Presence Of Cortisone Research Articles

Role of Calcitriol and Cortisol on Human Adipocyte Proliferation and Oxidative and Inflammatory Stress: A Microarray Study

Dietary calcium inhibits adiposity, and a key underlying mechanism is suppression of calcitriol, which modulates Ca²⁺ signaling and mitochondrial uncoupling in adipocytes. We demonstrated that calcitriol directly regulates adipocyte 11β-HSD-1 expression and cortisol production in human adipocytes in vitro and dietary calcium inhibits visceral adipose tissue 11β-HSD-1 expression in mice, indicating an interaction of calcitriol and cortisol in obesity. Consequently, we have evaluated the gene expression profile of human subcutaneous adipocytes treated with calcitriol and/or cortisone. Data analysis demonstrated significant calcitriol modulation of gene expression toward inhibition of the adipocyte apoptosis (e.g., VEGF and STC-2) and promotion of adipocyte proliferation (e.g., IGF-1 and IGF-1R). Calcitriol also up-regulated oxidative stress and inflammatory genes such as NOX-4 and TLR-3. The calcitriol/cortisone combination resulted in significant additional up-regulation of 11β-HSD-1 and down-regulation of adiponectin expression, while cortisone exerted little independent effect in the absence of calcitriol. Overall, calcitriol stimulated a pattern of adipocyte gene expression which favored adipocyte proliferation, oxidative and inflammatory stress and visceral adiposity, and these effects were amplified in the presence of cortisone; however, this conclusion must be tempered by the adipocyte source (subcutaneous) and requires confirmation in visceral adipocytes.

Microarray analysis of the effects of calcitriol and cortisone on human adipocyte gene expression

Dietary calcium inhibits adiposity in both mice and humans. A key underlying mechanism is suppression of calcitriol, which modulates Ca2+ signaling and mitochondrial uncoupling in adipocytes. We also demonstrated that calcitriol directly regulates adipocyte 11β-HSD 1 expression and cortisol production in human adipocytes in vitro and dietary calcium inhibits visceral adipose tissue 11β-HSD 1 expression in mice, indicating an interaction of calcitriol and cortisol in obesity. Consequently, we have evaluated the gene expression profile of human adipocytes treated with calcitriol and/or cortisone for 48 h. Analysis of the Affymetrix microarray data demonstrated significant calcitriol modulation of gene expression toward inhibition of the adipocyte apoptosis (e.g., VEGF and STC 2) and promotion of adipocyte proliferation (e.g., IGF 1 and IGF 1R). Calcitriol also upregulated genes involved in oxidative stress and inflammation such as NOX 4 and TLR-3. The calcitriol/cortisone combination resulted in significant additional upregulation of 11β-HSD 1 and downregulation of adiponectin expression, while cortisone exerted little independent effect in the absence of calcitriol. Overall, calcitriol stimulated a pattern of adipocyte gene expression which favored adipocyte proliferation, oxidative and inflammatory stress and visceral adiposity, and these effects were amplified in the presence of cortisone.

1, 25-Dihydroxyvitamin D3Modulation of Adipocyte Glucocorticoid Function

1,25-Dihydroxyvitamin D3 dose dependently increases intracellular calcium in human adipocytes. We have demonstrated that suppression of circulating 1,25-dihydroxyvitamin D3 levels by increasing dietary calcium reduces adipocyte intracellular calcium and reduces adiposity in both humans and rodents, with preferential loss of trunk fat. Autocrine production of cortisol by adipocytes of mice overexpressing 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD 1) in adipose tissue increases visceral adiposity, whereas knockout of 11beta-HSD 1 appears to attenuate truncal obesity. Accordingly, our objective was to investigate the role of 1,25-dihydroxyvitamin D3 in the modulation of adipocyte glucocorticoid metabolism. We examined the effect of 1,25-dihydroxyvitamin D3 or angiotensin II on cortisol production and expression using real-time reverse transcriptase-polymerase chain reaction of 11beta-HSD 1, angiotensin II receptor type 1 (AT1), and AT2 receptor in human adipocytes. Adipocytes produced negligible cortisol in the absence of substrate (cortisone). In the presence of cortisone (1 to 10 nM), there was significant cortisol production, which was dose dependently augmented (2- to 6-fold, p < 0.001) by 1,25-dihydroxyvitamin D3 (0.1 to 10 nM). 1,25-Dihydroxyvitamin D3 dose dependently increased 11beta-HSD 1 expression up to 2-fold (p < 0.01) in both the presence and absence of cortisone. In contrast, 1,25-dihydroxyvitamin D3 dose dependently decreased adipocyte AT1 expression (by 30% to 50%, p < 0.001) in both the presence and absence of cortisone, suggesting compensatory down-regulation of AT(1). We conclude that 1,25-dihydroxyvitamin D3 directly regulates adipocyte 11beta-HSD 1 expression and, consequently, local cortisol levels and that this may contribute to the preferential loss of visceral adiposity by high-calcium diets.

11β-Hydroxysteroid Dehydrogenase Expression and Glucocorticoid Synthesis Are Directed by a Molecular Switch during Osteoblast Differentiation

11beta-Hydroxysteroid dehydrogenase type 1 (11beta-HSD1) plays an important role in the prereceptor regulation of corticosteroids by locally converting cortisone into active cortisol. To investigate the impact of this mechanism on osteoblast development, we have characterized 11beta-HSD1 activity and regulation in a differentiating human osteoblast cell line (SV-HFO). Continuous treatment with the synthetic glucocorticoid dexamethasone induces differentiation of SV-HFO cells during 21 d of culture. Using this cell system, we showed an inverse relationship between 11beta-HSD1 activity and osteoblast differentiation. 11beta-HSD1 mRNA expression and activity were low and constant in differentiating osteoblasts. However, in the absence of differentiation (no dexamethasone), 11beta-HSD1 mRNA and activity increased strongly from d 12 of culture onward, with a peak around d 19. Promoter reporter studies provided evidence that specific regions of the 11beta-HSD1 gene are involved in this differentiation controlled regulation of the enzyme. Functional implication of these changes in 11beta-HSD1 is shown by the induction of osteoblast differentiation in the presence of cortisone. The current study demonstrates the presence of an intrinsic differentiation-driven molecular switch that controls expression and activity of 11beta-HSD1 and thereby cortisol production by human osteoblasts. This efficient mechanism by which osteoblasts generate cortisol in an autocrine fashion to ensure proper differentiation will help to understand the complex effects of cortisol on bone metabolism.

Stress cracking of polyurethanes by absorbed steroids

Circumferentially stretched polyurethanes, Pellethane 2363-80A and 2363-55D, were studied following exposure to three steroids of differing polarity; cholesterol, cholesteryl acetate and cortisone. Equilibrium uptakes, diffusion kinetics and environmental stress cracking (ESC) were investigated. Cholesterol reached equilibrium uptake by 16 h, whereas cortisone and cholesteryl acetate did not attain equilibrium uptake until 96 h, suggesting different processes. All uptakes were less than 0.6%. The cortisone diffusion curves showed an inflection at 36 h suggesting some sort of structural change in 55D. The glass transition temperature, T g , of the soft segment as a function of exposure time to cortisone and water shows a steady increase with cortisone content until after 36 h at which time the T g falls almost to the value for water equilibrated samples. This shows that cortisone is able to affect the mobility of soft segments. The relationship between circumferential strain, steroid uptake and ESC was investigated. Samples were stretched at 50% increments from 0 to 250%; after 12days exposure steroid uptakes were determined and the samples examined with SEM. Cholesterol promoted ESC in 80A and 55D at strains of 200 and 150%, respectively. Pellethane 55D was resistant to ESC in the presence of cortisone at all strains, while 80A cracked only at strains above 200%. The equilibrium uptake of cholesterol acetate was most sensitive to strain, particularly for 55D. Cholesterol acetate was the most effective ESC agent, cracking both 80A and 55D after 12days exposure with an uptake under 0.25% and a strain of only 50%.

Culture of thymic epithelial cells from mice and agerelated studies on the growing cells

Culture of epithelial cells from the thymus of mice was achieved in a medium modified to favor epithelial growth while inhibiting proliferation of fibroblasts. Epithelial cells were identified by the presence of desmosomes in electron microscopic preparations and by antibody to intermediate filaments containing keratin. Morphologically, the cells thus positively identified displayed two main patterns: carpets of large flat cells resembling paving stones which are confluent along the length of their membranes, and networks of cells interconnected by long cytoplasmic processes. These two types of cells were dominant in cultures derived from mice of all ages tested (newborn to nine months) but the relative proportion of each type appeared to change with the age of the donor mice and also with the concentrations of cortisone in the culture medium. Autoradiography revealed that the cultured cells were dividing, and that (in the presence of cortisone) the rate of DNA synthesis was decreased in a portion of the epithelial cells derived from mice in which thymic involution was already underway.

Angiogenesis Inhibition and Tumor Regression Caused by Heparin or a Heparin Fragment in the Presence of Cortisone

Heparin or a heparin fragment administered with cortisone inhibited angiogenesis, caused regression of large tumor masses, and prevented metastases. Oral administration of heparin resulted in the release of non-anticoagulant heparin fragments in the serum which, in the presence of cortisone, had similar anti-angiogenic and antitumor effects. Of all the heparin fragments tested, the most potent inhibition of angiogenesis in the presence of cortisone was provided by a hexasaccharide with a molecular weight of about 1600.

Stimulation of human chorionic gonadotropin secretion by glucocorticoids

The effects of glucocorticoids on hormone secretion by human placenta in organ culture were studied. The addition of cortisol resulted in a fourfold increase in human chorionic gonadotropin (hCG) secretion over that in untreated cultures after 144 hours' incubation (P < 0.05), and a twofold increase in hCG was observed in the presence of cortisone (P < 0.01). Dexamethasone stimulated hCG secretion in a dose-response manner (r = 0.9542; P < 0.01). Progesterone, which suppresses hCG under these conditions, decreased the cortisol-enhanced secretion of hCG (r = −0.9794; P < 0.01). No change in the secretion of human chorionic somatomammotropin was observed, but glucocorticoids increased heat-stable alkaline phosphatase activity (P < 0.001). The physiologic significance of glucocorticoid effects on placental hormone synthesis is ducussed.

The effects of cortisone treatment on the protein turnover of the soleus muscle after immobilization.

The glucocorticoid hormones appear to play an important role in mobilizing body proteins to facilitate gluconeogenesis in response to falling concentrations of plasma glucose. It has previously been shown (Goldberg & Goodman, 1969) that denervated skeletal muscle is more susceptible to the catabolic effects of cortisone. However, denervation is a rather drastic means by which to render a muscle inactive. In addition t o the deprivation of possible neurochemical influences, the presence of spontaneous electrical activity (Thesleff, 1963) and possible increased passive stretching of the tissue (Sola et al., 1973; Goldspink, 1978) make the denervated preparation a complicated system for studying inactivity. In the present investigation we have rendered the soleus muscle of young rats (approx. 40g pre-operatively, CD strain, Charles River U.K. Ltd.) inactive without depriving the muscle of its innervation. This was achieved by placing a plaster-of-Paris cast around the ankle joint of one hind limb. With thefoot restrained in the extended position the soleus muscle was immobilized in a fully shortened and unstretched state (Goldspink, 1977). We then examined changes in the size and protein turnover of the soleus muscle after 4 days of such immobilization. The changes in these NaCI-injected animals were compared with those in similar animals that had received two injections of cortisone acetate (0.25mg/day per lOOg body wt.) on each of the 4 days of immobilization. With this less complicated system, we re-examined the question of whether contractile activity protects muscle from the actions of catabolic hormones. In addition we have attempted to elucidate some of the mechanisms involved in such a response. The excess cortisone did not affect the growth rate of these young rats, as evidenced by similar increases in the body weights (approx. 2g/day) of both NaC1-treated and hormone-treated animals. Immobilization of the soleus muscle caused a significant decrease in the size of the tissue, compared with non-restrained controls (Table 1). These findings are in excellent agreement with a previous study (Goldspink, 1977), where it was shown that the immobilized soleus in such young rats undergoes a true atrophy and not simply decreased rates of growth. Since protein remained 16-17% of the muscle mass, changes in total protein closely paralleled the changes in muscle weight after immobilization (Table 1) . The presence of cortisone did not, however, appear t o have any additional influence on the weight of the immobilized muscle, nor indeed on the internal, non-restrained controls (Table 1). However, after cortisone treatment the protein composition fell to only 12% of the wet weight. Hence in the presence of excess cortisone, there was some fluid retention (Forsham, 1968) contributing to the unchanged wet weight of the immobilized tissue. This latter finding was not, however, true in the case of the control tissue on exposure to the hormone. So although cortisone had not affected the wet weight of the immobilized muscle, it had caused a greater net loss of protein (Table 1). Average rates of protein synthesis and protein breakdown were measured in vitro by the slightly modified method (Goldspink, 1978) of Fulks et al. (1975) to explain the changes in protein concentrations after immobilization, in the presence or absence of cortisone. Immobilization in the absence of excess hormone caused both a significant decrease in protein synthesis and an increase in protein degradation (Table 1). Thesecomplemen-

Factors involved in the uptake of corticosterone by rat liver cells

Isolated rat liver cells take up corticosterone rapidly; the initial rates increase with increasing temperature. A plot of the initial rates against the concentration of corticosterone indicated the presence of saturable and nonsaturable uptake systems. The Eadie-Hofstee plot showed the presence of two saturable and one nonsaturable uptake components. The apparent K t values of the saturable systems were 64 ± 40 nM ( n =3) and 1085 ± 313 nM ( n =12). The nonsaturable system, probably diffusion, contributed 12% to the total uptake between 15 and 72 nM corticosterone, the physiological concentration of the free corticosterone in rat serum. Metabolic inhibitors did not influence the uptake of corticosterone. N-Ethylmaleimide, 1-fluor-2,4-dinitrobenzene and sodium ethyl mercurithiosalicylate (1 mM each) decreased the uptake by 40%. Iodoacetate did not have any influence. Treatment of cells with phospholipase A inhibited the uptake 35–45%. In the presence of cortisone, cortisol, dexamethasone, aldosteron, testosterone, estradiol-17β and estrone (2 μM each) the uptake decreased 30–50%. The presence of serum proteins in the external medium inhibits the uptake of corticosterone. These results suggest that corticosterone is transported into the cell and is accumulated. Only the free hormone is available for uptake which in turn may be regulated by protein and lipid components in the plasma membrane of the liver cell.

The effect of 1alpha-hydroxyvitamin D3 and cortisone on the development of chick duodenal alkaline phosphatase in organ culture.

The development of alkaline phosphatase influenced by 1 alpha-OH-D3 (a synthetic active form of vitamin D3) and cortisone was studied in chick duodenal organ cultures. The administration of cortisone to the embryo in ovo on the 14th day of incubation resulted in a precocious increase in alkaline phosphatase after six days (20-day embryo). When duodena from 14-, 18- and 20-day embryos were cultured in the presence of cortisone, there was no significant enhancement of alkaline phosphatase activity except for a marginal effect observed in the 18-day duodenum. On the other hand, the alkaline phosphatase activity in cultured duodena from 20-day chick embryos was significantly stimulated by the addition of 1 alpha-OH-D3. The effects of cortisone and 1 alpha-OHD-3 were not additive. The activity of maltase, another intestinal enzyme, was not influenced by 1 alpha-OH-D3. Studies on inactivation of alkaline phosphatase by EDTA suggest that the observed increase in alkaline phosphatase activity induced by the administration of 1 alpha-OH-D in vitro may be related to the qualitative changes in the enzyme that take place during development in vivo.

Experimental Infection of Rabbits and Monkeys with Herpesvirus cuniculi

Summary Intradermal inoculation of rabbits with Herpesvirus cuniculi resulted in local erythematous lesions. Intracerebral inoculation produced histologically detectable inflammatory lesions in the nervous system of some rabbits and virus could be reisolated from the brain of one third of the animals, but no clinical symptoms were observed. Monkeys inoculated by various routes, in the presence of cortisone, failed to show any clinical symptoms or significant histological lesions attributable to the virus.

Colloid Concentration and Albumin Synthesis.

s1 November 1965Colloid Concentration and Albumin Synthesis.Marcus A. Rothschild, M.D., F.A.C.P., Murray Oratz, Ph.D., Sidney S. Schreiber, M.D., F.A.C.P.Marcus A. Rothschild, M.D., F.A.C.P.Search for more papers by this author, Murray Oratz, Ph.D.Search for more papers by this author, Sidney S. Schreiber, M.D., F.A.C.P.Search for more papers by this authorAuthor, Article, and Disclosure Informationhttps://doi.org/10.7326/0003-4819-63-5-915_2 SectionsAboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinkedInRedditEmail ExcerptIn the presence of hypergammaglobulinemia, and after the administration of gamma globulin and dextran, albumin levels decrease due to a decrease in albumin production. The extravascular albumin pool is conserved. However, in the presence of cortisone and following the administration of thyroid, albumin synthesis increases with a decrease in the extravascular albumin pool. These observations coupled with the oft observed reciprocal changes in albumin and globulin levels in the serum suggested the presence of a colloid osmotic regulatory system affecting control of albumin production and also that such a system might be located in an extravascular site. Albumin metabolism and... This content is PDF only. To continue reading please click on the PDF icon. Author, Article, and Disclosure InformationAffiliations: New York, New York PreviousarticleNextarticle Advertisement FiguresReferencesRelatedDetails Metrics 1 November 1965Volume 63, Issue 5Page: 915-915KeywordsAlbuminsAttentionDextranGlobulinsHypergammaglobulinemiaSerum albuminThyroid Issue Published: 1 November 1965 PDF downloadLoading ...

Sulfate fixation by embryonic chick lung in tissue culture.

SummaryChick embryonic lung tissue cultivated in vitro in the presence of sulfate labeled with S35 fixed sulfate in the protein of the tissue. Lung tissue which was not proliferating did not incorporate sulfate. Cultures of lung older than 2 weeks had a greatly reduced ability to fix sulfate. The presence of cortisone in the medium did not alter the incorporation of sulfate into the tissue.

Influenza Virus in Allantoic Sac Tissue Culture

Summary The use of allantoic sacs in suspended cell tissue cultures for the growth of influenza viruses has been investigated. Influenza A and B viruses grow readily in such cultures, with either Hanks-Simms' solution or Tyrode's solution as diluents. In the presence of the former solution, viral titers (influenza A) in tissue cultures reach a maximum slightly more slowly and decline more slowly than in Tyrode's solution. Step-wise curves of multiplication of virus are readily demonstrable without recourse to dilution or interference techniques. A complete cycle of multiplication of influenza A virus in this system requires approximately 18 hours, and increases the titer about 100-fold. Frequent changes of the nutrient fluid and the addition of embryo extracts do not increase the capacity of the tissue to support viral growth. The titer of hemagglutinin in such cultures is affected by the amount of tissue in the cultures and by the size of the inoculum. Quantitative study of the pentose and desozy-pentose nucleic acids of the cultures showed a progressive decrease in the former during the incubation of a tissue culture and insignificant alterations in the latter. Viral growth in such cultures may have increased the rate of decline of PNA more rapidly than in uninfected cultures, but had no consistent effect on DNA. Tyrode's and Hanks-Simms' solutions exerted similar effects on nucleic acid concentrations. The incorporation of radioactive phosphorus into nucleic acids of both types by the tissue cultures was rapid. There were no consistent effects on the pattern of uptake of phosphate by the tissue cultures as a consequence of viral multiplication in the cultures. The data indicate that multiplication of tissue cells was not prominent in these cultures, and that a net degradative effect predominated. Uptake of radioactive phosphorus offers a more reliable indication of sufficient viability of tissue cells to support viral growth in suspended cell cultures than do increases in nucleic acid concentration. In the presence of cortisone, both infected and uninfected cultures were characterized by the appearance of numerous cystic blebs of tissue, arising from the culture fragments of allantoic sac. Such outpouchings were less common in control cultures. No consistent effect of cortisone on the concentrations of pentose nucleic acid (PNA) or of desoxypentose nucleic acid (DNA) in the cultured tissues was demonstrable. The rate of incorporation of radioactive phosphate into PNA was slightly accelerated both in infected and uninfected cultures, but incorporation of phosphate into DNA was not significantly altered by the hormone. Cortisone did not affect viral multiplication significantly.

Epinephrine eosinopenia in surgically induced Addison's disease.

Abstract Five patients with carcinoma of the prostate had both their testes and adrenal glands removed, and were maintained in a eu-adrenal state by therapy with oral cortisone. Direct eosinophil counts were made before and four hours after injection of epinephrine, saline, and ACTH. There was a significant fall in eosinophils after injection of epinephrine, but ACTH or saline had no effect on the eosinophil level. The data indicate that epinephrine eosinopenia occurs in the presence of cortisone, despite complete interruption of the pituitary-adrenal system. Because of this finding, and other observations in the literature, the four-hour eosinophil cell response to parenteral epinephrine should not be used to test the function or integrity of the pituitary-adrenal system in man.