Insulin-stimulated Glucose Transport In Skeletal Muscle Research Articles

Improvements of insulin resistance in ovariectomized rats by a novel phytoestrogen from Curcuma comosa Roxb

BackgroundCurcuma comosa Roxb. (C. comosa) is an indigenous medicinal herb that has been used in Thailand as a dietary supplement to relieve postmenopausal symptoms. Recently, a novel phytoestrogen, (3R)-1,7-diphenyl-(4E,6E)-4,6-heptadien-3-ol or compound 049, has been isolated and no study thus far has investigated the role of C. comosa in preventing metabolic alterations occurring in estrogen-deprived state. The present study investigated the long-term effects (12 weeks) of C. comosa hexane extract and compound 049 on insulin resistance in prolonged estrogen-deprived rats.MethodsFemale Sprague-Dawley rats were ovariectomized (OVX) and treated with C. comosa hexane extract (125 mg, 250 mg, or 500 mg/kg body weight (BW)) and compound 049 (50 mg/kg BW) intraperitoneally three times per week for 12 weeks. Body weight, food intake, visceral fat weight, uterine weight, serum lipid profile, glucose tolerance, insulin action on skeletal muscle glucose transport activity, and GLUT-4 protein expression were determined.ResultsProlonged ovariectomy resulted in dyslipidemia, impaired glucose tolerance and insulin-stimulated skeletal muscle glucose transport, as compared to SHAM. Treatment with C. comosa hexane extract and compound 049, three times per week for 12 weeks, markedly reduced serum total cholesterol and low-density lipoprotein levels, improved insulin sensitivity and partially restored uterine weights in ovariectomized rats. In addition, compound 049 or high doses of C. comosa hexane extract enhanced insulin-mediated glucose uptake in skeletal muscle and increased muscle GLUT-4 protein levels.ConclusionsTreatment with C. comosa and its diarylheptanoid derivative improved glucose and lipid metabolism in estrogen-deprived rats, supporting the traditional use of this natural phytoestrogen as a strategy for relieving insulin resistance and its related metabolic defects in postmenopausal women.

Rapid Reversal of Insulin-Stimulated AS160 Phosphorylation in Rat Skeletal Muscle after Insulin Exposure

Increased phosphorylation of Akt substrate of 160 kDa (AS160) is essential to trigger the full increase in insulin-stimulated glucose transport in skeletal muscle. The primary aim of this study was to characterize the time course for reversal of insulin-stimulated AS160 phosphorylation in rat skeletal muscle after insulin removal. The time courses for reversal of insulin effects both upstream (Akt phosphorylation) and downstream (glucose uptake) of AS160 were also determined. Epitrochlearis muscles were incubated in vitro using three protocols which differed with regard to insulin exposure: no insulin (never exposed to insulin), transient insulin (30 min with 1.8 nmol/l insulin, then incubation without insulin for 10, 20 or 40 min), or sustained insulin (continuously incubated with 1.8 nmol/l insulin). After removal of muscles from insulin, Akt and AS160 phosphorylation reversed rapidly, each with a half-time of <10 min and essentially full reversal by 20 min. Glucose uptake reversed more slowly (half time between 10 and 20 min with essentially full reversal by 40 min). Removal of muscles from insulin resulted in a rapid reversal of the increase in AS160 phosphorylation which preceded the reversal of the increase in glucose uptake, consistent with AS160 phosphorylation being essential for maintenance of insulin-stimulated glucose uptake.

Modulation of insulin resistance and hypertension by voluntary exercise training in the TG(mREN2)27 rat.

Hypertension is often accompanied by insulin resistance of skeletal muscle glucose transport. The male heterozygous TG(mREN2)27 rat, which harbors a mouse transgene for renin, displays local elevations in the renin-angiotensin system and exhibits markedly elevated systolic blood pressure (SBP). The present study was undertaken to characterize insulin-stimulated skeletal muscle glucose transport in male heterozygous TG(mREN2)27 rats and to evaluate the effect of voluntary exercise training on SBP and skeletal muscle glucose transport. Compared with normotensive Sprague-Dawley rats, TG(mREN2)27 rats displayed a 53% elevation (P < 0.05) in SBP, a twofold increase in plasma free fatty acid levels, and an exaggerated insulin response during an oral glucose tolerance test. Moreover, insulin-mediated glucose transport (2-deoxyglucose uptake) in isolated epitrochlearis and soleus muscles of TG(mREN2)27 animals was 33 and 43% less, respectively, than in Sprague-Dawley controls. TG(mREN2)27 rats ran voluntarily for 6 wk and achieved daily running distances of 6-7 km over the final 3 wk. Training caused a 36% increase in peak aerobic capacity and a 16% reduction in resting SBP. Fasting plasma insulin (21%) and free fatty acid (34%) levels were reduced in the trained TG(mREN2)27 rats. Whole body glucose tolerance was improved in the trained TG(mREN2)27 rats and was associated with increases of 39 and 50% in insulin-mediated glucose transport in epitrochlearis and soleus muscles, respectively. Whole muscle GLUT-4 protein was increased in the soleus (23%), but not in the epitrochlearis, of trained TG(mREN2)27 rats. These data indicate that the male heterozygous TG(mREN2)27 rat is a model of both hypertension and insulin resistance. Importantly, both of these defects can be beneficially modified by voluntary exercise training.

Impaired insulin-receptor autophosphorylation is an early defect in fat-fed, insulin-resistant rats.

High-fat feeding results in impaired insulin signaling in skeletal muscle, but the role of the insulin receptor (IR) remains controversial. In the present study, female Fischer 344 rats were fed diets either low in fat [low fat, complex carbohydrate (LFCC)] or high in fat and sucrose (HFS). Insulin-stimulated skeletal muscle glucose transport, measured in purified sarcolemmal vesicles, was lower in rats consuming the HFS diet for 2 and 8 wk compared with LFCC controls (72.9 +/- 3.5, 67.6 +/- 3.5, and 86.1 +/- 3.5 pmol x mg(-1) x 15 s(-1), respectively; P < 0.05). Muscle IR content was unchanged in 2-wk HFS animals but was 50% lower in the 8-wk HFS group (P < 0.001). However, compared with LFCC, insulin-stimulated IR autophosphorylation was 26% lower in 2-wk HFS and 40% lower in 8-wk HFS animals (P < 0.005). Total muscle content of the proposed IR inhibitors cytokine tumor necrosis factor-alpha and membrane glycoprotein PC-1 was not significantly changed in HFS animals at either 2 or 8 wk. These results demonstrate that high-fat feeding induces insulin resistance in muscle concomitant with a diminished IR signaling capacity, although the mechanism remains unknown.

LEPTIN TREATMENT INCREASES GLUT4 BUT NOT GLUT1 PROTEIN CONCENTRATION IN INSULIN-RESISTANT MUSCLE

We have recently shown chronic leptin administration to improve insulin-stimulated skeletal muscle glucose transport in insulin-resistant skeletal muscle (Yaspelkis et al., AJP, in press), but did not address if basal glucose transport was affected. Therefore, the aim of this investigation was to determine if non-insulin stimulated (basal) glucose transport is improved in insulin-resistant skeletal muscle in response to chronic leptin administration. Twenty-four male Sprague Dawley rats were divided into one of two groups: 1) Normal diet (n = 8) or 2) High-Fat diet (n = 16). Normal diet (CON) animals received standard rat chow for 12 weeks while the other animals were provided a high fat diet to induce skeletal muscle insulin-resistance. Twelve days prior to hind limb perfusions, the high fat diet animals were further subdivided: High Fat (HF) (n = 8) or High Fat-Leptin (HF-LEP) (n = 8). During this 12 d period, HF-LEP animals were injected with leptin (5 mg leptin/kg) s.c. at 0800 and 1800 h daily while the CON and HF animals received vehicle (PBS). All animals were prepared for and subjected to bind limb perfusion following the 12 d treatment period to determine rates of basal skeletal muscle 3-O-methyl-D-glucose (3-MG) transport. Leptin administration did not alter rates of 3-MG transport under basal conditions in CON, HF-LEP or HF groups. Although total and plasma membrane (PM) GLUT4 protein concentration were significantly (p < 0.05) reduced in the skeletal muscle of the HF animals, this did not affect rates of basal glucose transport. It appeared that rates of 3-MG transport were unaffected due to the basal glucose transporter protein concentration, GLUT1, being similar among groups. Of interest, however, was that 12d of leptin treatment normalized both total and PM GLUT4 protein concentration in the skeletal muscle of the HF-LEP animals. The results of this investigation indicate that while a high fat diet and subsequent leptin treatment alter expression of the insulin-responsive glucose transporter (GLUT4) in insulin-resistant skeletal muscle, no effect is observed on non-insulin-stimulated glucose transport or GLUT1 protein concentration in response to a high fat diet or leptin administration. This suggests the expression of these two glucose transporter isoforms may be differentially regulated. Supported by NIH GM 48680

The β 2-Adrenergic modulator celiprolol reduces insulin resistance in obese zucker rats

Essential hypertension is associated with an increased incidence of insulin resistance of skeletal muscle glucose transport. The present study determined if celiprolol, an antihypertensive agent with selective β 1-adrenoceptor antagonist and additional β 2-agonistic properties, administered by gavageeither acutely (3 hr) or chronically (14 d), had a direct effect on improving glucose tolerance and insulin-stimulated glucose transport activity (using 2-deoxyglucose (2-DG) uptake) in isolated epitrochlearis muscles of the insulin-resistant obese Zucker rat. The effects of a selective β 1-blocker, metoprolol, were also assessed. Acute administration of celiprolol, but not metoprolol, increased insulin-stimulated 2-DG uptake in muscle by 22% (p < 0.05). Chronic celiprolol treatment significantly lowered fasting plasma insulin (22%) and free fatty acids (40%) in comparison to obese control values. Moreover, chronic celiprolol administration decreased the glucose-insulin index (calculated as the product of the glucose and insulin areas under the curve during an oral glucose tolerance test), by 32% (p < 0.05) compared to obese controls, indicating that peripheral insulin action was increased. Indeed, insulin-stimulated skeletal muscle 2-DG uptake was enhanced by 49% (p < 0.05) in these celiprolol-treated obese animals. Metoprolol was without significant effect on any of these variables following chronic administration. These findings indicate that, in this animal model of insulin resistance, the β 1-antagonist β 2-agonist celiprolol has a specific effect of improving insulin-stimulated skeletal muscle glucose transport that is independent of any hemodynamic alterations.

Effect of chronic bradykinin administration on insulin action in an animal model of insulin resistance.

The nonapeptide bradykinin (BK) has been implicated as the mediator of the beneficial effect of angiotensin-converting enzyme inhibitors on insulin-stimulated glucose transport in insulin-resistant skeletal muscle. In the present study, the effects of chronic in vivo BK treatment of obese Zucker (fa/fa) rats, a model of glucose intolerance and severe insulin resistance, on whole body glucose tolerance and skeletal muscle glucose transport activity stimulated by insulin or contractions were investigated. BK was administered subcutaneously (twice daily at 40 microg/kg body wt) for 14 consecutive days. Compared with a saline-treated obese group, the BK-treated obese animals had significantly (P < 0.05) lower fasting plasma levels of insulin (20%) and free fatty acids (26%), whereas plasma glucose was not different. During a 1 g/kg body wt oral glucose tolerance test, the glucose and insulin responses [incremental areas under the curve (AUC)] were 21 and 29% lower, respectively, in the BK-treated obese group. The glucose-insulin index, the product of the glucose and insulin AUCs and an indirect index of in vivo insulin action, was 52% lower in the BK-treated obese group compared with the obese control group. Moreover, 2-deoxyglucose uptake in the isolated epitrochlearis muscle stimulated by a maximally effective dose of insulin (2 mU/ml) was 52% greater in the BK-treated obese group. Contraction-stimulated (10 tetani) 2-deoxyglucose uptake was also enhanced by 35% as a result of the BK treatment. In conclusion, these findings indicate that in the severely insulin-resistant obese Zucker rat, chronic in vivo treatment with BK can significantly improve whole body glucose tolerance, possibly as a result of the enhanced insulin-stimulated skeletal muscle glucose transport activity observed in these animals.

Diet-induced insulin resistance precedes other aspects of the metabolic syndrome.

This study was designed to examine the effects of a high-fat refined-sugar (HFS) or a low-fat complex-carbohydrate (LFCC) diet on insulin-stimulated skeletal muscle glucose transport, plasma insulin, blood pressure, plasma triglycerides, plasma glycerol, body weight, and body fat in female Fischer rats. Insulin-stimulated glucose transport was significantly reduced in the HFS group at 2 wk, 2 mo, and 2 yr, whereas serum insulin was significantly elevated at all time points. Blood pressure was not significantly elevated in the HFS group until 12 mo, and all HFS animals were hypertensive by 18 mo. Glycerol, triglycerides, and abdominal fat cell size were not significantly different at 2 wk but were significantly elevated in the HFS rats at 2 and 6 mo. Body weight was similar in both groups until 20 wk on the diet, when the HFS rats started to gain more weight. These results demonstrate that insulin resistance and hyperinsulinemia occur before the other manifestations of the metabolic syndrome and that diet, not obesity, is the underlying cause.

Dexamethasone inhibits insulin-stimulated recruitment of GLUt4 to the cell surface in rat skeletal muscle

To test the hypothesis that glucocorticoids reduce insulin-stimulated skeletal muscle glucose transport by inhibiting the recruitment of GLUT4 glucose transporters to the cell surface, we determined the effect of glucocorticoid treatment on cell-surface GLUT4 using the impermeant glucose transporter photolabel, 2- N-4-(1-azi-2,2,2-trifluoroethyl)benzoyl-[2- 3 H]1,3-bis-( d-mannos-4-yloxy)-2-propylamine (ATB-[2- 3H]BMPA), and GLUT4 immunoprecipitation. Male Sprague-Dawley rats were treated with dexamethasone ([Dex] 0.9 mg/kg for 2 days) and compared against pair-fed controls. 2-[ 3H]deoxyglucose (2-[ 3H]DG) uptake in isolated soleus muscles was measured under conditions in which uptake reflects glucose transport activity. In control muscles, 2-[ 3H]DG uptake was stimulated eightfold by insulin (20 nmol/L). Dex treatement reduced maximal insulin-stimulated 2-[ 3H]DG uptake by 48% ± 4% (mean ± SEM) and decreased cell-surface (ATB-[2- 3H]BMPA—photolabeled) GLUT4 by 48% ± 3%, despite an increase in total muscle GLUT4 content of 26% ± 7%. These findings indicate that glucocorticoid-induced inhibition of insulin-stimulated glucose transport in muscle is due to impaired recruitment of GLUT4 to the cell surface.

Antihypertensive agent moxonidine enhances muscle glucose transport in insulin-resistant rats.

The sympatholytic antihypertensive agent moxonidine, a centrally acting selective I1-imidazoline receptor modulator (putative agonist), may be beneficial in hypertensive patients with insulin resistance. In the present study, the effects of chronic in vivo moxonidine treatment of obese Zucker rats--a model of severe glucose intolerance, hyperinsulinemia and insulin resistance, and dyslipidemia--on whole-body glucose tolerance, plasma lipids, and insulin-stimulated skeletal muscle glucose transport activity (2-deoxyglucose uptake) were investigated. Moxonidine was administered by gavage for 21 consecutive days at 2, 6, or 10 mg/kg body weight. Body weights in control and moxonidine-treated groups were matched, except at the highest dose, at which final body weight was 17% lower in the moxonidine-treated animals compared with controls. The moxonidine-treated (6 and 10 mg/kg) obese animals had significantly lower fasting plasma levels of insulin (17% and 19%, respectively) and free fatty acids (36% and 28%, respectively), whereas plasma glucose was not altered. During an oral glucose tolerance test, the glucose response (area under the curve) was 47% and 67% lower, respectively, in the two highest moxonidine-treated obese groups. Moreover, glucose transport activity in the isolated epitrochlearis muscle stimulated by a maximally effective insulin dose (13.3 nmol/L) was 39% and 70% greater in the 6 and 10 mg/kg moxonidine-treated groups, respectively (P<.05 for all effects). No significant alterations in muscle glucose transport were elicited by 2 mg/kg moxonidine. These findings indicate that in the severely insulin-resistant and dyslipidemic obese Zucker rat, chronic in vivo treatment with moxonidine can significantly improve, in a dose-dependent manner, whole-body glucose tolerance, possibly as a result of enhanced insulin-stimulated skeletal muscle glucose transport activity and reduced circulating free fatty acids.

Treadmill Running Increases Phosphatidylinostol 3-Kinase Activity in Rat Skeletal Muscle

Exercise has been shown to increase insulin-stimulated skeletal muscle glucose transport. Activation of phosphatidylinostol 3-kinase (PI 3-kinase) is required for insulin to stimulate glucose transport. The present study was designed to investigate whether treadmill running (60 min, 8% grade, 30 m/min) augments insulin-stimulated activation of PI 3-kinase. Insulin dramatically increased insulin receptor substrate-1 (IRS-1) tyrosine phosphorylation (p < 0.05). Treadmill running did not induce IRS-1 tyrosine phosphorylation, not did it alter insulin-stimulated IRS-1 tyrosine phosphorylation. Insulin increased PI 3-kinase activity by 3.0-fold (over basal activity) in white muscle and 5.2-fold in red muscle (p < 0.05). Exercise did not alter basal PI 3-kinase activity in either white or red muscle. However, in response to exercise, insulin-stimulated PI 3-kinase activity was significantly increased in both muscle fibers (p < 0.05). These results suggest that increased insulin responsiveness induced by exercise may be due, in part, to enhanced insulin-stimulated PI 3-kinase activity.