy-, mitophagy- and mitochondria-related markers. The fibrosis and apoptosis in the ischemic area were evaluated for Masson staining and TUNEL respectively. We also used western blot to analyze the expression of VE-Cadherin in lymphatic endothelial cells.

Results

We firstly exhibited a specific mechanism by which HN mitigates MI-reperfusion injury. We demonstrated that HN effectively reduces such injury in vivo and also inhibits dysfunction in lymphatic endothelial cells in vitro. Importantly, this inhibitory effect is mediated through BNIP3-associated mitophagy.

Conclusions

In conclusion, HN alleviates myocardial infarction-reperfusion injury by inhibiting lymphatic endothelial cells dysfunction, primarily through BNIP3-mediated mitophagy.

Keywords:

• Myocardial infarction-reperfusion injury
• humanin
• mitophagy
• lymphatic endothelial cells

Authors’ contributions

Cao Zou conceived and designed the study. Lu Chen, Xiaohua Yang and Kai Wang performed the experiments and wrote the paper. Lina Guo reviewed and edited the manuscript. All authors had read and approved the manuscript.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Additional information

Funding

This work was supported by Soochow University horizontal subject (No.H220694), Nurture projects for basic research of Shanghai Chest Hospital (No.2020YNJCQ08), Interdisciplinary Program of Shanghai Jiao Tong University (No.YG2021QN122), 2021 Shanghai "Rising Stars of Medical Talents" Youth Development Program (No.SHWSRS(2022)_065) and the Parallel Research Topic of Soochow university (No.H190426)