RESUMEN
CD146/MCAM has garnered significant attention for its potential contribution to cardiovascular disease; however, the transcriptional regulation and functions remain unclear. To explore these processes regarding cardiomyopathy, we employed doxorubicin, a widely used stressor for cardiomyocytes. Our in vitro study on H9c2 cardiomyoblasts highlights that, besides impairing the fatty acid uptake in the cells, doxorubicin suppressed the expression of fatty acid binding protein 4 (Fabp4) along with the histone deacetylase 9 (Hdac9), bromodomain and extra-terminal domain proteins (BETs: Brd2 and Brd4), while augmented the production of CD146/MCAM. Silencing and chemical inhibition of Hdac9 further augmented CD146/MCAM and deteriorated fatty acid uptake. In contrast, chemical inhibition of BETs as well as silencing of MCAM/CD146 ameliorated fatty acid uptake. Moreover, protein kinase C (PKC) inhibition abrogated CD146/MCAM, particularly in the nucleus. Taken together, our results suggest that epigenetic dysregulation of Hdac9, Brd2, and Brd4 alters CD146/MCAM expression, deteriorating fatty acid uptake by downregulating Fabp4. This process depends on the PKC-mediated nuclear translocation of CD146. Thus, this study highlights a pivotal role of CD146/MCAM in doxorubicin-induced cardiomyopathy.
Asunto(s)
Cardiomiopatías , Factores de Transcripción , Humanos , Antígeno CD146/genética , Antígeno CD146/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Epigénesis GenéticaRESUMEN
Gicerin/CD146 is a cell adhesion molecule which belongs to the immunoglobulin (Ig) superfamily. We have reported the existence of gicerin/CD146 in the nervous system, heart, lung and smooth muscles of blood vessels. In this study, we make a cardiac hypertrophy model rat by constricting the rat aorta (AAC, ascending aortic constriction) and examined the effect on the expression of gicerin/CD146 in the heart. We found that the expression level of gicerin/CD146 was increased by the AAC treatment. Next, stretch stimulation was applied to myocardial cell line H9c2 cells to confirm that gicerin/CD146 may participate in the cellular hypertrophy model. We also treated the cells with inhibitors of MAP pathway enzymes. In cultured myocardial cells, the expression level of gicerin/CD146 was increased by the stretch stimulation and decreased by inhibiting the MAP pathway. Based on the above findings, it is suggested that the expression of gicerin/CD146 is involved in cardiac hypertrophy, and that the MAP pathway may be involved in the expression of gicerin/CD146 RNA in the cardiomyocyte. In addition, the expression level of gicerin/CD146 RNA in neonatal rats was upregulated after birth. Therefore, it is suggested that gicerin/CD146 might participate in the increase of myocardial cell volume both in the pathway of cardiac hypertrophy and in the developmental growth of heart.
Asunto(s)
Antígeno CD146/metabolismo , Cardiomegalia/metabolismo , Regulación de la Expresión Génica , Corazón/crecimiento & desarrollo , Miocardio/metabolismo , Animales , Cardiomegalia/patología , Línea Celular , Masculino , Ratas , Ratas Sprague-DawleyRESUMEN
Although the role of interventional radiology in the field of obstetrical hemorrhage has been widely reported upon recently, the rate of procedure-related complications has not been fully determined. We present the case of a patient who developed an external iliac artery thrombosis, a rarely reported complication associated with prophylactic common iliac artery balloon occlusion (CIABO). After CIABO, we found that the dorsalis pedis artery of the right foot was weak and the foot was cold, despite the fact that the patient had no complaints. Computed tomography demonstrated a linear thrombus in the right external iliac artery. We managed the patient conservatively using a heparin drip without the need for thromboembolectomy. Our experience suggests that it is important to consider the risk of thrombosis formation after CIABO. Physical examination post-procedure is key to identifying this complication early.
Asunto(s)
Oclusión con Balón/efectos adversos , Pérdida de Sangre Quirúrgica/prevención & control , Cesárea/efectos adversos , Histerectomía/efectos adversos , Arteria Ilíaca/diagnóstico por imagen , Placenta Accreta/cirugía , Trombosis/etiología , Adulto , Oclusión con Balón/métodos , Cesárea/métodos , Femenino , Humanos , Histerectomía/métodos , Imagen por Resonancia Magnética , Placenta Accreta/diagnóstico por imagen , Embarazo , Trombosis/diagnóstico por imagen , Ultrasonografía PrenatalRESUMEN
PURPOSE: To evaluate the safety and efficacy of benzalkonium chloride (BAK)-optimized tafluprost (with a BAK concentration reduced from 0.01% to 0.001%) in glaucoma patients with existing superficial punctate keratitis (SPK). PATIENTS AND METHODS: A prospective, multicenter, open-label study was designed to compare BAK-optimized tafluprost administered over 12 weeks relative to other preserved prostaglandin analogs previously administered in Japanese glaucoma patients. Thirty patients with SPK graded at <6 points by area density (AD) scoring in 1 eye were recruited. The primary outcome measure was change in AD score at 12 weeks after the switch in treatment compared with that at baseline. Secondary outcome measures included changes in tear film breakup time (TBUT), hyperemia score, and intraocular pressure (IOP). Four patients were excluded from analysis because of treatment discontinuation. RESULTS: Mean AD score±SD decreased significantly from 3.4±0.9 to 1.8±1.8 after the switch (P<0.0001). Mean TBUT increased significantly from 6.3±3.3 to 8.0±4.2 seconds (P<0.01). Mean hyperemia score remained unchanged, whereas mean IOP decreased significantly from 15.6±2.6 to 14.4±2.0 mm Hg (P<0.01). For patients previously treated with BAK-preserved latanoprost (n=17) or bimatoprost (n=2), mean AD score decreased significantly from 3.4±0.9 to 1.8±1.8 (P<0.01) and mean TBUT increased significantly from 6.4±3.6 to 8.2±4.3 seconds (P<0.01); no such changes were apparent for patients previously treated with sofZia-preserved travoprost (n=7). CONCLUSIONS: BAK-optimized tafluprost is a treatment option to improve the condition of the ocular surface and to maintain IOP control in glaucoma patients with existing SPK who have been previously treated with other BAK-preserved prostaglandin analogs.
Asunto(s)
Antihipertensivos/uso terapéutico , Compuestos de Benzalconio/uso terapéutico , Glaucoma/tratamiento farmacológico , Queratitis/complicaciones , Conservadores Farmacéuticos/uso terapéutico , Prostaglandinas F/uso terapéutico , Anciano , Anciano de 80 o más Años , Antihipertensivos/efectos adversos , Pueblo Asiatico , Compuestos de Benzalconio/efectos adversos , Femenino , Humanos , Presión Intraocular/efectos de los fármacos , Masculino , Persona de Mediana Edad , Hipertensión Ocular/tratamiento farmacológico , Conservadores Farmacéuticos/efectos adversos , Estudios Prospectivos , Prostaglandinas F/efectos adversos , Tonometría OcularRESUMEN
1. Transgenic (TG) mice overexpressing an arg120gly missense mutation in heat shock protein B5 (HSPB5; i.e. R120G TG mice) exhibit desmin-related cardiomyopathy. Recently, the cardioprotective effect of nicorandil has been shown to prolong the survival of R120G TG mice. However, whether the TG mice exhibit ventricular arrhythmias and whether nicorandil can inhibit these arrhythmias remain unknown. In the present study we examined the effects of chronic nicorandil administration on ventricular electrical remodelling and arrhythmias in R120G TG mice. 2. Mice were administered nicorandil (15 mg/kg per day) or vehicle (water) orally from 5 to 30 weeks of age. Electrocardiograms (ECG) and optical action potentials were recorded from R120G TG mouse hearts. In addition, the expression of ventricular connexin 43 and the cardiac Na(+) channel Nav1.5 was examined in TG mice. 3. All ECG parameters tested were prolonged in R120G TG compared with non-transgenic (NTG) mice. Nicorandil improved the prolonged P, PQ and QRS intervals in R120G TG mice. Interestingly, impulse conduction slowing and increases in the expression of total and phosphorylated connexin 43 and Nav1.5 were observed in ventricles from R120G TG compared with NTG mice. Nicorandil improved ventricular impulse conduction slowing and normalized the increased protein expression levels of total and phosphorylated connexin 43, but not of Nav1.5, in R120G TG mouse hearts. Electrical rapid pacing at the ventricle induced ventricular tachyarrhythmias (VT) in six of eight R120G TG mouse hearts, but not in any of the eight nicorandil-treated R120G TG mouse hearts (P < 0.05). 4. These findings demonstrate that nicorandil inhibits cardiac electrical remodelling and that the prevention of VT by nicorandil is associated with normalization of connexin 43 expression in this model.
Asunto(s)
Antiarrítmicos/farmacología , Cardiomiopatías/fisiopatología , Desmina/fisiología , Nicorandil/farmacología , Taquicardia Ventricular/prevención & control , Anestesia , Animales , Western Blotting , Peso Corporal/efectos de los fármacos , Conexina 43/biosíntesis , Ecocardiografía , Estimulación Eléctrica , Electrocardiografía , Fenómenos Electrofisiológicos/efectos de los fármacos , Inmunohistoquímica , Técnicas In Vitro , Ratones , Ratones Transgénicos , Canal de Sodio Activado por Voltaje NAV1.5/biosíntesis , Tamaño de los Órganos/efectos de los fármacosRESUMEN
Early-onset epileptic encephalopathies (EOEE) are severe neurological disorders characterized by frequent seizures accompanied by developmental regression or retardation. Whole-exome sequencing of 12 patients together with five pairs of parents and subsequent Sanger sequencing in additional 328 EOEE patients identified two de novo frameshift and one missense mutations in SLC35A2 at Xp11.23, respectively. The three patients are all females. X-inactivation analysis of blood leukocyte DNA and mRNA analysis using lymphoblastoid cells derived from two patients with a frameshift mutation indicated that only the wild-type SLC35A2 allele was expressed in these cell types, at least in part likely as a consequence of skewed X-inactivation. SLC35A2 encodes a UDP-galactose transporter (UGT), which selectively supplies UDP-galactose from the cytosol to the Golgi lumen. Transient expression experiments revealed that the missense mutant protein was correctly localized in the Golgi apparatus. In contrast, the two frameshift mutant proteins were not properly expressed, suggesting that their function is severely impaired. Defects in the UGT can cause congenital disorders of glycosylation. Of note, no abnormalities of glycosylation were observed in three serum glycoproteins, which is consistent with favorably skewed X-inactivation. We hypothesize that a substantial number of neurons might express the mutant SLC35A2 allele and suffer from defective galactosylation, resulting in EOEE.
Asunto(s)
Proteínas de Transporte de Monosacáridos/genética , Mutación , Espasmos Infantiles/diagnóstico , Espasmos Infantiles/genética , Edad de Inicio , Animales , Encéfalo/patología , Encéfalo/fisiopatología , Línea Celular , Niño , Análisis Mutacional de ADN , Electroencefalografía , Exoma , Facies , Femenino , Expresión Génica , Orden Génico , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Lactante , Recién Nacido , Imagen por Resonancia Magnética , Ratones , Proteínas de Transporte de Monosacáridos/química , Fenotipo , Transporte de Proteínas , Isoformas de ARNRESUMEN
Microphthalmia with limb anomalies (MLA), also known as Waardenburg anophthalmia syndrome or ophthalmoacromelic syndrome, is a rare autosomal recessive disorder. Recently, we and others successfully identified SMOC1 as the causative gene for MLA. However, there are several MLA families without SMOC1 abnormality, suggesting locus heterogeneity in MLA. We aimed to identify a pathogenic mutation in one Lebanese family having an MLA-like condition without SMOC1 mutation by whole-exome sequencing (WES) combined with homozygosity mapping. A c.683C>T (p.Thr228Met) in FNBP4 was found as a primary candidate, drawing the attention that FNBP4 and SMOC1 may potentially modulate BMP signaling.
Asunto(s)
Proteínas Portadoras/genética , Mutación , Síndrome de Waardenburg/genética , Exones , Familia , Femenino , Homocigoto , Humanos , Péptidos y Proteínas de Señalización Intracelular , Masculino , Osteonectina/genética , Linaje , Análisis de Secuencia/métodosRESUMEN
Static encephalopathy of childhood with neurodegeneration in adulthood (SENDA) is a recently established subtype of neurodegeneration with brain iron accumulation (NBIA). By exome sequencing, we found de novo heterozygous mutations in WDR45 at Xp11.23 in two individuals with SENDA, and three additional WDR45 mutations were identified in three other subjects by Sanger sequencing. Using lymphoblastoid cell lines (LCLs) derived from the subjects, aberrant splicing was confirmed in two, and protein expression was observed to be severely impaired in all five. WDR45 encodes WD-repeat domain 45 (WDR45). WDR45 (also known as WIPI4) is one of the four mammalian homologs of yeast Atg18, which has an important role in autophagy. Lower autophagic activity and accumulation of aberrant early autophagic structures were demonstrated in the LCLs of the affected subjects. These findings provide direct evidence that an autophagy defect is indeed associated with a neurodegenerative disorder in humans.
Asunto(s)
Autofagia , Proteínas Portadoras/genética , Exoma/genética , Discapacidad Intelectual/etiología , Mutación/genética , Enfermedades Neurodegenerativas/etiología , Espasmos Infantiles/etiología , Adulto , Niño , Femenino , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Hierro/metabolismo , Síndrome de Lennox-Gastaut , Imagen por Resonancia Magnética , FenotipoRESUMEN
PURPOSE: Congenital cataract is one of the most frequent causes of visual impairment and childhood blindness. Approximately one quarter to one third of congenital cataract cases may have a genetic cause. However, phenotypic variability and genetic heterogeneity hamper correct genetic diagnosis. In this study, we used whole-exome sequencing (WES) to identify pathogenic mutations in two Korean families with congenital cataract. METHODS: Two affected members from each family were pooled and processed for WES. The detected variants were confirmed with direct sequencing. RESULTS: WES readily identified a CRYAA mutation in family A and a CRYGC mutation in family B. The c.61C>T (p.R21W) mutation in CRYAA has been previously reported in a family with congenital cataract and microcornea. The novel mutation, c.124delT, in CRYGC may lead to a premature stop codon (p.C42Afs*60). CONCLUSIONS: This study clearly shows the efficacy of WES for rapid genetic diagnosis of congenital cataract with an unknown cause. WES will be the first choice for clinical services in the near future, providing useful information for genetic counseling and family planning.
Asunto(s)
Catarata/congénito , Catarata/genética , Cristalinas/genética , Mutación , gamma-Cristalinas/genética , Secuencia de Aminoácidos , Pueblo Asiatico/genética , Secuencia de Bases , Codón sin Sentido , Análisis Mutacional de ADN , Exoma , Femenino , Mutación del Sistema de Lectura , Estudio de Asociación del Genoma Completo , Humanos , Masculino , Mutación Missense , Linaje , República de Corea , Eliminación de SecuenciaRESUMEN
Moyamoya disease (MMD) is characterized by severe stenoses of the arteries in the circle of Willis, which predispose the patient to brain ischemia and intracranial hemorrhage. We performed a retrospective chart review of 22 pregnancies in 16 patients with MMD at the Kyushu University Hospital. An uncomplicated Cesarean delivery was performed in nearly all patients. In the 20 pregnancies in patients with pre-existing MMD, two had transient ischemic symptoms in the postpartum period. Two patients not previously known to have MMD developed transient ischemic symptoms postpartum. One of these patients was initially diagnosed and managed as pre-eclampsia. For patients diagnosed with MMD, a good perinatal outcome can be expected with appropriate management. Neurological events, however, may still occur postpartum even in well-managed patients. MMD may mimic the signs and symptoms of other neurological or psychiatric disorders, thereby complicating diagnosis and management.
Asunto(s)
Enfermedad de Moyamoya/diagnóstico , Periodo Posparto , Complicaciones del Embarazo , Adolescente , Adulto , Niño , Preescolar , Femenino , Humanos , Embarazo , Estudios Retrospectivos , Adulto JovenRESUMEN
We describe a 33-year-old male patient with mental retardation and cerebellar ataxia whose brain magnetic resonance imaging (MRI) showed diffuse central hypomyelination. The associated hypogonadotropic hypogonadism and hypodontia were consistent with the clinical diagnosis of 4H syndrome. Two compound heterozygous mutations in POLR3A were found: p.Met852Val and p.Asn1249His. MRI of the brain showed cerebellar atrophy, atrophy of the corpus callosum, and diffuse hypomyelination extending as far as the U-fibers, with preservation of the basal ganglia. T2 hyperintensity was observed in the bilateral middle cerebellar peduncles. The patient showed almost normal development until 4-5years of age. After 25years of age, the patient showed a gradual but consistent motor and cognitive deterioration. We demonstrated the involvement of the corticospinal tract electrophysiologically, but peripheral nerve conduction was normal. Although this disease may start very early in life, the clinical course in the present case suggests that brains that initially appear to have developed normally may show dysfunction later in life, although the pathophysiological bases for this dysfunction may not be evident on MRIs.
Asunto(s)
Anodoncia/genética , Ataxia Cerebelosa/genética , Enfermedades Desmielinizantes/genética , Hipogonadismo/genética , Discapacidad Intelectual/genética , Tractos Piramidales/fisiopatología , ARN Polimerasa III/genética , Adulto , Anodoncia/complicaciones , Anodoncia/patología , Anodoncia/fisiopatología , Atrofia , Ataxia Cerebelosa/complicaciones , Ataxia Cerebelosa/patología , Ataxia Cerebelosa/fisiopatología , Cerebelo/patología , Cuerpo Calloso/patología , Enfermedades Desmielinizantes/complicaciones , Enfermedades Desmielinizantes/patología , Enfermedades Desmielinizantes/fisiopatología , Progresión de la Enfermedad , Humanos , Hipogonadismo/complicaciones , Hipogonadismo/patología , Hipogonadismo/fisiopatología , Discapacidad Intelectual/complicaciones , Discapacidad Intelectual/patología , Discapacidad Intelectual/fisiopatología , Imagen por Resonancia Magnética/métodos , Masculino , Conducción Nerviosa/fisiología , Neuroimagen/métodos , SíndromeRESUMEN
PURPOSE: Ohtahara syndrome (OS) is one of the most severe and earliest forms of epilepsy. STXBP1 and ARX mutations have been reported in patients with OS. In this study, we aimed to identify new genes involved in OS by copy number analysis and whole exome sequencing. METHODS: Copy number analysis and whole exome sequencing were performed in 34 and 12 patients with OS, respectively. Fluorescence in situ hybridization, quantitative polymerase chain reaction (PCR), and breakpoint-specific and reverse-transcriptase PCR analyses were performed to characterize a deletion. Immunoblotting using lymphoblastoid cells was done to examine expression of CASK protein. KEY FINDINGS: Genomic microarray analysis revealed a 111-kb deletion involving exon 2 of CASK at Xp11.4 in a male patient. The deletion was inherited from his mother, who was somatic mosaic for the deletion. Sequencing of the mutant transcript expressed in lymphoblastoid cell lines derived from the patient confirmed the deletion of exon 2 in the mutant transcript with a premature stop codon. Whole exome sequencing identified another male patient who was harboring a c.1A>G mutation in CASK, which occurred de novo. Both patients showed severe cerebellar hypoplasia along with other congenital anomalies such as micrognathia, a high arched palate, and finger anomalies. No CASK protein was detected by immunoblotting in lymphoblastoid cells derived from two patients. SIGNIFICANCE: The detected mutations are highly likely to cause the loss of function of the CASK protein in male individuals. CASK mutations have been reported in patients with intellectual disability with microcephaly and pontocerebellar hypoplasia or congenital nystagmus, and those with FG syndrome. Our data expand the clinical spectrum of CASK mutations to include OS with cerebellar hypoplasia and congenital anomalies at the most severe end.
Asunto(s)
Enfermedades Cerebelosas/genética , Epilepsia/genética , Guanilato-Quinasas/genética , Enfermedades Cerebelosas/patología , Preescolar , Femenino , Eliminación de Gen , Humanos , Immunoblotting , Hibridación Fluorescente in Situ , Imagen por Resonancia Magnética , Masculino , Neuroimagen , Análisis de Secuencia por Matrices de Oligonucleótidos , Reacción en Cadena en Tiempo Real de la Polimerasa , SíndromeRESUMEN
Oculofaciocardiodental syndrome (OFCD) is an X-linked dominant disorder associated with male lethality, presenting with congenital cataract, dysmorphic face, dental abnormalities and septal heart defects. Mutations in BCOR (encoding BCL-6-interacting corepressor) cause OFCD. Here, we report on a Korean family with common features of OFCD including bilateral 2nd-3rd toe syndactyly and septal heart defects in three affected females (mother and two daughters). Through the mutation screening and copy number analysis using genomic microarray, we identified a novel heterozygous mutation, c.888delG, in the BCOR gene and two interstitial microduplications at Xp22.2-22.13 and Xp21.3 in all the three affected females. The BCOR mutation may lead to a premature stop codon (p.N297IfsX80). The duplication at Xp22.2-22.13 involved the NHS gene causative for Nance-Horan syndrome, which is an X-linked disorder showing similar clinical features with OFCD in affected males, and in carrier females with milder presentation. Considering the presence of bilateral 2nd-3rd toe syndactyly and septal heart defects, which is unique to OFCD, the mutation in BCOR is likely to be the major determinant for the phenotypes in this family.
Asunto(s)
Catarata/genética , Cardiopatías Congénitas/genética , Microftalmía/genética , Mutación , Proteínas Nucleares/genética , Proteínas Proto-Oncogénicas/genética , Proteínas Represoras/genética , Translocación Genética , Adulto , Secuencia de Bases , Encéfalo/anomalías , Catarata/diagnóstico , Niño , Puntos de Rotura del Cromosoma , Variaciones en el Número de Copia de ADN , Femenino , Orden Génico , Haplotipos , Cardiopatías Congénitas/diagnóstico , Defectos de los Tabiques Cardíacos , Heterocigoto , Humanos , Proteínas de la Membrana , Microftalmía/diagnóstico , Linaje , Isoformas de Proteínas/genética , Inactivación del Cromosoma XAsunto(s)
Inmunosupresores/administración & dosificación , Metotrexato/administración & dosificación , Uveítis/tratamiento farmacológico , Síndrome Uveomeningoencefálico/tratamiento farmacológico , Adulto , Enfermedad Crónica , Femenino , Humanos , Masculino , Persona de Mediana Edad , Resultado del Tratamiento , Uveítis/complicaciones , Síndrome Uveomeningoencefálico/complicacionesRESUMEN
Congenital hypomyelinating disorders are a heterogeneous group of inherited leukoencephalopathies characterized by abnormal myelin formation. We have recently reported a hypomyelinating syndrome characterized by diffuse cerebral hypomyelination with cerebellar atrophy and hypoplasia of the corpus callosum (HCAHC). We performed whole-exome sequencing of three unrelated individuals with HCAHC and identified compound heterozygous mutations in POLR3B in two individuals. The mutations include a nonsense mutation, a splice-site mutation, and two missense mutations at evolutionally conserved amino acids. Using reverse transcription-PCR and sequencing, we demonstrated that the splice-site mutation caused deletion of exon 18 from POLR3B mRNA and that the transcript harboring the nonsense mutation underwent nonsense-mediated mRNA decay. We also identified compound heterozygous missense mutations in POLR3A in the remaining individual. POLR3A and POLR3B encode the largest and second largest subunits of RNA Polymerase III (Pol III), RPC1 and RPC2, respectively. RPC1 and RPC2 together form the active center of the polymerase and contribute to the catalytic activity of the polymerase. Pol III is involved in the transcription of small noncoding RNAs, such as 5S ribosomal RNA and all transfer RNAs (tRNA). We hypothesize that perturbation of Pol III target transcription, especially of tRNAs, could be a common pathological mechanism underlying POLR3A and POLR3B mutations.
Asunto(s)
Codón sin Sentido , Predisposición Genética a la Enfermedad/genética , Enfermedades Desmielinizantes del Sistema Nervioso Central Hereditarias/genética , Mutación Missense , ARN Polimerasa III/genética , Adolescente , Adulto , Secuencia de Bases , Cuerpo Calloso/patología , Exoma/genética , Femenino , Genes Recesivos/genética , Enfermedades Desmielinizantes del Sistema Nervioso Central Hereditarias/patología , Humanos , Masculino , Modelos Moleculares , Datos de Secuencia Molecular , Degradación de ARNm Mediada por Codón sin Sentido/genética , Linaje , Sitios de Empalme de ARN , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Secuencia de ADNRESUMEN
AIMS: The objectives of this study were to elucidate the effects of a potent dipeptidyl peptidase (DPP)-IV inhibitor, TS-021, combined with/without metformin on glycemic control and pathological changes in pancreatic islets in high-fat diet and streptozotocin-induced (HFD-STZ) diabetic mice. MAIN METHODS: The anti-diabetic effects of TS-021 and/or metformin in HFD-STZ mice were examined in both acute and chronic treatment studies. In addition, we performed immunohistochemical analysis after repeated administration of TS-021 and/or metformin to HFD-STZ mice twice a day for 5 weeks. KEY FINDINGS: In the acute treatment study, TS-021 and/or metformin significantly improved glucose tolerance and glucagon-like peptide-1 (GLP-1) level, and TS-021 alone or in combination with metformin significantly increased the plasma insulin level after nutrient ingestion. In the chronic treatment study, TS-021 in combination with metformin significantly lowered the glycosylated hemoglobin level, plasma insulin level, and α-cell-to-ß-cell area ratio in pancreatic islets. In particular, the combined treatment synergistically increased the insulin-positive area in pancreatic islets from 32.3% in diabetic mice treated with the vehicle to 51.1% (TS-021 alone, 35.3%; metformin alone, 30.6%). SIGNIFICANCE: The present study demonstrated that the coadministration of TS-021 and metformin synergistically improved the islet morphology by increasing the circulating level of biologically active GLP-1, which is thought to result from two different mechanisms (namely, an increase in GLP-1 secretion and DPP-IV inhibition). These findings strongly support the rationale for combined treatment with DPP-IV inhibitors plus metformin in clinical practice by clearly demonstrating an anti-diabetic effect associated with the remarkable improvement in pancreatic ß-cell morphology.
Asunto(s)
Bencenosulfonatos/uso terapéutico , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Hiperglucemia/tratamiento farmacológico , Hipoglucemiantes/uso terapéutico , Células Secretoras de Insulina/efectos de los fármacos , Metformina/uso terapéutico , Pirrolidinas/uso terapéutico , Animales , Bencenosulfonatos/farmacología , Diabetes Mellitus Tipo 2/sangre , Diabetes Mellitus Tipo 2/metabolismo , Diabetes Mellitus Tipo 2/patología , Dipeptidil Peptidasa 4/metabolismo , Sinergismo Farmacológico , Péptido 1 Similar al Glucagón/sangre , Hemoglobina Glucada/análisis , Hiperglucemia/sangre , Hiperglucemia/metabolismo , Hiperglucemia/patología , Hipoglucemiantes/farmacología , Insulina/sangre , Células Secretoras de Insulina/metabolismo , Células Secretoras de Insulina/patología , Masculino , Metformina/farmacología , Ratones , Ratones Endogámicos C57BL , Pirrolidinas/farmacologíaRESUMEN
We examined the toxicity of methamphetamine and dopamine in CATH.a cells, which were derived from mouse dopamine-producing neural cells in the central nervous system. Use of the quantitative real-time polymerase chain reaction revealed that transcripts of the endoplasmic reticulum stress related gene (CHOP/Gadd153/ddit3) were considerably induced at 24-48 h after methamphetamine administration (but only under apoptotic conditions), whereas dopamine slightly induced CHOP/Gadd153/ddit3 transcripts at an early stage. We also found that dopamine and methamphetamine weakly induced transcripts for the glucose-regulated protein 78 gene (Grp78/Bip) at the early stage. Analysis by immunofluorescence microscopy demonstrated an increase ofãCHOP/Gadd153/ddit3 and Grp78/Bip proteins at 24 h after methamphetamine administration. Treatment of CATH.a cells with methamphetamine caused a re-distribution of dopamine inside the cells, which mimicked the presynaptic activity of neurons with cell bodies located in the ventral tegmental area or the substantia nigra. Thus, we have demonstrated the existence of endoplasmic reticulum stress in a model of presynaptic dopaminergic neurons for the first time. Together with the recent evidence suggesting the importance of presynaptic toxicity, our findings provide new insights into the mechanisms of dopamine toxicity, which might represent one of the most important mechanisms of methamphetamine toxicity and addiction.
Asunto(s)
Retículo Endoplásmico/efectos de los fármacos , Metanfetamina/toxicidad , Factor de Transcripción CHOP/genética , Animales , Dopamina/biosíntesis , Dopaminérgicos/farmacología , Neuronas Dopaminérgicas/efectos de los fármacos , Neuronas Dopaminérgicas/metabolismo , Chaperón BiP del Retículo Endoplásmico , Regulación de la Expresión Génica/efectos de los fármacos , Ratones , Receptores de Dopamina D1/metabolismo , Factor de Transcripción CHOP/biosíntesisRESUMEN
PURPOSE: To characterize the mechanism of corneal ulceration associated with viral keratitis, the authors investigated the effects of polyinosinic-polycytidylic acid [poly(I:C)], a synthetic analog of viral double-stranded RNA, on the expression of matrix metalloproteinases (MMPs) in human corneal fibroblasts. METHODS: The expression of MMPs in human corneal fibroblasts cultured in the absence or presence of poly(I:C) was examined by immunoblot analysis, gelatin zymography, and quantitative reverse transcription-polymerase chain reaction analysis. The phosphorylation of the NF-κB inhibitor protein IκB-α was assessed by immunoblot analysis, and the concentration of interleukin (IL)-1ß in culture supernatants was measured by enzyme-linked immunosorbent assay. RESULTS: Poly(I:C) increased the expression of MMP-1 and MMP-3 in corneal fibroblasts at the mRNA and protein levels. It also induced the phosphorylation of IκB-α and the secretion of IL-1ß in these cells. The poly(I:C)-induced expression of MMP-1 and MMP-3 was attenuated by a synthetic inhibitor of NF-κB signaling and by IL-1 receptor antagonist; the latter also inhibited poly(I:C)-induced phosphorylation of IκB-α. CONCLUSIONS: Poly(I:C) induces the expression of MMP-1 and MMP-3 in human corneal fibroblasts in a manner dependent on activation of the transcription factor NF-κB and on IL-1ß secretion. These effects may play an important role in corneal ulceration associated with viral keratitis.
Asunto(s)
Antivirales/farmacología , Córnea/efectos de los fármacos , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Interleucina-1beta/metabolismo , Metaloproteinasas de la Matriz/genética , FN-kappa B/metabolismo , Poli I-C/farmacología , Células Cultivadas , Córnea/enzimología , Ensayo de Inmunoadsorción Enzimática , Fibroblastos/efectos de los fármacos , Fibroblastos/enzimología , Humanos , Proteínas I-kappa B/metabolismo , Immunoblotting , Metaloproteinasa 1 de la Matriz/genética , Metaloproteinasa 1 de la Matriz/metabolismo , Metaloproteinasa 3 de la Matriz/genética , Metaloproteinasa 3 de la Matriz/metabolismo , Metaloproteinasas de la Matriz/metabolismo , Inhibidor NF-kappaB alfa , Fosforilación , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Regulación hacia ArribaRESUMEN
Cardiac hypertrophy in the recipient fetus of twin-twin transfusion syndrome (TTTS) has been reported previously. We encountered an unusual set of monochorionic twins in which one twin had cardiac hypertrophy without TTTS while the other fetus had selective growth restriction. In this case, the diagnosis of selective growth restriction was made at 17 weeks of gestation, and right ventricular hypertrophy was identified in the co-twin at 21 weeks; however, no signs of TTTS were observed. At 29 weeks we concluded that the fetal circulation had deteriorated based on echocardiographic findings that included hydrops fetalis and an elevated preload index. Emergency cesarean section was performed. Our experience suggests that hypertrophic cardiomyopathy-like change in a monochorionic twin pregnancy may arise in settings outside of TTTS, including growth restriction of a co-twin. We believe our case will assist the discussion surrounding the etiology of cardiac hypertrophy in monochorionic twins.
Asunto(s)
Cardiomegalia/diagnóstico por imagen , Enfermedades en Gemelos/diagnóstico por imagen , Retardo del Crecimiento Fetal/diagnóstico por imagen , Transfusión Feto-Fetal/diagnóstico por imagen , Adulto , Cesárea , Femenino , Humanos , Hidropesía Fetal/diagnóstico por imagen , Embarazo , Gemelos Monocigóticos , UltrasonografíaRESUMEN
PURPOSE: The synthesis of chemokines and adhesion molecules by corneal fibroblasts contributes to the development of corneal lesions in severe ocular allergy. The effects of the antiallergy drug tranilast on the expression of such molecules were examined in human corneal fibroblasts. METHODS: The release of chemokines into culture supernatants and the expression of vascular endothelial cell adhesion molecule (VCAM)-1 on the cell surface were determined with enzyme-linked immunosorbent assays. The intracellular abundance of mRNAs was quantitated by reverse transcription and real-time polymerase chain reaction analysis. The phosphorylation of signaling proteins was examined by immunoblot analysis. RESULTS: Tranilast inhibited the release of the chemokines eotaxin-1 and TARC and the surface expression of VCAM-1, induced by the combination of TNF-alpha and IL-4 in corneal fibroblasts. Dexamethasone, but not cyclosporine A or tacrolimus, mimicked these effects of tranilast. Tranilast also inhibited the cytokine-induced upregulation of eotaxin-1 and TARC mRNAs in corneal fibroblasts. Tranilast inhibited the cytokine-induced phosphorylation of the NF-kappaB inhibitor IkappaBalpha and of mitogen-activated protein kinases (ERK, JNK, p38), without affecting that of STAT6, in corneal fibroblasts. CONCLUSIONS: Inhibition by tranilast of the cytokine-induced expression of eotaxin-1, TARC, and VCAM-1 in human corneal fibroblasts suggests that this drug might prove effective for treatment of the corneal manifestations of ocular allergic inflammation by targeting corneal fibroblasts directly.