Insulin increases glucose disposal into muscle. In addition, in vivo insulin elicits distinct nitric oxide synthase-dependent vascular responses to increase total skeletal muscle blood flow and to recruit muscle capillaries (by relaxing resistance and terminal arterioles, respectively). In the current study, we compared the temporal sequence of vascular and metabolic responses to a 30-min physiological infusion of insulin (3 mU ⅐ min ؊1 ⅐ kg ؊1 , euglycemic clamp) or saline in rat skeletal muscle in vivo. We used contrast-enhanced ultrasound to continuously quantify microvascular volume. Insulin recruited microvasculature within 5-10 min (P < 0.05), and this preceded both activation of insulin-signaling pathways and increases in glucose disposal in muscle, as well as changes in total leg blood flow. Moreover, L-NAME (N -nitro-L-arginine-methyl ester), a specific inhibitor of nitric oxide synthase, blocked this early microvascular recruitment (P < 0.05) and at least partially inhibited early increases in muscle glucose uptake (P < 0.05). We conclude that insulin rapidly recruits skeletal muscle capillaries in vivo by a nitric oxidedependent action, and the increase in capillary recruitment may contribute to the subsequent glucose uptake.

. Inhibiting NOS blocks microvascular recruitment and blunts muscle glucose uptake in response to insulin. Am J Physiol Endocrinol Metab 285: E123-E129, 2003; 10.1152/ajpendo.00021.2003.-We examined the effects of inhibiting nitric oxide synthase with N -nitro-L-arginine-methyl ester (L-NAME) on total hindlimb blood flow, muscle microvascular recruitment, and hindlimb glucose uptake during euglycemic hyperinsulinemia in vivo in the rat. We used two independent methods to measure microvascular perfusion. In one group of animals, microvascular recruitment was measured using the metabolism of exogenously infused 1-methylxanthine (1-MX), and in a second group contrast-enhanced ultrasound (CEU) was used. Limb glucose uptake was measured by arterial-venous concentration differences after 2 h of insulin infusion. Saline alone did not alter femoral artery flow, glucose uptake, or 1-MX metabolism. Insulin (10 mU ⅐ min Ϫ1 ⅐ kg Ϫ1 ) significantly increased hindlimb total blood flow (0.69 Ϯ 0.02 to 1.22 Ϯ 0.11 ml/min, P Ͻ 0.05), glucose uptake (0.27 Ϯ 0.05 to 0.95 Ϯ 0.08 mol/min, P Ͻ 0.05), 1-MX uptake (5.0 Ϯ 0.5 to 8.5 Ϯ 1.0 nmol/min, P Ͻ 0.05), and skeletal muscle microvascular volume measured by CEU (10.0 Ϯ 1.6 to 15.0 Ϯ 1.2 video intensity units, P Ͻ 0.05). Addition of L-NAME to insulin completely blocked the effect of insulin on both total limb flow and microvascular recruitment (measured using either 1-MX or CEU) and blunted glucose uptake by 40% (P Ͻ 0.05). We conclude that insulin specifically recruits flow to the microvasculture in skeletal muscle via a nitric oxide-dependent pathway and that this may be important to insulin's overall action to regulate glucose disposal. capillary recruitment; nitric oxide; nitric oxide synthase; muscle blood flow THERE IS ABUNDANT EVIDENCE that insulin augments total limb blood flow in humans (2, 29, 33) and experimental animals (20, 22) in a time-and dose-dependent fashion. It has been suggested that this action of insulin could, by facilitating the delivery of glucose and itself to muscle, contribute to insulin's overall action on glucose disposal (3), although this remains controversial (33).Substantial evidence suggests that nitric oxide (NO) is involved in insulin's action to increase limb blood flow in humans (5,8,25,26,29). NO in muscle is produced by nitric oxide synthase (NOS), located in both vascular endothelium (28) and myocytes (16). Inhibition of NO production by N -mono-methyl-L-arginine (L-NMMA) can fully abolish the effect of insulin to increase limb total blood flow in humans (4,5,26,29). In one study, this agent partially blocked (ϳ25%) insulin-mediated glucose uptake as well (5). Baron et al. (6) have also reported that, in the rat, L-NMMA increases mean arterial pressure and reduces whole body glucose infusion rate in a dose-dependent fashion during a euglycemic insulin clamp (12 mU⅐min Ϫ1 ⅐kg Ϫ1). Using intravital microscopy, Chen and Messina (8) demonstrated that insulin induces vasodilation of firstorder arterioles in rat cremaster and that the addition...

The vascular system controls the delivery of nutrients and hormones to muscle, and a number of hormones may act to regulate muscle metabolism and contractile performance by modulating blood flow to and within muscle. This review examines evidence that insulin has major hemodynamic effects to influence muscle metabolism. Whole body, isolated hindlimb perfusion studies and experiments with cell cultures suggest that the hemodynamic effects of insulin emanate from the vasculature itself and involve nitric oxide-dependent vasodilation at large and small vessels with the purpose of increasing access for insulin and nutrients to the interstitium and muscle cells. Recently developed techniques for detecting changes in microvascular flow, specifically capillary recruitment in muscle, indicate this to be a key site for early insulin action at physiological levels in rats and humans. In the absence of increases in bulk flow to muscle, insulin may act to switch flow from nonnutritive to the nutritive route. In addition, there is accumulating evidence to suggest that insulin resistance of muscle in vivo in terms of impaired glucose uptake could be partly due to impaired insulin-mediated capillary recruitment. Exercise training improves insulin-mediated capillary recruitment and glucose uptake by muscle.

We have previously shown that skeletal muscle capillaries are rapidly recruited by physiological doses of insulin in both humans and animals. This facilitates glucose and insulin delivery to muscle, thus augmenting glucose uptake. In obese rats, both insulin-mediated microvascular recruitment and glucose uptake are diminished; however, this action of insulin has not been studied in obese humans. Here we used contrast ultrasound to measure microvascular blood volume (MBV) (an index of microvascular recruitment) in the forearm flexor muscles of lean and obese adults before and after a 120-min euglycemic-hyperinsulinemic (1 mU ⅐ min ؊1 ⅐ kg ؊1 ) clamp. We also measured brachial artery flow, fasting lipid profile, and anthropomorphic variables. Fasting plasma glucose (5.4 ؎ 0.1 vs. 5.1 ؎ 0.1 mmol/l, P ‫؍‬ 0.05), insulin (79 ؎ 11 vs. 38 ؎ 6 pmol/l, P ‫؍‬ 0.003), and percent body fat (44 ؎ 2 vs. 25 ؎ 2%, P ‫؍‬ 0.001) were higher in the obese than the lean adults. After 2 h of insulin infusion, whole-body glucose infusion rate was significantly lower in the obese versus lean group (19.3 ؎ 3.2 and 37.4 ؎ 2.6 mol ⅐ min ؊1 ⅐ kg ؊1 respectively, P < 0.001). Compared with baseline, insulin increased MBV in the lean (18.7 ؎ 3.3 to 25.0 ؎ 4.1, P ‫؍‬ 0.019) but not in the obese group (20.4 ؎ 3.6 to 18.8 ؎ 3.8, NS). Insulin increased brachial artery diameter and flow in the lean but not in the obese group. We observed a significant, negative correlation between ⌬MBV and BMI (R ‫؍‬ ؊0.482, P ‫؍‬ 0.027) in response to insulin. In conclusion, obesity eliminated the insulin-stimulated muscle microvascular recruitment and increased brachial artery blood flow seen in lean individuals.

Intense exercise and insulin each increases total limb blood flow and recruits muscle capillaries, presumably to facilitate nutrient exchange. Whether mixed meals or light exercise likewise recruits capillaries is unknown. We fed 18 (9 M, 9 F) healthy volunteers a 480-kcal liquid mixed meal. Plasma glucose, insulin, brachial artery flow, and forearm muscle microvascular blood volume were measured before and after the meal. Brachial artery flow and microvascular volume were also examined with light (25% max), moderate (50%), and heavy (80%) forearm contraction every 20 s in 5 (4 M, 1 F) healthy adults. After the meal, glucose and insulin rose modestly (to ϳ7 mM and ϳ270 pM) and peaked by 30 min, whereas brachial artery blood flow (P Ͻ 0.05) and the microvascular volume (P Ͻ 0.01) each increased significantly by 60 min, and microvascular flow velocity did not change. For exercise, both 50 and 80%, but not 25% maximal handgrip, increased average forearm and brachial artery blood flow (P Ͻ 0.01). Flow increased immediately after each contraction and declined toward basal over 15 s. Exercise at 25% max increased microvascular volume threefold (P Ͻ 0.01) without affecting microvascular flow velocity or total forearm blood flow. Forearm exercise at 80% maximal grip increased both microvascular volume and microvascular flow velocity (P Ͻ 0.05 each). We conclude that light exercise and simple meals each markedly increases muscle microvascular volume, thereby expanding the endothelial surface for nutrient exchange, and that capillary recruitment is an important physiological response to facilitate nutrient/hormone delivery in healthy humans. INSULIN (1, 7, 18) AND EXERCISE (8, 13) each increase total limb blood flow, and each recruits flow to muscle capillaries (13,18). These vascular responses facilitate the delivery of oxygen, insulin, and glucose to muscle cells. Flow to individual capillary networks is controlled by small, ϳ20 m diameter terminal arterioles (21). We previously showed that during insulin infusion, relaxation of larger-resistance arterioles (which contribute to the regulation of total blood flow) and terminal arterioles can occur independently in rat (25, 29) and human (7) muscle.Muscle microvascular responses to physiological stimuli have been difficult to assess in humans due to the lack of noninvasive techniques suitable for assessing microvascular blood volume. We have developed two independent techniques for determining muscle microvascular volume noninvasively.The first technique is based on the metabolism of exogenously infused 1-methylxanthine (1-MX) to 1-methylurate by capillary endothelial xanthine oxidase (18,29). This has worked well in rodent models but has not been adapted to human studies. The second technique, contrast-enhanced ultrasound (CEU), relies on the contrast introduced by intravenously infused microbubbles (27). To measure microvascular volume, CEU takes advantage of the fact that during their continuous infusion it is possible to disrupt microbubbles by a single pulse of high...

Supraphysiological doses of insulin enhance total limb blood flow and recruit capillaries in skeletal muscle. Whether these processes change in response to physiological hyperinsulinemia is uncertain. To examine this, we infused either saline (n ‫؍‬ 6) or insulin (euglycemic clamp, 3.0 mU ⅐ min ؊1 ⅐ kg ؊1 , n ‫؍‬ 9) into anesthetized rats for 120 min.

. Vascular recruitment in skeletal muscle during exercise and hyperinsulinemia assessed by contrast ultrasound. Am J Physiol Endocrinol Metab 282: E714-E720, 2002. First published November 13; 10.1152/ ajpendo.00373.2001.-The purpose of this study was to noninvasively quantify the effects of insulin on capillary blood volume (capBV) and RBC velocity (V RBC) in skeletal muscle in vivo with the use of contrast-enhanced ultrasound. We performed contrast ultrasound of the rat hindlimb adductor muscles at baseline and after 2-h infusions of either insulin (3 or 40 mU ⅐ kg Ϫ1 ⅐ min Ϫ1 ) or saline. Saline-treated animals were also studied during contractile exercise. V RBC and capBV were calculated from the relation between pulsing interval and video intensity. Femoral artery blood flow, measured by a flow probe, increased with both contractile exercise and insulin. Contractile exercise increased capBV more than twofold and V RBC fivefold. Insulin also increased capBV more than twofold in a dose-dependent fashion but did not significantly alter V RBC. Saline infusion did not significantly alter capBV, V RBC, or femoral artery blood flow. We conclude that physiological changes in skeletal muscle capillary perfusion can be assessed in vivo with the use of contrastenhanced ultrasound. Exercise increases both V RBC and capBV, whereas hyperinsulinemia selectively increases only capBV, which may enhance skeletal muscle glucose uptake. blood volume; capillaries; insulin THE NUMBER OF PERFUSED CAPILLARIES (N cap ) in skeletal muscle can change in response to various physiological stimuli. Increases in N cap during skeletal muscle exercise are thought to enhance capillary surface area for oxygen exchange and to decrease oxygen diffusion distance (4, 6, 7). Similarly, it has been proposed that insulin increases N cap , thereby enhancing glucose delivery to skeletal muscle by augmenting surface area for diffusion. Experimental evidence for insulin-mediated changes in microvascular perfusion in humans has been based largely on increases in limb blood flow and skeletal muscle blood volume that occur in response to hyperinsulinemia (1,3,12,14,17). The extent of capillary recruitment in skeletal muscle remains uncertain because previous studies have used techniques that provide a measure of total blood volume, most of which resides in larger conduit and resistance vessels (16). Hence, capillary responses to various physiological stimuli have been difficult to assess in intact skeletal muscle due to a paucity of noninvasive methods capable of quantifying capillary blood volume (capBV).Recently, changes in myocardial and renal capBV have been measured with the use of contrast-enhanced ultrasound imaging (2,22,23). This technique also provides information on microbubble velocity within capillaries by measuring their reappearance rate in tissue after their destruction with acoustic energy (2,22). Because the microvascular rheology of microbubbles is similar to that of red blood cells (8), this method can be used to study microvascular re...

Insulin has important vascular actions that regulate blood flow, in addition to its classical actions to coordinate glucose homeostasis. Insulin-stimulated production of nitric oxide in vascular endothelium results in capillary recruitment and vasodilation that diverts and increases blood flow to skeletal muscle and consequently increases glucose disposal. Thus, vascular actions of insulin may be essential for coupling hemodynamic and metabolic homeostasis. A complete biochemical signaling pathway linking the insulin receptor to activation of endothelial nitric oxide synthase in vascular endothelium has recently been elucidated. Moreover, the time course and dose response for capillary recruitment in response to physiologic concentrations of insulin parallels that of insulin-mediated glucose uptake in vivo. Taken together, these observations suggest a molecular mechanism that may help to explain how insulin resistance contributes to cardiovascular components of the metabolic syndrome and vascular complications of diabetes.