Effect of endothelin on sex-dependent regulation of tone in coronary resistance vessels.

BACKGROUND/AIMS: Sex dependent differences in coronary artery vasoregulation may be due to variations in responses to endogenous vasoactive compounds including endothelin (ET-1) and nitric oxide (NO). METHODS: Septal coronary arteries (<200 µm) from healthy, sexually mature male, female and ovariectomized (i.e. surgical menopause) Sprague-Dawley rats were used. Myogenic tone, measured by pressure myography, was initially determined for all vessel segments studied before and after exposure to the nonselective ETA/ETB receptor blocker, bosentan (1 µM). Vasoconstrictor responses (vascular endothelium intact) to cumulative ET-1 (10-12 - 10-9 M) were assessed in a separate set of septal coronary vessels. Additional studies, examined the vasoconstrictor effects of ET-1 after NO blockade with L-NAME (200 µM). RESULTS: Myogenic tone was 26 ± 7% in male, 20 ± 7% in female (p = 0.04 versus male) and 24 ± 3% in ovariectomized (p = NS versus male/female) vessels. Antagonism of ET-1 receptors produced a greater reduction in myogenic tone in male, compared to female rats over a similar range of intraluminal pressure (20-80 mmHg). Robust constrictor responses to cumulative concentrations of ET-1 were observed in all vessels; however, male rats exhibited greater sensitivity to vasoconstrictor effects of ET-1. After exposure to L-NAME vessel responses to ET-1 were normalized in male and female (not studied in ovariectomized) groups. CONCLUSIONS: These findings confirm marked sex differences for myogenic tone and vessel constrictor responses to ET-1 in coronary resistance vessels. Results also suggest greater sensitivity to vasoconstrictor effects of ET-1 in male coronary resistance vessels.

Effect of Platelet GPIIb/IIIa Receptor Blockade With MK383 on Infarct Size and Myocardial Blood Flow in a Canine Reocclusion Model.

Platelet activation and aggregation during ischemia influence reperfusion-related myocyte necrosis, myocardial perfusion at the microvascular level, and thereby eventual recovery of cardiac performance. Inhibition of platelet activity therefore represents a worthwhile target to reduce cellular injury. The current study examined the effects of MK383 (tirofiban), a potent inhibitor of platelet aggregation, on infarct size and myocardial perfusion in canine subjects to either reocclusion (ie, 120-minute + 60-minute ischemia with intervening reperfusion) or prolonged occlusion (ie, 3 hours) followed by reperfusion (180 minutes). Platelet aggregation, infarct size (tetrazolium staining), coronary blood flow (flow probe), coronary vascular reserve, and myocardial perfusion (microspheres) were evaluated. MK383, administered at the time of reperfusion, produced a modest reduction of tissue necrosis (compared to saline-treated controls) in the reocclusion and prolonged occlusion studies. Blood flow in the infarct-related artery after coronary occlusion was comparable between treatment groups, as was myocardial perfusion in the deeper layers of the ischemic region; coronary vascular reserve decreased progressively during reperfusion. Of note, compensatory changes in blood flow within the adjacent nonischemic myocardium were not observed. In conclusion, we report that that limiting platelet aggregation during reperfusion impacted infarct development. Continued investigation into the mechanisms by which inhibition of platelet activity protects myocardium against ischemia-reperfusion injury and improves clinical outcomes is necessary.

Influence of cardiac nerve status on cardiovascular regulation and cardioprotection.

Neural elements of the intrinsic cardiac nervous system transduce sensory inputs from the heart, blood vessels and other organs to ensure adequate cardiac function on a beat-to-beat basis. This inter-organ crosstalk is critical for normal function of the heart and other organs; derangements within the nervous system hierarchy contribute to pathogenesis of organ dysfunction. The role of intact cardiac nerves in development of, as well as protection against, ischemic injury is of current interest since it may involve recruitment of intrinsic cardiac ganglia. For instance, ischemic conditioning, a novel protection strategy against organ injury, and in particular remote conditioning, is likely mediated by activation of neural pathways or by endogenous cytoprotective blood-borne substances that stimulate different signalling pathways. This discovery reinforces the concept that inter-organ communication, and maintenance thereof, is key. As such, greater understanding of mechanisms and elucidation of treatment strategies is imperative to improve clinical outcomes particularly in patients with comorbidities. For instance, autonomic imbalance between sympathetic and parasympathetic nervous system regulation can initiate cardiovascular autonomic neuropathy that compromises cardiac stability and function. Neuromodulation therapies that directly target the intrinsic cardiac nervous system or other elements of the nervous system hierarchy are currently being investigated for treatment of different maladies in animal and human studies.

The Physiopathology of Cardiorenal Syndrome: A Review of the Potential Contributions of Inflammation.

Inter-organ crosstalk plays an essential role in the physiological homeostasis of the heart and other organs, and requires a complex interaction between a host of cellular, molecular, and neural factors. Derangements in these interactions can initiate multi-organ dysfunction. This is the case, for instance, in the heart or kidneys where a pathological alteration in one organ can unfavorably affect function in another distant organ; attention is currently being paid to understanding the physiopathological consequences of kidney dysfunction on cardiac performance that lead to cardiorenal syndrome. Different cardiorenal connectors (renin-angiotensin or sympathetic nervous system activation, inflammation, uremia, etc.) and non-traditional risk factors potentially contribute to multi-organ failure. Of these, inflammation may be crucial as inflammatory cells contribute to over-production of eicosanoids and lipid second messengers that activate intracellular signaling pathways involved in pathogenesis. Indeed, inflammation biomarkers are often elevated in patients with cardiac or renal dysfunction. Epigenetics, a dynamic process that regulates gene expression and function, is also recognized as an important player in single-organ disease. Principal epigenetic modifications occur at the level of DNA (i.e., methylation) and histone proteins; aberrant DNA methylation is associated with pathogenesis of organ dysfunction through a number of mechanisms (inflammation, nitric oxide bioavailability, endothelin, etc.). Herein, we focus on the potential contribution of inflammation in pathogenesis of cardiorenal syndrome.

Impact of chronic kidney disease on myocardial blood flow regulation in dogs.

BACKGROUND/AIMS: Chronic kidney disease (CKD) increases cardiovascular risk possibly due to coronary microvessel dysfunction and impaired myocardial flow reserve. This study investigated the effects of CKD on the regulation and transmural distribution of myocardial blood flow along with oxygen demand during intravenous dobutamine-induced increases in cardiac work. METHODS: CKD was produced in dogs by a two-stage subtotal nephrectomy (kidney ablation-infarction model). Serum creatinine and blood urea nitrogen were evaluated during the development of CKD along with systemic blood pressure (tail-cuff plethysmography). After 5 weeks, the CKD dogs were staged according to the International Renal Interest Society staging system; all dogs were anesthetized and surgically prepared for blood flow studies. Data analyses were performed between sham control (CTR) and stage 1 and 2 CKD dogs. RESULTS: At baseline, myocardial blood flow and diastolic aortic pressure were similar for all groups. During intravenous dobutamine, myocardial blood flow was markedly higher than CTR even though hematocrit levels declined with the severity of CKD. In the CTR dogs, myocardial blood flow increased in direct relation to cardiac work. However, in the CKD dogs (stage 1 and 2), maximum blood flow was achieved with low-dose dobutamine, indicating that coronary autoregulation is more readily exhausted with minimal increases in cardiac work during CKD. CONCLUSION: We report that CKD markedly impairs coronary vascular reserve and myocardial blood flow regulation which could contribute to greater cardiovascular risk and poor clinical outcomes in CKD patients.

Influence of cardiac decentralization on cardioprotection.

The role of cardiac nerves on development of myocardial tissue injury after acute coronary occlusion remains controversial. We investigated whether acute cardiac decentralization (surgical) modulates coronary flow reserve and myocardial protection in preconditioned dogs subject to ischemia-reperfusion. Experiments were conducted on four groups of anesthetised, open-chest dogs (n = 32): 1- controls (CTR, intact cardiac nerves), 2- ischemic preconditioning (PC; 4 cycles of 5-min IR), 3- cardiac decentralization (CD) and 4- CD+PC; all dogs underwent 60-min coronary occlusion and 180-min reperfusion. Coronary blood flow and reactive hyperemic responses were assessed using a blood volume flow probe. Infarct size (tetrazolium staining) was related to anatomic area at risk and coronary collateral blood flow (microspheres) in the anatomic area at risk. Post-ischemic reactive hyperemia and repayment-to-debt ratio responses were significantly reduced for all experimental groups; however, arterial perfusion pressure was not affected. Infarct size was reduced in CD dogs (18.6 ± 4.3; p = 0.001, data are mean ± 1 SD) compared to 25.2 ± 5.5% in CTR dogs and was less in PC dogs as expected (13.5 ± 3.2 vs. 25.2 ± 5.5%; p = 0.001); after acute CD, PC protection was conserved (11.6 ± 3.4 vs. 18.6 ± 4.3%; p = 0.02). In conclusion, our findings provide strong evidence that myocardial protection against ischemic injury can be preserved independent of extrinsic cardiac nerve inputs.

Blood flow to the heart from noncoronary arteries: an intriguing but challenging research field.

INTRODUCTION: It has been proven that blood supply to the heart can include blood from noncoronary collateral circulation. Whether this network can somehow be augmented to provide an alternative therapy for ischemic patients is an intriguing hypothesis with no clear answer yet due to the challenging nature of this research field. In an attempt to enhance noncoronary collateral blood flow, we experimented with left internal thoracic artery ligature and angiogenic growth factors in a canine model. MATERIALS AND METHODS: Eight dogs weighing between 20 and 29 kg were scheduled for three operations each. Neutron-activated microspheres were used for blood flow measurement throughout the study. The first operation was a left thoracotomy for ameroid constrictor positioning at the proximal segment of the circumflex coronary artery. Three weeks later the left internal thoracic artery was ligatured distally. A micro-pump was positioned in the mediastinum, connected with a small catheter inserted in the proximal segment of the left internal thoracic artery for slow delivery of vascular endothelial growth factor. The protocol called for a coronary angiography 4 weeks later, sacrifice of the animals, and explantation of the heart for microsphere analysis. RESULTS: Four of the dogs failed to tolerate ameroid constrictor positioning by thoracotomy and died after the first operation. The remaining four dogs underwent a second operation to ligate their left internal thoracic arteries and implant micro-pumps with catheters. The implant was a success, but only one dog survived in the postoperative period. When the last dog was voluntarily sacrificed after a third operation, coronary angiography showed no neovascularization, nor did heart slice analysis reveal microspheres. CONCLUSIONS: Despite the technical challenge of building research protocols to exactly quantify blood from noncoronary arteries, there are many historical, anatomical, physiopathological, clinical, radiological, and surgical indications suggesting that blood flow from extracardiac structures may play an important role in ischemic heart disease. Our preliminary investigation combining internal thoracic artery occlusion and angiogenic growth factors was unsuccessful for several reasons, the main one being it was too heavy a protocol for the dogs. Despite this, we think that noncoronary collateral blood flow represents an intriguing research field worthy of debate and further study.

Role of the autonomic nervous system in cardioprotection by remote preconditioning in isoflurane-anaesthetized dogs.

AIMS: Remote ischaemic preconditioning (rIPC) protects cardiac and non-cardiac tissues against ischaemic injury. Although there is increased demand to investigate its potential clinical applicability, fundamental mechanisms responsible for rIPC-mediated protection remain unresolved. We examined in isoflurane-anaesthetized dogs whether an intact cardiac nervous system was necessary to mediate rIPC protection against ischaemic injury. METHODS AND RESULTS: Dogs were randomly allocated to six groups: 1, control (CON, no-rIPC); 2, rIPC (4 × 5 min renal artery occlusion/reperfusion); 3, autonomic ganglionic blockade with hexamethonium (HEX, no-rIPC; 20 mg/kg iv); 4, HEX + rIPC; 5, cardiac decentralization by surgical ablation of extracardiac nerves (DCN, no-rIPC); and 6, DCN + rIPC. All dogs underwent 60 min coronary occlusion and 180 min reperfusion; cardiac haemodynamic parameters were monitored. Regional blood flow (microspheres) in the heart and kidneys was assessed. Necrotic tissue was visualized using triphenyltetrazolium staining and related to anatomic risk zone size (area at risk; P = NS between groups) and coronary collateral blood flow. Infarct size (% AAR) was 29 ± 5 (mean ± 1 SD) in CON and 15 ± 4 in rIPC dogs (P = 0.001 vs. CON); 24 ± 3 in HEX vs. 12 ± 2 in HEX + rIPC (P = 0.001 vs. HEX); and 20 ± 2 in DCN vs. 12 ± 4 in DCN + rIPC (P = 0.001 vs. DCN). In CON dogs, infarct size was inversely related to coronary collateral flow; this relation was shifted downwards in all groups pre-treated with rIPC. CONCLUSION: We report robust myocardial protection by rIPC against ischaemic injury in canines that was not abrogated by either pharmacological or surgical decentralization of cardiac nerves.

Enhancement of noncoronary collateral circulation: the hypothesis of an alternative treatment for ischemic heart disease.

The internal thoracic arteries (ITAs) are a source of "noncoronary collateral circulation" (NCCC), or "noncoronary collateral blood flow" (NCCBF). The hypothesis herein is that enhancement of NCCC may represent an alternative means of myocardial blood supply: (1) Ligature of the ITAs creates a local hypertensive status and increases the perfusion pressure within the channels leading to the heart; (2) Myocardial ischemic stimulus diverts most ITA-related collateral flow to the heart rather than to the chest wall; (3) The ITAs may develop neo-collaterals owing to their ischemia-related plastic potential; (4) Angiogenic growth factor administration within the ITAs enhances neo-collateral development. These elements may pave the way for a new field of cardiovascular research aimed at enhancing NCCC as a new therapeutic option for ischemic heart disease.

Coronary vasoregulation in health and disease.

The present article reviews pertinent contributions from the Coronary Physiology Research Group at the Quebec Heart Institute to the understanding of coronary physiology in health and disease. Mechanisms that contribute to regulation of coronary blood flow and its distribution across the ventricular wall are discussed. Data from animal studies of ischemia-reperfusion injury are also presented and discussed in the context of current concepts regarding postischemic myocardial protection strategies. Future research directions regarding the cardiac nervous system and its importance in the regulation of coronary blood flow, cardiac function and myocyte injury during acute myocardial infarction are also discussed.

Long-term impact of diabetes and its comorbidities in patients undergoing isolated primary coronary artery bypass graft surgery.

BACKGROUND: The objective of this study was to identify the impact of diabetes and related comorbidities, namely chronic renal failure, peripheral vascular disease, and low ejection fraction (<35%), on long-term survival of patients undergoing coronary artery bypass graft surgery. METHODS AND RESULTS: A unicenter study was conducted on 9125 survivors of isolated coronary artery bypass graft surgery between 1992 and 2002. There were 6581 nondiabetic patients and 2544 diabetics, including 1809 patients with noninsulin-dependent diabetes mellitus and 735 patients with insulin-dependent diabetes mellitus. Cardiac-specific survival at 5 and 10 years was lower in insulin-dependent diabetes mellitus compared with both nondiabetic mellitus patients and patients with noninsulin-dependent diabetes mellitus (P<0.0001). However, freedom from cardiac-related death was similar for patients with noninsulin-dependent diabetes mellitus and nondiabetes mellitus patients up to 6 years (P=0 0.08) after surgery and was significantly lower thereafter (P=0.004). Cardiac-specific survival after coronary artery bypass graft surgery in patients with one or more comorbidities was comparable (P=0.4) for both nondiabetes mellitus patients and patients with noninsulin-dependent diabetes mellitus, but was significantly lower for those requiring insulin therapy (P<0.0001). Noninsulin-dependent diabetes mellitus was not an independent predictor of long-term cardiac death (hazard ratio: 1.09, P=0.41); however, insulin-dependent diabetes mellitus, chronic renal failure, peripheral vascular disease, and low ejection fraction were all independent risk factors for late cardiac death (all P<0.0001). The impact of comorbidities on the long-term risk of cardiac death was similar for the 3 groups. CONCLUSIONS: Noninsulin-dependent diabetes is not an independent predictor of late cardiac death after coronary artery bypass graft surgery, because cardiac-related survival is similar to that of nondiabetic patients for 6 years after surgery. In diabetic and nondiabetic patients, cardiac survival is adversely affected by the need for insulin therapy and/or the presence and number of comorbidities such as chronic renal failure, peripheral vascular disease, and low ejection fraction.

Influence of acute renal failure on coronary vasoregulation in dogs.

Impaired renal function is associated with an increased risk for cardiovascular events and death, but the pathophysiology is poorly defined. The hypothesis that coronary blood flow regulation and distribution of ventricular blood flow could be compromised during acute renal failure (ARF) was tested. In two separate groups (n = 14 each) of dogs with ARF, (1) coronary autoregulation (pressure-flow relations), vascular reserve (reactive hyperemia), and myocardial blood flow distribution (microspheres) and (2) coronary vessel responses to intracoronary infusion of select endothelium-dependent and -independent vasodilators were evaluated. In addition, coronary pressure-flow relations and vascular reserve after inhibition of nitric oxide and prostaglandin release were evaluated. Under resting conditions, myocardial oxygen consumption increased in dogs with ARF compared with no renal failure (NRF; 11.8 +/- 9.2 versus 5.0 +/- 1.5 ml O(2)/min per 100 g; P = 0.01), and the autoregulatory break point of the coronary pressure-flow relation was shifted to higher diastolic coronary pressures (60 +/- 17 versus 52 +/- 8 mmHg in NRF; P = 0.003); the latter was shifted further rightward after inhibition of both nitric oxide and prostaglandin release. The endocardial/epicardial blood flow ratio was comparable for both groups, suggesting preserved ventricular distribution of blood flow. In dogs with ARF, coronary vascular conductance also was reduced (P = 0.001 versus NRF), but coronary zero-flow pressure was unchanged. Vessel reactivity to each endothelium-dependent/independent compound also was blunted significantly. In conclusion, under resting conditions, coronary vascular tone, reserve, and vessel reactivity are markedly diminished with ARF, suggesting impaired vascular function. Consequently, during ARF, small increases in myocardial oxygen demand would induce subendocardial ischemia as a result of a limited capacity to increase oxygen supply and thereby contribute to higher risk for adverse coronary events and mortality.

Comparison of neutron activated and radiolabeled microsphere methods for measurement of transmural myocardial blood flow in dogs.

BACKGROUND: The 'gold standard' radioactive microsphere (RM) technique for measurement of organ blood flow under various experimental conditions is inaccessible to many researchers due to increasing environmental concerns regarding safety and disposal of low-level radioactive waste materials. A new method using neutron activated microspheres (NAM) has recently been described. METHODS: We compared regional myocardial blood flows using the new formulation STERIspheres (NAM; 15.0 +/- 0.1 [SD] microm; density 1.5 gr/mL) with RM (15.0 +/- 0.1 [SD] microm; density 1.5 gr/mL) under different experimental conditions during acute ischemia-reperfusion injury in dogs. Random paired combinations of four different RM and NAM were co-injected into the left atrium during autoregulation, coronary occlusion and flow-mediated hyperemia (reperfusion) in the same animal. The left ventricle was divided into non-ischemic and ischemic regions and further subdivided into endocardial, mid-myocardial and epicardial portions. After gamma-counting, blood and myocardial tissue samples (n = 180) were dried and then shipped to a core facility for neutron activation and analysis. NAM-RM blood flow data were directly compared by ANOVA and regression analysis; Bland and Altman analysis was also performed to assess mean differences in blood flow with NAM-RM. RESULTS: A direct relation for blood flow between NAM-RM was observed; the slope of the relation (1.17 RM +/- 0.04 [SEE]) was different from unity but the intercept (0.06 +/- 0.06 [SEE]) was not different from the origin. Intermethod mean differences were minimal between NAM-RM in the low to normal range of blood flow and were increased at the higher blood flow levels the latter being of minor physiological consequence. A direct relation for endo/epicardial blood flow ratios between NAM-RM was also observed; the slope of the relation (0.98 RM +/- 0.04 [SEE]) and the intercept (0.03 +/- 0.06 [SEE]) were not different from unity or the origin, respectively. CONCLUSIONS: RESULTS show that in addition to limiting production of radioactive waste materials, NAM accurately measure myocardial blood flow, endocardial/epicardial and ischemic/non-ischemic blood flow distributions over a wide range. We compared myocardial blood flows using paired combinations of neutron activated (NAM) and the 'gold standard' radiolabeled microspheres (RM) co-injected during autoregulation, coronary occlusion and flow-mediated hyperemia in an in situ canine ischemia-reperfusion preparation. A direct relation for blood flow and endo/epicardial blood flow ratios between NAM-RM was observed; intermethod mean differences between NAM-RM were minimal in the low to normal blood flow range but increased at higher blood flow levels. These results indicate that NAM accurately measure myocardial blood flow and its transmural distribution in addition to limiting unnecessary production of radioactive laboratory waste products.

Angiotensin inhibition and coronary autoregulation in a canine model of LV hypertrophy.

In humans with hypertension and LV hypertrophy, beneficial effects of angiotensin inhibition may be associated with preserved autoregulatory capacity. We studied the effect of acute angiotensin converting enzyme (ACE) inhibition on coronary autoregulatory pressure-flow relations and transmural distribution of blood flow in sham and LV hypertrophy dogs. Heart/body weight ratio increased (p = 0.001) from 5.5 +/- 0.7 in sham to 6.9 +/- 0.5 in LV hypertrophy dogs. The lower coronary pressure limit (LPL) on the pressure-flow relation was 47 +/- 2 mmHg in sham and 57 +/- 6 mmHg (p = 0.001) in LV hypertrophy dogs; after acute ACE-inhibition the LPL was reduced to 40 +/- 5 mmHg and 49 +/- 6 mmHg (p = 0.001), respectively. Transmural distribution of blood flow was preserved at the LPL in both groups before and after acute ACE-inhibition. Concomitant blockade of prostaglandin and nitric oxide release and bradykinin catabolism had no additional effects on the LPL and distribution of blood flow. After acute ACE-inhibition in LV hypertrophy dogs, distribution of blood flow across the LV wall was preserved and subendocardial vascular reserve was maintained even though the LPL was significantly lower. Preservation of autoregulatory capacity by ACE inhibitors contributes to beneficial outcome in patients with hypertension and LV hypertrophy.

Myocardial blood flow after chronic cardiac decentralization in anesthetized dogs: effects of ACE-inhibition.

Coronary blood flow regulation was studied in dogs with an intact or chronically decentralized intrinsic cardiac nervous system. We also examined the effect of angiotensin-converting enzyme inhibition (ACEI) on coronary autoregulatory pressure-flow relations and distribution of blood flow since the renin-angiotensin system may play a critical role in vasoregulation. Myocardial oxygen demand was reduced in the chronic decentralized dogs compared to the control dogs. The lower pressure limit of the autoregulatory pressure-flow relation was similar for the control and chronic decentralized dogs (47+/-2 and 44+/-7 mm Hg, respectively; p = NS). After ACEI, the lower pressure limit shifted leftward to 40 mm Hg (p=0.001) in both groups. Concomitant blockade of cyclooxygenase, bradykinin catabolism and nitric oxide synthase had no further effect on the lower pressure limit. Total myocardial blood flow was lower (p=0.001) in the chronic decentralized dogs compared to the control dogs, while transmural distribution of blood flow was preserved in both groups. The results show that even though myocardial oxygen requirements are lower in the chronically decentralized heart compared to controls, coronary autoregulation is maintained at levels observed in normally innervated hearts. The present findings indicate that intrinsic cardiac neurons contribute to coronary autoregulatory control and myocardial blood flow distribution even in the absence of cardiac connections to the central nervous system. In addition, in the chronic decentralized dog, ACEI allows the heart to work at lower coronary perfusion pressures while myocardial blood flow distribution is preserved.