ABSTRACT
Long-term effects of breastfeeding on maternal bone are not fully understood. Excessive maternal bone loss stimulated by serotonin signaling during lactation may increase bone fragility later in life. We hypothesized that inhibiting nonneuronal serotonin activity by feeding a small-molecule inhibitor of the rate-limiting enzyme in serotonin synthesis [tryptophan hydroxylase 1 (TPH1)] would preserve maternal bone postweaning without affecting neonatal bone. Chow supplemented with the small-molecule TPH1 inhibitor LP778902 (~100 mg/kg) or control chow was fed to C57BL/6 dams throughout pregnancy and lactation, and blood was collected on days 1 and 21 of lactation. Dams returned to a common diet postweaning and were aged to 3 or 9 mo postweaning. Pups were euthanized at weaning. The effect of TPH1 inhibition on dam and pup femoral bone was determined by micro-computed tomography. Peripartum dietary supplementation with LP778902 decreased maternal serum serotonin concentrations ( P = 0.0007) and reduced bone turnover, indicated by serum NH2-terminal propeptide of type I collagen ( P = 0.01) and COOH-terminal collagen cross-links ( P = 0.02) concentrations, on day 21 of lactation. Repressed bone turnover from TPH1 inhibition was not associated with structural changes in maternal femur at 3 or 9 mo postweaning. By contrast, neonates exposed to peripartum LP778902 demonstrated differences in trabecular and cortical femoral bone compared with pups from control dams, with fewer ( P = 0.02) and thinner ( P = 0.001) trabeculae as well as increased trabecular spacing ( P = 0.04). Additionally, cortical porosity was increased ( P = 0.007) and cortical tissue mineral density was decreased ( P = 0.005) in pups of LP778902-treated dams. Small-molecule TPH1 inhibitors should be carefully considered in pregnant and lactating women, given potential risks to neonatal bone development.
Subject(s)
Bone Density/drug effects , Bone Remodeling/drug effects , Femur/diagnostic imaging , Maternal Nutritional Physiological Phenomena/drug effects , Serotonin/blood , Tryptophan Hydroxylase/antagonists & inhibitors , Animals , Biomarkers/blood , Collagen Type I/blood , Dietary Supplements , Female , Lactation/drug effects , Mice , Peptides/blood , Pregnancy , X-Ray MicrotomographyABSTRACT
Pulmonary arterial hypertension (PAH) is a progressive disease defined by a chronic elevation in pulmonary arterial pressure with extensive pulmonary vascular remodeling and perivascular inflammation characterized by an accumulation of macrophages, lymphocytes, dendritic cells, and mast cells. Although the exact etiology of the disease is unknown, clinical as well as preclinical data strongly implicate a role for serotonin (5-HT) in the process. Here, we investigated the chronic effects of pharmacological inhibition of tryptophan hydroxylase 1 (TPH1), the rate-limiting enzyme in peripheral 5-HT biosynthesis, in two preclinical models of pulmonary hypertension (PH), the monocrotaline (MCT) rat and the semaxanib (SUGEN, Medinoah, Suzhou, China)-hypoxia rat. In both PH models, ethyl (S)-8-(2-amino-6-((R)-1-(5-chloro-[1,1'-biphenyl]-2-yl)-2,2,2-trifluoroethoxy)pyrimidin-4-yl)-2,8-diazaspiro[4.5]decane-3-carboxylate and ethyl (S)-8-(2-amino-6-((R)-1-(3',4'-dimethyl-3-(3-methyl-1 H-pyrazol-1-yl)-[1,1'-biphenyl]-4-yl)-2,2,2-trifluoroethoxy)pyrimidin-4-yl)-2,8-diazaspiro[4.5]decane-3-carboxylate, novel orally active TPH1 inhibitors with nanomolar in vitro potency, decreased serum, gut, and lung 5-HT levels in a dose-dependent manner and significantly reduced pulmonary arterial pressure, and pulmonary vessel wall thickness and occlusion in male rats. In the MCT rat model, decreases in lung 5-HT significantly correlated with reductions in histamine levels and mast cell number (P < 0.001, r2 = 0.88). In contrast, neither ambrisentan nor tadalafil, which are vasodilators approved for the treatment of PAH, reduced mast cell number or 5-HT levels, nor were they as effective in treating the vascular remodeling as were the TPH1 inhibitors. When administered in combination with ambrisentan, the TPH1 inhibitors showed an additive effect on pulmonary vascular remodeling and pressures. These data demonstrate that in addition to reducing vascular remodeling, TPH1 inhibition has the added benefit of reducing the perivascular mast cell accumulation associated with PH.
Subject(s)
Enzyme Inhibitors/pharmacology , Hypertension, Pulmonary/physiopathology , Pulmonary Artery/drug effects , Pulmonary Artery/physiopathology , Tryptophan Hydroxylase/antagonists & inhibitors , Vascular Remodeling/drug effects , Animals , Cell Proliferation/drug effects , Disease Models, Animal , Gene Expression Regulation, Enzymologic/drug effects , HEK293 Cells , Humans , Hypertension, Pulmonary/metabolism , Hypertension, Pulmonary/pathology , Lung/drug effects , Lung/metabolism , Male , Myocytes, Smooth Muscle/cytology , Myocytes, Smooth Muscle/drug effects , Myocytes, Smooth Muscle/pathology , Pulmonary Artery/metabolism , Pulmonary Artery/pathology , Rats , Rats, Sprague-Dawley , Serotonin/blood , Serotonin/metabolism , Tryptophan Hydroxylase/metabolismABSTRACT
BACKGROUND: Matrix metalloproteinase (MMP)-9, a member of the MMP superfamily is consistently implicated in the pathophysiology of atherosclerosis and plaque rupture, the most common mechanism responsible for acute coronary syndrome (ACS). AIM: To summarize the role of MMP-9 in atherosclerosis and its potential implications in assessment and treatment of coronary artery disease (CAD). METHODS: We reviewed the PubMed database for relevant data regarding the role of MMP-9 in the pathophysiology of atherosclerosis. In the light of these data, we postulate potential implications of MMP-9 in the management and treatment of CAD. RESULTS AND CONCLUSIONS: Existing data strongly support the role of MMP-9 in plaque destabilization and rupture. Based on the current knowledge, MMP-9 can potentially serve as a diagnostic biomarker in ACS and a prognostic biomarker in ACS and chronic CAD patients. MMP-9 is reduced by therapies that are associated with favourable outcome in atherosclerosis and thus may serve as a surrogate biomarker of treatment efficacy. However, large morbidity and mortality trials are still required to confirm that MMP-9 reduction is associated with improved outcome independent of the traditional risk factors (i.e. low-density lipoprotein cholesterol). Given its role in plaque rupture, inhibition of MMP-9 may promote plaque stabilization and consequently reduce cardiovascular events. Yet, the efficacy and safety of MMPs inhibitors should be first studied in preclinical models of atherosclerosis.
Subject(s)
Coronary Artery Disease/enzymology , Matrix Metalloproteinase 9/metabolism , Animals , Coronary Artery Disease/diagnosis , Coronary Artery Disease/drug therapy , Humans , Matrix Metalloproteinase 9/blood , Matrix Metalloproteinase Inhibitors , Mice , Protease Inhibitors/pharmacology , Risk FactorsABSTRACT
OBJECTIVE: Studies in bone marrow transplanted from ATP-binding cassette transporter A1 (ABCA1)-deficient mice into normal mice provides direct evidence that the absence of leukocyte ABCA1 exerts a marked proatherogenic effect independent of changes in plasma lipids, suggesting that ABCA1 plays a key role in the regulation of cholesterol homeostasis and function of macrophages. Therefore, we examined whether the absence of ABCA1 affects the morphology, properties, and functional activities of macrophages that could be related to the development of atherosclerosis. METHODS AND RESULTS: We conducted a series of experiments in macrophages isolated from Abca1-deficient and wild-type mice and compared several of their properties that are thought to be related to the development of atherosclerosis. Macrophages isolated from Abca1-deficient mice have an increase in cholesterol content, expression of scavenger receptors, and secretion of chemokines, growth factors, and cytokines, resulting in an increased ability to respond to a variety of chemotactic factors. CONCLUSIONS: Our studies indicate that the absence of ABCA1 leads to significant changes in the morphology, properties, and functional activities of macrophages. These changes, together with the proinflammatory condition present in ABCA1-deficient mice and increased reactivity of macrophages to chemotactic factors, play a key role in the development and progression of atherosclerosis.
Subject(s)
ATP-Binding Cassette Transporters/genetics , Atherosclerosis/physiopathology , Cholesterol/metabolism , Macrophages, Peritoneal/metabolism , Receptors, Scavenger/genetics , ATP Binding Cassette Transporter 1 , Animals , Atherosclerosis/immunology , Atherosclerosis/pathology , Cells, Cultured , Chemokines/metabolism , Chemotaxis, Leukocyte/physiology , Female , Lipopolysaccharides/pharmacology , Macrophages, Peritoneal/pathology , Macrophages, Peritoneal/ultrastructure , Male , Mice , Mice, Inbred C57BL , Mice, Inbred DBA , Mice, Mutant Strains , Microscopy, Electron , Shock, Septic/immunology , Shock, Septic/pathology , Shock, Septic/physiopathology , Tumor Necrosis Factor-alpha/metabolism , Vacuoles/metabolism , Vacuoles/ultrastructureABSTRACT
Studies with ATP-binding cassette transporter (ABCA1)-deficient mice have been critical in demonstrating the relation between ABCA1 expression, cellular lipid efflux, and HDL metabolism. The phenotype of the ABCA1-deficient mouse parallels the phenotype observed in human Tangier disease, including substantial reductions in both apolipoprotein B and apolipoprotein AI with confounding affects on atherosclerosis.
Subject(s)
ATP-Binding Cassette Transporters/physiology , ATP Binding Cassette Transporter 1 , ATP-Binding Cassette Transporters/chemistry , ATP-Binding Cassette Transporters/genetics , Animals , Apolipoproteins B/metabolism , Arteriosclerosis/blood , Arteriosclerosis/genetics , Cholesterol, HDL/metabolism , Disease Models, Animal , Female , Foam Cells/pathology , Humans , Infertility/genetics , Liver/metabolism , Male , Mice , Mice, Inbred C57BL , Mice, Inbred DBA , Mice, Knockout , Phenotype , Structure-Activity Relationship , Tangier Disease/genetics , Tangier Disease/metabolismABSTRACT
Leukotriene B4 (LTB4) is a potent chemotactic agent that activates monocytes through the LTB4 receptor (BLTR). We tested the hypothesis that LTB4 receptor blockade would slow atherosclerotic progression by inhibiting monocyte recruitment. Homozygous low-density receptor knockout (LDLr(-/-)) mice and apolipoprotein E deficient (apoE(-/-)) mice were treated with a specific LTB4 receptor antagonist, CP-105,696, for 35 days. In apoE(-/-)mice, treatment with the LTB4 antagonist did not affect plasma lipid concentrations but significantly reduced CD11b levels both in vascular lesions and whole blood. Compared with age-matched controls, lipid accumulation and monocyte infiltration were significantly reduced in treated apoE(-/-) mice at all time points tested. Lesion area reduction was also demonstrated in LDLr(-/-) mice maintained on a high-fat diet. LTB4 antagonism had no significant effect on lesion size in mice possessing the null alleles for another chemotactic agent, monocyte chemoattractant protein-1 (MCP-1(-/-)xapoE(-/-)), suggesting MCP-1 and LTB4 may either interact or exert their effects by a common mechanism. These results demonstrate that in a preclinical model of atherosclerosis LTB4 receptor blockade reduces lesion progression and further suggest a previously unrecognized role for LTB4 or other oxidized lipids recognized by the BLTR receptor in the pathogenesis of this disease.
Subject(s)
Arteriosclerosis/etiology , Benzopyrans/pharmacology , Carboxylic Acids/pharmacology , Foam Cells , Leukotriene Antagonists/pharmacology , Receptors, Leukotriene B4/antagonists & inhibitors , Animals , Apolipoproteins E/genetics , Arteriosclerosis/metabolism , Arteriosclerosis/pathology , Chemokine CCL2/genetics , Disease Progression , Immunohistochemistry , Leukotriene B4/physiology , Lipids/blood , Macrophage-1 Antigen/immunology , Macrophage-1 Antigen/metabolism , Mice , Mice, Inbred C57BL , Mice, Knockout , Monocytes/cytology , Monocytes/metabolism , Receptors, CCR2 , Receptors, Chemokine/metabolism , Receptors, LDL/geneticsABSTRACT
The ATP-binding cassette transporter A1 (ABCA1) encodes a membrane protein that promotes cholesterol and phospholipid efflux from cells. Mutations in ABCA1 lead to HDL deficiency and tissue accumulation of macrophages in patients with homozygous Tangier disease. In this study, we examined whether the complete absence of ABCA1 or selected inactivation in macrophages is accompanied by an increase in atherosclerotic lesion progression in hypercholesterolemic apolipoprotein E-deficient (apoE(-/-)) mice and LDLR receptor-deficient (LDLr(-/-)) mice. The absence of ABCA1 led to reduced plasma cholesterol levels in both the apoE(-/-) and LDLr(-/-) mice, along with severe skin xanthomatosis characterized by marked foamy macrophages and cholesterol ester accumulation. However, the complete absence of ABCA1 did not affect the development, progression, or composition of atherosclerotic lesions in either the LDLr(-/-) or the apoE(-/-) mice fed a chow or atherogenic diet. In contrast, bone marrow transplantation studies demonstrated that the selective inactivation of ABCA1 in macrophages markedly increased atherosclerosis and foam cell accumulation in apoE(-/-). Taken together, these findings demonstrate that the complete absence of ABCA1 has a major impact on plasma lipoprotein homeostasis, and the proposed antiatherogenic effect resulting from ABCA1 deficiency is compensated by a less atherogenic profile. ABCA1 deficiency in macrophages, however, demonstrates the antiatherogenic properties of ABCA1 independent of plasma lipids and HDL levels.
Subject(s)
ATP-Binding Cassette Transporters/metabolism , Apolipoproteins E/deficiency , Arteriosclerosis/etiology , Cholesterol/blood , Hyperlipidemias/complications , Macrophages/metabolism , Receptors, LDL/deficiency , ATP Binding Cassette Transporter 1 , Animals , Arteriosclerosis/metabolism , Arteriosclerosis/pathology , Bone Marrow Transplantation , Crosses, Genetic , Female , Hyperlipidemias/metabolism , Lipids/blood , Lipoproteins/blood , Macrophages/pathology , Male , Mice , Mice, Inbred C57BL/genetics , Mice, Inbred DBA/genetics , Xanthomatosis/pathologyABSTRACT
Since the identification of mutations in the ATP-binding cassette transporter (ABCA1), the relationship between ABCA1 expression, cholesterol efflux, high-density lipoprotein (HDL) biosynthesis and cholesterol homeostasis has been a subject of intense investigation. Several studies have provided significant new information with regards to pathways controlling ABCA1 expression and activity and established that this transporter facilitates the efflux of cholesterol and phospholipids to apoprotein acceptors, leading to the formation of nascent HDL particles. Although ABCA1 appears to play a critical role in cholesterol flux from tissues, and despite a considerable interest in developing pharmacological agents that increase ABCA1 activity, the role of ABCA1 in preventing atherosclerosis remains unclear.
Subject(s)
ATP-Binding Cassette Transporters/physiology , Arteriosclerosis/physiopathology , ATP Binding Cassette Transporter 1 , ATP-Binding Cassette Transporters/drug effects , ATP-Binding Cassette Transporters/genetics , ATP-Binding Cassette Transporters/metabolism , Animals , Cholesterol, HDL/biosynthesis , Cholesterol, HDL/genetics , HumansABSTRACT
Hyperglycemia resulting from type 2 diabetes mellitus (T2DM) is the main cause of diabetic complications such as retinopathy and neuropathy. A reduction in hyperglycemia has been shown to prevent these associated complications supporting the importance of glucose control. Glucokinase converts glucose to glucose-6-phosphate and determines glucose flux into the ß-cells and hepatocytes. Since activation of glucokinase in ß-cells is associated with increased risk of hypoglycemia, we hypothesized that selectively activating hepatic glucokinase would reduce fasting and postprandial glucose with minimal risk of hypoglycemia. Previous studies have shown that hepatic glucokinase overexpression is able to restore glucose homeostasis in diabetic models; however, these overexpression experiments have also revealed that excessive increases in hepatic glucokinase activity may also cause hepatosteatosis. Herein we sought to evaluate whether liver specific pharmacological activation of hepatic glucokinase is an effective strategy to reduce hyperglycemia without causing adverse hepatic lipids changes. To test this hypothesis, we evaluated a hepatoselective glucokinase activator, PF-04991532, in Goto-Kakizaki rats. In these studies, PF-04991532 reduced plasma glucose concentrations independent of changes in insulin concentrations in a dose-dependent manner both acutely and after 28 days of sub-chronic treatment. During a hyperglycemic clamp in Goto-Kakizaki rats, the glucose infusion rate was increased approximately 5-fold with PF-04991532. This increase in glucose infusion can be partially attributed to the 60% reduction in endogenous glucose production. While PF-04991532 induced dose-dependent increases in plasma triglyceride concentrations it had no effect on hepatic triglyceride concentrations in Goto-Kakizaki rats. Interestingly, PF-04991532 decreased intracellular AMP concentrations and increased hepatic futile cycling. These data suggest that hepatoselective glucokinase activation may offer glycemic control without inducing hepatic steatosis supporting the evaluation of tissue specific activators in clinical trials.
Subject(s)
Diabetes Mellitus, Experimental/complications , Enzyme Activators/pharmacology , Glucokinase/metabolism , Hyperglycemia/complications , Hyperglycemia/drug therapy , Imidazoles/pharmacology , Liver/drug effects , Liver/enzymology , Nicotinic Acids/pharmacology , Animals , Enzyme Activators/adverse effects , Enzyme Activators/therapeutic use , Glucose/metabolism , Hepatocytes/drug effects , Hepatocytes/metabolism , Imidazoles/adverse effects , Imidazoles/therapeutic use , Liver/pathology , Male , Nicotinic Acids/adverse effects , Nicotinic Acids/therapeutic use , Non-alcoholic Fatty Liver Disease/chemically induced , Organ Specificity , RatsABSTRACT
OBJECTIVE: The CCR2 receptor plays a crucial role in monocyte recruitment and has been implicated as a contributing factor to atherosclerosis. CCR2 receptor deletion leads to significant inhibition of lesion development. Our objective was to determine if CCR2 receptor blockade with a small molecule would have a beneficial effect of decreasing established lesions. METHODS AND RESULTS: We demonstrated that CCR2 blockade had no significant effect on advanced lesions or the progression of fatty streaks. CCR2 blockade in mice resulted in elevations in plasma CCL2 levels and a significant reduction in the plasma Ly-6C(hi) subpopulations of monocytes expressing the CCR2 receptor. Neither CCL2 elevation nor margination of the Ly-6C(hi) population was observed in CCR2(-/-) mice. CONCLUSIONS: CCR2 receptor blockade with a small molecule antagonist at dose levels showing efficacy in several inflammatory models did not show a beneficial effect in murine models of atherosclerosis. Elevations in CCL2 and margination of Ly-6C(hi) cells demonstrate that the role of CCR2 in controlling monocyte levels goes beyond the control of monocyte emigration.