人生长激素的体外重组及在原代鼠成肌细胞内的表达

目的 :探讨原代鼠骨骼肌成肌细胞异位表达重组人生长激素 (rhGH)的可行性。方法 :将目的基因人生长激素 (hGH)亚克隆至真核表达载体pcDNA3上 ,构建重组质粒pcDNA hGH。分别用酶切电泳及DNA测序的方法对重组质粒进行鉴定。用阳离子聚合物SofastTM介导的基因转移技术将重组质粒导入体外培养的原代大鼠骨骼肌成肌细胞。收集细胞培养上清液 ,用放射免疫分析 (RIA)的方法检测rhGH。结果 :酶切电泳和DNA测序显示hGHcDNA成功地插入到空载体中 ;rhGH表达持续 3周以上。结论 :该实验成功构建了能在真核细胞内表达的重组质粒pcDNA hGH ,为进一步研究以组织进行的基因治疗奠定了基础。     

Human growth hormone and prolactin during pregnancy (author's transl)]

In order to know the secretory behaviors of human growth hormone (hGH) and human prolactin (hPRL) during pregnancy, the following studies were undertaken. Twenty three normal pregnant women of every period of gestation, eighteen women of postpartum and five nonpregnant subjects volunteered for this study. After fasting overnight, these volunteers were placed at complete bed rest, and a fasting antecubital venous blood sample was drawn at 8:00 a.m. Then L-arginine, 30 g, was infused intravenously over a 30 minute period, and venous blood samples were drawn at 30, 45, 60, 90, 120, minutes after infusion. Serum hGH level was detected by hGH radioimmunoassay Kit (Dainabot) and serum hPRL concentration was measured by double-antibody radioimmunoassay system (NIH-NIAMDD). In addition, serum hCS level was measured by hCS-Kobe double-antibody radioimmunoassay system in comparison with the secretory behaviors of hGH and hPRL. 1. Serum hGH, hPRL and hCS concentrations during pregnancy. HGH concentration remained almost unchanged through the course of pregnancy, but hPRL and hCS concentrations increased with the programs of pregnancy. 2. Serum hGH and hPRL concentrations in puerperium. HGH level did not change as compared to that of nonpregnant or pregnant women. HPRL concentration maintained high level in 1-3 postpartum weeks. 3. Effect of arginine on the concentrations of serum hGH, hPRL and hCS during pregnancy. The hGH response decreased, but that of hPRL increased along with the progress of pregnancy. During the arginine loading test there was no significant change in hCS concentration. 4. Effect of arginine on the concentrations of serum hGH and hPRL in puerperium. The HGH response was suppressed at the first week of the postpartum. The response of hPRL was lower than that of late pregnancy. To summarize, hGH and hPRL have some similar biological characters, but there was a difference in secretion pattern of the two hormones during pregnancy. Reserve function of hGH secretion was suppressed, but that of hPRL secretion increased along with the progress of pregnancy. And in the third trimester of pregnancy, the difference of the secretory behavior and secretory reserve function between hGH and hPRL was prominent.     

Effect of administered human growth hormone on growth retardation in inflammatory bowel disease

In a previous study of 22 severely growth-retarded children with inflammatory bowel disease (IBD), endocrinologic evaluation revealed hypogonadism and low growth hormone (HGH) levels. This was attributed to a secondary hypopituitarism. In order to assess this hypothesis, two individuals from this group and another similar child who was not so severely growth-retarded as to be included in the initial study were given HGH replacement in an acute trial and for a 6-month interval. No significant height increment could be attributed to the HGH administration although a definite anabolic response was present in each patient during the acute trial. It appears that the youngsters with IBD may have a relative end-organ resistance to the metabolic and growth-promoting effects of HGH and that their growth problems are not related to hypopituitarism.     

The UK Sport perspective on detecting growth hormone abuse

Background and objectivesHuman growth hormone (hGH) is seen as a doping risk in sport because of its possible anabolic and lipolytic effects. As a result of this hGH is prohibited for athletes to use both in and out-of-competition by the World Anti-Doping Agency (WADA) requiring Anti-Doping Organisations to work with research teams in identifying ways to detect hGH abuse.ApproachThis paper reviews and discusses the UK Sport perspective on the challenges faced in detecting hGH and in particular draws upon the experiences gained during the collaborative efforts with the GH-2004 research team in achieving the implementation of the Marker Method for hGH detection.ConclusionsIn 2008 significant progress has been made; there is one test for detecting HGH approved for use in anti-doping and a second detection method pending. This is a strong reflection of the ongoing research efforts in anti-doping and the progress being made by the Anti-Doping Organisations in reducing the risk that doping poses to sport.     

Effect of alcohol administration on plasma growth hormone response to insulin-induced hypoglycemia.

Plasma growth hormone (HGH) response to insulin-induced hypoglycemia was assessed in a group of normal adult volunteers, with and without prior consumption of ethanol. A significant difference was found in peak HGH concentrations on the two occasions, indicating that prior consumption of ethanol attenuates the normal HGH response to insulin-induced hypoglycemia. It is suggested that ethanol may deplete catecholamine stores in the neurons of the ventromedial nucleus of the hypothalamus and may thereby impair secretion of growth hormone releasing factor. Under certain circumstances this could be of importance in the pathogenesis of alcohol-induced hypoglycemia.     

Human growth hormone as a regulator of blood glucose concentration and as a diabetogenic substance

Summary Human growth hormone (HGH) has recently been shown to play a prominent role in the control of blood glucose homeostasis. Furthermore, it has long been known that administration of growth hormone in animals can induce a diabetes-like state. In human subjects, exogenous administration of HGH or hypersecretion of the endogenous hormone in acromegaly is accompanied by glucose intolerance in only about 25 per cent of the cases. — In this paper, data are presented which give a more diversified picture of the so-called diabetogenic action of HGH. It is suggested that HGH, although decreasing the peripheral utilization of glucose, is not a primary diabetogenic factor, since its insulinogenic action causes a compensatory hyperinsulinism, with normal glucose tolerance as the result. HGH is diabetogenic only in prediabetic subjects whose pancreas is unable to respond to the insulinogenic effect of the hormone. In such subjects, the diabetogenic action of HGH not being counterbalanced by a compensatory hyperinsulinism, glucose intolerance may result. Thus, HGH may be regarded as anadditional factor for the development of diabetes, the major prerequisite being a preëxisting prediabetic state.Presented as an invited lecture at the VI Acta Endocrinologica Congress, Helsinki, Finland, August 8th–12th, 1967.     

Determination of growth hormone in plasma of psoriatic arthritis, psoriasis vulgaris and seronegative spondylarthritis

By means of radio-immunoassay the concentration of human growth hormone (HGH) was measured in the blood plasma of 61 patients with psoriasis (21 suffering from psoriasis vulgaris and 40 with psoriatic arthritis), 30 patients with ankylosing spondylitis, 9 with atypical spondylarthritis and 34 patients with diseases of the soft tissue or degenerative joint and spinal column disease. No connection was found between the HGH concentration and the skin lesions in psoriasis. On the other hand a correlation between HGH and the sacroiliitis in psoriatic arthritis and seronegative spondyloarthropathies may be possible. In contrast to the plasma of psoriatics, the mean HGH concentration was higher in the plasma of patients with degenerative joint diseases. Therefore the results of this paper confirm those opinions in the literature which deny increased HGH concentrations in psoriatics. The beneficial effect of the therapeutic administration of somatostatin, an inhibitor of the release of HGH, in psoriasis vulgaris and psoriatic arthritis is - if indeed it occurs - attributable to other hitherto unidentified mechanisms.     

Heterogeneity of human growth hormone and prolactin secreted in vitro: Immunoassay and radioreceptor assay correlations.

Gel filtration of sera and of in vitro pituitary incubation media from a normal subject and from subjects with pituitary tumors or physiologically elevated growth hormone (hGH) and/or prolactin (hPRL) levels, has been performed. Heterogeneous immunoreactive forms of both hormones with comparable elution profiles in sera and in in vitro incubation media were identified. In each instance the most retarded component ('little hPRL' and 'little hGH') migrated identically with the respective radioiodinated pituitary standard and constituted the major component identified. Several components of intermediate mobility were identified and characterized as 'big hPRL', 'big hGH,' and 'big big hGH'. In addition, a void volume immunoreactive peak was often identified. No interconversion was demonstrated on refiltration of prolactin tumor sera or normal pituitary incubation media. Radioreceptor activity utilizing pregnant rabbit liver membranes for hGH and rabbit mammary membranes for hPRL indicates comparable immunologic and receptor activity in all forms identified following gel filtration of prolactin tumor sera (for hPRL) and normal pituitary incubation media (for hGH). Only for the larger species of HGH, identified by gel filtration of clinical grade hGH, was diminished receptor activity demonstrated. These data suggest that the human pituitary synthesizes and secretes protein hormones of different molecular size according to gel filtration elution volume profiles but with comparable receptor and immunologic assay reactivity. These results support but do not establish the proposal that pituitary hormones are also secreted as prohormones.     

Human pituitary growth hormone (hGH) therapy in growth hormone deficiency

This review has attempted to answer a number of questions regarding human growth hormone therapy in growth hormone deficiency. I believe that the available data support several conclusions which form a suggested current approach to the clinical use of hGH. While these conclusions are derived from data obtained using pituitary growth hormone, it is likely that they are applicable to growth hormone manufactured by recombinant DNA technology, as well. Treatment should be begun as early as the diagnosis can be made in anticipation of a better initial and long-term response in younger patients. Growth hormone should be administered on the basis of body weight in an initial dose of 0.06-0.10 unit/kg 3 times a week. Growth hormone may be administered either intramuscularly or subcutaneously. Therapy should be continuous whenever possible. Treatment should be given until there is no further response which generally will reflect closure of the epiphyses. Associated hormone deficiencies should be adequately treated, and patients should be periodically evaluated for the development of additional deficiencies. Concomitant therapy is not indicated unless deficiencies are clearly demonstrated. Thyroid replacement should be at full dosage, while glucocorticoid replacement should probably not exceed 10-15 mg/m2 x day. Gonadal steroids should be used at the bone age when puberty is expected, and hGH should be continued during pubertal development. There is no general indication for giving anabolic/androgenic steroid in combination with hGH in prepubertal patients. If a waning effect of therapy is observed, the dose of hGH should be incrementally increased, and/or the addition of anabolic/androgenic steroid therapy should be considered. While most reports have focused on the effect of hGH on linear growth, changes in weight, bone age, body proportions, and body composition have also been observed. The effect on bone age is variable, but there is greater enhancement of linear growth than of epiphyseal development in the majority of treated patients. Bone age must be monitored during hGH administration whether or not anabolic/androgenic steroids are used concurrently. Growth hormone administration is remarkably free of side effects. However, neutralizing antibodies to hGH may develop and they should be sought in patients in whom an unexplained decrease in response is observed. Certainly the available incomplete data allow for different conclusions. The expanding supply of hGH should lead to a more systematic evaluation and provide more definite answers to the questions which this review has considered.     

Monoclonal antibodies recognize conformational alterations leading to decreased bioactivity of human growth hormone

By means of monoclonal antibodies to five different antigenic determinants on human growth hormone (hGH) and polyclonal antisera from mice and rabbits, the immunoactivity of native hGH was compared with that of reduced and S-carboxymethylated hGH, which has an unstable conformation. Native and reduced and S-carboxymethylated hGH were also tested in a radioreceptor assay which reflects the bioactivity of the hormone. The IM-9 cell line was used as a receptor source in this assay. All five monoclonal antibodies were superior in discriminating between the native active form of hGH and its reduced and S-carboxymethylated form, which has a markedly reduced receptor binding activity.     

EFFECT OF l-DOPA ON THE SECRETION OF PLASMA GROWTH HORMONE IN CHILDREN AND ADOLESCENTS

The stimulation of growth hormone release in children by L-dopa has been studied. An oral dose of 0.5 g L-dopa was administered to fourteen children with, and to fifteen children without, hypothalamic-pituitary insufficiency. In the control group, L-dopa induced a release of pituitary growth hormone, the peak of which occurred from 30 to 60 min after ingestion. Nine out of fifteen control subjects showed peak levels of plasma growth hormone greater than 8 ng/ml. None of the patients with hypothalamic-pituitary insufficiency showed levels greater than 5 ng/ml. In five out of six children with measurable amounts of plasma HGH, and in fourteen children with a lack of HGH, there was a good correlation between the HGH response after L-dopa, insulin hypoglycaemia and arginine infusion. It is concluded that the administration of one oral dose of L-dopa can be used as a provocative test of growth hormone secretion.     

Transitory growth hormone deficiency successfully treated with human growth hormone

This study was carried out in order to determine whether children with a transitory type of growth hormone deficiency showed an accelerated growth in height velocity on treatment with human growth hormone (HGH). Following careful diagnostic routine procedures 13 extremely short children were diagnosed as having isolated growth hormone deficiency, and were successfully treated with HGH. A true isolated growth hormone deficiency was present in 5 of the children, whereas 8 showed a normal increase in serum growth hormone on repeated growth hormone stimulation tests after their development of puberty and termination of HGH treatment.Three boys with bone ages of 5.5, 8.0 and 9.5 years showed an undisputable effect following HGH administration. They showed an initial growth at the start of treatment, and a second growth spurt during development of puberty. Two of the boys reached final statures of 14 cm taller than the predicted heights. The other patients, including the children with true isolated growth hormone deficiency showed an initial spurt of growth at the start of the HGH treatment immediately followed by a pubertal growth spurt. The mean accleration of height velocity for the children with true isolated growth hormone deficiency was from 3.4 cm during the year before treatment to 7.0 cm during the first year on treatment, as compared to 2.8 and 7.4 cm, respectively, for the children with transitory growth hormone deficiency. A girl with severe anorexia nervosa who had a transitory growth hormone deficiency, showed an accelerated high velocity from 1.1 cm to 7.6 cm during the first year following treatment with HGH.     

Urinary hGH, albumin, alpha 1 MG and beta 2 MG in normal and diabetic children

I measured urinary hGH level in children with insulin dependent diabetes mellitus (DM) and studied the relation with urinary albumin alpha 1-microglobulin (alpha 1 MG), beta 2-microglobulin (beta 2MG) and plasma HbA1. 24-hour urine or night-time urine was collected in 54 normal children (NC) and 61 DM. Urinary hGH was estimated by using a sensitive sandwich enzyme immunoassay. Urinary albumin, alpha 1 MG and beta 2MG were measured by RIA. HGH concentration in 24-hour urine in NC significantly correlated with urinary albumin and beta 2MG concentrations (p less than 0.01). In DM, hGH concentration in 24-hour urine also correlated with urinary albumin concentration (p less than 0.05). To avoid the effects of posture and exercise, night-time urine was used in the following study. Urinary hGH concentration was significantly higher in DM for all age (p less than 0.05-0.01) than that in NC. Urinary alpha 1MG and beta 2MG concentrations were also significantly higher in DM than those in NC. HGH concentration in night-time urine in NC has not correlated with urinary albumin, alpha 1MG nor beta 2MG. In DM, however hGH significantly correlated with urinary albumin and beta 2MG. Even in DM with normal concentrations of urinary albumin and beta 2MG, urinary hGH was significantly higher than in NC. Urinary hGH in poorly controlled DM was higher than in well controlled DM. Urinary hGH concentration showed positive correlation with plasma HbA1 in DM. We conclude that urinary hGH relates to not only serum hGH concentration but also renal function. In DM with normal renal function, hGH concentration in night-time urine was higher than in NC, which suggested high serum hGH concentration in DM.     

Kyolic and Pycnogenol increase human growth hormone secretion in genetically-engineered keratinocytes.

The amount of human growth hormone (HGH) decreases significantly after the age of 30. This decrease has been implicated as one of the major causes in the signs of aging, such as thinning of the skin and bones, a decrease in lean muscle mass and an increase in adipose tissue. Supplementing the body's dwindling supply with recombinant human growth hormone (rHGH) has been shown to reverse the signs and symptoms of aging. However, drawbacks in rHGH replacement therapy include prohibitively high cost, the need for repeated injection and side effects such as carpel tunnel syndrome, gynecomastia and insulin resistance. The purpose of this study was to establish an in vitro model using genetically-engineered keratinocytes to screen natural compounds for the ability to stimulate HGH secretion. We now report that a combination of equal amounts of L-arginine and L-lysine, aged garlic extract (Kyolic), S-allyl cysteine and Pycnogenol significantly increased secretion of HGH in this in vitro model. The data indicate that this in vitro model may be used to screen for other secretagogues.     

Discordant immune and growth response to pituitary and biosynthetic growth hormone in siblings with isolated growth hormone deficiency type IA

Two brothers with familial isolated growth hormone deficiency type IA homozygous for the same 6.7 kb deletion on chromosome 17 including the growth hormone gene were intermittently treated with various forms of hGH for more than 7 years. While the elder brother (Patient 1) showed a good growth response to pituitary hGH, the younger one (Patient 2) developed high titre growth blocking hGH antibodies early in the course of treatment and grew only 2.2-3.9 cm/year on a hGH dose of 12-26 IU/m2 per week. When the younger brother was changed to a higher dose (33 IU/m2 per week) of biosynthetic methionyl hGH he had striking catch-up growth and he has subsequently maintained a height velocity of 10.0 cm/year for the last 2 years. During this time his antibody titres have decreased over 1000-fold. These findings demonstrate that therapy with biosynthetic methionyl hGH may provide an effective form of treatment for subjects with isolated growth hormone deficiency type IA who do not grow in response to native hGH, and imply that biosynthetic methionyl hGH may be less antigenic than pituitary derived hGH.     

Direct effect of human growth hormone to inhibit glucose uptake in cultured human fibroblasts

We have examined the effect of human growth hormone (HGH) on both basal and insulin stimulated glucose uptake in human fibroblasts. High concentrations of HGH (100 μg/mL) depressed both basal and insulin-stimulated glucose uptake by 25%. Significant inhibition was not seen at concentrations below 50 μg/mL of HGH. HGH-related hormones like human prolactin and ovine growth hormone had no effect on glucose uptake while high concentrations of ovine prolactin reduced basal glucose uptake, albeit to a lesser degree than HGH. In conclusion, high concentrations of HGH act independently of insulin to inhibit glucose uptake in human fibroblasts. These data may help explain the glucose imbalance and insulin resistance that is characteristic of acromegaly.     

Binding characteristics of a biologically active variant of human growth hormone (20K) to growth hormone and lactogen receptors

The dose-dependent displacement characteristics of a biologically active 20,000 molecular weight human pituitary growth hormone variant (20K) and of human growth hormone (hGH) were compared using hGH liver plasma membrane receptors both from 20-30 day pregnant rabbits and from normal female rats and also using mammary gland plasma membrane receptors from 5-6 day lactating rabbits. Different preparations of 20K were found to be only 3-20% as potent as hGH when compared at the dose necessary to cause 50% displacement of (125I)iodo-hGH from liver receptor and was 22-53% as effective as hGH in the mammary receptor assay system. These findings suggest that 20K and hGH may have separate receptors or that the binding characteristics of the two hormones may be quite different.     

Disappearance rates of plasma growth hormone after intravenous somatostatin in renal and liver disease.

Plasma immunoreactive growth hormone (hGH) was measured before, during and after the administration of intravenous somatostatin to 3 patients with chronic renal failure and 4 with severe liver disease who had elevation of basal hGH. During somatostatin infusion, the hGH levels declined acutely in a linear fashion when log hGH was plotted against time. Rather surprisingly, the plasma hGH half disappearance time (t 1/2) was 27 min and 18 min in liver and renal disease respectively. These values do not differ from data obtained on normal subjects using exogenous hGH, labelled or unlabelled. Control data on normal subjects using this technique are not available as it was not possible to measure subnormal levels of plasma hGH with the required precision. It is possible that our findings of plasma hGH T 1/2 in liver and renal disease within the normal range reported using exogenous hGH might suggest that high levels of plasma hGH found in these two diseases are primarily caused by hypersecretion rather than impaired clearance.     

Receptor-active growth hormone in Laron dwarfism.

The hepatic radioreceptor assay for hGH has been applied to the detection of hGH in the sera of patients with high growth hormone dwarfism (Laron dwarfism). Substantial quantities of receptor-active hGH were found in the sera of all 7 patients studied. In one patient, arginine infusion elicited a prompt increase in both immunoactive and receptor-active hGH. These observations suggest that circulating hGH in Laron dwarfism is biologically active and support the concept that the disease may be caused by a generalized defect in hGH receptors.     

Reversible hypothyroidism in growth hormone-deficient children treated with human growth hormone.

Six children with human growth hormone (hGH) deficiency became hypothyroid during the course of their therapy with hGH. This was accompanied by a decreasing growth rate, clinical symptoms of hypothyroidism and decreased serum T4 concentrations. Three of the 6 patients returned to the euthyroid state, both clinically and biochemically, with cessation of hGH therapy, and reinstitution of hGH precipitated hypothyroidism again in 2 of the three. The patients who remained hypothyroid have evidence of multiple pituitary trophic hormone deficiencies while those who reverted to euthyroidism appear to have isolated hGH deficiency. Evaluation of thyroid function while on hGH showed low T4, free T4 and T3 concentrations. The serum thyrotropin (TSH) response to thyrotropin-releasing hormone (TRH) was absent or markedly blunted in 4 of 6 patients while receiving long-term hGH therapy but was normal or exaggerated in all patients when tested before or after only 5 days of hGH therapy. These data indicate that exogenous hGH results in an inhibition of the TSH response to TRH. The mechanism of this inhibition is unclear, but we postulate that it may be mediated by somatostatin secretion in response to pulse doses of hGH.