ABSTRACT
OBJECTIVE@#To explore the genetic basis for an infant with permanent neonatal diabetes mellitus (PNDM).@*METHODS@#Clinical data of the child was collected. Targeted capture-next generation sequencing was carried out to identify the potential variants. Candidate variant was verified by Sanger sequencing of her family members.@*RESULTS@#The child was a 4-month-and-26-day female featuring onset of ketoacidosis accompanied with fasting blood glucose of 24.4 mmol/L, positive urine glucose, decreased serum C-peptide, HbA1c of 9.58%, and negative diabetes autoantibody. Genetic testing revealed that she has carried a heterozygous c.314T>G (p.L105R) variant of the INS gene. Sanger sequencing verified that neither of her parents has carried the same variant, which was also unreported in the literature. The variant was classified as likely pathogenic based on the ACMG guidelines.@*CONCLUSION@#The c.314T>G (P.L105R) variant of the INS gene probably underlay the genetic etiology in this child. Genetic testing should be conducted for children with suspected PNDM for early diagnosis and appropriate treatment.
Subject(s)
Humans , Infant , Child , Infant, Newborn , Female , Mutation , Insulin/genetics , Diabetes Mellitus/genetics , Genetic TestingABSTRACT
A 2-year-old boy was admitted to Shanghai Children's Medical Center, Shanghai Jiao Tong University School of Medicine in Nov 30th, 2018, due to polydipsia, polyphagia, polyuria accompanied with increased glucose levels for more than 2 weeks. He presented with symmetrical short stature [height 81 cm (-2.2 SD), weight 9.8 kg (-2.1 SD), body mass index 14.94 kg/m2 (P10-P15)], and with no special facial or physical features. Laboratory results showed that the glycated hemoglobin A1c was 14%, the fasting C-peptide was 0.3 ng/mL, and the islet autoantibodies were all negative. Oral glucose tolerance test showed significant increases in both fasting and postprandial glucose, but partial islet functions remained (post-load C-peptide increased 1.43 times compared to baseline). A heterozygous variant c.1366C>T (p.R456C) was detected in GATA6 gene, thereby the boy was diagnosed with a specific type of diabetes mellitus. The boy had congenital heart disease and suffered from transient hyperosmolar hyperglycemia after a patent ductus arteriosus surgery at 11 months of age. Insulin replacement therapy was prescribed, but without regular follow-up thereafter. The latest follow-up was about 3.5 years after the diagnosis of diabetes when the child was 5 years and 11 months old, with the fasting blood glucose of 6.0-10.0 mmol/L, and the 2 h postprandial glucose of 17.0-20.0 mmol/L.
Subject(s)
Male , Child , Humans , Child, Preschool , Infant , Diabetes Mellitus, Type 2/complications , Mutation, Missense , C-Peptide/genetics , China , Insulin/genetics , Glucose , Blood Glucose , GATA6 Transcription Factor/geneticsABSTRACT
ABSTRACT BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) is an increasing global health concern defined by excessive hepatic fat content in the absence of excessive alcohol consumption. OBJECTIVE: Given the pivotal role of insulin resistance in NAFLD, we hypothesized that insulin (INS) and insulin receptor (INSR) gene polymorphisms may be associated with NAFLD risk. METHODS: A total of 312 subjects, including 153 cases with biopsy-proven NAFLD and 159 controls were enrolled in this case-control study. Four polymorphisms in INS (rs3842752, rs689) and INSR (rs1052371, rs1799817) genes were genotyped using PCR-RFLP method. RESULTS: The cases with NAFLD were older and had higher BMI, systolic blood pressure, diastolic blood pressure, as well as higher serum levels of aspartate aminotransferase, alanine aminotransferase, and gamma glutamyl transferase than the controls (P<0.001). The "TT" genotype of INSR rs1799817 compared with "CC" genotype occurred more frequently in the controls than the cases with NAFLD and the difference remained significant after adjustment for confounding factors (P=0.018; OR=0.10, 95%CI=0.02-0.76). However, no significant difference was found for INS rs3842752, INS rs689, and INSR rs1052371 gene polymorphisms between the cases with NAFLD and the controls either before or after adjustment for the confounders. CONCLUSION: These findings corroborate the hypothesis that genetic polymorphisms related to insulin resistance play a role in NAFLD susceptibility. Specifically, the INSR rs1799817 "TT" genotype had a protective effect for NAFLD. However, our results remain to be validated in other studies.
RESUMO CONTEXTO: A doença hepática gordurosa não alcoólica (NAFLD) é uma preocupação global crescente da saúde definida pelo excesso de teor de gordura hepática na ausência de consumo excessivo de álcool. OBJETIVO: Dado o papel crucial da resistência à insulina no NAFLD, criou-se a hipótese de que os polimorfismos genéticos da insulina (INS) e do receptor de insulina (INSR) podem estar associados ao risco de NAFLD. MÉTODOS: Um total de 312 indivíduos, incluindo 153 casos com NAFLD comprovado por biópsia e 159 controles foram inscritos neste estudo de caso-controle. Quatro polimorfismos em genes INS (rs3842752, rs689) e INSR (rs1052371, rs1799817) foram genotipados utilizando o método PCR-RFLP. RESULTADOS: Os casos com NAFLD foram mais idosos e apresentaram maior IMC, pressão arterial sistólica, pressão arterial diastólica, bem como níveis séricos mais elevados de aspartato aminotransferase, de alanina aminotransferase e de gama glutamil transpeptidase do que os controles (P<0,001). O genótipo "TT" de INSR rs1799817 em comparação com o genótipo "CC" ocorreu com mais frequência nos controles do que os casos com NAFLD e a diferença permaneceu significativa após ajuste para fatores de confusão (P=0,018; OR=0,10, IC95%=0,02-0,76). No entanto, não foi encontrada diferença significativa para INS rs3842752, INS rs689 e INSR rs1052371 polimorfismos genéticos entre os casos com NAFLD e os controles antes ou depois do ajuste para os fatores de confusão. CONCLUSÃO: Esses achados corroboram a hipótese de que os polimorfismos genéticos relacionados à resistência à insulina desempenham um papel na suscetibilidade do NAFLD. Especificamente, o genótipo INSR rs1799817 "TT" teve um efeito protetor para o NAFLD. No entanto, nossos resultados necessitam ser validados em outros estudos.
Subject(s)
Humans , Adult , Aged , Receptor, Insulin/genetics , Genetic Predisposition to Disease , Non-alcoholic Fatty Liver Disease/genetics , Polymorphism, Genetic , Case-Control Studies , Insulin/genetics , Middle AgedABSTRACT
Inhibition of CD36, a fatty acid transporter, has been reported to prevent glucotoxicity and ameliorate high glucose induced beta cell dysfunction. Ezetimibe is a selective cholesterol absorption inhibitor that blocks Niemann Pick C1-like 1 protein, but may exert its effect through suppression of CD36. We attempted to clarify the beneficial effect of ezetimibe on insulin secreting cells and to determine whether this effect is related to change of CD36 expression. mRNA expression of insulin and CD36, intracellular peroxide level and glucose stimulated insulin secretion (GSIS) under normal (5.6 mM) or high glucose (30 mM) condition in INS-1 cells and primary rat islet cells were compared. Changes of the aforementioned factors with treatment with ezetimibe (20 μM) under normal or high glucose condition were also assessed. mRNA expression of insulin was decreased with high glucose, which was reversed by ezetimibe in both INS-1 cells and primary rat islets. CD36 mRNA expression was increased with high glucose, but decreased by ezetimibe in INS-1 cells and primary rat islets. Three-day treatment with high glucose resulted in an increase in intracellular peroxide level; however, it was decreased by treatment with ezetimibe. Decrease in GSIS by three-day treatment with high glucose was reversed by ezetimibe. Palmitate uptake following exposure to high glucose conditions for three days was significantly elevated, which was reversed by ezetimibe in INS-1 cells. Ezetimibe may prevent glucotoxicity in pancreatic β-cells through a decrease in fatty acid influx via inhibition of CD36.
Subject(s)
Animals , Male , Rats , Anticholesteremic Agents/pharmacology , CD36 Antigens/antagonists & inhibitors , Cells, Cultured , Ezetimibe/pharmacology , Flow Cytometry , Glucose/toxicity , Insulin/genetics , Insulin-Secreting Cells/cytology , Palmitic Acid/metabolism , RNA, Messenger/metabolism , Rats, Sprague-Dawley , Reactive Oxygen Species/metabolism , Real-Time Polymerase Chain ReactionABSTRACT
OBJECTIVE: To assess bone thickness for miniscrew placement in the mandible during mixed dentition by using digital volumetric tomograph (DVT). MATERIAL AND METHODS: A total of 15 healthy patients aged 8-10 years old, with early exfoliated mandibular second deciduous molar, were included. DVT images of one quadrant of the mandible were obtained using Kodak extraoral imaging systems and analyzed by Kodak dental imaging software. The error of the method (EM) was calculated using Dahlberg's formula. Mean and standard deviation were calculated at 6 and 8 mm from the cementoenamel junction (CEJ).Paired t-test was used to analyze the measurements. RESULTS: Buccal cortical bone thickness, mesiodistal width and buccolingual bone depth at 6 mm were found to be 1.73 + 0.41, 2.15 + 0.49 and 13.18 + 1.22 mm, respectively; while at 8 mm measurements were 2.42 + 0.34, 2.48 + 0.33 and 13.65 + 1.25 mm, respectively. EM for buccal cortical bone thickness, mesiodistal width and buccolingual bone depth was 0.58, 0.40 and 0.48, respectively. The difference in measurement at 6 and 8 mm for buccal cortical plate thickness (P < 0.05) and buccolingual bone thickness (P < 0.05) was found to be significant, whereas for mesiodistal width it was insignificant (P > 0.05). CONCLUSION: Bone thickness measurement has shown promising evidence for safe placement of miniscrews in the mandible during mixed dentition. The use of miniscrew is the best alternative, even in younger patients. .
OBJETIVO: avaliar, por meio de tomografia volumétrica digital (TVD), a espessura óssea necessária para a instalação de mini-implante na arcada inferior durante a fase de dentição mista. MÉTODOS: um total de 15 pacientes saudáveis, com idades entre 8 e 10 anos, com segundo molar inferior decíduo irrompido recentemente, foram incluídos no presente estudo. Imagens de TVD da hemiarcada inferior foram obtidas utilizando sistemas de imagens extrabucais Kodak. As imagens foram analisadas por meio do programa de imagens Kodak. O erro do método (EM) foi calculado utilizando a fórmula de Dahlberg. Médias e desvios-padrão foram calculados de 6 a 8mm aquém da junção amelocementária. O teste t foi utilizado para a análise das medidas. RESULTADOS: a espessura do osso cortical vestibular, largura mesiodistal e profundidade óssea vestibulolingual, a 6mm, foram de 1,73 + 0,41; 2,15 + 0,49; e 13,18 + 1,22 mm, respectivamente. Já a 8mm, os valores foram de 2,42 + 0,34; 2,48 + 0,33; e 13,65 + 1,25mm. O EM para a espessura do osso cortical vestibular, largura mesiodistal e profundidade óssea vestibulolingual foi de 0,58, 0,40 e 0,48mm, respectivamente. A diferença entre as medidas a 6 e 8mm para a espessura do osso cortical vestibular (p < 0,05) e a espessura óssea vestibulolingual (p < 0,05) foi significativa, embora não tenha sido significativa para a largura mesiodistal (p < 0,05). CONCLUSÃO: a mensuração da espessura óssea demonstra evidências promissoras para a segura instalação de mini-implantes na arcada inferior e na fase de dentição mista. O uso de mini-implantes tem se mostrado a melhor alternativa, mesmo nos casos de pacientes mais jovens. .
Subject(s)
Humans , Male , Middle Aged , /genetics , /metabolism , Islets of Langerhans/metabolism , Alleles , Fasting/metabolism , Genome-Wide Association Study/methods , Glucose/genetics , Glucose/metabolism , Insulin Resistance/genetics , Insulin/genetics , Insulin/metabolism , Polymorphism, Single Nucleotide/genetics , Receptor, Insulin/genetics , Receptor, Insulin/metabolism , Signal Transduction/geneticsABSTRACT
As células-tronco mesenquimais (CTM) constituem uma ferramenta promissora para o campo de terapia celular. Além de seu potencial de diferenciação em diferentes tipos celulares, as CTM apresentam a habilidade de secretar moléculas bioativas e, assim, exercer múltiplos efeitos biológicos, tais como indução da regeneração de tecidos lesionados, redução de fibrose e modulação do sistema imune. A superexpressão dos fatores de crescimento G-CSF e IGF-1, conhecidos por seus efeitos sobre os processos de imunomodulação, sobrevivência celular e reparo tecidual, pode ampliar as ações terapêuticas das CTM. O objetivo deste trabalho consiste em gerar e caracterizar linhagens de CTM de camundongo superexpressando hGCSF ou hIGF-1. Um sistema lentiviral de segunda geração foi utilizado para modificação de CTM para expressão ectópica dos genes de interesse. As sequências codificantes de hG-CSF e hIGF-1 foram amplificadas por PCR e subclonadas em um vetor lentiviral de transferência, contendo um promotor constitutivo. As partículas lentivirais foram produzidas a partir da cotransfecção de células da linhagem HEK293FT...
Mesenchymal stem cells (MSCs) are a promising tool for the cell therapy field. In addition to their potential for differentiation into different cell types, MSCs have the ability to secrete bioactive molecules and thus exert multiple biological effects such as induction of the injured tissue regeneration, fibrosis reduction and modulation of the immune system. The overexpression of the growth factors G-CSF and IGF-1, known for their effects on immune modulation processes, cell survival and tissue repair, can result in a magnification of MSCs' therapeutic actions. The objective of this work is to generate and characterize mouse MSCs lines overexpressing hG-CSF or hIGF-1. A second generation lentiviral system was used to modify MSCs derived from mice for the ectopic expression of the genes of interest. The coding sequences of hG-CSF and hIGF-1 were amplified by PCR and subcloned into a lentiviral transfer vector containing a constitutive promoter. The lentiviral particles were produced from the co-transfection of HEK293FT...
Subject(s)
Animals , Stem Cells/classification , Stem Cells/immunology , Stem Cells/pathology , Insulin , Insulin/analysis , Insulin/genetics , Insulin/blood , Insulin/urineABSTRACT
This study evaluated the bone regeneration process in rabbit calvaria induced by three types of biomaterials: two xenogenous, consisting of deproteinized bovine bone, while the other was alloplastic, based on biphasic calcium phosphate. Five New Zealand white rabbits weighing between 2,900 and 3,500 g were submitted to four standard 8 mm-diameter perforations at the parietal bone. Three perforations were filled with three grafts and biomaterials, two of them received bovine Bio-Oss(r) and Endobon(r) Xenograft Granules, and the other consisted of fully alloplastic Straumann(r) Bone Ceramic. The fourth remaining cavity was used as control with coagulum. After eight weeks, the animals were sacrificed, and the samples were prepared for morphometric and qualitative analysis. The cavities filled with alloplastic biomaterials showed higher percentages of newly formed bone (p<0.05), while the cavities with xenogenous biomaterials showed higher amount of residual graft (p<0.05). Although the results showed greater bone formation with Straumann(r) Bone Ceramic, further studies are required to prove which is the more effective biomaterial for bone induction process.
Este estudo avaliou o processo de reparação óssea induzida por três biomateriais, dois de origem xenógena constituído de osso bovino desproteinizado e um aloplástico à base de fosfato de cálcio bifásico, em calota craniana de coelhos. Em cinco coelhos brancos da Nova Zelândia com peso entre 2.900 e 3.500 g, foram realizadas quatro perfurações padronizadas de 8 mm de diâmetro nos ossos parietais e enxertados dois biomateriais de origem bovina: Bio-Oss(r) e Endobon(r) Xenograft Granules e um totalmente aloplástico: Straumann(r)Bone Ceramic. Uma cavidade permaneceu com coágulo e foi utilizado como controle. Após oito semanas os animais foram sacrificados e as amostras preparadas para análise morfométrica e qualitativa. Os resultados mostraram que as cavidades preenchidas com o biomaterial aloplástico apresentaram percentualmente maior quantidade de osso neoformado (p<0,05). Apesar dos resultados mostrarem maior neoformação óssea pelo Straumann(r)Bone Ceramic, há a necessidade de mais estudos para se comprovar qual biomaterial é mais efetivo no processo de indução óssea.
Subject(s)
Humans , Cell Line , Doxycycline/pharmacology , Gene Expression Regulation , Islets of Langerhans , Insulin/metabolism , /physiopathology , /therapy , Insulin/genetics , Islets of Langerhans/metabolism , TransfectionABSTRACT
Objective Zinc transporter 8 autoantibodies (ZnT8A) have been poorly studied in non-Caucasian individuals. We aimed to investigate the prevalence of ZnT8 autoantibodies in patients with T1D and their first degree relatives (FDR) from a multiethnic population, as well as its relation with the insulin (INS) or the protein tyrosine phosphatase non-receptor 22 (PTPN22) gene polymorphisms. Subjects and methods ZnT8A were analyzed in sera from T1D patients (n = 72, mean age of 30.3 ± 11.4 years) of variable duration (15.7 ± 11.8 years) and their FDR (n = 78, mean age of 18.3 ± 9.1 years) by a triple mix Radioligand Binding Assay (RBA) for the ZnT8 autoantibody (ZnT8-RWQ) variants. SNP (single nucleotide polymorphism) for INS and PTPN22 were genotyped. Results The prevalence of ZnT8A was higher in T1D patients than FDR, for ZnT8TripleA (24% vs. 4%,p = 0.001), ZnT8RA (24% vs. 4%, p < 0.001) and ZnT8QA (15% vs. 3%, p = 0.004). All FDR with ZnT8A (n = 3) had at least another positive antibody. Heterozygosis for PTPN22 was associated with a higher frequency of ZnT8TripleA (p = 0.039) and ZnT8RA (p = 0.038). Conclusions ZnT8A is observed in non-Caucasian patients with T1D, even years after the disease onset, as well as in their FDR. In those, there was an overlap between ZnT8A and other T1D antibodies. ZnT8A was associated with PTPN22 polymorphisms. Further longitudinal studies are necessary to elucidate the importance of these findings in the natural history of T1D patients with multiethnic background. .
Objetivo Os autoanticorpos transportadores de zinco 8 (ZnT8A) foram pouco estudados em indivíduos não caucasianos. Nosso objetivo foi investigar a prevalência de autoanticorpos ZnT8 em pacientes com T1D e seus parentes de primeiro grau (PPG) em uma população multiétnica, assim como a sua relação com os polimorfismos genéticos da insulina (INS) ou proteína tirosina fosfatase não receptora tipo 22 (PTPN22). Sujeitos e métodos ZnT8A foram analisados no soro de pacientes com T1D (n = 72, idade média de 30,3 ± 11,4 anos) de duração variável (15,7 ± 11,8 anos) e seus PPG (n = 72, idade média de 30,3 ± 11,4 anos) usando-se um ensaio de competição com radioligantes (RBA) para variantes dos autoanticorpos ZnT8 (ZnT8-RWQ). Os polimorfismos de nucleotídeo único para a INS e PTPN22 foram genotipados. Resultados A prevalência de ZnT8A foi mais alta em pacientes T1D do que nos PPG, para ZnT8TriploA (24% contra 4%, p = 0,001), ZnT8RA (24% contra 4%, p < 0,001) e ZnT8QA (15% contra 3%, p = 0,004). Todos os PPG com ZnT8A (n = 3) apresentaram positividade para pelo menos outro anticorpo. A heterozigose para PTPN22 foi associada a uma frequência mais alta de ZnT8TriploA (p = 0,039) e de ZnT8RA (p = 0,038). Conclusões Os ZnT8A foram observados em pacientes não caucasianos com T1D, mesmo depois de anos do início da doença, assim como em seus PPG. Nos parentes, houve uma sobreposição entre os ZnT8A e outros anticorpos para T1D. Os ZnT8A mostraram-se associados aos polimorfismos PTPN22. São necessários outros estudos longitudinais para se elucidar a importância desses achados na história natural de pacientes com T1D com antecedentes étnicos variados. .
Subject(s)
Adolescent , Adult , Child , Female , Humans , Male , Young Adult , Autoantibodies/immunology , Cation Transport Proteins/immunology , Diabetes Mellitus, Type 1/immunology , Family/ethnology , Autoantibodies/genetics , Brazil/epidemiology , Brazil/ethnology , Cation Transport Proteins/blood , Cation Transport Proteins/genetics , Diabetes Mellitus, Type 1/epidemiology , Diabetes Mellitus, Type 1/ethnology , Diabetes Mellitus, Type 1/genetics , Genotype , Insulin/genetics , Prevalence , Polymorphism, Genetic/genetics , /genetics , Radioligand AssayABSTRACT
OBJECTIVE: To investigate the influence of (CA)n repeats in the insulin-like growth factor 1 gene and a variable number of tandem repeats of the insulin gene on birth size in children who are small or adequate-sized for gestational age and to correlate these polymorphisms with serum insulin-like growth factor 1 levels and insulin sensitivity in children who are small for gestational age, with and without catch-up growth. PATIENTS AND METHODS: We evaluated 439 infants: 297 that were adequate-sized for gestational age and 142 that were small for gestational age (66 with and 76 without catch-up). The number of (CA)n repeat in the insulin-like growth factor 1 gene and a variable number of tandem repeats in the insulin gene were analyzed using GENESCAN software and polymerase chain reaction followed by enzymatic digestion, respectively. Clinical and laboratory data were obtained from all patients. RESULTS: The height, body mass index, paternal height, target height and insulin-like growth factor 1 serum levels were higher in children who were small for gestational age with catch-up. There was no difference in the allelic and genotypic distributions of both polymorphisms between the adequate-sized and small infants or among small infants with and without catch-up. Similarly, the polymorphisms were not associated with clinical or laboratory variables. CONCLUSION: Polymorphisms of the (CA)n repeats of the insulin-like growth factor 1 gene and a variable number of tandem repeats of the insulin gene, separately or in combination, did not influence pre- or postnatal growth, insulin-like growth factor 1 serum levels or insulin resistance. .
Subject(s)
Female , Humans , Infant, Newborn , Male , Infant, Small for Gestational Age , Insulin-Like Growth Factor I/genetics , Insulin/genetics , Polymorphism, Genetic , Tandem Repeat Sequences/genetics , Adenosine , Brazil , Birth Weight/genetics , Blood Glucose/genetics , Body Height/genetics , Body Weight/genetics , Cytosine , Insulin Resistance/genetics , Insulin-Like Growth Factor I/analysis , Risk FactorsABSTRACT
Type 1 diabetes (T1D) results from autoimmune-mediated destruction of the pancreatic beta cells, a process that is conditioned by multiple genes and environmental factors. The process that destroys the pancreatic b cells in T1D is mediated by T cells and leads to a complex phenotype influenced by multiple factors. It has been more than 30 years since the publication of the first evidence suggesting the involvement of a specific chromosomal region, HLA, in modulating the risk for T1D. HLA locus has been known for decades to contribute strongly with the attributable to genetic risk. In addition to HLA, many proposed candidate loci have been described that are associated with risk of developing the disease, including the insulin gene (INS), PTPN22,CTLA-4, PD-1, IL2-RA and IFIH1 which together do not contribute more than 15 percent of the risk. This review compiled the data on T1D genes and discusses the major genetic impact of these genetic aspects in T1D etiology.
Subject(s)
Humans , Diabetes Mellitus, Type 1/genetics , Genetic Markers , DEAD-box RNA Helicases/genetics , /genetics , HLA Antigens/genetics , Genetic Predisposition to Disease , Insulin/genetics , /genetics , /geneticsABSTRACT
Glucagon-like peptide-1 (GLP-1) is a potent glucoincretin hormone and an important agent for the treatment of type 2 diabetes. Here we demonstrate that B-cell translocation gene 2 (BTG2) is a crucial regulator in GLP-1-induced insulin gene expression and insulin secretion via upregulation of pancreatic duodenal homeobox-1 (PDX-1) in pancreatic beta-cells. GLP-1 treatment significantly increased BTG2, PDX-1 and insulin gene expression in pancreatic beta-cells. Notably, adenovirus-mediated overexpression of BTG2 significantly elevated insulin secretion, as well as insulin and PDX-1 gene expression. Physical interaction studies showed that BTG2 is associated with increased PDX-1 occupancy on the insulin gene promoter via a direct interaction with PDX-1. Exendin-4 (Ex-4), a GLP-1 agonist, and GLP-1 in pancreatic beta-cells increased insulin secretion through the BTG2-PDX-1-insulin pathway, which was blocked by endogenous BTG2 knockdown using a BTG2 small interfering RNA knockdown system. Finally, we revealed that Ex-4 and GLP-1 significantly elevated insulin secretion via upregulation of the BTG2-PDX-1 axis in pancreatic islets, and this phenomenon was abolished by endogenous BTG2 knockdown. Collectively, our current study provides a novel molecular mechanism by which GLP-1 positively regulates insulin gene expression via BTG2, suggesting that BTG2 has a key function in insulin secretion in pancreatic beta-cells.
Subject(s)
Animals , Humans , Male , Mice , Rats , Gene Expression Regulation/drug effects , Glucagon-Like Peptide 1/pharmacology , Homeodomain Proteins/genetics , Immediate-Early Proteins/genetics , Insulin/genetics , Insulin-Secreting Cells/drug effects , Mice, Inbred C57BL , Peptides/pharmacology , Promoter Regions, Genetic/genetics , Protein Binding/drug effects , Trans-Activators/genetics , Tumor Suppressor Proteins/genetics , Venoms/pharmacologyABSTRACT
Objective: The high global incidence of type 2 diabetes has challenged researchers to establish animal models that resemble the chronic stage observed in type 2 diabetes patients. One such model is induced by neonatal streptozotocin (n-STZ) administration to rat pups at 0, 2, or 5 days after birth. In this study, we assessed lns-1 gene expression and tissue insulin levels as well as serum concentration of glucose and insulin, insulin resistance, and histological changes of the islets of Langerhans in n5-STZ rats after 20-weeks post-induction. Methods: Wistar rat pups were randomly distributed into a control group and a streptozotocin-induced group. Experimental induction involved a single intraperitoneal injection of streptozotocin (150 mg/kg) into neonates at five days after birth. Results: At 20 weeks post-induction, streptozotocin-induced rats exhibited increased serum glucose levels, reduced serum insulin levels, impaired glucose metabolism and insulin resistance compared to control rats. Histologically, streptozotocin-induced rats exhibited atrophic islets, vacuolization, and significantly fewer insulin-positive cells. lns-1 gene expression was significantly decreased in n5-STZ rats in comparison to the control group. Conclusion: Our findings support that the n5-STZ model 20 weeks post-induction represents an appropriate experimental tool to study T2D and to evaluate novel therapeutic agents and targets that involve insulin gene expression and secretion, as well as complications caused by chronic diabetes.
Subject(s)
Animals , Female , Rats , Diabetes Mellitus, Experimental/metabolism , Gene Expression Regulation , Insulin/genetics , Islets of Langerhans/metabolism , Animals, Newborn , Disease Models, Animal , Diabetes Mellitus, Experimental/chemically induced , Immunohistochemistry , Insulin Resistance , Insulin/metabolism , Random Allocation , Rats, Wistar , Streptozocin , Time FactorsABSTRACT
Las enfermedades cardiovasculares y el síndrome metabólico son patologías que afectan a gran parte de la población mundial, de etiología poligénica y multifactorial, resultado de la interacción entre factores genéticos, ambientales, sociales y culturales. Los principales genes candidatos a estudiar son aquellos relacionados con la regulación de la homeostasis de la glucosa, del metabolismo lipídico y/o de la secreción y acción de la insulina, como son el gen de los Receptores de los Proliferadores Perixosomales Activados gamma 2 (PPARγ2), la Proteína Enlazante de Ácidos Grasos Intestinal (FABP2) y la Apolipoproteína E (ApoE). Evaluar la relación entre los polimorfismos Pro12Ala del gen PPARγ2, Ala54Thr del gen FABP2 y del gen de ApoE en habitantes del Sector Los Eucaliptos de la Parroquia San Juan. 308 individuos de dicha comunidad, 98 hombres y 210 mujeres, clasificados en hipercolesterolémicos, hipertrigliceridémicos, resistentes a la insulina y controles de acuerdo a sus niveles de colesterol total, triglicéridos e índice HOMA. Extracción de 10 mL de sangre venosa para la determinación de química sanguínea y extracción de ADN, amplificación mediante PCR de un fragmento de 102pb del gen de PPARγ2, uno de 180pb del gen de FABP2 y otro de 244pb del gen de ApoE, y posterior RFLP. Se encontró una frecuencia alélica de 0,91 para el alelo Pro y 0,09 para el Ala del gen de PPARγ2; 0,70 para el alelo Ala del gen FABP2 y 0,30 para el Thr, mientras que para los alelos del gen de ApoE la frecuencia fue de ε2=0,05, ε3=0,80 y ε4=0,15. Se encontró relación entre el alelo ε4 de ApoE y la hipercolesterolemia, además del alelo ε2 como factor protector ante el desarrollo de hipercolesterolemia, hipertrigliceridemia y resistencia a la insulina, no encontrándose asociación alguna entre los polimorfismos de los restantes genes y las patologías mencionadas.
Cardiovascular disease and metabolic syndrome are diseases that affect worldwide, with multiple genetic and environmental components contributing to susceptibility. The main candidate genes to study are those related to the regulation of glucose homeostasis, lipid metabolism, insulin secretion and action and obesity, these include the genes for Peroxisome proliferator-activated receptor gamma 2 (PPARγ2), fatty acid-binding protein 2 (FABP2) and Apolipoprotein E (Apo E). The aim of this study was to investigate the relationship between polymorphisms of Pro12Ala PPARγ2 gene, Ala54Thr FABP2 gene and the ApoE gene in residents from the The Eucaliptus of the Parroquia San Juan. 308 subjects, 98 men and 210 women, classified as hypercholesterolemic, hypertriglyceridemic, insulin resistant and controls according to their levels of total cholesterol, triglycerides and HOMA index. Extraction of 10 mL whole blood for determination of chemistry and DNA extraction, PCR amplification of a 102 bp fragment PPARγ2 gene, a 180 bp FABP2 gene and a 244 bp of ApoE gene, and subsequent RFLP. An allele frequency for allele Pro 0.91 and 0.09 for gene PPARγ2 Ala and 0.70 for the allele of the gene FABP2 Ala and 0.30 for Thr, while for the different alleles of ApoE gene frequency was ε2=0.05, ε3=0.80 and ε4=0.15. We found a relationship between the ApoE ε4 allele and hypercholesterolemia, in the other hand, Apo E ε2 allele was found as a protective factor against the development of hypercholesterolemia, hypertriglyceridemia and insulin resistance, we did not found association between polymorphisms of the other genes and the pathologies mentioned above.
Subject(s)
Humans , Male , Adult , Female , Cardiovascular Diseases/diagnosis , Cardiovascular Diseases/etiology , Insulin/analysis , Insulin/genetics , Insulin/chemistry , Polymorphism, Genetic , Metabolic Syndrome/complications , Metabolic Syndrome/diagnosis , Blood Chemical Analysis , Glucose Metabolism Disorders , Hematology , Lipid Metabolism DisordersABSTRACT
Transgenic mice carrying the human insulin gene driven by the K-cell glucose-dependent insulinotropic peptide (GIP) promoter secrete insulin and display normal glucose tolerance tests after their pancreatic p-cells have been destroyed. Establishing the existence of other types of cells that can process and secrete transgenic insulin would help the development of new gene therapy strategies to treat patients with diabetes mellitus. It is noted that in addition to GIP secreting K-cells, the glucagon-like peptide 1 (GLP-1) generating L-cells share/ many similarities to pancreatic p-cells, including the peptidases required for proinsulin processing, hormone storage and a glucose-stimulated hormone secretion mechanism. In the present study, we demonstrate that not only K-cells, but also L-cells engineered with the human preproinsulin gene are able to synthesize, store and, upon glucose stimulation, release mature insulin. When the mouse enteroendocrine STC-1 cell line was transfected with the human preproinsulin gene, driven either by the K-cell specific GIP promoter or by the constitutive cytomegalovirus (CMV) promoter, human insulin co-localizes in vesicles that contain GIP (GIP or CMV promoter) or GLP-1 (CMV promoter). Exposure to glucose of engineered STC-1 cells led to a marked insulin secretion, which was 7-fold greater when the insulin gene was driven by the CMV promoter (expressed both in K-cells and L-cells) than when it was driven by the GIP promoter (expressed only in K-cells). Thus, besides pancreatic p-cells, both gastrointestinal enteroendocrine K-cells and L-cells can be selected as the target cell in a gene therapy strategy to treat patients with type 1 diabetes mellitus.
Subject(s)
Animals , Humans , Mice , Enteroendocrine Cells/physiology , Gastric Inhibitory Polypeptide/pharmacology , Glucagon-Like Peptide 1/pharmacology , Glucose/pharmacology , Insulin-Secreting Cells/metabolism , Insulin , Protein Precursors/genetics , Diabetes Mellitus, Type 1/therapy , Enteroendocrine Cells/drug effects , Genetic Engineering , Genetic Therapy/methods , Hypoglycemic Agents/pharmacology , Insulin-Secreting Cells/cytology , Insulin/genetics , Mice, TransgenicABSTRACT
El ambiente intrauterino y los primeros años de vida, son fundamentales para la programación de una serie de situaciones en nuestro organismo. Son muchas las teorías que indican que la lactancia materna (LM) puede tener beneficio en la prevención de la obesidad. Esto hace referencia a que la LM posee una serie de componentes hormonales, que favorecen la maduración o la forma de interpretación de ciertas áreas del cerebro, que tienen que ver con los mecanismos de saciedad y hambre. Existen estudios bastante clásicos que demuestran cómo ésta se regula en el niño alimentado con LM, y la impor tancia que el lo representa, en consideración del efecto de la insulina en la formación de tejido adiposo. La variedad metodológica de los estudios sobre LM hace compleja una buena homologación de los resultados, e incluso hasta indicar que la LM es perjudicial. En conclusión, la LM parece disminuir el riesgo de obesidad hacia la vida adulta; sin que los mecanismos biológicos de estas asociaciones estén totalmente definidos; y además del efecto directo de los nutrientes, existen otros mecanismos que podrían explicar esta asociación.
The intrauterine environment and early life are fundamental to programming a series of situations in our body. There are many theories that suggest that breastfeeding (BF) can have benefits in preventing obesity. This refers to the BF has a number of hormonal components that promote maturation or manner of interpretation of certain brain areas, which have to do with the mechanisms of satiety and hunger. There are enough classic studies showing how it is regulated in the BF, and the importance it represents, in consideration of the effect of insulin in adipose tissue formation. The variety methodological studies complicates BF approval good results, and even indicate that the BF is harmful. In conclusion, the BF seems to decrease the risk of obesity into adulthood, but the biological mechanisms of these associations to be fully defined, and in addition to the direct effect of nutrients, there are other mechanisms that could explain this association.
Subject(s)
Humans , Male , Female , Infant, Newborn , Infant , Milk, Human/immunology , Obesity/classification , Obesity/complications , Obesity/diet therapy , Obesity/epidemiology , Obesity/genetics , Obesity/metabolism , Obesity/pathology , Insulin/classification , Insulin/deficiency , Insulin/adverse effects , Insulin/genetics , Obesity/prevention & control , Obesity/psychology , Obesity/therapyABSTRACT
Oxidative stress induced by chronic hyperglycemia in type 2 diabetes plays a crucial role in progressive loss of beta-cell mass through beta-cell apoptosis. Glucagon like peptide-1 (GLP-1) has effects on preservation of beta-cell mass and its insulin secretory function. GLP-1 possibly increases islet cell mass through stimulated proliferation from beta-cell and differentiation to beta-cell from progenitor cells. Also, it probably has an antiapoptotic effect on beta-cell, but detailed mechanisms are not proven. Therefore, we examined the protective mechanism of GLP-1 in beta-cell after induction of oxidative stress. The cell apoptosis decreased to ~50% when cells were treated with 100 microM H2O2 for up to 2 hr. After pretreatment of Ex-4, GLP-1 receptor agonist, flow cytometric analysis shows 41.7% reduction of beta-cell apoptosis. This data suggested that pretreatment of Ex-4 protect from oxidative stress-induced apoptosis. Also, Ex-4 treatment decreased GSK3beta activation, JNK phosphorylation and caspase-9, -3 activation and recovered the expression of insulin2 mRNA in beta-cell lines and secretion of insulin in human islet. These results suggest that Ex-4 may protect beta-cell apoptosis by blocking the JNK and GSK3beta mediated apoptotic pathway.
Subject(s)
Animals , Cricetinae , Humans , Apoptosis , Caspase 3/metabolism , Caspase 9/metabolism , Cells, Cultured , Flow Cytometry , Glucagon-Like Peptide 1/pharmacology , Glycogen Synthase Kinase 3/metabolism , Hydrogen Peroxide/toxicity , Insulin/genetics , Insulin-Secreting Cells/drug effects , JNK Mitogen-Activated Protein Kinases/metabolism , Oxidative Stress , Peptides/pharmacology , Phosphorylation , Receptors, Glucagon/agonists , Signal Transduction , Venoms/pharmacologyABSTRACT
INTRODUCTION:Type 1A diabetes mellitus (T1ADM) is a multifactorial disease in which genetic and environmental aspects are important to its development. The association of genetic variations with disease has been demonstrated in several studies; however, the role of some gene loci has not yet been fully elucidated. OBJECTIVE:To compare the frequency of HLA alleles and polymorphism in CTLA-4 and insulin genes in Brazilians with T1ADM and individuals without the disease, as well as to identify genetic markers that are able to discriminate between diabetic and non-diabetic individuals. METHODS: The presence of HLA DQB1, DQA1 and DRB1 alleles, as well as the -2221 MspI polymorphism in the insulin gene and 49 A/G in the CTLA-4 gene were identified by the "Time-resolved fluorometer" technique after hybridization with probes labeled with Eu (III) / Sm (III) and Tb (III). RESULTS: The DQB1 *0302 and DQA1 *03 alleles were identified as predisposed to T1ADM, and the DQB1 *0301 allele presented a protective effect against the disease.The DQA1 label proved to be able to differentiate between 71.13 percent of the diabetic and non-diabetic individuals.This value increased to 82.47 percent when the DQB1 label was added. No significant difference in the frequency of polymorphisms in the insulin and CTLA-4 genes was observed between the two groups. CONCLUSIONS: The genetic markers that best characterized and discriminated diabetic and non-diabetic individuals were the HLA DQA1 and DQB1.alleles.
INTRODUÇÃO: O diabetes melito tipo 1 (T1ADM) é uma doença multifatorial em que os aspectos genéticos e ambientais são importantes para o seu desenvolvimento. A associação das variações genéticas com a doença tem sido demonstrada em vários trabalhos, no entanto, o papel de alguns locos gênicos não foi ainda completamente elucidado. OBJETIVOS: Comparar a frequência de alelos do HLA e polimorfismos nos genes CTLA-4 e insulina em brasileiros com T1ADM e indivíduos sem a doença, além de identificar marcadores gênicos que sejam capazes de discriminar indivíduos diabéticos e não diabéticos. MÉTODOS: A presença dos alelos de HLA DQB1, DQA1 e DRB1, bem como dos polimorfismos -2221 MspI no gene da insulina e 49 A/G no gene CTLA-4, foram identificados por meio da técnica Time-resolved fluorometer, após hibridização com sondas marcadas com Eu (III)/Sm (III) e Tb (III). RESULTADOS: Os alelos DQB1*0302 e DQA1*03 foram identificados como sendo de predisposição ao T1ADM, e o alelo DQB1*0301 mostrou um efeito protetor à doença. Analisando somente o marcador DQA1, este mostrou ser capaz de diferenciar 71,13 por cento dos indivíduos entre diabéticos e não diabéticos, cujo valor aumentou para 82,47 por cento quando adicionado o marcador DQB1. A frequência dos polimorfismos nos genes da insulina e CTLA-4 não mostrou diferença significativa entre os dois grupos estudados. CONCLUSÕES: Os marcadores genéticos que melhor caracterizaram e discriminaram diabéticos e não diabéticos foram os alelos de HLA DQA1 e DQB1.
Subject(s)
Adolescent , Adult , Humans , Antigens, CD/genetics , Diabetes Mellitus, Type 1/genetics , HLA-D Antigens/genetics , Insulin/genetics , Polymorphism, Genetic/genetics , Alleles , Case-Control Studies , Discriminant Analysis , Gene Frequency , Genotype , Genetic Predisposition to Disease/geneticsABSTRACT
O diabetes melito tipo 1 auto-imune (DM1A) resulta da destruição auto-imune seletiva das células-beta pancreáticas produtoras de insulina. O principal determinante genético de suscetibilidade para o DM1A está em genes do complexo principal de histocompatibilidade, no cromossomo 6p211.3 (locus IDDM1), responsável por 40 por cento ou mais da agregação familiar dessa doença. O maior risco é conferido pelo genótipo do antígeno leucocitário humano HLA-DR3-DQA1* 0501-DQB1*0201/DR4-DQA1*0301-QB1*0302, e o haplótipo HLA-DR15-DQA1* 0102-DQB1*0602 é associado à proteção. Três outros loci relacionados à predisposição a DM1A são o número variável de freqüências repetidas (VNTR) do gene da insulina (IDDM2), que confere 10 por cento da suscetibilidade genética, o antígeno-4 associado ao linfócito T citotóxico (CTLA-4) e o protein tyrosine phosphatasis nonreceptor-type 22 (PTPN22). Muitos outros genes suspeitos de predispor à auto-imunidade estão sendo investigados. O DM1A é freqüentemente associado com doença auto-imune tiroidiana, doença celíaca, doença de Addison e várias outras doenças auto-imunes, caracterizadas por auto-anticorpos órgãos-específicos, relacionados aos mesmos determinantes genéticos. Esses anticorpos são úteis na detecção de auto-imunidade órgão-específica antes do aparecimento da doença clínica, prevenindo comorbidades.
Type 1 A diabetes mellitus (T1AD) results from the autoimmune destruction of the insulin producing pancreatic beta-cells. The largest contribution to genetic susceptibility comes from several genes located in the major histocompatibility complex on chromosome 6p21.3 (IDDM1 locus), accounting for at least 40 percent of the family aggregation of this disease. The highest-risk human leukocyte antigen HLA genotype for T1AD is DR3-DQA1*0501-DQB1*0201/DR4-DQA1*0301-DQB1*0302, whereas -DR15-DQA1*0102-DQB1*0602 haplotype is associated with dominant protection. Three other T1D loci associated with predisposition are the Variable Number for Tandem Repeats (VNTR) near the insulin gene (IDDM2), which accounts to 10 percent of genetic susceptibility, the Cytotoxic T-Lymphocyte-associated Antigen (CTLA-4)(IDDM 12) and the Protein Tyrosine Phosphatasis Nonreceptor-type 22 (PTPN22). Many other gene suspected to predispose to autoimmunity have been investigated. T1AD is frequently associated with autoimmune thyroid disease, celiac disase, Addison´s disease and many other autoimmune diseases, characterized by organ-specific autoantibodies and related to the same genetic background. Using these autoantibodies, organ specific autoimmunity may be detected before the development of clinical disease preventing significant morbidity.
Subject(s)
Female , Humans , Male , Autoimmunity/genetics , Diabetes Mellitus, Type 1/genetics , Diabetes Mellitus, Type 1/immunology , Genetic Predisposition to Disease/genetics , Age of Onset , Autoimmunity/immunology , HLA-DQ Antigens/genetics , HLA-DQ Antigens/immunology , HLA-DR Antigens/genetics , HLA-DR Antigens/immunology , Hypoglycemic Agents/immunology , Insulin/genetics , Insulin/immunologyABSTRACT
O diabetes neonatal (DN) é uma condição rara caracterizada por hiperglicemia, que necessita de tratamento com insulina, diagnosticado nos primeiros meses de vida. Clinicamente pode ser classificado em DN transitório quando ocorre remissão da doença em poucos meses, podendo haver recorrência posterior; ou permanente quando, como o nome indica, não ocorre remissão. Ambas as condições são geneticamente heterogêneas; entretanto a maioria dos casos de DN transitório é decorrente de anormalidades da região de imprinted no cromossomo 6q24. Mutações ativadoras em heterozigose no gene KCNJ11, que codifica a subunidade Kir6.2 do canal de potássio ATP-sensível, são a causa mais comum de DN permanente. No presente artigo, discutimos as características clínicas do DN, os mecanismos moleculares envolvidos e suas implicações terapêuticas.
Neonatal diabetes is a rare condition characterized by hyperglycemia, requiring insulin treatment, diagnosed within the first months of life. The disorder may be either transient, resolving in infancy or early childhood with possible relapse later, or permanent in which case lifelong treatment is necessary. Both conditions are genetically heterogeneous; however, the majority of the cases of transient neonatal diabetes are due to abnormalities of an imprinted region of chromosome 6q24. For permanent neonatal diabetes, the most common causes are heterozygous activating mutations of KCNJ11, the gene encoding the Kir6.2 sub-unit of the ATP-sensitive potassium channel. In this article we discuss the clinical features of neonatal diabetes, the underlying genetic defects and the therapeutic implications.
Subject(s)
Humans , Infant, Newborn , Diabetes Mellitus/genetics , Mutation , Diabetes Mellitus/drug therapy , Homeodomain Proteins/genetics , Hypoglycemic Agents/therapeutic use , Insulin/genetics , Insulin/therapeutic use , KATP Channels/genetics , Potassium Channels, Inwardly Rectifying/genetics , Potassium Channels, Inwardly Rectifying/therapeutic use , Tolbutamide/therapeutic use , Trans-Activators/geneticsABSTRACT
PURPOSE: The role of insulin 3-like (Insl3) hormone signaling in the testicular descent process has been demonstrated. The purpose of the present study was to evaluate epididymal development in Insl3-deficient mice. MATERIALS AND METHODS: Heterozygous and homozygous Insl3 mutants of a mixed CD1 X 129/Sv genetic background were generated by breeding Insl3-/- females with Insl3+/- males, and their genotypes were determined by polymerase chain reaction. On the first postnatal day, newborn males were sacrificed, embedded in paraffin, and cut in 4 µm sections. Sections were stained with hematoxylin/eosin and immunoreacted with anti-± actin antibody. RESULTS: An analysis of stained sections indicated an arrest in the development of the epididymis in all homozygous mice. The cauda and corpus of the epididymis were undersized. Compared to the heterozygous epididymis, the homozygous epididymis had fewer peritubular layers and dwarfish musculature. We confirmed this with immunostaining with monoclonal antibodies against ± -smooth muscle actin. CONCLUSION: Defective development of the smooth musculature in the epididymis of Insl3 homozygous mutant mice, combined with its high intraabdominal undescended position, supports previous observations regarding the importance of intact epididymis morphology and function for descent of the epididymo-testicular unit.