Review article
Endothelium-specific CYP2J2 overexpression improves cardiac dysfunction by promoting angiogenesis via Jagged1/Notch1 signaling

https://doi.org/10.1016/j.yjmcc.2018.08.027Get rights and content

Highlights

  • Endothelium-specific overexpression of CYP2J2 in rats improved cardiac function in post-MI.

  • CYP2J2 increased the density of EETs in plasma, and EETs promoted angiogenesis via Jagged1/Notch1 signaling pathway.

  • EETs promoted angiogenesis, and the neovessels performed functionally, thus increasing myocardial blood flow.

  • After AMI in patients, the expression of CYP2J2 was increased, while decreased in OMI.

Abstract

Background

Myocardial infarction (MI) contributes to the development of cardiac remodeling and heart failure. Insufficient post-MI myocardial angiogenesis has been identified as a non-negligible event which precipitates heart failure progression. Previous studies reported that cytochrome P450 epoxygenase and its metabolites exerted beneficial effects on cardiovascular diseases. However, the role of cytochrome P450 2J2 (CYP2J2) in post-MI heart failure is incompletely understood.

Methods and results

First, western blot and real-time PCR analyses showed that CYP2J2 expression increased clearly in patients with acute MI and old MI, compared to control. Second, echocardiography and histological studies showed that transgenic (TG) rats had relatively preserved cardiac function, as well as attenuated remodeling, and reduced scar formation, compared to the wild-type (WT) littermates after MI eight weeks. Importantly, the cardioprotective effect induced by CYP2J2 overexpression was abrogated by VEGFR2 inhibitor—cediranib. More intriguingly, positron emission computed Tomography (PET) analyses showed that TG rats displayed better myocardial perfusion than WT rats. We found that these effects were linked to increasing circulating EETs and enhancing myocardial angiogenesis. Additionally, in vitro study demonstrated that 11, 12-epoxyeicosatrienoic acid (11, 12-EET) induced more robust tube formation and markedly increased VEGF-A and bFGF expression in hypoxia and normoxia. Finally, western blot analyses uncovered that CYP2J2 and 11, 12-EET promoted angiogenesis via the Jagged1/Notch1 signaling pathway.

Conclusions

Our findings demonstrate that CYP2J2 improves cardiac function by increasing the concentration of circulating EETs, and boosting angiogenesis via the Jagged1/Notch1 signaling pathway in MI-induced heart failure.

Introduction

Heart failure following myocardial infarction (MI) remains one of the major causes of death worldwide and is a worsening public health problem [1,2]. During MI, inadequate blood flow to an ischemic heart muscle leads to cardiomyocytes death and pathological remodeling, which head to heart failure over time [3,4]. Although ischemia induces endogenous myocardial angiogenesis, this process is insufficient for maintaining a normal myocardial blood supply. The application of advanced revascularization strategies, such as primary percutaneous coronary intervention (PCI) and surgical bypass, have led to a remarkable decrease in mortality rates. However, these strategies have inherent limitation, such as the ‘no-reflow’ phenomenon. In addition, a large proportion of patients with MI who received these treatments showed inadequate myocardial perfusion, owing to dysfunction of the microcirculation or lack of myocardial angiogenesis [5]. Hence, therapeutic angiogenesis which aims to restore blood flow to the ischemic myocardial zone is considered as a promising treatment to salvage ischemic myocardium and prevents the transition to heart failure.

The term“angiogenesis” refers to the process of blood vessels formation from pre-existing vascular bed [6], angiogenic growth factors act as fundamental role in this process. Vascular endothelial growth factor A (VEGF-A) and basic fibroblast growth factor (bFGF) are the major and well-studied growth factors in ischemic tissues among the current known pro-angiogenic growth factors [7]. VEGF-A can initiate multiple signaling pathways that orchestrate a variety of complex biological effects, such as endothelial cells maturation, angiogenesis and arteriogenesis in response to hypoxia or healing, and it stimulates angiogenesis through vascular endothelial growth factor receptor-2 (VEGFR2) signaling mainly [8].

Cytochrome P450 (CYP) epoxygenases metabolize arachidonic acid (AA) to four biologically and cardioprotective epoxyeicosatrienoic acids (EETs): 5, 6-EET, 8, 9-EET, 11, 12-EET, and 14, 15-EET. The primary products are 11, 12- and 14, 15-EET. Via the activity of soluble epoxide hydrolase (sEH), EETs are metabolized to dihydroxyeicosatrienoic acid (DHET), a compound that is more stable and less bioactive [9]. The human cytochrome P450 epoxygenase, CYP2J2, is the only member of the human CYP2J family, which expresses abundantly in coronary artery endothelial cells, smooth muscle cells, and cardiomyocytes [[10], [11], [12]]. CYP2J2 and EETs have pleotropic effects in cardioprotection, including alleviating inflammatory reactions [13], inhibiting apoptosis [14], and attenuating cardiac remodeling and hypertrophy [15,16]. Previous studies also revealed that cardiac-specific overexpression of human CYP2J2 or EETs administration directly enhanced cardiac recovery of left ventricular function during ischemia/reperfusion involved activation of p42/p44-MAPK signaling pathway [17,18]. However, these studies mainly focused the cardioprotection of CYP2J2 and EETs on cardiomyocytes, little was known about the cardioprotection on endothelial cells-induced angiogenesis. Vascular endothelial cells play a prominent role in cardiovascular physiology and pathobiology. Recent studies have gradually recognized the crucial role of endothelium-specific CYP2J2 overexpression in ischemia disease [19,20]. Thus, the present study demonstrated that endothelium-specific CYP2J2 overexpression protected against post-MI heart failure in rats, and this cardioprotective role was associated with increasing the level of circulating EETs and promoting angiogenesis via Jaggged1/Notch1 signaling.

Section snippets

Origin of human heart samples

Human heart tissue samples diagnosed with MI were obtained from patients who underwent cardiac transplantation in our hospital, Union Hospital (Wuhan, China) and organ donors from 2013―2016. Control hearts were obtained from sex- and age-matched subjects who died in traffic accidents or had been diagnosed with aortic regurgitation. These patients did not have ischemic heart diseases, their family members donated their organs voluntarily. The study was approved by the Ethics Review Board of

RNA interference

The small interfering RNA (siRNA) oligonucleotides of Notch1 and the corresponding control were synthesized by RuiBo (China). Transfection was performed when HUVECs reached the density of approximately 30%―40% confluent. HUVECs were transfected with the siRNA of Notch1 using Lipofectamine2000 (Invitrogen, USA) 48 h prior to treatment with the scheduled conditions. The siRNA was diluted to a final concentration of 50 nM in Opti-Mem I (Invitrogen). After that, transfection reagent (Invitrogen,

CYP2J2 and sEH expression are up-regulated in patients with heart failure post MI

We detected CYP2J2 and sEH protein and messenger RNA expression in human heart samples (Information pertaining to these heart samples is presented in Supplementary Table S1). Western blot analysis showed that CYP2J2 expression was upregulated clearly in patients with AMI and OMI, compared to control patients (Fig. 1A and B). Interestingly, sEH expression tended to be increased in patients with AMI, while did not alter in patients with OMI, compared control patients (Fig. 1A and C). Next, qPCR

Discussion

The main finding of our work is that endothelium-specific CYP2J2 overexpression favourably affects the performance of failing hearts (as evaluated by echocardiography and cardiac catheterization). Specifically, CYP2J2 overexpression improved the LV ejection fraction and lowered LV wall stress. We also demonstrated that gain-of-function in CYP2J2 decreased mortality rates and attenuated LV remodeling, as assessed by analyses of fibrosis, LV wall thickness, and HW-to-BW ratios. These effects were

Sources of funding

This research was supported by the following grants: NIH 31130031 and NIH 91439203.

Acknowledgements

We are grateful to Riverine Technology Co., Ltd. (Wuhan, China) for providing technical assistance with the small-animal PET imaging analysis. We thank the Department of Pulmonary and Critical Care Medicine (TongJi Hospital, TongJi Medical College, Huazhong University of Science and Technology) for providing three gas incubator and thank Department of Physiology and Tianjin Medical University (Tianjin, China) for performing LC/MS/MS analysis.

Nonstandard abbreviations and acronyms

    MI

    Myocardial infarction

    HF

    Heart failure

    EETs

    Epoxyeicosatrienoic acids

    CYP

    Cytochrome P450

    sEH

    Soluble epoxide hydrolase

    AMI

    Acute myocardial infarction

    OMI

    Old myocardial infarction

    TG

    Transgenic

    WT

    Wild-type

    VEGF

    Vascular endothelial growth factor

    PET

    Positron emission computed tomography

    IZ

    Infarct Zone

    BZ

    Border Zone

    RZ

    Remote Zone

    LC/MS/MS

    Liquid chromatography tandem mass spectroscopy

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    These authors contributed equally to this work.

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