Lack of prostaglandin involvement in the mitogenic effect of TSH on canine thyroid cells in primary culture

The production of prostaglandin E₂ (PGE₂) by cultured dog thyroid cells was high in a serum-containing medium and low in a serum-free, completely defined medium. Thyrotropin (TSH) and epidermal growth factor (EGF), two mitogenic factors for these cells, did not stimulate PGE₂ release. Indomethacin, at a concentration which completely inhibited PGE₂ production, had no effect on thyroid cell multiplication and DNA synthesis stimulated by TSH and EGF. It is concluded that cyclooxygenase products are not involved in the proliferation of canine thyroid cells and its control by TSH.     

Prostacyclin stimulates the adenylate cyclase system of human thyroid tissue

Since Prostacyclin (PGI₂) is a major product of arachidonic acid metabolism in the human thyroid, we have studied the effects of PGI₂ on cAMP accumulation in human thyroid slices and cultured thyrocytes. In both systems, PGI₂ caused a dose- and time-dependent increase of cAMP accumulation with higher potency and efficacy than PGE₂. Two optically active isomers of 5,6-dihydro-PGI₂, i.e. stable synthetic analogs of PGI₂, had qualitatively similar effects to PGI₂. The relative potency ratio between the α- and β- isomer as well as their potency compared to PGI₂ were substantially similar to their potency in inhibiting human platelet aggregation. In thyroid slices, PGI₂ and its stable analogs had a greater than TSH in causing cAMP accumulation; however, in contrast to TSH, this effect was not associated with increased iodothyronine release except at maximal PGI₂ concentrations. TSH had no detectable effect on thyroidal PGI₂ synthesis and release. In cultured thyrocytes the effects of PGI₂ and its stable analogs were considerably less than those obtained with TSH and required higher concentrations. Such a discrepancy was not found in the case of PGE₂. These findings suggest the existence of a specific PGI₂-responsive adenylate cyclase system in human significance.     

In vitro effects of arachidonic acid and of inhibitors of its metabolism on the dog thyroid gland

Incubation of dog thyroid tissue with arachidonic acid (10 to 200 μM) led to the following events:

- low conversion to prostaglandins E₂ and F_2α: 0.07% and 0.02% per hour and 100 mg tissue, respectively

- inhibition of the stimulatory effect of low concentrations of TSH on thyroid secretion: the secretory effect of supra-maximal concentrations of TSH and of dB-cAMP was unaffected

- inhibition of the cyclic AMP accumulation induced by TSH: this effect was inhibited neither by indomethacin nor by ETYA; cyclic AMP accumulation in response to cholera toxin or PGE₁ was unaffected

- no effect on cyclic GMP level

- stimulation of thyroid proteins iodination.

ETYA, but not indomethacin, depressed the iodination of thyroid proteins in resting and stimulated tissue. These data show that arachidonic acid-or a metabolite-can modulate thyroid responsiveness to TSH and suggest that lipoxygenase-products of arachidonic acid metabolism could be involved in thyroid proteins iodination.     

Effects of thyrotropin and N6,O2′-dibutyryl cyclic 3′,5′-adenosine monophosphate on prostaglandin levels in thyroid

We have shown that TSH increases PG levels in isolated bovine thyroid cells. We now report that TSH also increases PG levels in rat and mouse thyroid, and that these effects may be mediated via cyclic AMP. PG and cyclic AMP levels in intact rat and mouse thyroid lobes were measured by radioimmunoassay. During 60-min incubations at 37°C, 25 mU/ml TSH effected a 75–83% increase in PGE₁ and PGF_2α ”equivalents“ in rat thyroid; parallel measurements of endogenous cyclic AMP in these intact thyroid lobes revealed that maximal TSH-induced increase in cyclic AMP also required 60-min incubations. In mouse thyroid, 5 mU/ml TSH increased PGE₁ and PGF_2α levels 38–82% above basal; this TSH effect was evident within 15 min of incubation, thus mimicking the time-course of TSH-induced increase in mouse thyroid cyclic AMP. Exogenous DBcAMP, 0.5 to 3 mM, effected a dose-related increase in mouse thyroid PG levels. The stimulatory effects of both TSH and DBcAMP on mouse thyroid PG levels were abolished by aspirin and indomethacin. These studies suggest that TSH-induced increase in endogenous PG levels in thyroid may be mediated by cyclic AMP.     

Stimulation by thyrotropin of horse thyroid plasma membranes adenylate cyclase: evidence of cooperativity

Horse thyroid plasma membranes were prepared by partition in an aqueous two phases system. The membrane fraction was enriched in adenylate cyclase and only slightly contaminated with mitochondria and lysosomes. Adenylate cyclase activity was stimulated by TSH and PGE₁. The TSH stimulatory effect was nearly immediate and occured in the same range of concentrations that activates intact cell metabolism. It was potentiated by GTP and ITP. A quantitative analysis of the data suggests that activation of thyroid adenylate cyclase by TSH is a cooperative process.     

Enhancement of human neutrophil migration by prostaglandin E2

Physiologic concentrations of prostaglandin E₂ but not A₁ or F₁ α enhance human neutrophil migration in response to a chemotactic stimulus while higher concentrations decrease the response. Enhanced migration requires the presence of PGE₂ and is not observed when PMNs are preincubated with optimal doses of PGE₂. Furthermore, maximal enhancement is only observed when low concentrations of chemotactic factor are utilized to stimulate PMNs. Experiments indicate that PGE₂ acts to enhance the chemokinetic response of human PMNs to chemotactic factors. These data support a role for physiologic concentrations of PGE₂ in the control of PMN function at an inflammatory site.     

Effects of PGE1 or PGE2 on luteal function in pseudopregnant rats

Effects of PGE₁ or PGE₂ on luteal function were studied in 163 pseudopregnant rats. PGE₁ (10, 100, or 300μg) given intrauterine every 6 hr did not shorten pseudopregnancy (P < 0.05), however, the same doses of PGE₂ given intrauterine every 6 hr advanced luteolysis (P < 0.05). PGE₁ (100 or 300μg) given every 4 hr intramuscular maintained levels of progesterone in peripheral blood above controls (P < 0.05) while 100 or 300μg of PGE₂ hastened the decline in progesterone (P < 0.05). The antiluteolytic effect of PGE₁ was not via an inhibition of PGF secretion (P < 0.05) by the uterus or by induction of ovulation in treated animals. Moreover, PGE₁ (100, 200, or 500μg) given intramuscular every 4 hr from day 4 of pseudopregnancy until the next proestrus delayed luteal regression around 3 days (P < 0.05). PGE₂ at doses of 100, 200, or 500μg every 4 hr given intramuscular consistently shortened pseudopregnancy (P < 0.05). Lower doses were without effect (P < 0.05). Based on the above data it is concluded that PGE₂ is consistently luteolytic whereas PGE₁ is not luteolytic in pseudopregnant rats and that PGE₁ may be an antiluteolysin.     

Functional activity of thyroid gland in male rats with acute and mild streptozotocin diabetes

Type 1 diabetes mellitus (DM1) and dysfunction of the thyroid gland (TG) are the most common endocrine diseases, which are interrelated. However, the molecular mechanisms of thyroid dysfunction in DM1 and the role of adenylyl cyclase signaling system (ACSS) in this process remain poorly understood. Typically for studying etiology and pathogenesis of thyroid diseases in DM1 the models of acute DM1 induced by high doses of streptozotocin (STZ) are used. At the same time, a suitable model for this purpose is the model of mild DM1 initiated by moderate doses of STZ, which more closely resembles human DM1. The aim of this study was a comparative study of the functional state of the thyroid gland in rats with 30-day acute DM1 induced by injection of STZ at a dose of 65 mg/kg, and in rats with 30- and 210-day mild DM1 induced by three consecutive injections of STZ at medium doses (30–40 mg/kg). For this purpose in diabetic animals the levels of thyroid hormones and TSH and the functional activity of hormone-sensitive ACSS in membranes isolated from thyroid gland were studied. It was shown that in blood of rats with acute DM1 the levels of fT₄, fT₃, and tT₃ were decreased by 45, 23 and 19%, respectively, while the level of TSH did not change significantly. In rats with the 30-day mild DM1 the concentration of fT₄ was decreased by 32%, while the levels of tT₄, tT₃, and TSH were similar to that in control. In rats with prolonged mild DM1 after 150 and 210 days following the first treatment with STZ the levels of tT₄, fT₄, and tT₃ were significantly reduced, but the concentration of TSH in rats with 210-day mild DM1 was increased by 119%. The results obtained in the study of thyroid status and TSH levels in rats with prolonged mild DM1 are in good agreement with the data obtained in the study of thyroid diseases in patients with DM1. It was found that the AC basal activity in the membranes isolated from the thyroid gland of diabetic rats did not change, except for the rats with the prolonged mild DM1 where this activity was increased by 21%. In all groups of diabetic rats the decrease of AC stimulating effects of GppNHp (10^?5 M) and TSH (10^?8 M) was found, and in the rats with prolonged mild DM1 the AC effect of PACAP-38 (10^?6 M) was also reduced. The decrease of AC effect of TSH varied among different groups of the diabetic animals: in the rats with acute DM1 this effect was reduced by 46% and in the rats with 30- and 210-day mild DM1-by 18 and 34%. Thus, it was concluded that the key cause of the thyroid resistance to TSH under conditions of DM1 is a weakening of the signal transduction generated by TSH via the ACSS.     

The hyperalgesic effects of prostacyclin and prostaglandin E2

Hyperalgesia induced in rat paws or dog knee joints by prostacyclin (PGI₂) and prostaglandin E₂ was measured by a modification of the Randall-Selitto method (1) of by the degree of incapacitation (2). In both species PGI₂ induced an immediate hyperalgesic effect but the effect of PGE₂ had a longer latency. Low doses of PGI₂ caused a short lasting effect but PGE₂, large doses of PGI₂ or successive administration of small doses of PGI₂ caused a long lasting effect.It is suggested that prostacyclin mediates rat paw hyperalgesia induced by carrageenin. The long lasting hyperalgesic effect of PGE₂ and high doses of PGI₂ is possibly an indirect effect caused by stimulation of a sensory nerve sensitising mechanism.     

Plasma concentration of 13,14-dihydro-15-keto-PGE2 (PGEM) after various ways of cervix ripening with PGE2

PGEM concentration was determined radioimmunologically in a non-pregnant woman, in whom PGE₂ was infused intravenously at increasing rates and in women, in whom labor was induced by various methods for local application of PGE₂. There was excellent correlation between the amount of PGE₂ infused intravenously and the levels of PGEM determined in the peripheral plasma. The following methods of local application of PGE₂ were included in the study: 0.4mg PGE₂ gel placed retroamnially by means of a balloon catheter, 0.4 and 0.5mg PGE₂ applied endocervically and 3mg PGE₂ placed intravaginally in form of a single vaginal tablet; also induced was a control-group, where only vaginal examination was performed. Bloods were drawn before, 30 minutes, 1, 2 and 3 hours after PGE₂ administration. Mean levels of PGEM in the maternal peripheral plasma did not change neither within nor between the various groups. It is concluded from the present study, that local application of doses currently used to soften the cervix and/or induce labor at term do not lead to the same PGEM-concentration in the maternal blood as after intravenous infusion of PGE₂ in doses normally used to induce labor.     

Nociceptive Response to Prostaglandins and Analgesic Actions of Aspirin and Morphine

VANE et al.^1–3 have proposed that aspirin and allied antiinflammatory drugs act by inhibiting the production of prostaglandins in the tissues. Because, however, prostaglandins E₁ and E₂ (PGE₁ and PGE₂) had been reported not to elicit pain in human skin at doses inducing inflammation^{4, 5}, Vane did not suggest that the inhibition of prostaglandin production fully explains the analgesic action of aspirin-like drugs. Nonetheless, PGE₁ PGE₂ or PGF_2α irritates pulmonary^{6, 7} or ocular⁸ mucous membrane and, when injected by the subcutaneous or intramuscular route, PGE₂ or PGF_2α causes pain⁹.     

Optimum Recombinant Human Thyrotropin Dose in Patients With Differentiated Thyroid Carcinoma and End-Stage Renal Disease

ObjectiveTo evaluate serum thyrotropin (TSH) concentrations after conventional (0.9 mg) or half-dose (0.45 mg) administration of recombinant human TSH (rhTSH) injections intramuscularly in patients with end-stage renal disease and differentiated thyroid cancer.MethodsIn this case series, we administered 2 doses of 0.9-mg rhTSH or 2 doses of 0.45-mg rhTSH to 3 patients with renal failure and differentiated thyroid cancer who were receiving hemodialysis. Basal serum TSH concentrations were assessed while the patients were taking thyroid hormone therapy. Serum TSH was measured on days 2, 3, 5, 8, 10, 14, and 17 of the study. Thyroglobulin and thyroglobulin antibodies were also measured on days 5 and 7. Patients were asked to report any adverse effects.ResultsPatient 1, who received 2 injections of 0.9- mg rhTSH administered on days 1 and 3, had persistently elevated serum TSH levels for approximately 11 days. Peak serum TSH measured on day 5 was 644 mIU/L. Self-limited diarrhea was the only reported adverse effect. Patients 2 and 3 received 0.45 mg of rhTSH on 2 consecutive days (days 1 and 2), and both exhibited persistently elevated serum TSH levels for 12 days. The peak serum TSH values on day 3 were 402 mIU/L in Patient 2 and 386 mIU/L in Patient 3. No adverse events were observed in these 2 patients. Patient 2 received thyrotropin alfa for injection to confirm disease status. Patient 3 also received a radioiodine dose because of presumed persistent disease.ConclusionHigh serum TSH levels achieved after conventional and half-dose administration of rhTSH suggest that a dose adjustment might be considered in patients with end-stage renal disease. (Endocr Pract. 2008;14: 961-966)     

Specificity of the stimulatory effect of prostaglandins on hormone release in rat anterior pituitary cells in culture

Specificity of the effect of prostaglandins (PGs) on hormone release by the anterior pituitary gland was studied using cells in primary culture. Growth hormone (GH) release is stimulated by all eight PGs studied, PGE₁ and E₂ being 1000-fold more potent than the corresponding PGFs. The release of luteinizing hormone (LH), follicle-stimulating hormone (FSH), and prolactin (PRL) remains unchanged upon addition of PGEs. While the basal release of thyrotropin (TSH) is only slightly stimulated by concentrations of PGEs above 10⁻⁶M, an important potentiation of the stimulatory effect of thyrotropin-releasing hormone on TSH release is observed. The release of GH, TSH and LH is stimulated equally well by PGAs and PGBs at concentrations higher than 10⁻⁶M, 3 × 10⁻⁶M, and 10⁻⁵M, respectively. PGFs do not affect the release of any of the measured pituitary hormones at concentrations below 10⁻⁴M. The stimulation of GH release by PGE₂ can be inhibited by the PG antagonist 7-oxa-13-prostynoic acid, a half-maximal inhibition being found at a concentration of 4 × 10⁻⁵M of the antagonist in the presence of 10⁻⁶M PGE₂. In the presence of somatostatin (10⁻⁸M), the inhibition of GH release cannot be reversed by PGE₂ at concentrations up to 10⁻⁴M. 8-bromo-cyclic AMP-induced GH release is additive with that produced by PGE₂.The present data show that 1) of the five pituitary hormones measured, only GH release is stimulated by prostaglandins at relatively low concentrations, 2) the PGE-induced GH release can be competitively inhibited by 7-oxa-13-prostynoic acid, 3) the inhibition of GH release by somatostatin cannot be reversed by PGE₂ and 4) the PGEs increase the responsiveness of the thyrotrophs to TRH.     

Prostaglandin E2 inhibits the proliferation of human gingival fibroblasts via the EP2 receptor and Epac

Elevated levels of prostaglandins such as PGE₂ in inflamed gingiva play a significant role in the tissue destruction caused by periodontitis, partly by targeting local fibroblasts. Only very few studies have shown that PGE₂ inhibits the proliferation of a gingival fibroblast (GF) cell line, and we expanded this research by using primary human GFs (hGFs) and looking into the mechanisms of the PGE₂ effect. GFs derived from healthy human gingiva were treated with PGE₂ and proliferation was assessed by measuring cell number and DNA synthesis and potential signaling pathways were investigated using selective activators or inhibitors. PGE₂ inhibited the proliferation of hGFs dose‐dependently. The effect was mimicked by forskolin (adenylate cyclase stimulator) and augmented by IBMX (a cAMP‐breakdown inhibitor), pointing to involvement of cAMP. Indeed, PGE₂ and forskolin induced cAMP generation in these cells. Using selective EP receptor agonists we found that the anti‐proliferative effect of PGE₂ is mediated via the EP₂ receptor (which is coupled to adenylate cyclase activation). We also found that the effect of PGE₂ involved activation of Epac (exchange protein directly activated by cAMP), an intracellular cAMP sensor, and not PKA. While serum increased the amount of phospho‐ERK in hGFs by ～300%, PGE₂ decreased it by ～50%. Finally, the PGE₂ effect does not require endogenous production of prostaglandins since it was not abrogated by two COX‐inhibitors. In conclusion, in human gingival fibroblasts PGE₂ activates the EP₂—cAMP—Epac pathway, reducing ERK phosphorylation and inhibiting proliferation. This effect could hamper periodontal healing and provide further insights into the pathogenesis of inflammatory periodontal disease. J. Cell. Biochem. 108: 207–215, 2009. © 2009 Wiley‐Liss, Inc.     

Inotropic effect of PGE1 and PGE2 on isolated rat atria. Influence of adrenergic mechanisms

The effects of a wide range of PGE₁ and PGE₂ concentrations on the isometric developed tension of isolated rat atria beating spontaneously or paced at a fixed rate, were explored. PGE₁ only produced a negative inotropic effect (NIE), whereas PGE₂ elicited a biphasic inotropic action; negative at low concentrations and positive (PIE) at higher ones. Phenoxybenzamine and phentolamine failed to modify either the NIE or the PIE, but subthreshold exogenous norepinephrine abolished the NIE, suggesting a presynaptic inhibitory effect of PGEs on the adrenergic neurotransmitter release. Auricles pretreated with subthreshold norepinephrine react with a PIE to PGE₁, but not to PGE₂. On the contrary in the presence of subthreshold methoxamine the PIE of PGE₂ was increased whereas the action of PGE₁ was not modified.     

Differential Effects of Prostaglandin E1 and Prostaglandin E2 on Growth and Differentiation of Murine Myeloid Leukemic Cell Line,M1

Effects of prostaglandins (PGs) of the E series on growth and differentiation of murine myeloid leukemic cell line M1 were studied. PGE₁, but not PGE₂, inhibited the growth of M1 cells. PGE₂ neither inhibited nor augmented the antiproliferative effect of PGE₁. PGE₁ augmented the differentiation of M1 cells into macrophage-like cells induced by interleukin 6. PGE₂, however, did not exhibit any effect on the differentiation. PGE₁ caused a marked increase in intracellular cAMP level in M1 cells, whereas PGE₂ had no effect. These results indicate that M1 cells are able to respond only to PGE₁. Radiolabeled PGE₁ binding experiments, however, revealed that there was no specific binding in M1 cells, suggesting that the cells express low numbers of receptors or very low affinity receptors specific for PGE₁. Stable agonists of PGI₂, iloprost, cicaprost or carbacyclin, also potently inhibited the growth of M1 cells. These findings suggest that PGE₁ as well as PGI₂ may play a role in the differentiation of monocyte-macrophage lineage cells.     

Potentiation of bradykinin-induced vascular permeability increase by prostaglandin E2 and arachidonic acid in rabbit skin

The activity of prostaglandins (PG) in producing vascular permeability was quantitated by dye extraction method in skin of anaesthetized rabbits. PGE₁ and PGE₂ (0.01–10 μg) produced increase in vascular permeability. Activity was approximately equal to that of histamine (Hist) and $\text{1}\text{20}$ of that of bradykinin (BK) on a weight basis. The activity of PGF_1α and PGF_2α was only $\text{1}\text{20}$ of that of PGE₁ or PGE₂.In spite of the relatively low potency of PGE₁ and PGE₂ in the rabbit, near threshold doses (0.1 or 1 μg) of PGE₂ could potentiate permeability responses to bradykinin (0.1 μg) by 10 or 100-fold, respectively. Equivalent doses (0.1 or 1 μg) of histamine could not potentiate the bradykinin responses. Arachidonic acid (AA) at 1 μg, produced a 10-fold potentiation in the permeability response to bradykinin (0.1 μg). Pretreatment of the rabbits with indomethacin (20 mg/kg, i.p.) reduced the responses of BK (0.1 μg) + AA (1 μg) down to a similar magnitude of those seen with bradykinin alone. However, indomethacin did not block responses to either, BK alone, BK + PGE₂, or BK + Hist. Various doses (1, 10, 100 and 300 μg) of arachidonic acid alone also produced increase in cutaneous vascular permeability, although its potency was only $\text{1}\text{3}$–$\text{1}\text{8}$ of that of PGE₂. This activity of arachidonic acid was attributed in part to its bioconversion to PGE₂, since its activity was significantly reduced by the prostaglandin antagonist, diphloretin phosphate (DPP) (60 mg/kg, i.v.) and by indomethacin (20 mg/kg, i.p.), which blocks conversion of arachidonic acid to prostaglandins. Arachidonic acid may owe some of its permeability increaseing effects to histamine release, since its effects were also reduced by the antihistamine, pyrilamine (2.5 mg/kg, i.v.).     

Uncoordinate regulation of collagenase,stromelysin, and tissue inhibitor of metalloproteinases genes by prostaglandin E2: Selective enhancement of collagenase gene expression in human dermal fibroblasts in culture

The degradative effects of interleukin-1 (IL-1) on the extracellular matrix of connective tissue are mediated primarily by metalloproteinases and prostaglandins. Clinical observations suggest that these effects can be prevented, to some extent, by the use of non-steroidal anti-inflammatory drugs. We have examined the role of prostaglandin E₂ (PGE₂) in IL-1-induced gene expression by human skin fibroblasts in culture. Incubation of confluent fibroblast cultures with varying concentrations (0.01–1.0 μg/ml) of PGE₂ led to a dose-dependent elevation of collagenase mRNA steady-state levels, the promoter activity, and the secretion of the protein, whereas relatively little effect was observed on stromelysin and TIMP gene expression. Exogenous PGE₂ had no additive or synergistic effect with IL-1 on collagenase gene expression. Furthermore, commonly used non-steroidal anti-inflammatory drugs (indomethacin, acetyl salicylic acid and ibuprofen), at doses which block prostaglandin synthesis in cultured fibroblasts, failed to counteract IL-1-induced collagenase and stromelysin gene expression, nor did they affect TIMP expression. Although the effects of PGE₂ did not potentiate those of IL-1 on collagenase gene expression in vitro, one could speculate that massive production of PGE₂ by connective tissue cells in vivo in response to inflammatory mediators such as IL-1 or tumor necrosis factor-α, could lead to sustained expression of collagenase in connective tissue cells after clearance of the growth factors.     

Marked inhibition of gastric secretion by two prostaglandin analogs given orally to man

Two prostaglandin analogs, 15(S)-15-methyl PGE₂, methyl ester, and 16, 16-dimethyl PGE₂ were administered to human volunteers for their possible effect in inhibiting gastric secretion. Both analogs, given orally, inhibited gastric secretion stimulated by pentagastrin, and the effect was dose dependent. The inhibition lasted for more than 4 hours, and no side-effects were noted at the doses used. When given intraduodenally, through a thin tube swallowed the night before, 15($\text{S}$)-15-methyl PGE₂, methyl ester was more active than 16, 16-dimethyl PGE₂. In view of their oral and prolonged activity, these analogs may have clinical potential in the treatment of peptic ulcer.