Original article
Elevated 20-HETE in metabolic syndrome regulates arterial stiffness and systolic hypertension via MMP12 activation

https://doi.org/10.1016/j.yjmcc.2018.02.005Get rights and content

Highlights

  • 20-HETE regulates large artery remodeling (arterial stiffness) via MMP12 activation and elastin degradation

  • 20-HETE antagonists reverse arterial stiffness and normalize systolic blood pressure in metabolic syndrome

  • Effects of 20-HETE antagonists on arterial stiffness are Ang II-independent; effects on systolic blood pressure are partially Ang II-dependent

  • 20-HETE antagonists may provide viable therapy for treatment of isolated systolic hypertension

Abstract

Arterial stiffness plays a causal role in development of systolic hypertension. 20-hydroxyeicosatetraeonic acid (20-HETE), a cytochrome P450 (CYP450)-derived arachidonic acid metabolite, is known to be elevated in resistance arteries in hypertensive animal models and loosely associated with obesity in humans. However, the role of 20-HETE in the regulation of large artery remodeling in metabolic syndrome has not been investigated. We hypothesized that elevated 20-HETE in metabolic syndrome increases matrix metalloproteinase 12 (MMP12) activation leading to increased degradation of elastin, increased large artery stiffness and increased systolic blood pressure. 20-HETE production was increased ~7 fold in large, conduit arteries of metabolic syndrome (JCR:LA-cp, JCR) vs. normal Sprague-Dawley (SD) rats. This correlated with increased elastin degradation (~7 fold) and decreased arterial compliance (~75% JCR vs. SD). 20-HETE antagonists blocked elastin degradation in JCR rats concomitant with blocking MMP12 activation. 20-HETE antagonists normalized, and MMP12 inhibition (pharmacological and MMP12-shRNA-Lnv) significantly improved (~50% vs. untreated JCR) large artery compliance in JCR rats. 20-HETE antagonists also decreased systolic (182 ± 3 mmHg JCR, 145 ± 3 mmHg JCR + 20-HETE antagonists) but not diastolic blood pressure in JCR rats. Whereas diastolic pressure was fully angiotensin II (Ang II)-dependent, systolic pressure was only partially Ang II-dependent, and large artery stiffness was Ang II-independent. Thus, 20-HETE-dependent regulation of systolic blood pressure may be a unique feature of metabolic syndrome related to high 20-HETE production in large, conduit arteries, which results in increased large artery stiffness and systolic blood pressure. These findings may have implications for management of systolic hypertension in patients with metabolic syndrome.

Introduction

Large artery stiffness is a major risk factor for hypertension, stroke and myocardial infarction [1]. Structural properties of large arteries, medial thickness and composition of the extracellular matrix (ECM), are the main determinants of large artery stiffness [2].

Despite the significant contribution of the ECM to increased blood pressure development and maintenance, no currently available anti-hypertensive therapy directly targets the ECM, although some (e.g. angiotensin converting enzyme inhibitors (ACEIs) and angiotensin receptor blockers (ARBs)) do affect it. Targeting elastin fiber integrity has been proposed as an effective therapy to specifically target systolic hypertension. Studies in elastin knockout mice have shown that its deficiency leads to a severe and fatal increase in systolic pressure, which is preceded by arterial stiffening, suggesting a causative role for elastin in maintenance of not only arterial compliance but also of normal systolic pressure [3].

Matrix metalloproteinase 12 (MMP12), one of the major elastin-degrading proteases, is secreted primarily from monocytes and neutrophils [4], but can also be made by synthetic vascular smooth muscle cells (VSMCs) [5]. Its upstream regulators are unknown. It has been implicated in chronic obstructive pulmonary disease and emphysema, atherosclerosis in type II diabetes, aneurysms, cancer and stiffening of femoral arteries in response to injury [[5], [6], [7], [8], [9]]. Its role in arterial stiffness in metabolic syndrome or hypertensive animals or humans has not been investigated. MMPs classically associated with the regulation of hypertension-induced arterial stiffness are MMP2 and MMP9, but their activity does not fully account for either large artery stiffness or systolic hypertension in animal models or humans [10].

20-hydroxyeicosatetraeonic acid (20-HETE) is synthesized from arachidonic acid by cytochrome P450 (CYP) [11]. In rats, CYP4A (A1, A2, A3, A8) and CYP4F (F1, F4) are the CYP isoforms which produce 20-HETE. 20-HETE is synthesized and released from many cell types including VSMCs [12], endothelial cells, neutrophils [13] and bone marrow cells [14].

The association with and causative role of 20-HETE in the development of hypertension is now well established. In the SHR rat, depletion of CYP4A decreased 20-HETE and significantly lowered mean arterial blood pressure [15]. Androgen-driven hypertension in mice and rats, the Cyp4a14 (−/−) androgen-dependent 20-HETE-mediated hypertension mouse model, the Cyp4a12 doxycycline inducible mouse model, and the Sprague-Dawley (SD) rat overexpressing CYP4A2 in the vascular endothelium exhibit increased 20-HETE production and hypertension [[16], [17], [18], [19]]. However, 20-HETE's role in the regulation of vascular structural remodeling is just emerging. Only recently, Schwartzman's group demonstrated that 20-HETE induced microvascular remodeling, which was only partially Ang II-dependent [20]. Furthermore, all previous studies evaluated the effect of 20-HETE manipulation only on mean arterial blood pressure (MABP). Moreover, because in normal, healthy animals 20-HETE is not produced in large arteries, all work focused on the role of small, resistance arteries and the contribution of myogenic tone to the regulation of systemic blood pressure. Therefore, nothing is known about the possible effect of 20-HETE in large arteries or on extracellular matrix (ECM) remodeling.

In this study, we explore a novel idea that, in the metabolic syndrome, 20-HETE specifically modulates systolic but not diastolic blood pressure, and that it does so by regulating large artery compliance, as a critical determinant of arterial stiffness, through regulating elastin degradation via MMP12 activation.

Section snippets

Animals

10–12 week old, male JCR:LA-cp (JCR; S. Proctor, University of Alberta, Edmonton, Canada) (650–700 g), spontaneously hypertensive obese (SHROB, Charles Rivers, Wilmington, MA) (650–700 g) and Sprague-Dawley (SD; Charles Rivers) (300–350 g) rats were used in all experiments. The JCR rat is a cross between the lean LA/N Zucker and the spontaneously hypertensive obese (SHROB) rat developed in the laboratory of Dr. Carl Hansen at the National Institutes of Health and sent to Drs. James C. Russell

CYP4A, CYP4F and 20-HETE are elevated in metabolic syndrome

CYP4A (4A1, 2, 3 and 8) and CYP4F (4F1 and 4) are the CYP isoforms expressed in rat which make 20-HETE; the rat expresses 4F5 and 6 but these two isoforms do not make 20-HETE [32]. Our results indicate that CYP4A and CYP4F are expressed in carotid arteries and aorta of JCR and SHROB but not of SD rats (~2 fold JCR vs. SD) (Figs. 1A and VIII, Supplement). By RT-PCR analysis, we have identified the CYP4A isoforms expressed in arteries of JCR and SD rats to be 4A2 and 4A3; whereas, 4A1 expression

Discussion

The most important findings in this study are: 1) that 20-HETE is highly elevated in large, conduit arteries of metabolic syndrome animals which correlates with increased elastin degradation, decreased large artery compliance and increased blood pressure in these animals, and 2) that 20-HETE antagonism reverses elastin degradation, restores large artery compliance and normalizes systolic but not diastolic blood pressure in the metabolic syndrome animals, and 3) that this effect is in part Ang

Sources of funding

NIH R01HL093052 (PR), NIH 1F31HL137356 (AS), P01HL034300 (MLS).

Disclosures

None.

Acknowledgments

The authors would like to acknowledge Dr. Alberto Nasjletti (Department of Pharmacology, New York Medical College) for critical review of the manuscript prior to submission.

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