RESUMO
BACKGROUND: Human growth hormone (hGH) is best known for influencing bone and muscle growth, as well as body composition, but the use of recombinant hGH is controversial. Amino acids are a potentially safer alternative; however, preliminary investigations of the effects of oral amino acids on hGH release have been inconclusive. Therefore, we tested the effects of a novel blend of amino acids optimized to increase hGH release. STUDY QUESTION: Does an investigational amino acid supplement affect hGH release? STUDY DESIGN: This was a randomized, placebo-controlled, double-blind, crossover study that included 16 (12 men, 4 women; age 32 ± 14 years; body mass index 26.4 ± 5.0 kg/m) healthy participants. All participants received both placebo and the amino acid supplement after an overnight fast and completed all study visits. Treatment order was randomized, and each treatment was separated by a 1-week washout period. MEASURES AND OUTCOMES: The primary outcomes were the percent change in hGH from baseline to 120 minutes and the area under the curve of hGH over baseline. Serum hGH was measured using enzyme-linked immunosorbent assay at baseline and 15, 30, 60, 90, and 120 minutes. RESULTS: At 120 minutes, hGH levels increased by 682% (8-fold) from baseline and were significantly higher than placebo (P = 0.01). In addition, a significantly higher mean area under the curve was observed for the amino acid supplement compared with the placebo [20.4 (95% confidence interval, 19.9-21.0 ng/mL) vs. 19.7 (95% confidence interval, 18.7-20.6 ng/mL); P = 0.04]. CONCLUSIONS: These results show that a single dose of the oral amino acid supplement was sufficient to significantly increase hGH levels in healthy adult men and women. CLINICAL TRIAL REGISTRY:: clinicaltrials.gov NCT01540773.
Assuntos
Aminoácidos/farmacologia , Suplementos Nutricionais , Hormônio do Crescimento Humano/biossíntese , Adulto , Estudos Cross-Over , Método Duplo-Cego , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
Most commercial production of recombinant pharmaceutical proteins involves the use of mammalian cell lines, E. coli or yeast as the expression host. However, recent work has demonstrated the potential of eukaryotic microalgae as platforms for light-driven synthesis of such proteins. Expression in the algal chloroplast is particularly attractive since this organelle contains a minimal genome suitable for rapid engineering using synthetic biology approaches; with transgenes precisely targeted to specific genomic loci and amenable to high-level, regulated and stable expression. Furthermore, proteins can be tightly contained and bio-encapsulated in the chloroplast allowing accumulation of proteins otherwise toxic to the host, and opening up possibilities for low-cost, oral delivery of biologics. In this commentary we illustrate the technology with recent examples of hormones, protein antibiotics and immunotoxins successfully produced in the algal chloroplast, and highlight possible future applications.
Assuntos
Produtos Biológicos/metabolismo , Clorófitas/genética , Cloroplastos/genética , Suplementos Nutricionais/provisão & distribuição , Genoma de Cloroplastos , Microalgas/genética , Antibacterianos/biossíntese , Antibacterianos/química , Produtos Biológicos/química , Chlamydomonas reinhardtii/genética , Chlamydomonas reinhardtii/metabolismo , Clorófitas/metabolismo , Cloroplastos/metabolismo , Expressão Gênica , Engenharia Genética/métodos , Vetores Genéticos/química , Vetores Genéticos/metabolismo , Hormônio do Crescimento Humano/biossíntese , Hormônio do Crescimento Humano/genética , Imunotoxinas/genética , Imunotoxinas/metabolismo , Microalgas/metabolismo , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/genética , Biologia Sintética/métodos , Transformação Genética , TransgenesRESUMO
In order to observe the nutrition state in the severe multiple trauma patients undergoing adjuvant recombinant human growth hormone (rhGH) nutritional support therapy, 45 patients with severe multiple traumas (ISS>25) were randomly divided into 3 groups. All the 3 groups had been supplied with nitrogen and caloricity according to the need of patients for 16 days. The rhGH therapy started 48 h after surgery and lasted for 14 days in two rhGH-treated groups in which rhGH was 0.2 and 0.4 U/(kg . d) respectively, and the resting group served as control one. The levels of nitrogen balance, prealbumin and safety variables (blood sugar, Na+, TT3 and TT4) were observed and compared among the three groups. The levels of nitrogen balance on the postoperative day (POD) 3 and 5 in the rhGH-treated groups were -1.28+/-3.19, 5.45+/-2.00 and -0.18+/-2.55, 6.11+/-1.60, respectively, which were significantly higher than those in the control group (-5.17+/-1.68 and -1.08+/-3.31, P<0.01). The values of prealbumin on the POD 3 and 5 in the rhGH-treated groups were 180.19+/-27.15, 194.44+/-50.82 and 194.94+/-29.65, 194.11+/-16.17, respectively, which were significantly higher than those in the control group (117.42+/-19.10 and 135.63+/-28.31, P<0.01). There was no significant difference between the rhGH 0.2 U/(kg . d) group and rhGH 0.4 U/(kg . d) group in both of the levels of nitrogen balance and prealbumin. It is concluded that the nutritional support therapy with adjuvant rhGH which starts 48 h after surgery improves the nutrition state of the patients with severe multiple trauma. It is safe for severe multiple trauma patients who accept rhGH at the dose of 0.2 and 0.4 U/(kg . d).
Assuntos
Hormônio do Crescimento Humano/uso terapêutico , Traumatismo Múltiplo/terapia , Nitrogênio/metabolismo , Apoio Nutricional , Proteínas Recombinantes/uso terapêutico , Adulto , Idoso , Estado Terminal , Feminino , Hormônio do Crescimento Humano/biossíntese , Hormônio do Crescimento Humano/genética , Humanos , Masculino , Pessoa de Meia-Idade , Traumatismo Múltiplo/cirurgia , Estado Nutricional , Período Pós-Operatório , Pré-Albumina/metabolismo , Proteínas Recombinantes/biossínteseRESUMO
Insulin-like growth factor I (IGF-I) exerts a negative feedback effect on GH secretion via either direct actions at the pituitary level or indirect ones at the hypothalamic level, through stimulation of somatostatin (SS) and/or inhibition of GHRH release. In fact, recombinant human IGF-I (rhIGF-I) in humans inhibits spontaneous GH secretion as well as the GH response to GHRH and even more to GH/GH-releasing peptides, whose main action is on the hypothalamus, antagonizing SS and enhancing GHRH activity. The aim of the present study was to further clarify in humans the mechanisms underlying IGF-I-induced inhibition of somatotroph secretion. In six normal young volunteers (all women; mean +/- SEM: age, 28.3+/-1.2 yr; body mass index, 21.3+/-1.2 kg/m2) we studied the GH response to GHRH (1 microg/kg, iv, at 0 min), both alone and combined with arginine (ARG; 0.5 g/kg, iv, from 0-30 min), which probably acts via inhibition of hypothalamic SS release, after pretreatment with rhIGF-I (20 microg/kg, sc, at -180 min) or placebo. rhIGF-I increased circulating IGF-I levels (peak at -60 vs. -180 min: 54.9+/-3.9 vs. 35.9+/-3.3 mmol/L; P < 0.05) to a reproducible extent, and these levels remained stable and within the normal range until 90 min. The mean GH concentration over 3 h (from -180 to 0 min) before ARG and/or GHRH was not modified by placebo or rhIGF-I. After placebo, the GH response to GHRH (peak, 23.6+/-2.9 microg/L) was strikingly enhanced (P < 0.05) by ARG coadministration (69.6+/-9.9 microg/L). rhIGF-I blunted the GH response to GHRH (13.1+/-4.5 microg/L; P < 0.05), whereas that to GHRH plus ARG was not modified (59.5+/-8.9 microg/L), although it occurred with some delay. Mean glucose and insulin concentrations were not modified by either placebo or rhIGF-I. In conclusion, ARG counteracts the inhibitory effect of rhIGF-I on somatotroph responsiveness to GHRH in humans. These findings suggest that the acute inhibitory effect of rhIGF-I on the GH response to GHRH takes place on the hypothalamus, possibly via enhancement of SS release, and that ARG overrides this action.
Assuntos
Arginina/farmacologia , Hormônio Liberador de Hormônio do Crescimento/farmacologia , Hormônio do Crescimento Humano/biossíntese , Hipotálamo/metabolismo , Fator de Crescimento Insulin-Like I/antagonistas & inibidores , Adulto , Área Sob a Curva , Arginina/efeitos adversos , Glicemia/metabolismo , Feminino , Hormônio do Crescimento Humano/efeitos adversos , Humanos , Hipotálamo/citologia , Hipotálamo/efeitos dos fármacos , Insulina/sangue , Fator de Crescimento Insulin-Like I/farmacologia , Proteínas Recombinantes/efeitos adversos , Proteínas Recombinantes/farmacologiaRESUMO
This review systematically analyses recent knowledge of the biology of the growth hormone-releasing peptides. Many years before native GHRH had been isolated and sequenced, the synthesis of an enkephalin analog, devoid of any opioid activity but capable of specifically releasing GH from in vitro pituitaries, prompted the design of a number of structurally interrelated GHRPs with improved GH-releasing activity. Nowadays, GHRPs are the most effective GH-secretagogues known and could be used profitably in humans with GH hyposecretory disturbances to promote a pattern of GH secretion that mimics physiology in a better way than the exogenously administered GH.
Assuntos
Substâncias de Crescimento/farmacologia , Hormônio do Crescimento Humano/efeitos dos fármacos , Oligopeptídeos/farmacologia , Animais , Canais de Cálcio/metabolismo , Desenho de Fármacos , Encefalinas/farmacologia , Hormônio Liberador de Hormônio do Crescimento/efeitos dos fármacos , Hormônio do Crescimento Humano/biossíntese , Humanos , Hipotálamo/efeitos dos fármacos , Hipotálamo/fisiologia , Adeno-Hipófise/efeitos dos fármacos , RNA Mensageiro/metabolismo , Receptores de Superfície Celular/metabolismo , Somatostatina/efeitos dos fármacos , Relação Estrutura-AtividadeRESUMO
Dwarf tyrosine hydroxylase-human GH (TH-hGH) transgenic mice carrying the hGH reporter gene targeted by the TH promoter express hGH in those regions of the hypothalamus responsible for regulation of pituitary GH secretion. Central expression of the hGH gene decreases GH-releasing hormone (GHRH) and increases somatostatin, which ultimately impacts on pituitary function by reducing the overall amount of GH produced. In the present study, we sought to determine if the reduction of pituitary GH in TH-hGH mice could be attributed to a decrease in somatotrope cell numbers and/or an impairment of somatotrope function. Pituitaries from TH-hGH or wild-type (WT) male and female mice were enzymatically dispersed, counted, and immunostained for GH, PRL, TSH, and ACTH. The total number of pituitary cells recovered from TH-hGH pituitaries was approximately one-half of that from WT controls. However, the proportion of cells that stained for GH and PRL were virtually identical (males, GH-TH-hGH, 58.1 +/- 1.0% [mean +/- SEM] vs. WT, 60.7 +/- 1.0%; PRL-TH-hGH, 43.4 +/- 2.2% vs. WT, 43.1 +/- 0.7%; females, GH-TH-hGH, 47.9 +/- 2.3% vs. WT, 41.5 +/- 3.5%; PRL-TH-hGH, 43.3 +/- 3.2% vs. WT, 47.1 +/- 3.3%). In contrast, percentages of both TSH- and ACTH-containing cells were increased in TH-hGH pituitaries relative to controls (males, TSH-TH-hGH, 15.1 +/- 2.3% vs. WT, 9.6 +/- 1.5%; ACTH-TH-hGH, 24.5 +/- 2.5% vs. WT, 10.9 +/- 0.9%; females: TSH-TH-hGH, 11.3 +/- 0.7% vs. WT, 7.5 +/- 0.6%; ACTH-TH-hGH, 19.8 +/- 1.6% vs. WT, 9.3 +/- 0.8%; P < 0.05). Calculation of the absolute number of each cell type per pituitary demonstrated TH-hGH mice to have about one-half the number of GH and PRL cells, whereas TSH and ACTH cell populations were comparable with that of their WT counterparts. Immunocytochemical localization of GH cells within pituitary sections from TH-hGH mice revealed that somatotropes were confined primarily to the lateral wings of the adenohypophysis, in contrast to the heterogeneous distribution of GH-immunostained cells in WT pituitaries. To assess the functional capacity of the somatotrope populations, pituitary cells from TH-hGH and WT mice were challenged with mouse GHRH (0.01-10 nM). The quantity of GH released (as assessed by both RIA and reverse hemolytic plaque assay) under basal and stimulated conditions did not differ among TH-hGH and WT pituitary cell cultures. Similarly, GHRH induced intracellular cAMP levels were comparable. These results indicate that proliferation of pituitary somatotropes and lactotropes is much more sensitive to changes in GHRH input than is the capability of developing regulated GH secretory function.
Assuntos
Hormônio do Crescimento/deficiência , Hormônio do Crescimento Humano/genética , Hipotálamo/fisiologia , Adeno-Hipófise/metabolismo , Regiões Promotoras Genéticas , Tirosina 3-Mono-Oxigenase/genética , Hormônio Adrenocorticotrópico/biossíntese , Animais , Peso Corporal , Células Cultivadas , Feminino , Hormônio do Crescimento Humano/biossíntese , Humanos , Hipotálamo/fisiopatologia , Masculino , Camundongos , Camundongos Transgênicos , Adeno-Hipófise/citologia , Prolactina/biossíntese , Caracteres Sexuais , Tireotropina/biossíntese , Tirosina 3-Mono-Oxigenase/biossínteseRESUMO
The control of mRNA stability is crucial to the regulation of cytokine expression. We describe here a novel, potent destabilizing element found in the 3' untranslated region of granulocyte colony-stimulating factor mRNA. This element, which appears to require at least one stem-loop structure, we term the stem-loop destabilizing element (SLDE). Functionally equivalent elements appear to also exist in the interleukin 2 and interleukin 6 mRNAs. The SLDE is functionally distinct from the A+U-rich elements, which are also present in these and other cytokine mRNAs, because it destabilizes a chimeric mRNA in a tumor cell line in which A+U-rich elements do not function. In addition, the effect of the SLDE is insensitive to calcium ionophore and is therefore regulated independently of A+U destabilizing elements. The existence of two distinct mRNA-destabilizing elements provides an additional mechanism for the differential regulation of cytokine expression.