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1.
Am J Physiol Gastrointest Liver Physiol ; 286(1): G95-G101, 2004 Jan.
Article in English | MEDLINE | ID: mdl-12881226

ABSTRACT

Luminal acidification provides the strongest physiological stimulus for duodenal HCO3- secretion. Various neurohumoral mechanisms are believed to play a role in acid-stimulated HCO3- secretion. Previous studies in the rat and human duodenum have shown that guanylin and Escherichia coli heat-stable toxin, both ligands of the transmembrane guanylyl cyclase receptor [guanylate cyclase C (GC-C)], are potent stimulators for duodenal HCO3- secretion. We postulated that the GC-C receptor plays an important role in acid-stimulated HCO3- secretion. In vivo perfusion studies performed in wild-type (WT) and GC-C knockout (KO) mice indicated that acid-stimulated duodenal HCO3- secretion was significantly decreased in the GC-C KO animals compared with the WT counterparts. Pretreatment with PD-98059, an MEK inhibitor, resulted in attenuation of duodenal HCO3- secretion in response to acid stimulation in the WT mice with no further effect in the KO mice. In vitro cGMP generation studies demonstrated a significant and comparable increase in cGMP levels on acid exposure in the duodenum of both WT and KO mice. In addition, a rapid, time-dependent phosphorylation of ERK was observed with acid exposure in the duodenum of WT mice, whereas a marked attenuation in ERK phosphorylation was observed in the KO animals despite equivalent levels of ERK in both groups of animals. On the basis of these studies, we conclude that transmembrane GC-C is a key mediator of acid-stimulated duodenal HCO3- secretion. Furthermore, ERK phosphorylation may be an important intracellular mediator of duodenal HCO3- secretion.


Subject(s)
Bicarbonates/metabolism , Duodenum/enzymology , Duodenum/metabolism , Guanylate Cyclase/physiology , Intestinal Mucosa/enzymology , Intestinal Mucosa/metabolism , Receptors, Peptide/physiology , Acids , Animals , Blotting, Western , Cyclic AMP/biosynthesis , Enzyme Inhibitors/pharmacology , Female , Guanylate Cyclase/genetics , Hydrochloric Acid/pharmacology , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Mitogen-Activated Protein Kinases/antagonists & inhibitors , Mitogen-Activated Protein Kinases/metabolism , Phenotype , Phosphorylation , Receptors, Enterotoxin , Receptors, Guanylate Cyclase-Coupled , Receptors, Peptide/genetics , Respiratory Mechanics/drug effects
2.
J Med Chem ; 46(13): 2683-96, 2003 Jun 19.
Article in English | MEDLINE | ID: mdl-12801232

ABSTRACT

Previous data have shown that RXR-selective agonists (e.g., 3 and 4) are insulin sensitizers in rodent models of non-insulin-dependent diabetes mellitus (NIDDM). Unfortunately, they also produce dramatic increases in triglycerides and profound suppression of the thyroid hormone axis. Here we describe the design and synthesis of new RXR modulators that retain the insulin-sensitizing activity of RXR agonists but produce substantially reduced side effects. These molecules bind selectively and with high affinity to RXR and, unlike RXR agonists, do not activate RXR homodimers. To further evaluate the antidiabetic activity of these RXR modulators, we have designed a concise and systematic structure-activity relationship around the 2E,4E,6Z-7-aryl-3-methylocta-2,4,6-trienoic acid scaffold. Selected compounds have been evaluated using insulin-resistant rodents (db/db mice) to characterize effects on glucose homeostasis. Our studies demonstrate the effectiveness of RXR modulators in lowering plasma glucose in the db/db mouse model.


Subject(s)
Caprylates/chemical synthesis , Diabetes Mellitus, Type 2/blood , Hypoglycemic Agents/chemical synthesis , Receptors, Retinoic Acid/drug effects , Transcription Factors/drug effects , Animals , Blood Glucose/analysis , Caprylates/chemistry , Caprylates/pharmacology , Hypoglycemic Agents/chemistry , Hypoglycemic Agents/pharmacology , Insulin Resistance , Male , Mice , Radioligand Assay , Rats , Rats, Sprague-Dawley , Receptors, Retinoic Acid/metabolism , Retinoid X Receptors , Structure-Activity Relationship , Transcription Factors/metabolism
3.
Metabolism ; 50(8): 963-71, 2001 Aug.
Article in English | MEDLINE | ID: mdl-11474486

ABSTRACT

Fibrates and thiazolidinediones are used clinically to treat hypertriglyceridemia and hyperglycemia, respectively. Fibrates bind to the peroxisome proliferator-activated receptor (PPAR)-alpha, and thiazolidinediones are ligands of PPAR-gamma. These intracellular receptors form heterodimers with retinoid X receptor to modulate gene transcription. To elucidate the target genes regulated by these compounds, we treated Zucker diabetic fatty rats (ZDF) for 15 days with a PPAR-alpha-specific compound, fenofibrate, a PPAR-gamma-specific ligand, rosiglitazone, and a PPAR-alpha/-gamma coagonist, GW2331, and measured the levels of several messenger RNAs (mRNAs) in liver by real-time polymerase chain reaction. All 3 compounds decreased serum glucose and triglyceride levels. Fenofibrate and GW2331 induced expression of acyl-coenzyme A (CoA) oxidase and enoyl-CoA hydratase and reduced apolipoprotein C-III and phosphoenolpyruvate carboxykinase mRNAs. Rosiglitazone modestly increased apolipoprotein C-III mRNA and had no effect on expression of the other 2 genes in the liver but increased the expression of glucose transporter 4 and phosphoenolpyruvate carboxykinase in adipose tissue. We identified a novel target in liver, mitogen-activated phosphokinase phosphatase 1, whose down-regulation by PPAR-alpha agonists may improve insulin sensitivity in that tissue by prolonging insulin responses. The results of these studies suggest that activation of PPAR-alpha as well as PPAR-gamma in therapy for type 2 diabetes will enhance glucose and triglyceride control by combining actions in hepatic and peripheral tissues.


Subject(s)
Gene Expression Regulation/physiology , Liver/metabolism , Receptors, Cytoplasmic and Nuclear/physiology , Transcription Factors/physiology , Animals , Apolipoprotein C-III , Apolipoproteins C/genetics , Base Sequence , Butyrates/pharmacology , DNA Primers , Diabetes Mellitus, Experimental/genetics , Diabetes Mellitus, Experimental/metabolism , Fenofibrate/pharmacology , Male , Phenylurea Compounds/pharmacology , Phosphoenolpyruvate Carboxykinase (ATP)/genetics , RNA, Messenger/genetics , Rats , Rats, Zucker , Receptors, Cytoplasmic and Nuclear/agonists , Receptors, Cytoplasmic and Nuclear/chemistry , Transcription Factors/agonists , Transcription Factors/chemistry , Triglycerides/blood
4.
Mol Pharmacol ; 59(4): 765-73, 2001 Apr.
Article in English | MEDLINE | ID: mdl-11259621

ABSTRACT

Both retinoid X receptor (RXR)-selective agonists (rexinoids) and thiazolidinediones (TZDs), PPAR (peroxisome proliferator-activated receptor)-gamma-specific ligands, produce insulin sensitization in diabetic rodents. In vitro studies have demonstrated that TZDs mediate their effects via the RXR/PPAR-gamma complex. To determine whether rexinoids lower hyperglycemia by activating the RXR/PPAR-gamma heterodimer in vivo, we compared the effects of a rexinoid (LG100268) and a TZD (rosiglitazone) on gene expression in white adipose tissue, skeletal muscle, and liver of Zucker diabetic fatty rats (ZDFs). In adipose tissue, rosiglitazone decreased tumor necrosis factor-alpha (TNF-alpha) mRNA and induced glucose transporter 4 (GLUT4), muscle carnitine palmitoyl-transferase (MCPT), stearoyl CoA desaturase (SCD1), and fatty acid translocase (CD36). In contrast, LG100268 increased TNF-alpha and had no effect or suppressed the expression of GLUT4, MCPT, SCD1, and CD36. In liver, the rexinoid increased MCPT, SCD1, and CD36 mRNAs, whereas rosiglitazone induced only a small increase in CD36. In skeletal muscle, rosiglitazone and LG100268 have similar effects; both increased SCD1 and CD36 mRNAs. The differences in the pattern of genes induced by the rexinoids and the TZDs in diabetic animals found in these studies suggests that these compounds may have independent and tissue-specific effects on metabolic control in vivo.


Subject(s)
Diabetes Mellitus, Experimental/metabolism , Hypoglycemic Agents/pharmacology , Nicotinic Acids/pharmacology , Tetrahydronaphthalenes/pharmacology , Thiazoles/pharmacology , Thiazolidinediones , Adipose Tissue/drug effects , Adipose Tissue/metabolism , Animals , Blood Glucose/drug effects , Diabetes Mellitus/drug therapy , Diabetes Mellitus/metabolism , Diabetes Mellitus, Experimental/complications , Diabetes Mellitus, Experimental/drug therapy , Disease Models, Animal , Gene Expression/drug effects , Glucose Tolerance Test , Hyperglycemia/blood , Hyperglycemia/drug therapy , Hyperglycemia/etiology , Hyperinsulinism/blood , Hyperinsulinism/drug therapy , Hyperinsulinism/etiology , Liver/drug effects , Liver/metabolism , Male , Muscle, Skeletal/drug effects , Muscle, Skeletal/metabolism , Obesity , RNA, Messenger/analysis , Rats , Rats, Zucker , Receptors, Cytoplasmic and Nuclear/metabolism , Receptors, Retinoic Acid/agonists , Receptors, Retinoic Acid/genetics , Receptors, Retinoic Acid/metabolism , Retinoid X Receptors , Rosiglitazone , Transcription Factors/agonists , Transcription Factors/genetics , Transcription Factors/metabolism
5.
Mol Pharmacol ; 59(2): 170-6, 2001 Feb.
Article in English | MEDLINE | ID: mdl-11160850

ABSTRACT

Hypertriglyceridemia is a frequent complication accompanying the treatment of patients with either retinoids or rexinoids, [retinoid X receptor (RXR)-selective retinoids]. To investigate the cellular and molecular basis for this observation, we have studied the effects of rexinoids on triglyceride metabolism in both normal and diabetic rodents. Administration of a rexinoid such as LG100268 (LG268) to normal or diabetic rats results in a rapid increase in serum triglyceride levels. LG268 has no effect on hepatic triglyceride production but suppresses post-heparin plasma lipoprotein lipase (LPL) activity suggesting that the hypertriglyceridemia results from diminished peripheral processing of plasma very low density lipoproteins particles. Treatment of diabetic rats with rexinoids suppresses skeletal and cardiac muscle but not adipose tissue LPL activity. This effect is independent of changes in LPL mRNA. In C2C12 myocytes, LG268 suppresses the level of cell surface (i.e., heparin-releasable) LPL activity without altering LPL mRNA. This effect is very rapid (t(1/2) = 2 h) and is blocked by the transcriptional inhibitor actinomycin D. These studies demonstrate that RXR ligands can have dramatic effects on the post-translational processing of LPL and suggest that skeletal muscle may be an important target of rexinoid action. In addition, these data underscore that the metabolic consequences of RXR activation are distinct from either retinoic acid receptor or peroxisome proliferator-activated receptor activation.


Subject(s)
Lipoprotein Lipase/metabolism , Nicotinic Acids/pharmacology , Receptors, Retinoic Acid/metabolism , Tetrahydronaphthalenes/pharmacology , Transcription Factors/metabolism , Animals , Cells, Cultured , Heart/drug effects , Hypertriglyceridemia/blood , Hypertriglyceridemia/chemically induced , Lipoprotein Lipase/drug effects , Lipoproteins, VLDL/drug effects , Lipoproteins, VLDL/metabolism , Muscle, Skeletal/drug effects , Muscle, Skeletal/enzymology , Myocardium/enzymology , Myocardium/metabolism , Nicotinic Acids/adverse effects , Rats , Rats, Sprague-Dawley , Rats, Zucker , Receptors, Retinoic Acid/drug effects , Retinoid X Receptors , Retinoids , Tetrahydronaphthalenes/adverse effects , Transcription Factors/drug effects , Triglycerides/blood
6.
Int J Obes Relat Metab Disord ; 24(8): 997-1004, 2000 Aug.
Article in English | MEDLINE | ID: mdl-10951538

ABSTRACT

OBJECTIVE: To investigate whether retinoid X receptor agonists act as insulin sensitizers and compare their effects with that of thiazolidinedione BRL 49653 in obese Zucker rats. DESIGN: In two independent studies, obese Zucker rats were dosed orally once daily for 14 days with one of the following treatments: LG 100268 (20 mg/kg), LG 100324 (20 mg/kg), BRL 49653 (3 mg/kg) or vehicle. MEASUREMENTS: Daily food intake and body weight gain, blood glucose, plasma and pancreatic insulin, whole body glucose disposal (by euglycaemic-hyperinsulinaemic clamp) and tissue glucose utilization. RESULTS: The retinoid X receptor agonists (rexinoids) LG 100268 and LG 100324 caused a reduction in the food intake of obese Zucker rats relative to controls and to rats receiving BRL 49653. The two rexinoids also produced a marked decrease in the body weight gain, whereas the growth rate of rats treated with BRL 49653 tended to increase. Both rexinoids and BRL 49653 reduced the plasma insulin concentration of fed rats. LG 100268 and LG 100324 also significantly lowered blood glucose concentrations after 1 week of treatment. The 5 h fasted plasma insulin concentration was significantly lower in the rexinoid-treated groups and the terminal insulin level (at the end of the clamp) tended to be lower in all treated groups compared with animals given the dosing vehicle. However, pancreatic insulin content was not affected by any of the treatments. Under euglycaemic-hyperinsulinaemic clamp conditions, there were no significant differences in the rate of hepatic glucose output and whole body glucose disposal, except that, in experiment 1, BRL 49653 caused significant increase in the glucose infusion rate and muscle glucose utilization. In experiment 2, a similar glucose infusion rate to the controls was achieved in all treatment groups but the steady-state insulin concentration in the treated animals was only about 50% of that in the control animals, despite the fact that all rats received a similar insulin infusion concentration. This suggests that both the rexinoids and BRL 49653 increased insulin clearance. CONCLUSIONS: Chronic administration of retinoid X receptor agonists LG 100268 and LG 100324 to Zucker fa/fa rats reduces food intake and body weight gain, lowers plasma insulin concentrations while maintaining normoglycaemia, indicating an improvement of insulin sensitivity.


Subject(s)
Hypoglycemic Agents/pharmacology , Insulin Resistance , Nicotinic Acids/pharmacology , Obesity/metabolism , Receptors, Retinoic Acid/agonists , Tetrahydronaphthalenes/pharmacology , Thiazoles/pharmacology , Thiazolidinediones , Transcription Factors/agonists , Analysis of Variance , Animals , Blood Glucose/drug effects , Body Weight/drug effects , Eating/drug effects , Insulin/blood , Male , Rats , Rats, Zucker , Retinoid X Receptors , Rosiglitazone
8.
J Med Chem ; 41(18): 3461-6, 1998 Aug 27.
Article in English | MEDLINE | ID: mdl-9719599

ABSTRACT

A series of 6-aryl-1,2-dihydro-2,2,4-trimethylquinolines was synthesized and tested for functional activity on the human progesterone receptor isoform B (hPR-B) in mammalian (CV-1) cells. The lead compound LG001447 (1,2-dihydro-2,2, 4-trimethyl-6-phenylquinoline) was discovered via directed high throughput screening of a defined chemical library utilizing an hPR-B cotransfection assay. Electron-withdrawing substituents at the meta position of the C(6) aryl group afforded substantial improvements in hPR modulatory activity. Several analogues were able to potently block the effects of progesterone in vitro. Two compounds, 10 (LG120753) and 11 (LG120830) with potencies comparable or equal to the steroidal hPR antagonist onapristone (ZK98,299), were demonstrated to act as antiprogestins in vivo after oral administration to rodents. This is the first disclosure of orally active nonsteroidal antiprogestins.


Subject(s)
Hormone Antagonists , Quinolines , Receptors, Progesterone/antagonists & inhibitors , Administration, Oral , Animals , Cell Line , Cercopithecus , Embryo Implantation/drug effects , Female , Gonanes/pharmacology , Hormone Antagonists/administration & dosage , Hormone Antagonists/chemical synthesis , Hormone Antagonists/chemistry , Hormone Antagonists/pharmacology , Humans , Mice , Pregnancy , Quinolines/administration & dosage , Quinolines/chemical synthesis , Quinolines/chemistry , Quinolines/pharmacology , Receptors, Progesterone/biosynthesis , Structure-Activity Relationship
9.
Gastroenterology ; 113(2): 533-41, 1997 Aug.
Article in English | MEDLINE | ID: mdl-9247473

ABSTRACT

BACKGROUND & AIMS: Duodenal bicarbonate secretion is an important factor in epithelial protection. The role of the cystic fibrosis transmembrane conductance regulator (CFTR) in acid-induced bicarbonate secretion is unknown. The aim of this study was to determine whether CFTR mediates acid-stimulated duodenal epithelial bicarbonate secretion. METHODS: Basal and stimulated bicarbonate secretion was examined in the cystic fibrosis murine model cftrm1UNC, which displays defective CFTR in various organs including chloride transport abnormalities in epithelia. After anesthesia, the proximal duodenum was cannulated and perfused with isotonic saline, and [HCO3-] was determined. RESULTS: Basal bicarbonate secretion was diminished in cystic fibrosis vs. normal mice, 2.8 +/- 0.7 vs. 4.7 +/- 1.7 mumol.cm-1.h-1, respectively (P < 0.001). Luminal acidification failed to elicit a bicarbonate secretory response in cystic fibrosis compared with normal littermates (peak response, 2.3 +/- 0.2 vs. 9.9 +/- 1.5 mumol.cm-1.h-1, respectively; P < 0.01). Prostaglandin E2- and vasoactive intestinal peptide-stimulated bicarbonate secretion were also significantly impaired in cystic fibrosis. Defective bicarbonate secretion in cystic fibrosis genotypes was due to decreased net fluid secretion and [HCO3-]. CONCLUSIONS: Basal and stimulated proximal duodenal bicarbonate secretion may involve a CFTR-mediated transport pathway. It is likely that CFTR, directly or indirectly, has a major functional role in mediating bicarbonate transport in the proximal duodenum.


Subject(s)
Bicarbonates/metabolism , Cystic Fibrosis Transmembrane Conductance Regulator/physiology , Cystic Fibrosis/metabolism , Duodenum/metabolism , Hydrochloric Acid/pharmacology , Animals , Bicarbonates/pharmacokinetics , Biological Transport, Active/physiology , Cystic Fibrosis Transmembrane Conductance Regulator/genetics , Dinoprostone/pharmacology , Disease Models, Animal , Duodenum/drug effects , Duodenum/physiology , Epithelium/drug effects , Epithelium/metabolism , Epithelium/physiology , Female , Genotype , Male , Mice , Mice, Inbred CFTR/genetics , Mice, Inbred CFTR/metabolism , Polymerase Chain Reaction , Vasoactive Intestinal Peptide/pharmacology
10.
Am J Physiol ; 272(4 Pt 1): G872-8, 1997 Apr.
Article in English | MEDLINE | ID: mdl-9142920

ABSTRACT

The role of the cystic fibrosis transmembrane conductance regulator (CFTR) in duodenal alkaline secretion has not been directly examined. The aims of this series of experiments were to determine if CFTR mediates basal and stimulated duodenal epithelial HCO3- secretion. Utilizing the cystic fibrosis murine model (cftr(m1UNC)), we compared normal [CFTR(+/+)] littermates (34-46 days old) with CFTR(-/-) animals (34-39 days old). Anesthesia was induced and maintained with intraperitoneal Hypnorm-midazolam. The proximal duodenum (4-7 mm) was cannulated and perfused with 154 mM NaCl. Either forskolin (10(-6)-10(-4) M) or carbachol (10(-6)-10(-3) M) was perfused intraluminally to activate adenosine 3',5'-cyclic monophosphate (cAMP)- and Ca2+-mediated HCO3- secretion, respectively. Effluent volumes were weighed and HCO3- quantitated by back titration. Basal HCO3- secretion was diminished significantly (P < 0.01) in CFTR(-/-)vs. normal CFTR(+/+) mice (2.8 +/- 0.5 vs. 5.3 +/- 0.4 micromol x cm(-1) x h(-1)). Moreover, in CFTR(-/-) mice, both forskolin- and carbachol-stimulated peak HCO3- secretions were fourfold less compared with those in CFTR(+/+) littermates (3.7 +/- 0.2 vs. 15.6 +/- 2.1 and 4.7 +/- 0.3 vs. 14.2 +/- 2.5 micromol x cm(-1) x h(-1), respectively; P < 0.01). In conclusion, CFTR plays a significant role in mediating basal, cAMP-, and Ca2+-activated duodenal epithelial HCO3- secretion.


Subject(s)
Bicarbonates/metabolism , Calcium/physiology , Cyclic AMP/physiology , Cystic Fibrosis Transmembrane Conductance Regulator/physiology , Duodenum/metabolism , Intestinal Mucosa/metabolism , Animals , Carbachol/pharmacology , Colforsin/pharmacology , Mice , Mice, Mutant Strains
11.
Nature ; 386(6623): 407-10, 1997 Mar 27.
Article in English | MEDLINE | ID: mdl-9121558

ABSTRACT

Retinoic acid receptors (RAR), thyroid hormone receptors (TR), peroxisome proliferator activated receptors (PPARs) and the orphan receptor, LXR, bind preferentially to DNA as heterodimers with a common partner, retinoid X receptor (RXR), to regulate transcription. We investigated whether RXR-selective agonists replicate the activity of ligands for several of these receptors? We demonstrate here that RXR-selective ligands (referred to as rexinoids) function as RXR heterodimer-selective agonists, activating RXR: PPARgamma and RXR:LXR dimers but not RXR:RAR or RXR:TR heterodimers. Because PPARgamma is a target for antidiabetic agents, we investigated whether RXR ligands could alter insulin and glucose signalling. In mouse models of noninsulin-dependent diabetes mellitus (NIDDM) and obesity, RXR agonists function as insulin sensitizers and can decrease hyperglycaemia, hypertriglyceridaemia and hyperinsulinaemia. This antidiabetic activity can be further enhanced by combination treatment with PPARgamma agonists, such as thiazolidinediones. These data suggest that the RXR:PPARgamma heterodimer is a single-function complex serving as a molecular target for treatment of insulin resistance. Activation of the RXR:PPARgamma dimer with rexinoids may provide a new and effective treatment for NIDDM.


Subject(s)
Diabetes Mellitus, Type 2/metabolism , Insulin Resistance , Insulin/pharmacology , Obesity/metabolism , Receptors, Retinoic Acid/agonists , Thiazolidinediones , Transcription Factors/agonists , Animals , Bexarotene , Blood Glucose/metabolism , Cells, Cultured , Diabetes Mellitus, Type 2/blood , Female , Glucose Tolerance Test , Hypoglycemic Agents/pharmacology , Insulin/blood , Ligands , Mice , Mice, Inbred C57BL , Mice, Obese , Nicotinic Acids/pharmacology , Obesity/blood , Receptors, Cytoplasmic and Nuclear/genetics , Receptors, Cytoplasmic and Nuclear/metabolism , Receptors, Retinoic Acid/genetics , Receptors, Retinoic Acid/metabolism , Retinoid X Receptors , Rosiglitazone , Tetrahydronaphthalenes/pharmacology , Thiazoles/pharmacology , Transcription Factors/genetics , Transcription Factors/metabolism , Transfection
14.
J Reprod Fertil ; 105(2): 215-20, 1995 Nov.
Article in English | MEDLINE | ID: mdl-8568763

ABSTRACT

Pseudopregnant mice were treated systemically with monoclonal anti-progesterone antibody (DB3) (model 1), or progesterone receptor antagonists RU486 or ZK98,299 (ZK299) (model 2) on day 3 post coitum. On day 4, sesame oil was administered intraluminally into one uterine horn to induce decidualization. On day 7, the average mass of the oil-injected horn was 335.2 +/- 52.4 mg, eight times greater than that of the non-injected horn (40.8 +/- 5.3 mg; P < 0.001). After treatment with DB3, RU486 or ZK299, the masses of the injected horns did not differ significantly from those of non-injected horns. In the control group, concentrations of progesterone receptors (ligand-binding assay) increased twofold in the decidualized (52.2 +/- 7.4 fmol mg-1) compared with the non-injected horn (26.0 +/- 7.6 fmol mg-1; P < 0.05), whereas oestrogen receptor content (ligand-exchange assay) decreased by 53% (104.9 +/- 18.2 versus 224.3 +/- 18.1 fmol mg-1; P < 0.001). In model 1, antibody-treated animals showed a tenfold increase in the concentration of progesterone receptors (261.7 +/- 81.1 fmol mg-1; P < 0.001), but there was no differential distribution of progesterone or oestrogen receptors in the oil-injected versus non-injected uterine horns. In model 2, uterine progesterone and oestrogen receptors again showed no differential response between injected and non-injected horns regardless of the route of administration (systemic or intraluminal). Concentrations of progesterone receptors in RU486-treated (35.8 +/- 9.4 fmol mg-1) and ZK299-treated (32.0 +/- 10.2 fmol mg-1) mice were comparable to those in non-injected horns (35.3 +/- 6.3 and 34.2 +/- 5.1 fmol mg-1, respectively) and were not significantly different from the control group (26.0 +/- 7.6 fmol mg-1). The results show that oil-induced decidualization is accompanied by increased concentrations of progesterone receptors and decreased concentrations of oestrogen receptors. When decidualization is blocked by anti-progesterone treatment (antibody against progesterone or progesterone receptor antagonist), there are differing effects on receptor responses with an increase in progesterone receptors and decrease in oestrogen receptors after passive immunization, and no change in progesterone receptors and a reduction in oestrogen receptors after anti-progestins. The anti-decidualization effect in the two models was therefore achieved via dissimilar uterine receptor responses.


Subject(s)
Decidua/growth & development , Pseudopregnancy/metabolism , Receptors, Estrogen/drug effects , Receptors, Progesterone/drug effects , Animals , Antibodies, Monoclonal/pharmacology , Decidua/drug effects , Female , Gonanes/pharmacology , Mice , Mice, Inbred Strains , Mifepristone/pharmacology , Progesterone/antagonists & inhibitors , Progesterone/immunology
15.
J Endocrinol ; 147(2): 331-7, 1995 Nov.
Article in English | MEDLINE | ID: mdl-7490563

ABSTRACT

Anti-progesterone treatment using specific anti-progesterone antibodies or a progesterone receptor (PR) antagonist during first pregnancy impairs postpartum maternal behaviour in mice. This effect is demonstrable only if the treatment is given during pregnancy but not immediately after parturition. The purpose of the present studies was to investigate if maternal behaviour is also impaired by anti-progesterone treatment in subsequent pregnancies. Studies with a monoclonal antibody to progesterone (DB3; 4.5 nmol/mouse) showed that injection of females on day 17 of second pregnancy did not cause maternal rejection but the latency of pup retrieval was prolonged especially during the first 3 days of lactation. This phenomenon was not observed in animals that had previous experience of full length lactation. Experiments were carried out with mifepristone (RU486; 10 micrograms/mouse) injected at day 17 of first, second or third pregnancies. Pup rejection (22.5% vs 12.3%) and prolongation of the retrieval latency (62.3 +/- 13.3 vs 19.7 +/- 6.5 s; P < 0.02) were observed following the first pregnancy. No abnormal behavioural effects were found in mothers treated in second or third pregnancy who had prior full length lactation experience. Control females subjected to only one pup retrieval test after first delivery rejected their pups if treated in their second pregnancy (27.3% vs 4.4%; P < 0.001) and displayed a marginal prolongation of the retrieval latency period (20.9 +/- 7.0 vs 7.4 +/- 2.6 s). Anti-progesterone treatment had no negative influence when administered during third pregnancy.(ABSTRACT TRUNCATED AT 250 WORDS)


Subject(s)
Antibodies, Monoclonal/administration & dosage , Behavior, Animal/drug effects , Maternal Behavior/drug effects , Progesterone/physiology , Animals , Female , Hormone Antagonists/pharmacology , Immunization, Passive , Lactation/physiology , Mice , Mice, Inbred BALB C , Mifepristone/pharmacology , Pregnancy , Progesterone/antagonists & inhibitors , Progesterone/immunology
16.
J Epidemiol Community Health ; 49(4): 373-8, 1995 Aug.
Article in English | MEDLINE | ID: mdl-7650460

ABSTRACT

STUDY OBJECTIVE: The study aimed to examine the concurrence in the variation of monthly numbers of deaths in summer and winter from the four main underlying causes - respiratory, circulatory, neoplastic, and all others - in four countries. In particular, the hypothesis that most non-respiratory concurrent deaths are miscoded respiratory deaths and that a large proportion of the winter mortality currently attributed to circulatory disorders should be attributed to respiratory causes was considered. DESIGN: Mortality data were analysed graphically in relation to cause. Each of the four series of monthly data underwent time series analysis to remove auto-correlation, seasonality, and secular trends. Associations between paired causes of death and between multiple series (using Kendall's coefficient of concordance) were then examined after modelling. SETTING: Monthly deaths (65 years and over) related to underlying cause were examined for England and Wales (nine years), The Netherlands (nine years), Denmark (10 years), and Portugal (10 years - all ages). Weekly data for England and Wales (51 weeks) were also analysed. MAIN RESULTS: All combinations of monthly deaths related to underlying cause were strongly associated in all four countries. This concurrence was evident down to the lowest monthly values so that all seasonally related deaths above the minimum monthly value can be used as an estimate of the "concurrent" proportion. Associations involving deaths from neoplasm were weakest. Concurrence was evident even on a weekly analysis (England and Wales). Concurrent deaths in England and Wales accounted for 31.1% of respiratory, 16.0% of circulatory, 3.5% of neoplastic, 14.1% of deaths from other causes and 14.2% for all deaths combined. The equivalent percentages for concurrent deaths from all causes were 8.4% in the Netherlands, 9.3% in Denmark, and 16.8% in Portugal. CONCLUSIONS: Concurrence, which was present in each of the underlying causal groups in each of the four national data sets examined, suggests a common cause separate from the underlying cause that has been used in the presentation of mortality statistics. If the person concerned had not died at that time, as a result of this cause, he would not have died from the recorded underlying cause. Most of these non-respiratory concurrent deaths are miscoded. As a consequence, a large proportion of winter mortality currently attributed to circulatory disorders should be attributed to other causes, probably respiratory. More intensive research into the contribution made by acute respiratory diseases is proposed. The proportion of concurrent deaths varied in the four countries thereby limiting the validity of simple comparisons of national mortality statistics.


Subject(s)
Cardiovascular Diseases/mortality , Neoplasms/mortality , Respiratory Tract Diseases/mortality , Adolescent , Adult , Aged , Causality , Child , Child, Preschool , Denmark/epidemiology , England/epidemiology , Humans , Infant , Infant, Newborn , Middle Aged , Netherlands/epidemiology , Portugal/epidemiology , Seasons , Wales/epidemiology
18.
J Endocrinol ; 145(2): 363-9, 1995 May.
Article in English | MEDLINE | ID: mdl-7616170

ABSTRACT

Passive transfer of a monoclonal antibody against progesterone produces a high incidence of maternal rejection in mice after recovery from antibody-induced infertility. To investigate the mechanisms involved in this reduction of maternal care, we have examined whether the effect is due to long-term exposure to antibody. Antibody was administered i.p. either on day 2 or day 17 of pregnancy. When a low dose (1.0 nmol) was given on day 2, pregnancy proceeded normally but 44.8% pups delivered at term were rejected compared with 12.7% in the control group. When a higher dose (4.5 nmol) of antibody was given on day 17, pregnancy continued normally to term and the rejection rate was 48.8% (control: 11.1%). When the same amount of antibody was injected after delivery (day 1 of lactation), no detrimental effect was found on subsequent maternal care to the young, the rejection rate being comparable between antibody-treated and control groups (5.3% vs 4.6%). To determine if the presence of antibody interfered with lactation or suckling, a bolus injection of 10 microCi [3H]H2O was given to mice treated at day 17 with antibody or saline. The levels of radioactivity present in both mothers and pups and the first 5-day pup growth curves showed identical patterns, indicating that milk availability and the suckling process were not affected. Crossfostering studies revealed that antibody-treated mothers rejected 25.5% of fostered pups compared with 8.5% found in the control females when antibody was administered on day 17 of pregnancy and the entire litters were crossfostered between the two groups immediately after delivery.(ABSTRACT TRUNCATED AT 250 WORDS)


Subject(s)
Maternal Behavior/physiology , Pregnancy, Animal , Progesterone/physiology , Animals , Antibodies, Monoclonal/administration & dosage , Female , Immunization, Passive , Lactation/physiology , Mice , Mice, Inbred BALB C , Postpartum Period , Pregnancy , Progesterone/immunology
19.
J Endocrinol ; 145(2): 371-7, 1995 May.
Article in English | MEDLINE | ID: mdl-7616171

ABSTRACT

The rapid onset of normal maternal behaviour after parturition in mice, consisting of cleaning, warming, feeding and protection of offspring, is primed by oestrogen, progesterone and oxytocin. Previous studies showed that passive transfer of monoclonal antibodies against progesterone significantly increases the incidence of maternal rejection of pups. To test the hypothesis that aberrant maternal behaviour is due to partial progesterone withdrawal leading to hormonal imbalance during late pregnancy, maternal rejection was assessed following treatment with a progesterone receptor antagonist. Mifepristone (RU486) was given subcutaneously on either day 2 (100 micrograms) or day 17 (50 micrograms) of pregnancy, or on the first day of lactation (100 micrograms). Maternal behaviour was monitored twice daily for the first 6 days of lactation and pup rejection recorded for a further 15 days. Maternal rejection was significantly greater after mifepristone administration on either day 2 or day 17 (28.6% and 38.3%) compared with controls (11.1% and 5.2% respectively). Rejection was negligible in both treated and control groups if mifepristone was given after parturition. When mothers were treated at day 17, the length of the latent period before pups were retrieved and returned to the nest was markedly increased in mifepristone-treated mothers (46.3 s) compared with controls (4.4 s) though the effect was transient. The results indicate that mifepristone interferes with the hormonal priming mechanism(s) necessary for the onset of normal maternal behaviour by a receptor-mediated effect. The similarity of the present results and those obtained with anti-progesterone antibodies implies that receptor antagonism or antibody scavenging of progesterone influence a common central nervous mechanism that is essential for the normal priming process.


Subject(s)
Maternal Behavior/drug effects , Mifepristone/pharmacology , Pregnancy, Animal/drug effects , Receptors, Progesterone/antagonists & inhibitors , Animals , Female , Lactation/drug effects , Mice , Mice, Inbred BALB C , Pregnancy
20.
J Steroid Biochem Mol Biol ; 49(2-3): 123-9, 1994 Jun.
Article in English | MEDLINE | ID: mdl-8031708

ABSTRACT

The brain isozyme of creatine kinase (CKB) is a major component of the estrogen-induced proteins in the rat uterus. Hormonal specificity of this response was studied in cotransfection assays using the rat CKB promoter linked to the bacterial chloramphenicol acetyltransferase gene. Response was specific for estrogen as 17 beta-estradiol in the presence of estrogen receptor dramatically stimulated the CKB promoter. This induction was completely blocked by the estrogen antagonist ICI 164,384. Nuclear receptors for progesterone, androgen, glucocorticoid and vitamin D did not significantly activate the CKB promoter in the presence of their respective ligands. Creatine kinase (CK) activity was analyzed in decidualized mouse uterus to assess estrogenic activity in vivo. Upon oil stimulation, uterine horns of day 4 pseudopregnant mice underwent a dramatic outgrowth in response to endogenous progesterone. This response was accompanied by a significant decrease in CK activity from a control value of 1.44 +/- 0.25 to 0.38 +/- 0.08 IU/mg protein (P < 0.001), indicating that the action of estrogen was suppressed. Treatment of females one day prior to oil-stimulation with progesterone receptor antagonists, RU486 (Mifepristone) or ZK299 (Onapristone), or with a monoclonal antibody to progesterone (DB3), abolished decidualization, and also restored the CK activity to the control value. These results suggest that CK can be used as a specific cellular marker to detect unopposed estrogen action in the mouse uterus associated with progesterone withdrawal or receptor blockade.


Subject(s)
Creatine Kinase/metabolism , Estradiol/pharmacology , Progesterone/antagonists & inhibitors , Uterus/enzymology , Animals , Brain/enzymology , Creatine Kinase/genetics , Decidua/drug effects , Decidua/physiology , Estradiol/analogs & derivatives , Female , Gene Expression/drug effects , HeLa Cells , Humans , Isoenzymes , Male , Mice , Mice, Inbred BALB C , Mifepristone/pharmacology , Polyunsaturated Alkamides , Progesterone/pharmacology , Promoter Regions, Genetic , Receptors, Estrogen/genetics , Receptors, Estrogen/physiology , Receptors, Progesterone/genetics , Receptors, Progesterone/physiology , Recombinant Fusion Proteins , Transfection , Uterus/drug effects
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