Beta-adrenergic Agonist Treatment Research Articles

Dynamic transcriptome changes during adipose tissue energy expenditure reveal critical roles for long noncoding RNA regulators.

Enhancing brown fat activity and promoting white fat browning are attractive therapeutic strategies for treating obesity and associated metabolic disorders. To provide a comprehensive picture of the gene regulatory network in these processes, we conducted a series of transcriptome studies by RNA sequencing (RNA-seq) and quantified the mRNA and long noncoding RNA (lncRNA) changes during white fat browning (chronic cold exposure, beta-adrenergic agonist treatment, and intense exercise) and brown fat activation or inactivation (acute cold exposure or thermoneutrality, respectively). mRNA–lncRNA coexpression networks revealed dynamically regulated lncRNAs to be largely embedded in nutrient and energy metabolism pathways. We identified a brown adipose tissue–enriched lncRNA, lncBATE10, that was governed by the cAMP-cAMP response element-binding protein (Creb) axis and required for a full brown fat differentiation and white fat browning program. Mechanistically, lncBATE10 can decoy Celf1 from Pgc1α, thereby protecting Pgc1α mRNA from repression by Celf1. Together, these studies provide a comprehensive data framework to interrogate the transcriptomic changes accompanying energy homeostasis transition in adipose tissue.

Molecular Signature of Human Brown Adipocyte-Like PAZ6 Cells

Background: Prevalence of metabolic syndrome is directly correlated with increasing occurrence of obesity characterized by accumulation of fat in visceral, lower body, and upper body subcutaneous depots. In a number of species, brown adipose tissue (BAT) converts triglycerides into heat by non-shivering thermogenesis, thus controlling the amount of white adipose tissue. Until recently, BAT was thought to be mostly absent in adult humans but now sizeable BAT depots have been visualized by Positron Emission Tomography (PET) and Computed Tomography (CT) scanning. Relatively little is known about the signaling pathways differentially regulated in brown adipocytes compared to white adipocytes. However, our progress of understanding is hindered by the paucity of well-characterized reliable in vitro model systems. Objectives: The aim of this study is to characterize the only in vitro model of human BAT cell line, and to identify signature genes for BAT, and compounds implicated in the enhancement of brown phenotype. Methods: A human BAT cell line, PAZ6, was previously created by immortalizing somatic cells using the large T antigen of Simian Virus 40. The expression of BAT associated markers has been verified by real time RT-PCR done on RNA and immunoblot using specific antibodies performed on cell lysates and supernatants, before and after differentiation and treatment with various compounds such as adrenergic receptor agonist (isoproterenol) and PPAR agonist (rosiglitazone). Results: We have identified several BAT associated markers, including PDRM16 and the 3 adrenergic receptor. This marker is highly expressed in PAZ6 cells compared to the white adipocyte. In addition, we demonstrate that these cells share a common precursor with myocytes but not with white adipocytes. Our results show that several genes are up-regulated after PPAR gamma activation. Moreover, we confirm that these cells respond to Beta adrenergic agonist treatment. Finally, we identify that the transcription factor farnesoid X Receptor (FXR) induces the expression of BAT associated marker following treatment by its agonist : CDCA. Conclusions: The PAZ6 cells constitute a readily available surrogate for human brown adipocytes to develop pharmacologic strategies to promote BAT expansion and activation.

Treatment of vascular smooth muscle cells with estradiol and beta-adrenergic agonists has an additive effect on cAMP levels, but no additive effect on inhibition of collagen synthesis

Several studies have suggested that increased cell levels of cAMP result in decreased rates of collagen synthesis. Oestrogen treatment of vascular smooth muscle cells (VSMCs) has been shown to cause increased levels of cAMP and decreased rates of collagen synthesis. Beta-adrenergic agonists are also known to increase cellular levels of cAMP in VSMCs, although the effect of beta-adrenergic agonists on collagen synthetic rates in VSMCs is unknown. Since beta-agonists and oestrogens are commonly used clinical agents these studies were conducted to determine the potential of these agents to have an additive effect on cell cAMP levels and inhibition of collagen synthetic rates. When VSMCs were treated with both oestrogen and isoproterenol there was an additive effect on cellular cAMP levels although the observed decrease in collagen synthetic rates was the same as observed in cells treated with just oestrogen. Treatment of VSMCs with propranolol inhibited isoproterenol-induced changes in cAMP but had no effect on either oestrogen-induced increases in cAMP levels or inhibition of collagen synthesis. The cellular location of cAMP following beta-adrenergic agonist treatment was different from the distribution of cAMP in control or oestrogen-treated VSMCs. This difference in cellular distribution of cAMP may partially explain the absence of collagen synthesis inhibition following beta-adrenergic agonist treatment of VSMCs.

Brown Fat UCP1 Is Specifically Expressed in Uterine Longitudinal Smooth Muscle Cells

Until now, uncoupling protein 1 (UCP1) was considered as unique to brown adipocytes. It supports a highly regulated uncoupling of oxidative phosphorylation that is associated with diet as well as with non-shivering thermogenesis. Here we report that UCP1 is not specific to brown adipocytes and can be expressed in longitudinal smooth muscle layers. In the uterus, this conclusion was drawn from different convergent data. A specific antibody against mouse UCP1 revealed, in mitochondrial fractions, a protein with the same molecular weight as brown fat UCP1. Sensitive and specific reverse transcriptase-polymerase chain reaction detected a mRNA whose sequence was totally homologous to that of brown fat UCP1 mRNA. Antibody against UCP1 as well as a UCP1 antisense probe specifically stained uterine longitudinal smooth muscles. UCP1 was also expressed in longitudinal smooth muscle of digestive and male reproductive tracts but was never expressed in other types of smooth muscle, including those of arterial vessels. In uterine tract, UCP1 content was increased after cold exposure or beta-adrenergic agonist treatment. It was also up-regulated during the postovulatory period after sexual cycle synchronization. Its content transiently increased during gestation and decreased markedly after birth. These regulations strongly argue about a role for UCP1 in thermogenesis as well as in relaxation of longitudinal smooth muscle layers.

Effects of dietary inclusion of the beta-adrenergic agonist, salbutamol, on growth performance and whole-body composition of rohuLabeo rohita(Hamilton) fingerlings fed diets containing two protein levels

A 60-day feeding experiment was conducted in the laboratory to evaluate the effect of oral administration of the beta-adrenergic agonist (BAA), salbutamol, on growth, nutrient utilization and whole-body composition of rohu Labeo rohita (Hamilton) fingerlings (average weight 5.51 ± 0.07 g) acting as a repartitioning agent in intermediary metabolism and redistributing nutrients for muscle synthesis. Two diets (diets 30/0 and 40/0) were formulated containing 30% and 40% crude protein to serve as basal diets containing the same ingredient composition. Another four diets (diets 30/3, 30/6, 40/3 and 40/6) were prepared in the same way to contain either 3 or 6 mg kg−1 BAA salbutamol at each protein level making the total of six experimental diets. Rohu fingerlings were fed with the experimental diets in three replicate treatments at a restricted feeding regime equivalent to 2% of body weight (BW) day−1. A significant (P < 0.05) interaction effect was found between dietary protein level and BAA treatment. Dietary incorporation of BAA at both protein levels significantly (P < 0.05) increased growth and nutrient utilization in terms of weight gain (%), specific growth rate (SGR), feed:gain ratio (FGR) and protein efficiency ratio (PER). BAA treatment at the 6 mg kg−1 concentration led to a 12% rise in growth with the 30% protein diet and a 6% rise with the 40% protein diet, above their respective controls (without BAA treatment). However, no significant differences were found by raising the BAA concentration from 3 to 6 mg kg−1. The apparent digestibility values for protein, lipid and energy were only higher with the 30% protein diets containing BAA. At both protein levels tested, BAA exerted a significant (P < 0.05) positive influence on protein retention and a negative influence on lipid retention efficiency, although the differences between the 3 and 6 mg kg−1 BAA diets were not significant. This indicated that BAA acted as a repartitioning agent in suppressing lipid deposition in favour of protein accretion. BAA induced significantly (P < 0.05) higher whole-body protein and lipid relative to the untreated groups. The condition factor improved significantly (P < 0.05) in the BAA-treated dietary groups. The fillet percentage displayed a direct correlation and the frame percentage a negative correlation with BAA concentration in the diets. The results indicate that the BAA, salbutamol, was effective in producing growth enhancement, improved body composition (higher protein and lower lipid accretion) and efficient nutrient utilization at the 3 mg kg−1 dietary incorporation level and thus has a potential for application in formulated diets for the Indian major carp, rohu, under culture conditions.

Differential effects of dexamethasone and clenbuterol on rat growth and on beta2-adrenoceptors in lung and skeletal muscle.

Beta-adrenergic agonists increase growth rate, but their efficacy is reduced over time as the number of beta2-adrenoceptors in muscle decreases. Dexamethasone increases beta2-adrenoceptor density in many tissues, but this effect has not been reported in skeletal muscle. In this study, male rats were treated daily for 10 d with either clenbuterol (4 mg/kg of feed), dexamethasone (.2 mg/kg BW, s.c.), or clenbuterol plus dexamethasone. Untreated rats served as controls. Dexamethasone caused a marked suppression of growth rate, which resulted in decreased (P < .001) body weight (-29%), carcass weight (-30%), hind-limb muscles (-22%), omental fat (-22%), and heart weight (-10%). Feed intake was reduced (-26%), but feed conversion efficiency was also impaired (P < .001). Clenbuterol caused a small increase in growth rate (+6%; P < .05), with an increase in leg muscle (+7%; P < .01) and heart mass (+8%; P < .05). Feed efficiency was improved (P < .001) by clenbuterol. Rats given the combined treatment still showed a reduction in growth rate (-81%). Clenbuterol caused only a mild attenuation of the effects of dexamethasone on feed intake, BW, and carcass weight, but reduced the catabolic effect of dexamethasone on hind-limb muscle to only -8%. Clenbuterol caused a slight increase in the affinity beta2-adrenoceptors in lung for binding to the radioligand (-)[125I]iodocyanopindolol. Relative to control values, the density of beta2-adrenoceptors in lung was +31% with dexamethasone treatment, -45% with clenbuterol, and -23% with the combined treatment. Clenbuterol also decreased beta2-adrenoceptors in skeletal muscle (-35%), but so did dexamethasone (-13%), so the effects of the beta-adrenergic agonist were not attenuated through use of the combined treatment (-40%). The results show that the inductive effect of glucocorticoids on beta2-adrenoceptors is tissue-specific and that glucocorticoid treatment is not a useful adjunct to beta-adrenergic agonist treatment in animal production.

Impact of beta-adrenergic agonist on Na+ channel and Na+-K+-ATPase expression in alveolar type II cells.

It has been shown that short-term (hours) treatment with beta-adrenergic agonists can stimulate lung liquid clearance via augmented Na+ transport across alveolar epithelial cells. This increase in Na+ transport with short-term beta-agonist treatment has been explained by activation of the Na+ channel or Na+-K+-ATPase by cAMP. However, because the effect of sustained stimulation (days) with beta-adrenergic agonists on the Na+ transport mechanism is unknown, we examined this question in cultured rat alveolar type II cells. Na+-K+-ATPase activity was increased in these cells by 10(-4) M terbutaline in an exposure time-dependent manner over 7 days in culture. This increased activity was also associated with an elevation in transepithelial current that was inhibited by amiloride. The enzyme's activity was also augmented by continuous treatment with dibutyryl-cAMP (DBcAMP) for 5 days. This increase in Na+-K+-ATPase activity by 10(-4) M terbutaline was associated with an increased expression of alpha1-Na+-K+-ATPase mRNA and protein. beta-Adrenergic agonist treatment also enhanced the expression of the alpha-subunit of the epithelial Na+ channel (ENaC). These increases in gene expression were inhibited by propranolol. Amiloride also suppressed this long-term effect of terbutaline and DBcAMP on Na+-K+-ATPase activity. In conclusion, beta-adrenergic agonists enhance the gene expression of Na+-K+-ATPase, which results in an increased quantity and activity of the enzyme. This heightened expression is also associated with augmented ENaC expression. Although the cAMP system is involved, the inhibition of enhanced enzyme activity with amiloride suggests that increased Na+ entry at the apical surface plays a role in this process.

Alveolar Epithelial Fluid Clearance Mechanisms Are Intact After Moderate Hyperoxic Lung Injury In Rats

The capacity of the alveolar epithelial barrier to remove excess alveolar fluid from the airspaces of the lung was studied in an experimental model of moderate hyperoxic lung injury. Rats were exposed to 100% oxygen for 40 h in an exposure chamber and compared with control animals exposed to room air. Extravascular lung water was calculated gravimetrically. Alveolar and lung liquid clearance were studied over 1 h by instillation of a 5% albumin solution with 1.5 microCi of 125I-labeled albumin (6 mL/kg into both lungs). The concentration of both the unlabeled and labeled albumin was used to calculate alveolar liquid clearance. Hyperoxic rats developed pulmonary edema, with a 33% increase in extravascular lung water to 5.3 +/- 0.1 g of water per gram of dry lung, compared with 4.0 +/- 0.2 g of water per gram of dry lung in control rats (p < 0.05). This degree of edema was associated with a significant increase in the alveolar-arterial oxygen difference (241 +/- 61 vs 124 +/- 14 mm Hg in control animals exposed to room air, p < 0.05). Despite this moderate degree of lung injury, alveolar fluid clearance was normal (30 +/- 3%) compared with control rats (33 +/- 6%). Furthermore, the hyperoxic injured rats responded normally to an exogenous beta-adrenergic agonist (terbutaline, 10(-4) mol/L) with a 67% increase in the rate of alveolar liquid clearance (50 +/- 5%). Thus, in the setting of moderate hyperoxic lung injury, the alveolar epithelial barrier is still capable of removing fluid at a normal rate and responding to beta-adrenergic agonist treatment. These experimental results have potential clinical implications for patients with acute lung injury.

Tocolytic therapy with fenoterol induces selective down-regulation of beta-adrenergic receptors in human myometrium.

Tocolytic therapy with beta-adrenergic receptor agonists is a standard regimen to prevent preterm birth. Agonists exposure of beta-adrenergic receptors causes receptor desensitization in other organs, and this may limit the therapeutic value of beta-adrenergic receptor agonists. To study the effects of prolonged beta-adrenergic agonist treatment in human myometrium, we obtained biopsies during Caesarean section of 14 pregnant patients who had received fenoterol for at least 5 days and 14 untreated pregnant controls. The densities of total beta-adrenergic receptors, which are mainly of the beta 2-subtype as assessed by [125I]iodo-cyanopindolol binding in crude membrane fractions, were more than 50% smaller in women receiving fenoterol, whereas alpha 2-adrenergic receptor densities were similar. Gs and Gi G-protein alpha-subunit densities were unaltered as assessed by Western blotting and pertussis toxin-catalyzed [32P]ADP-ribosylation. beta-Adrenergic receptor kinase (beta ARK) activity, as determined using bovine rhodopsin as the substrate, was the same in the two groups. Adenylyl cyclase activities in the presence of guanine nucleotides, NaF, forskolin, or Mn+2 were also not altered by fenoterol treatment. The messenger RNA (mRNA) concentrations of beta 2-adrenergic receptors, beta ARK-I and glyceraldehyde-3-phosphate dehydrogenase (as a reference), as determined by quantitative PCR, were unaffected by fenoterol treatment. We conclude that tocolysis with fenoterol results in a selective down-regulation of myometrial beta-adrenergic receptors, which is not associated with a reduction in the respective mRNA concentrations or alterations of alpha 2-adrenergic receptors, Gs and Gi alpha-subunits, or beta ARK activity or mRNA.

Temporal response of rabbits to beta-adrenergic agonist feeding: tissue weight, calpains and calpastatin activities, and nucleic acid and protein concentrations.

Forty-eight crossbred rabbits were used in three replications of a 2 x 4 factorial arrangement to investigate the short-term responses of tissue accretion, calpains and calpastatin activity, and nucleic acid and protein concentrations to beta-adrenergic agonist (BAA) feeding. Rabbits were fed a 17% CP diet with or without 7 ppm of L644,969 and slaughtered after 1, 4, 8, or 16 d of treatment. Empty body dressing percentage and biceps femoris weight (as a percentage of empty body weight [EBW]) were significantly higher in the treated rabbits than in the controls after 16 d of treatment. Heart and liver weights (as a percentage of EBW) were higher (P < .05) after 1 d and liver weight (as a percentage of EBW) was lower (P < .05) after 16 d in treated vs controls. Except for an elevation of skeletal muscle m-calpain after 16 d, BAA-supplementation did not affect the calpain-calpastatin system. Muscle RNA concentrations and RNA:DNA ratios were higher (P < .05) in treated rabbits after 1 d and remained higher thereafter. Protein:RNA ratios were lower (P < .01) in treated than in control rabbits after 4 d and remained lower throughout the trial. Muscle DNA content was lower after 4 d and higher after 16 d; RNA content was higher after 4, 8, and 16 d; and protein content was higher after 16 d in treated vs control rabbits. Liver nucleic acid and protein concentrations were not affected by BAA treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects over time of feeding a beta-adrenergic agonist to wether lambs on animal performance, muscle growth, endogenous muscle proteinase activities, and meat tenderness.

Forty wether lambs were used in a 2 x 4 factorial arrangement to determine the response of animal performance, muscle growth, proteinase activity, and meat tenderness to beta-adrenergic agonist (BAA) supplementation. Lambs were fed a finishing diet with or without 4 ppm of L644,969 and slaughtered after 0, 2, 4, and 6 wk of treatment. The ADG was higher (P < .05) in the treated than in the control lambs after 2 wk and returned to control levels thereafter. Semitendinosus weight and calpastatin activity were higher and mu-calpain activity was lower in the treated than in the control lambs after 2, 4, and 6 wk. Cathepsin B activity was higher (P < .01) and cystatin-like activity was lower (P < .05) after 2 wk in treated than in control lambs but returned to control levels thereafter. Longissimus protein:DNA was higher after 4 (P < .05) and 6 (P < .01) wk in the treated lambs than in the controls. The concentration of RNA and RNA:DNA ratio were higher (P < .01) in the longissimus and semitendinosus muscles in the treated lambs after 2 wk and remained higher throughout the study. Semitendinosus protein and RNA content were higher after 2, 4, and 6 wk and DNA content was higher after 2 and 6 wk in the treated than in the control lambs. Longissimus shear-force values were higher (P < .001) in the treated than in the control lambs at all slaughter end points. These data indicate a rapid alteration of muscle growth, activity of the calpain-calpastatin system, and meat tenderness during BAA treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Efficacy of anticholinergic and beta-adrenergic agonist treatment of maximal cholinergic bronchospasm in tracheally intubated rabbits.

Cholinergically induced bronchoconstriction is thought to be a major cause of bronchospasm during anesthesia. We used tracheally intubated rabbits (4-mm endotracheal tube) stimulated with methacholine to assess the efficacy of beta-adrenergic agonist and anticholinergic treatment in reversing the increases in respiratory system resistance. Four groups were compared: (a) inhaled metaproterenol, 20 puffs via metered dose inhaler (0.65 mg/puff); (b) inhaled ipratropium bromide, 20 puffs from a metered dose inhaler (18 micrograms/puff); (c) 2 mg of intravenous atropine; and (d) no treatment after methacholine challenge as a control group. Methacholine increased respiratory system resistance from 0.041 +/- 0.001 (mean +/- SEM) to 0.098 +/- 0.006 cm H2O.mL-1.s-1 (P < 0.001). Whereas beta-adrenergic agonist treatment was ineffective in ameliorating bronchoconstriction, inhaled ipratropium bromide and atropine were highly effective, causing an 86%-88% reversal in the methacholine-induced increase in respiratory system resistance. Both these agents were also effective in improving dynamic compliance. We conclude that inhaled ipratropium bromide is effective in treating cholinergic bronchospasm even when administered via a small endotracheal tube and that the beta-adrenergic agonist metaproterenol is ineffective in rabbits in the face of maximal cholinergic stimulation.

Adrenergic control of circulating lymphocyte subpopulations. Effects of congestive heart failure, dynamic exercise, and terbutaline treatment.

The current studies were undertaken to explore the relationship between enhanced sympathetic nervous activity and lymphocyte subset distribution in three settings: congestive heart failure, dynamic exercise, and beta-adrenergic agonist treatment. We compared the number and subset distribution of circulating lymphocytes in 36 patients with congestive heart failure and 31 age-matched control subjects. The number of circulating lymphocytes was lower in heart failure than in control. This was due to a reduction in Tsuppressor/cytotoxic and natural killer cells without significant alteration of Thelper cells. The extent of the alteration was similar in patients with idiopathic and ischemic heart failure, but the reduction was more pronounced in patients with New York Heart Association class III-IV than in class I-II. The plasma catecholamine elevation in heart failure was also independent of etiology but more pronounced in the more severely ill patients. We also assessed lymphocyte subsets after acute stimulation of sympathetic activity by dynamic exercise and after treatment with the beta-adrenergic agonist terbutaline. Dynamic exercise until exhaustion increased the number of circulating lymphocytes in healthy controls and heart failure patients in a subset-selective manner. By contrast, a 7-d treatment with terbutaline caused a reduction in the circulating number of lymphocytes in some subsets that was identical to that seen in heart failure patients. We conclude that prolonged sympathetic activity reduces the number of circulating lymphocytes by a beta-adrenergic mechanism. Such alterations might be involved in the pathophysiology of heart failure and other disease states involving increased activity of the sympathetic nervous system.

Growth, feed efficiency and carcass composition of finishing Friesian steers fed the beta-adrenergic agonist L-644,969.

The beta-adrenergic agonist L-644,969 was evaluated to determine its effects on growth performance and carcass composition of Friesian steers. L-644,969 is the R,R isomer of 6-amino [[(1-methyl-3-phenylpropyl) amino] methyl]-3-pyridine methanol dihydrochloride. Four groups of 18 steers, averaging 380 kg body weight, were individually given ad libitum access to a pelleted concentrate diet that contained either 0, .25, 1.0 or 4.0 ppm L-644,969 for the final 12 wk of the finishing period. Live weight gain was not affected by L-644,969, but feed consumption was linearly reduced (5.5, 6.3 and 15.7%; P less than .01) and feed conversion efficiency was linearly increased (16, 25 and 31%; P less than .01) relative to unmedicated controls, respectively. In addition, L-644,969 quadratically increased carcass weight (3.7, 9.3 and 8.5%; P less than .01) and dressing percentage (2.7, 7.9 and 7.9%; P less than .001). The proportion of trimmed fat in the carcass was quadratically reduced (14.5, 29 and 36%; P less than .001) and yield of lean meat quadratically increased (6.7, 13 and 15.6%; P less than .001). beta-adrenergic agonist treatment altered the distribution of lean meat such that a greater (P less than .001) proportion of the total lean was in the hind portion of carcasses from treated animals. Based on these findings, we suggest that L-644,969 may have utility as an agent to improve efficiency of production of lean beef.

Interactions between response to inhaled prostaglandin E2 and chronic beta-adrenergic agonist treatment.

Cumulative inhalation dose-response curves for the response to prostaglandin E2 (PGE2) have been constructed in normal subjects and patients with mild, stable asthma. In normal subjects cumulative inhalation dose-response curves were also constructed for salbutamol. In normal subjects dose-related bronchodilatation occurred in response to both PGE2 and salbutamol, although both the within-subject and the between-subject variation was significantly greater with salbutamol. Most asthmatic subjects gave a biphasic response to PGE2 on at least one occasion, PGE2 being a bronchoconstrictor above a certain level of specific airways conductance (sGaw) and a bronchodilator below. Chronic treatment with inhaled salbutamol (400 micrograms four times a day) had no effect on the normal subjects' response to salbutamol but there was a significant shift of the PGE2 dose-response curve to the left, indicating increased bronchodilatation (p less than 0.02). Stabilisation of the asthmatics' dose-response curve in the direction of bronchodilatation also occurred and was more pronounced (p less than 0.005). In the normal subjects PGE2 may be concerned in the control of airway smooth-muscle tone and in limiting bronchoconstriction induced by mediators such as histamine, and chronic salbutamol treatment may be important in enhancing these effects of PGE2. 80 mg oral propranolol given one and a half hours before had no effect on PGE2-induced bronchodilatation; but the question whether chronic treatment with beta-blockers has any effect needs investigation.