ABSTRACT
OBJECTIVES: This study evaluated how deproteinization using sodium hypochlorite (6% NaOCl) or hypochlorous acid (50 ppm HOCl) with or without the subsequent use of an arylsulfinate salt-containing agent (Clearfil DC Activator; DCA; Kuraray Noritake Dental) affects the micro-tensile bond strength (µTBS) and formation of an acid-base resistant zone (ABRZ) of a two-step self-etch adhesive on eroded dentin. METHODS: Coronal dentin surfaces of sound human molars were exposed to 48 cycles of demineralization (1% citric acid; 5 minutes) and remineralization (buffer solution with pH=6.4; 3.5 hours). They were then assigned to experimental groups according to the pretreatment used: none (negative control), NaOCl, NaOCl+DCA, HOCl, and HOCl+DCA. Sound dentin surfaces with no pretreatment were used as a positive control. The dentin surfaces were bonded with Clearfil SE Bond 2 (Kuraray Noritake Dental), and µTBS was measured either after 24 hours or 20,000 thermal cycles (TC). The µTBS data were statistically analyzed using a mixed-model analysis of variance (ANOVA) and t-tests with Bonferroni correction. Failure mode was determined with scanning electron microscopy (SEM), which was also used for the observation of ABRZ. RESULTS: Among experimental groups, there was no significant difference between the negative control, HOCl, and HOCl+DCA after 24 hours, but the HOCl-pretreated groups exhibited significantly higher µTBS than the negative control after TC (p<0.01). Pretreatment with NaOCl and NaOCl+DCA resulted in significantly higher µTBS (p<0.001), but the highest µTBS was measured on sound dentin (p<0.001). TC decreased µTBS significantly in all groups (p<0.001) except for sound dentin and NaOCl+DCA (p>0.05). Adhesive failures prevailed in eroded groups, whereas cohesive failures were predominant on sound dentin. ABRZ was recognized in all groups but marked morphological differences were observed. CONCLUSIONS: The combined use of 6% NaOCl and the arylsulfinate salt-containing agent partially reversed the compromised bonding performance on eroded dentin, while the effect of 50 ppm HOCl was negligible.
Subject(s)
Dental Bonding , Dental Cements , Humans , Dental Bonding/methods , Dentin-Bonding Agents/pharmacology , Dentin-Bonding Agents/chemistry , Dentin , Materials Testing , Resin Cements/pharmacology , Resin Cements/chemistry , Tensile StrengthABSTRACT
In this laboratory study, the microtensile bond strengths (µTBS) of resin-modified glass ionomer cement (RM-GIC) to sound and artificial caries-affected bovine root dentin (ACAD) using three different conditioning agents were evaluated after 24 hours and three months. The fractured interface was examined with a scanning electron microscope (SEM). Specimens were created on bovine root dentin that was embedded in epoxy resin. For the ACAD specimens, artificial carious lesions were created. The RM-GIC (Fuji II LC) was applied either directly (no treatment), after application of self conditioner, cavity conditioner, or 17% ethylenediamine tetraacetic acid (EDTA) applied for 60 seconds, on sound dentin and ACAD, then light cured. They were stored in artificial saliva for 24 hours or three months. Following this, the specimens were cut into sticks for the µTBS test, and the failure mode of the debonded specimens was examined by using SEM. Pretest failures were excluded from the statistical analysis of the µTBS values because of their high incidence in some groups. Results showed that the µTBS values were significantly affected by the dentin substrate as well as the conditioning agent. Self conditioner provided the highest and most stable µTBS values, while cavity conditioner showed stable µTBS values on sound dentin. Both self conditioner and cavity conditioner had significantly higher µTBS values than the no treatment groups. EDTA conditioning reduced the µTBS after three months to sound dentin, while it showed 100% pretest failure with ACAD for both storage periods.
Subject(s)
Dental Bonding/methods , Dental Caries/therapy , Dentin/metabolism , Glass Ionomer Cements/therapeutic use , Tooth Root/metabolism , Animals , Cattle , Dental Stress Analysis , In Vitro Techniques , Microscopy, Electron, Scanning , Tensile StrengthABSTRACT
Nerve growth factor (NGF) functions to modulate osteoarthritis (OA)-associated pain. Although recent studies suggest that tumour necrosis factor (TNF)-α and interleukin (IL)-1ß mediate NGF activity in human synovial fibroblasts, the regulation of NGF expression in human synovial macrophages remains unclear. Here, we examined the role of macrophages in the production and regulation of synovial (SYN) NGF in osteoarthritic knee joints by examining the mRNA expression of TNF-α and IL-1ß in freshly isolated CD14-positive (macrophage-rich fraction) and CD14-negative cells (fibroblast-rich fraction) in synovial tissue from OA patients by quantitative polymerase chain reaction. We also examined the effects of IL-1ß and TNF-α on NGF mRNA expression in cultured CD14-positive (macrophage-rich fraction) and CD14-negative cells (fibroblast-rich fraction). In addition, to examine the contribution of macrophages to NGF, TNF-α and IL-1ß expression, we injected clodronate liposomes systemically into STR/Ort mice, an osteoarthritis animal model, to deplete macrophages. TNF-α and IL-1ß mRNA levels in CD14-positive cells from the SYN of OA patients was significantly higher than that in CD14-negative cells, while NGF expression did not differ markedly between the two cell fractions. In addition, treatment of human cultured CD14-positive and -negative cells with IL-1ß and TNF-α enhanced NGF mRNA and protein levels. Expression of NGF, IL-1ß and TNF-α was also reduced significantly in STR/Ort mice upon macrophage depletion. These findings suggest that IL-1ß and TNF-α regulate NGF expression and production in synovial macrophages and fibroblasts in osteoarthritic joints.
Subject(s)
Macrophages/metabolism , Nerve Growth Factor/biosynthesis , Nerve Growth Factor/genetics , Osteoarthritis, Knee/metabolism , Osteoarthritis/metabolism , Synovial Membrane/immunology , Aged , Aged, 80 and over , Animals , Cells, Cultured , Clodronic Acid/administration & dosage , Disease Models, Animal , Female , Fibroblasts/drug effects , Gene Expression Regulation , Humans , Interleukin-1beta/genetics , Interleukin-1beta/metabolism , Interleukin-1beta/pharmacology , Lipopolysaccharide Receptors/immunology , Liposomes , Macrophages/drug effects , Macrophages/immunology , Male , Mice , Nerve Growth Factor/immunology , Osteoarthritis/immunology , Osteoarthritis, Knee/immunology , Polymerase Chain Reaction , Synovial Membrane/metabolism , Tumor Necrosis Factor-alpha/genetics , Tumor Necrosis Factor-alpha/metabolism , Tumor Necrosis Factor-alpha/pharmacologyABSTRACT
Recent studies have reported that calcitonin gene-related peptide (CGRP) contributes to joint pain. However, regulation of the CGRP/CGRP receptor signalling in osteoarthritis (OA) is not fully understood. To investigate the regulation of CGRP/CGRP receptor signalling by macrophages in the synovial tissue (ST) of OA joints, we characterized the gene expression profiles of CGRP and CGRP receptors in the ST of OA mice (STR/Ort). In addition, we examined whether macrophage depletion by the systemic injection of clodronate-laden liposomes affected the expression of CGRP and CGRP receptors in ST. CD11c(+) macrophages in the ST of STR/Ort and C57BL/6J mice were analysed by flow cytometry. Real-time polymerase chain reaction (PCR) was used to evaluate the expression of interleukin (IL)-1ß, CGRP, calcitonin receptor-like receptor (CLR) and receptor activity-modifying protein 1 (RAMP1) in F4/80(+) and F4/80(-) cells. The effects of IL-1ß on the expression of CGRP and CLR by cultured synovial cells were also examined. The percentage of CD11c(+) macrophages in the ST of STR/Ort was higher than that in C57/BL6J mice. Notably, the F4/80(+) cell fraction expressed IL-1ß highly, whereas the F4/80(-) cell fraction expressed CGRP, CLR, and RAMP1 highly. In addition, expression of the IL-1ß and CLR genes was increased in ST, but was decreased upon macrophage depletion, and the IL-1ß treatment of cultured synovial cells up-regulated CLR. Taken together, the present findings suggest that synovial macrophages are the major producers of IL-1ß and regulators of CLR in OA mice. Therefore, macrophages and IL-1ß may be suitable therapeutic targets for treating OA pain.
Subject(s)
Calcitonin Gene-Related Peptide/metabolism , Calcitonin Receptor-Like Protein/metabolism , Interleukin-1beta/metabolism , Macrophages/immunology , Osteoarthritis/immunology , Receptor Activity-Modifying Protein 1/metabolism , Receptors, Calcitonin/metabolism , Animals , Antigens, Differentiation/metabolism , Calcitonin Gene-Related Peptide/genetics , Calcitonin Receptor-Like Protein/genetics , Cells, Cultured , Clodronic Acid/administration & dosage , Disease Models, Animal , Gene Expression Regulation/drug effects , Humans , Interleukin-1beta/genetics , Macrophages/drug effects , Male , Mice , Mice, Inbred C57BL , Mice, Transgenic , Receptor Activity-Modifying Protein 1/genetics , Receptors, Calcitonin/genetics , Signal Transduction/drug effects , Synovial Membrane/immunologyABSTRACT
To understand more clearly the link between osteoarthritis and hyperlipidaemia, we investigated the inflammatory macrophage subsets and macrophage-regulated matrix metalloprotease-3 (MMP-3) and A disintegrin and metalloprotease with thrombospondin motifs-4 (ADAMTS4) in synovial (ST) and adipose tissues (AT) of osteoarthritic mice with hyperlipidaemia (STR/Ort). CD11c(+) F4/80(+) CD11b(+) macrophage populations in the ST and AT of 9-month-old STR/Ort and C57BL/6J mice were characterized and compared by flow cytometry and real-time polymerase chain reaction (PCR) analyses. Expression of tumour necrosis factor (TNF)-α, MMP-3 and ADAMTS4, and the response of these factors to anionic liposomal clodronate induced-macrophage depletion were also evaluated by real-time PCR. Expression of TNF-α in CD11c(+) cells, which were isolated by magnetic beads, was compared to CD11c(-) cells. In addition, the effect of TNF-α on cultured synovial fibroblasts and adipocytes was investigated. CD11c(+) F4/80(+) CD11b(+) macrophages were increased in ST and AT of STR/Ort mice. The CD11c(+) cell fraction highly expressed TNF-α. Expression of TNF-α and MMP3 was increased in ST and AT, and was decreased upon macrophage depletion. TNF-α treatment of cultured synovial fibroblasts and adipocytes markedly up-regulated MMP-3. CD11c(+) F4/80(+) CD11b(+) macrophages were identified as a common inflammatory subset in the AT and ST of STR/Ort mice with hyperlipidaemia. The induction of inflammation in AT and ST may be part of a common mechanism that regulates MMP3 expression through TNF-α. Our findings suggest that increased numbers of CD11c(+) macrophages and elevated levels of TNF-α and MMP-3 in AT and ST may explain the relationship between hyperlipidaemia and OA.
Subject(s)
Adipose Tissue/metabolism , CD11c Antigen/metabolism , Macrophages/metabolism , Matrix Metalloproteinase 3/metabolism , Osteoarthritis/metabolism , Synovial Membrane/metabolism , Tumor Necrosis Factor-alpha/metabolism , ADAM Proteins/genetics , ADAM Proteins/metabolism , ADAMTS4 Protein , Animals , CD11c Antigen/genetics , Disease Models, Animal , Fibroblasts/metabolism , Gene Expression , Hyperlipidemias/complications , Macrophages/immunology , Male , Matrix Metalloproteinase 3/genetics , Mice , Osteoarthritis/complications , Osteoarthritis/genetics , Procollagen N-Endopeptidase/genetics , Procollagen N-Endopeptidase/metabolism , Synovial Membrane/cytology , Tumor Necrosis Factor-alpha/geneticsABSTRACT
Disturbance of blood supply to the femoral head is a risk factor for corticosteroid-associated osteonecrosis. The aim was to measure blood supply of the proximal femur during corticosteroid therapy in systemic lupus erythematosus (SLE) patients. We repeatedly performed 78 dynamic MRIs of 19 hip joints in 19 SLE patients after initiation of corticosteroid administration for one year. Blood supply of the femoral head (epiphysis, growth plate, and metaphysis), the femoral neck, and the medial circumflex femoral artery were measured in terms of peak percent enhancement. At the first month, blood supply of the growth plate was significantly higher in the pediatric group (<15 years old) than in the adolescent and adult group (>15 years old). At the fourth month, blood supply in every part of the femoral head (epiphysis, growth plate, and metaphysis) was significantly higher in the pediatric group than in the adolescent and adult group. Multiple regression analysis revealed that blood supply to the femoral head depended on the number of days after initiation of corticosteroid administration and the age at the time of dynamic MRI. Blood supply to the femoral head is abundant in pediatric patients and is a function of the number of days after initiation of corticosteroid administration.
Subject(s)
Femur Head/blood supply , Glucocorticoids/adverse effects , Lupus Erythematosus, Systemic/drug therapy , Adolescent , Adult , Age Factors , Child , Female , Femur Head/drug effects , Femur Neck/blood supply , Femur Neck/drug effects , Glucocorticoids/administration & dosage , Glucocorticoids/therapeutic use , Growth Plate/blood supply , Growth Plate/drug effects , Hip Joint/blood supply , Hip Joint/drug effects , Humans , Magnetic Resonance Imaging , Male , Middle Aged , Osteonecrosis/chemically induced , Prospective Studies , Regression Analysis , Risk Factors , Time Factors , Young AdultABSTRACT
BACKGROUND AND PURPOSE: DTI can provide valuable structural information that may become an innovative tool in evaluating lumbar foraminal nerve root entrapment. The purpose of this study was to visualize the lumbar nerve roots and to measure their FA in healthy volunteers and patients with lumbar foraminal stenosis by using DTI and tractography with 3T MR imaging. MATERIALS AND METHODS: Eight patients with lumbar foraminal stenosis and 8 healthy volunteers underwent 3T MR imaging. In all subjects, DTI was performed with echo-planar imaging at a b-value of 800 s/mm(2) and the lumbar nerve roots were visualized with tractography. Mean FA values in the lumbar nerve roots were quantified on DTI images. RESULTS: In all subjects, the lumbar nerve roots were clearly visualized with tractography. In all patients, tractography also showed abnormalities such as tract disruption, nerve narrowing, and indentation in their course through the foramen. Mean FA values were significantly lower in entrapped roots than in intact roots. CONCLUSIONS: We demonstrated that DTI and tractography of human lumbar nerves can visualize and quantitatively evaluate lumbar nerve entrapment with foraminal stenosis. We believe that DWI is a potential tool for the diagnosis of lumbar nerve entrapment.
Subject(s)
Diffusion Tensor Imaging/methods , Nerve Compression Syndromes/pathology , Spinal Nerve Roots/injuries , Spinal Nerve Roots/pathology , Adult , Aged , Female , Humans , Image Enhancement/methods , Lumbar Vertebrae/pathology , Male , Middle Aged , Pilot Projects , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
OBJECTIVES: The present study evaluated the effects of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) and CPP-ACP with 900 ppm fluoride (CPP-ACPF) pastes on inhibition of enamel demineralization over time, using polychromatic micro-computed tomography (micro-CT). METHODS: Enamel blocks were prepared from bovine teeth. The specimens were each treated by one of the following agents, 30 min daily for 7 days: deionized water (negative control); CPP-ACP paste; CPP-ACPF paste; and NaF solutions (positive controls) (90, 900, and 9000 ppm F). After treatment, the specimens were immersed in a demineralizing solution (pH 4.5) for 24, 72, and 120 h. Mean mineral loss (ML) and lesion depth (LD) after each period were determined from mineral density profiles obtained using micro-CT. RESULTS: ML values in all the treatment groups were significantly smaller than those in the control group after 72 and 120 h of demineralization (p < 0.05, two-way ANOVA and t-test with Bonferroni correction). ML values in CPP-ACPF and NaF solution groups were significantly smaller compared to CPP-ACP group after 72 h (p < 0.05). LD values in the CPP-ACPF and all the NaF solutions groups were significantly smaller compared to the control group after 120 h (p < 0.05). The 9000 ppm F group showed the lowest nominal ML and LD values. CONCLUSIONS: The application of CPP-ACP or CPP-ACPF pastes to sound enamel surfaces resulted in inhibition of enamel demineralization, and a better effect was noted for the latter paste. Quantitative assessment using polychromatic micro-CT demonstrated to be useful for detecting mineral density changes.
Subject(s)
Cariostatic Agents/pharmacology , Caseins/pharmacology , Dental Enamel/drug effects , Sodium Fluoride/pharmacology , Tooth Demineralization/pathology , X-Ray Microtomography/methods , Animals , Cariostatic Agents/administration & dosage , Caseins/administration & dosage , Cattle , Hydrogen-Ion Concentration , Imaging, Three-Dimensional/methods , Materials Testing , Minerals/analysis , Sodium Fluoride/administration & dosage , Solutions , Suspensions , Temperature , Time Factors , Tooth Demineralization/prevention & controlABSTRACT
OBJECTIVE: The aim of this study was to evaluate twice-daily injections of biphasic insulin lispro vs. basal-bolus (BB) therapy with regard to quality-of-life (QOL) and glycaemic control in insulin-naïve type 2 diabetic patients. METHODS: Twenty-eight patients with type 2 diabetes were randomized to receive either twice-daily 50/50 premixed insulin lispro (Mix50 group) or BB (NPH insulin at bedtime and preprandial insulin lispro) therapy (BB group) for 12 weeks. Glycated haemoglobin (HbA1C), 1,5-anhydroglucitol (1,5-AG), blood plasma glucose level, body mass index (BMI), daily total insulin dosage and insulin therapy-related QOL (ITR-QOL) were studied. RESULTS: ITR-QOL was significantly better in the Mix50 than in the BB group (103.1 +/- 9.8 vs. 90.6 +/- 19.4; p < 0.05). HbA(1c) improved in both groups (from 11.1 +/- 2.1 to 6.9 +/- 1.0% with Mix50 vs. from 11.0 +/- 2.3 to 6.6 +/- 0.8% with BB therapy). CONCLUSION: These results might suggest that twice-daily injections of premixed rapid-acting insulin analogue therapy could achieve good glycaemic control and better QOL compared with BB therapy in insulin-naïve type 2 diabetes.
Subject(s)
Blood Glucose/metabolism , Diabetes Mellitus, Type 2/drug therapy , Hypoglycemic Agents/administration & dosage , Insulin, Isophane/administration & dosage , Insulin/analogs & derivatives , Quality of Life , Adult , Aged , Aged, 80 and over , Body Mass Index , Diabetes Mellitus, Type 2/blood , Female , Glycated Hemoglobin/metabolism , Humans , Insulin/administration & dosage , Insulin Lispro , Male , Middle Aged , Treatment Outcome , Young AdultABSTRACT
Using a rat model the characteristics of the sensory neurones of the dorsal-root ganglia (DRG) innervating the hip were investigated by retrograde neurotransport and immunohistochemistry. Fluoro-Gold solution (FG) was injected into the left hip of ten rats. Seven days later the DRG from both sides between T12 and L6 were harvested. The number of FG-labelled calcitonin gene-related peptide-immunoreactive or isolectin B4-binding neurones were counted. The FG-labelled neurones were distributed throughout the left DRGs between T13 and L5, primarily at L2, L3, and L4. Few FG-labelled isolectin B4-binding neurones were present in the DRGs of either side between T13 and L5, but calcitonin gene-related peptide-immunoreactive neurones made up 30% of all FG-labelled neurones. Our findings may explain the referral of pain from the hip to the thigh or lower leg corresponding to the L2, L3 and L4 levels. Since most neurones are calcitonin gene-related peptide-immunoreactive peptide-containing neurones, they may have a more significant role in the perception of pain in the hip as peptidergic DRG neurones.
Subject(s)
Calcitonin Gene-Related Peptide/metabolism , Ganglia, Spinal/physiopathology , Hip/innervation , Lumbar Vertebrae/innervation , Neurons, Afferent/physiology , Pain Measurement/methods , Animals , Biomechanical Phenomena , Fluorescent Dyes , Ganglia, Spinal/metabolism , Immunohistochemistry , Male , Rats , Rats, Sprague-Dawley , StilbamidinesABSTRACT
Buffer-gas pressure broadening for the nu(1)+nu(3) band of H(2)O at 1.34-1.44 mum for a variety of buffer gases was investigated at room temperature using continuous-wave cavity ring-down spectroscopy. The effective interaction energy of water dimer under room temperature conditions was evaluated from the pressure broadening coefficients for rare gases using Permenter-Seaver's relation. Monte Carlo simulations were performed using ab initio molecular orbital calculations to evaluate the interaction energies for the water dimer at 300 K. In this theoretical calculation, the orientations of the two water molecules were statistically treated.
Subject(s)
Models, Chemical , Models, Molecular , Water/chemistry , Computer Simulation , Dimerization , Energy Transfer , Macromolecular Substances/chemistry , Molecular Conformation , Pressure , TemperatureABSTRACT
OBJECTIVE: To address the clinical implications of leptin and to re-examine the relationship between leptin and its potential humoral regulators such as insulin, nonesterified fatty acids (NEFA) and triiodothyronine (T3) in low-calorie diet (LCD) for obese humans. DESIGN: Longitudinal study. SETTING: University and foundation hospitals. SUBJECTS: Ten obese men and 10 premenopausal obese women. INTERVENTIONS: Five men and five women took 800 kcal/day LCD and another five men and five women took 1400 kcal/day balanced deficit diet (BDD) during 4 weeks. RESULTS: Plasma leptin levels in the LCD group decreased more markedly (46.2+/-14.6 to 13.2+/-3.6 ng/ml) than that expected for the decrement in percentage fat (39.0+/-1.7 to 35.9+/-1.7%) and body mass index (BMI; 35.4+/-2.4 to 33.1+/-2.2 kg/m(2)), while that in the BDD group did not decrease significantly (14.9+/-3.5 to 13.4+/-2.8 ng/ml). The ratio of the decrease in leptin levels to that of BMI during the first week was significantly greater than that during the following 3 weeks (39.5+/-2.7 vs 29.3+/-2.1%, P=0.017). The plasma insulin and T3 levels also fell substantially in the first week and continued to decrease during the entire course. Plasma leptin levels measured weekly in each subject were correlated well with insulin (r=0.586, P=0.0003) and T3 (r=0.785, P=0.0004). Multiple regression analyses after adjustment for the time course and BMI revealed that serum levels of T3 were independently correlated with plasma leptin levels (r=0.928, P<0.0001). The plasma NEFA level was markedly elevated during the first 2 weeks and decreased thereafter. CONCLUSIONS: A rapid fall in leptin during the first week of LCD, coordinated by insulin, T3 and NEFA, should be beneficial for responding to decreased energy intake. Inversely, in view of the powerful effect of leptin on energy dissipation, the present findings suggest the potential usefulness of leptin in combination with caloric restriction for the treatment of obesity. SPONSORSHIP: The Ministry of Education, Culture, Sports, Science and Technology of Japan and the Ministry of Health, Labour and Welfare of Japan.
Subject(s)
Diet, Reducing , Fatty Acids, Nonesterified/blood , Leptin/blood , Obesity/diet therapy , Triiodothyronine/blood , Adolescent , Adult , Blood Glucose/metabolism , Body Mass Index , Energy Intake , Female , Humans , Insulin/blood , Longitudinal Studies , Male , Middle Aged , Obesity/blood , Time FactorsABSTRACT
There have been several reports on the use of extracorporeal shock waves in the treatment of pseudarthrosis, calcifying tendinitis, and tendinopathies of the elbow. However, the pathomechanism of pain relief has not been clarified. To investigate the analgesic properties of shock wave application, we analyzed whether it produces morphologic changes in cutaneous nerve fibres. In normal rat skin, the epidermis is heavily innervated by nerve fibres immunoreactive for protein gene product (PGP) 9.5 and by some fibres immunoreactive for calcitonin gene-related peptide (CGRP). There was nearly complete degeneration of epidermal nerve fibres in the shock wave-treated skin, as indicated by the loss of immunoreactivity for PGP 9.5 or CGRP. Reinnervation of the epidermis occurred 2 weeks after treatment. These data show that relief of pain after shock wave application to the skin results from rapid degeneration of the intracutaneous nerve fibres.
Subject(s)
High-Energy Shock Waves , Nerve Fibers/radiation effects , Skin/radiation effects , Animals , Calcitonin Gene-Related Peptide/metabolism , Epidermis/innervation , Epidermis/radiation effects , Hindlimb , Immunohistochemistry , Male , Nerve Degeneration , Nerve Fibers/metabolism , Pain Measurement , Rats , Rats, Sprague-Dawley , Skin/innervation , Thiolester Hydrolases/metabolism , Ubiquitin ThiolesteraseABSTRACT
Uncoupling protein 3 (UCP3), which uncouples electron transport from ATP synthesis, is expressed at high levels in the skeletal muscle, an important organ in glucose and lipid metabolism. Because several reports proposed that fatty acids induced UCP3 gene expression in skeletal muscle in vivo, in the present study we examined the regulation of UCP3 gene expression by various fatty acids using L6 myotubes. UCP3 gene expression was increased in L6 myotubes by various fatty acids or by alpha-bromopalmitate, a nonmetabolized derivative of palmitic acid. Because fatty acids are also known as agonists for PPARs, we examined the involvement of PPARs in the regulation of the UCP3 gene expression. L-165041, a PPAR delta agonist, increased UCP3 gene expression in L6 myotubes, whereas neither Wy 14,643, a PPAR alpha agonist, nor Pioglitazone, a PPAR gamma agonist, increased it. Therefore, we conclude that UCP3 gene expression is increased by the activation of PPAR delta in L6 myotubes and postulate that PPAR delta mediates at least some part of the increased UCP3 gene expression by fatty acids in skeletal muscle in vivo.
Subject(s)
Carrier Proteins/genetics , Muscle, Skeletal/physiology , Receptors, Cytoplasmic and Nuclear/physiology , Transcription Factors/physiology , Animals , Cell Line , Fatty Acids/pharmacology , Ion Channels , Mitochondrial Proteins , Rats , Receptors, Cytoplasmic and Nuclear/agonists , Signal Transduction/drug effects , Transcription Factors/agonists , Uncoupling Protein 3 , Up-Regulation/drug effectsABSTRACT
Thiazolidinediones, insulin-sensitizing agents, have been reported to increase glucose uptake along with the expression of glucose transporters in adipocytes and cardiomyocytes. Recently, we have further suggested that the translocation of GLUT4 is stimulated by thiazolidinediones in L6 myocytes. However, the direct effects of thiazolidinediones on translocation of glucose transporters have not yet been determined. In this study, using hemagglutinin epitope-tagged GLUT4 (GLUT4-HA), we provide direct evidence of the effect of troglitazone on the translocation of GLUT4 in rat epididymal adipocytes. Primary cultures of rat adipocytes were transiently transfected with GLUT4-HA and overexpressed eightfold compared with endogenous GLUT4 in transfected cells. A total of 24 h of treatment with troglitazone (10(-4) mol/l) increased the cell surface level of GLUT4-HA by 1.5 +/- 0.03-fold (P < 0.01) without changing the total amount of GLUT4-HA, whereas it increased the protein level of endogenous GLUT4 (1.4-fold) without changing that of GLUT1. Thus, the direct effect on the translocation can be detected apart from the increase in endogenous GLUT4 content using GLUT4-HA. Troglitazone not only increased the translocation of GLUT4-HA on the cell surface in the basal state but also caused a leftward shift in the dose-response relations between GLUT4-HA translocation and insulin concentration in the medium (ED(50): from approximately 0.1 to 0.03 nmol/l). These effects may partly contribute to the antidiabetic activity of troglitazone in patients with obesity and type 2 diabetes.
Subject(s)
Adipocytes/metabolism , Chromans/pharmacology , Monosaccharide Transport Proteins/metabolism , Muscle Proteins , Thiazoles/pharmacology , Thiazolidinediones , Animals , Biological Transport/drug effects , Epitopes , Glucose Transporter Type 1 , Glucose Transporter Type 4 , Hemagglutinins/immunology , Hemagglutinins/metabolism , Insulin/pharmacology , Male , Rats , Rats, Sprague-Dawley , TroglitazoneABSTRACT
A 24-year-old female suffered from acute pancreatitis, followed by simultaneous onset of painless goiter, elevation of thyroid hormones and diabetic ketoacidosis. Two months later, her insulin secreting function was severely decreased and positive for anti-GAD and anti-islet cell antibodies, whereas the serum glucagon level was normal, suggesting an autoimmune-related destruction specifically of beta cells. In addition, the initial hyperthyroid state was followed by a hypothyroid phase which later recovered to an euthyroid state, suggesting an initial destruction of thyroid cells. Because anti-thyroidal antibodies were positive, it is likely that the thyroidal destruction was also autoimmune-related. This case implies common pathogenic mechanisms in the autoimmunity related destruction of beta cells and thyroid cells.
Subject(s)
Diabetes Mellitus, Type 1/etiology , Pancreatitis/complications , Thyroiditis/etiology , Acute Disease , Adult , Diabetes Mellitus, Type 1/complications , Female , Humans , Thyroiditis/complications , Time FactorsABSTRACT
A number of studies have demonstrated that insulin resistance in the skeletal muscle plays a pivotal role in the insulin resistance associated with obesity and type 2 diabetes. A decrease in GLUT4 translocation from the intracellular pool to the plasma membranes in skeletal muscles has been implicated as a possible cause of insulin resistance. Herein, we examined the effects of an insulin-sensitizing drug, troglitazone (TGZ), on glucose uptake and the translocation of GLUT4 in L6 myotubes. The prolonged exposure (24 h) of L6 myotubes to TGZ (10(-5) mol/l) caused a substantial increase in the 2-deoxy-[3H]D-glucose (2-DG) uptake without changing the total amount of the glucose transporters GLUT4, GLUT1, and GLUT3. The TGZ-induced 2-DG uptake was completely abolished by cytochalasin-B (10 micromol/l). The ability of TGZ to translocate GLUT4 from light microsomes to the crude plasma membranes was greater than that of insulin. Both cycloheximide treatment (3.5 x 10(-6) mol/l) and the removal of TGZ by washing reversed the 2-DG uptake to the basal level. Moreover, insulin did not enhance the TGZ-induced 2-DG uptake additively. The TGZ-induced 2-DG uptake was only partially reversed by wortmannin to 80%, and TGZ did not change the expression and the phosphorylation of protein kinase B; the expression of protein kinase C (PKC)-lambda, PKC-beta2, and PKC-zeta; or 5'AMP-activated protein kinase activity. a-Tocopherol, which has a molecular structure similar to that of TGZ, did not increase 2-DG uptake. We conclude that the glucose transport in L6 myotubes exposed to TGZ for 24 h is the result of an increased translocation of GLUT4. The present results imply that the effects of troglitazone on GLUT4 translocation may include a new mechanism for improving glucose transport in skeletal muscle.
Subject(s)
Chromans/pharmacology , Monosaccharide Transport Proteins/genetics , Monosaccharide Transport Proteins/metabolism , Muscle Proteins , Muscle, Skeletal/metabolism , Nerve Tissue Proteins , Thiazoles/pharmacology , Thiazolidinediones , Animals , Biological Transport/drug effects , Cell Line , Cell Membrane/metabolism , Cycloheximide/pharmacology , Deoxyglucose/metabolism , Gene Expression Regulation/drug effects , Glucose Transporter Type 1 , Glucose Transporter Type 3 , Glucose Transporter Type 4 , Insulin/pharmacology , Kinetics , Muscle, Skeletal/cytology , Muscle, Skeletal/drug effects , Protein Transport/drug effects , RNA, Messenger/genetics , Rats , Receptors, Cytoplasmic and Nuclear/genetics , Rosiglitazone , Transcription Factors/genetics , Transcription, Genetic/drug effects , TroglitazoneABSTRACT
Lipoatrophic diabetes is caused by a deficiency of adipose tissue and is characterized by severe insulin resistance, hypoleptinemia, and hyperphagia. The A-ZIP/F-1 mouse (A-ZIPTg/+) is a model of severe lipoatrophic diabetes and is insulin resistant, hypoleptinemic, hyperphagic, and shows severe hepatic steatosis. We have also produced transgenic "skinny" mice that have hepatic overexpression of leptin (LepTg/+) and no adipocyte triglyceride stores, and are hypophagic and show increased insulin sensitivity. To explore the pathophysiological and therapeutic roles of leptin in lipoatrophic diabetes, we crossed LepTg/+ and A-ZIPTg/+ mice, producing doubly transgenic mice (LepTg/+:A-ZIPTg/+) virtually lacking adipose tissue but having greatly elevated leptin levels. The LepTg/+:A-ZIPTg/+ mice were hypophagic and showed improved hepatic steatosis. Glucose and insulin tolerance tests revealed increased insulin sensitivity, comparable to LepTg/+ mice. These effects were stable over at least 6 months of age. Pair-feeding the A-ZIPTg/+ mice to the amount of food consumed by LepTg/+:A-ZIPTg/+ mice did not improve their insulin resistance, diabetes, or hepatic steatosis, demonstrating that the beneficial effects of leptin were not due to the decreased food intake. Continuous leptin administration that elevates plasma leptin concentrations to those of LepTg/+:A-ZIPTg/+ mice also effectively improved hepatic steatosis and the disorder of glucose and lipid metabolism in A-ZIP/F-1 mice. These data demonstrate that leptin can improve the insulin resistance and diabetes of a mouse model of severe lipoatrophic diabetes, suggesting that leptin may be therapeutically useful in the long-term treatment of lipoatrophic diabetes.
Subject(s)
Diabetes Mellitus, Lipoatrophic/drug therapy , Diabetes Mellitus, Lipoatrophic/physiopathology , Insulin Resistance , Leptin/therapeutic use , Animals , Blood Glucose/analysis , Body Weight/drug effects , Diabetes Mellitus, Lipoatrophic/pathology , Eating , Gene Expression , Infusion Pumps , Injections , Leptin/administration & dosage , Leptin/blood , Leptin/genetics , Lipids/blood , Mice , Mice, Inbred Strains , Mice, Transgenic/genetics , Organ Size , Transgenes/geneticsABSTRACT
Ghrelin, an endogenous ligand for growth hormone secretagogue (GHS) receptor originally isolated from the stomach, occurs in the hypothalamic arcuate nucleus and may play a role in energy homeostasis. Synthetic GHSs have activated the hypothalamic arcuate neurons containing neuropeptide Y (NPY), suggesting the involvement of NPY in some of ghrelin actions. This study was designed to elucidate the role of ghrelin in the regulation of food intake. A single intracerebroventricular (ICV) injection of ghrelin (5-5,000 ng/rat) caused a significant and dose-related increase in cumulative food intake in rats. Ghrelin (500 ng/rat) was also effective in growth hormone-deficient spontaneous dwarf rats. Hypothalamic NPY mRNA expression was increased in rats that received a single ICV injection of ghrelin (500 ng/rat) (approximately 160% of that in vehicle-treated groups, P < 0.05). The ghrelin's orexigenic effect was abolished dose-dependently by ICV co-injection of NPY Y1 receptor antagonist (10-30 microg/rat). The leptin-induced inhibition of food intake was reversed by ICV co-injection of ghrelin in a dose-dependent manner (5-500 ng/rat). Leptin reduced hypothalamic NPY mRNA expression by 35% (P < 0.05), which was abolished by ICV co-injection of ghrelin (500 ng/rat). This study provides evidence that ghrelin is an orexigenic peptide that antagonizes leptin action through the activation of hypothalamic NPY/Y1 receptor pathway.
Subject(s)
Growth Hormone/metabolism , Hypothalamus/metabolism , Leptin/antagonists & inhibitors , Neuropeptide Y/physiology , Peptide Hormones , Peptides/physiology , Animals , Drug Combinations , Ghrelin , Growth Hormone/deficiency , Hypothalamus/drug effects , Injections, Intraventricular , Leptin/pharmacology , Male , Neuropeptide Y/genetics , Peptides/pharmacology , RNA, Messenger/antagonists & inhibitors , RNA, Messenger/metabolism , Rats , Rats, Sprague-DawleyABSTRACT
A case of a volar periscapholunate dislocation is reported. Treatment by closed reduction and Kirschner wire fixation gave a good clinical result.