Steven K. Nishiyama

Ultrasound Assessment of Flow-Mediated Dilation

Developed in 1992, the flow-mediated dilation test is now the most commonly used noninvasive assessment of vascular endothelial function in humans. Since its inception, scientists have refined their understanding of the physiology, analysis, and interpretation of this measurement. Recently, a significant growth of knowledge has added to our understanding and implementation of this clinically relevant research methodology. Therefore, this tutorial provides timely insight into recent advances and practical information related to the ultrasonic assessment of vascular endothelial function in humans.

Acute Reversal of Endothelial Dysfunction in the Elderly After Antioxidant Consumption

Aging is associated with a pro-oxidant state and a decline in endothelial function. Whether acute, enteral antioxidant treatment can reverse this decrement in vascular function is not well known. Flow-mediated vasodilation and reactive hyperemia were evaluated after consumption of either placebo or an oral antioxidant cocktail (vitamin C, 1000 mg; vitamin E, 600 IU; &agr;-lipoic acid, 600 mg) in 87 healthy volunteers (42 young: 25±1 years; 45 older: 71±1 years) using a double-blind, crossover design. Blood velocity and brachial artery diameter (ultrasound Doppler) were assessed before and after 5-minute forearm circulatory arrest. Serum markers of lipid peroxidation, total antioxidant capacity, endogenous antioxidant activity, and vitamin C were assayed, and plasma nitrate, nitrite, and 3-nitrotyrosine were determined. In the placebo trial, an age-related reduction in brachial artery vasodilation was evident (young: 7.4±0.6%; older: 5.2±0.4%). After antioxidant consumption, flow-mediated vasodilation improved in older subjects (placebo: 5.2±0.4%; antioxidant: 8.2±0.6%) but declined in the young (placebo: 7.4±0.6%; antioxidant: 5.8±0.6%). Reactive hyperemia was reduced with age, but antioxidant administration did not alter the response in either group. Together, these data demonstrate that antioxidant consumption acutely restores endothelial function in the elderly while disrupting normal endothelium-dependent vasodilation in the young and suggest that this age-related impairment is attributed, at least in part, to free radicals.

Antioxidants and aging: NMR-based evidence of improved skeletal muscle perfusion and energetics

We sought to examine the potential role of oxidative stress on skeletal muscle function with advancing age. Nuclear magnetic resonance (NMR) was employed to simultaneously assess muscle perfusion (arterial spin labeling) and energetics ((31)P NMR spectroscopy) in the lower leg of young (26 + or - 5 yr, n = 6) and older (70 + or - 5 yr, n = 6) healthy volunteers following the consumption of either placebo (PL) or an oral antioxidant (AO) cocktail (vitamins C and E and alpha-lipoic acid), previously documented to decrease plasma free radical concentration. NMR measurements were made during and after 5 min of moderate intensity (approximately 5 W) plantar flexion exercise. AO administration significantly improved end-exercise perfusion (AO, 50 + or - 5, and PL, 43 + or - 4 ml x 100 g(-1) x min(-1)) and postexercise perfusion area under the curve (AO, 1,286 + or - 236, and PL, 866 + or - 144 ml/100 g) in older subjects, whereas AO administration did not alter hemodynamics in the young group. Concomitantly, muscle oxidative capacity (time constant of phosphocreatine recovery, tau) was improved following AO in the older (AO, 43 + or - 1, and PL, 51 + or - 7 s) but not the young (AO, 54 + or - 5, and PL, 48 + or - 7 s) group. These findings support the concept that oxidative stress may be partially responsible for the age-related decline in skeletal muscle perfusion during physical activity and reveal a muscle metabolic reserve capacity in the elderly that is accessible under conditions of improved perfusion.

Angiotensin II in the Elderly: Impact of Angiotensin II Type 1 Receptor Sensitivity on Peripheral Hemodynamics

Exercise hyperemia is attenuated in the elderly, which may be attributed to local vasoregulatory pathways within the skeletal muscle vasculature. Therefore, we sought to determine whether healthy aging is associated with changes in angiotensin II (Ang II) receptor sensitivity through measurements of leg blood flow in resting and exercising skeletal muscle. In 12 (n=6 young, 24±1 years; n=6 older, 68±3 years) healthy volunteers, we determined changes in leg blood flow (ultrasound Doppler) before and during intra-arterial infusion of Ang II (0.8 ng/mL of leg blood flow per minute). Heart rate, arterial blood pressure, common femoral artery diameter, and mean blood velocity were measured at rest and during knee-extensor exercise at 20% and 40% of the maximal work rate (WRmax). At rest, Ang II infusion decreased leg blood flow to a greater extent in older (−61±8%) subjects compared with younger subjects (−31±5%). Compared with rest, Ang II–mediated vasoconstriction (leg blood flow) during exercise was diminished in both older and younger subjects at 20% (older: −7±5%; younger: −21±2%) and 40% WRmax (older: −5±4%; younger: −9±3%). These data identify a clear age-related hypersensitivity to Ang II in the resting leg, which may contribute to the recognized decrement in leg blood flow in this cohort. However, the diminished vasoconstriction to Ang II during exercise suggests that the elevation in Ang II type 1 receptor sensitivity documented at rest does not contribute significantly to the blunted exercise hyperemia experienced with advancing age.

The paradox of oxidative stress and exercise with advancing age.

Aging, vascular function, and exercise are thought to have a common link in oxidative stress. Both antioxidant supplementation and exercise training have been identified as interventions that may reduce oxidative stress, but their interaction in older humans is not well understood.

Vascular function and endothelin-1: tipping the balance between vasodilation and vasoconstriction

Endothelin-1 (ET-1), a potent vasoconstrictor secreted by vascular endothelial cells, has been implicated in the pathophysiology of numerous cardiovascular diseases, yet the direct impact of ET-1 on vascular function remains unclear. Therefore, in seven young (23 ± 1 yr) healthy subjects, we investigated the effect of an intra-arterial infusion of ET-1 on reactive hyperemia (RH) and flow-mediated dilation (FMD) in the popliteal artery following 5 min of suprasystolic cuff occlusion. ET-1 infusion significantly attenuated basal leg blood flow (control: 62 ± 4 ml/min, ET-1: 47 ± 9 ml/min), RH [area-under-curve (AUC); control: 162 ± 15 ml, ET-1: 104 ± 16 ml], and peak RH (control: 572 ± 51 ml/min, ET-1: 412 ± 32 ml/min) (P < 0.05). Administration of ET-1 also reduced FMD (control: 2.4 ± 0.3%, ET-1: 0.5 ± 0.5%) and FMD normalized for shear rate (control: 10.5 × 10-4 ± 2.0 × 10-4%/s-1, ET-1: 0.9 × 10-4 ± 2.8 ×10-4%/s-1). These findings reveal that elevated levels of ET-1 have a significant impact on vascular function, indicating that studies employing RH and FMD as markers of microvascular function and nitric oxide bioavailability, respectively, should exercise caution, as ET-1 can impact these assessments by tipping the balance between vasodilation and vasoconstriction, in favor of the latter.NEW & NOTEWORTHY Endothelin-1 (ET-1) is recognized as the bodys most potent endogenous vasoconstrictor, but the impact of this peptide on vascular function is not well understood. The present study revealed that the intra-arterial administration of ET-1 impaired both microvascular and conduit vessel function of the leg in young, healthy, humans. Studies employing vascular testing in patient cohorts that experience a disease-related increase in ET-1 should thus exercise caution, as ET-1 clearly impairs vascular function.

Response to Antioxidants and Endothelial Dysfunction in Young and Elderly People: Is Flow-Mediated Dilation Useful to Assess Acute Effects?

Recently, our group published a study in Hypertension that used flow-mediated vasodilation (FMD) and reactive hyperemia to assess the impact of an enteral antioxidant (AO) mixture, of known efficacy, on the endothelial dysfunction associated with aging.1 In a Letter to the Editor, Tsikas et al2 expressed some concern regarding the divergent FMD and assay results between the young and old subjects in our study and also challenged the informational value of FMD. Regarding the results of the assay-based assessments of NO and oxidative stress performed in our study, it should be noted that AO consumption did not actually alter biomarkers of oxidative stress or NO in the young (Table 2 in Reference 1), and, thus, the statement that our findings “show even increases in oxidative stress and decreases in [FMD] in young people” is, in fact, incorrect. In addition, previous studies from our group using electron paramagnetic …