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OBJECTIVE: In our previous study, we identified a notable increase in miR-548ag content after obesity, which contributes to the progression of Type 2 diabetes Mellitus(T2DM) through the up-regulation of Dipeptidyl Peptidase-4(DPP4) expression within the liver. However, the precise molecular mechanisms underlying the upregulation of DPP4 by miR-548ag remain elusive. Mature miRNAs rich in GU sequences can activate the TLR(7/8)/NF-κB signalling pathway, which transcriptionally activates DPP4 expression. Notably, the proportion of GU sequences in hsa-miR-548ag was found to be 47.6%. The study proposes a hypothesis suggesting that miR-548ag could potentially increase DPP4 expression in hepatocytes by activating the TLR(7/8)/NF-κB signalling pathway. METHODS: Male C57BL/6J mice were fed normal chow diet (NCD, n = 16) or high-fat diet (HFD, n = 16) for 12 weeks. For a duration of 6 weeks, NCD mice received intraperitoneal injections of a miR-548ag mimic, while HFD mice and db/db mice (n = 16) were administered intraperitoneal injections of a miR-548ag inhibitor. qRT-PCR and Western Blot were used to detect the expression level of miR-548ag, DPP4 and the activation of TLR(7/8)/NF-κB signalling pathway. HepG2 and L02 cells were transfected with miR-548ag mimic, miR-548ag inhibitor, TLR7/8 interfering fragment, and overexpression of miR-548ag while inhibiting TLR7/8, respectively. RESULTS: (1) We observed elevated levels of miR-548ag in the serum, adipose tissue, and liver of obese mice, accompanied by an upregulation of TLR7/8, pivotal protein in the NF-κB pathway, and DPP4 expression in the liver. (2) miR-548ag promotes DPP4 expression in hepatocytes via the TLR(7/8)/NF-κB signalling pathway, resulting in a reduction in the glucose consumption capacity of hepatocytes. (3) The administration of a miR-548ag inhibitor enhanced glucose tolerance and insulin sensitivity in db/db mice. CONCLUSIONS: MiR-548ag promotes the expression of DPP4 in hepatocytes by activating the TLR(7/8)/NF-κB signalling pathway. MiR-548ag may be a potential target for the treatment of T2DM.
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Dipeptidil Peptidasa 4 , Hepatocitos , Ratones Endogámicos C57BL , MicroARNs , FN-kappa B , Transducción de Señal , Animales , Ratones , Masculino , MicroARNs/metabolismo , MicroARNs/genética , Hepatocitos/metabolismo , Dipeptidil Peptidasa 4/metabolismo , Dipeptidil Peptidasa 4/genética , FN-kappa B/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Humanos , Obesidad/metabolismo , Obesidad/genética , Dieta Alta en Grasa , Modelos Animales de Enfermedad , Regulación hacia Arriba , Receptor Toll-Like 7/metabolismo , Receptor Toll-Like 7/genéticaRESUMEN
Dissimilatory arsenate-respiring prokaryotes (DARPs) are considered to be a key impetus of the reductive dissolution of solid-phase arsenic. However, little is known about the interaction between nitrate and DARPs so far. In this study, we showed that nitrate either inhibited or promoted the DARP population-catalyzed reductive mobilization of As in sediments. Metagenomic analysis of the microbial communities in the microcosms after seven days of As release assays suggested that microbes mainly consisted of: Type-I DARPs having potential to reduce NO3- into NO2- and Type-II DARPs having potential to reduce NO3- to NH4+. We further isolated two cultivable DARPs, Neobacillus sp. A01 and Paenibacillus sp. A02, which represent Type-I and -II DARPs, respectively. We observed that nitrate suppressed A01-mediated release of As(III) but promoted A02-mediated release of As(III). Furthermore, we demonstrated that this observation was due to the fact that nitrite, the end product of incomplete denitrification by Type-I DARPs, suppressed the arrA gene expression per cell and growth of all DARPs, whereas ammonium, the end product of dissimilatory nitrate reduction to ammonium (DNRA) by Type-II DARPs, enhanced the arrA gene expression per cell and significantly promoted the growth of all DARPs. These findings suggest that the actual effects of nitrate on DARP population-catalyzed reductive mobilization of arsenic, largely depend on the ratio of Type-I to Type-II DARPs in sediments.
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Arsénico , Nitratos , Arseniatos , NitritosRESUMEN
OBJECTIVE: Our previous results have shown that obesity-induced excessive palmitic acid (PA) can promote the expression of KLF7, which plays a vital role in regulation of inflammation, glucose metabolism. But the exact mechanism of PA up-regulating the expression of KLF7 is not clear yet. This study is intend to explore whether PA promoting KLF7 expression through GPRs/NF-κB signaling pathway, causing inflammation and glucose metabolism disorders. METHODS: Cells were blocked GPRs/NF-κB under PA stimulation in vitro to demonstrate the molecular mechanism of PA up-regulates KLF7 expression. The regulatory effect of p65 on KLF7 was detected by luciferase reporter gene assay. Blocking GPRs/NF-κB in diet-induced obesity mice to detect the expression of KLF7, inflammatory cytokines and glucose metabolism related factors, clarifying the effects of GPRs/NF-κB on KLF7 in vivo. RESULTS: In 3T3-L1 adipocytes and HepG2 cells, PA could up-regulate the expression of KLF7 by promoting the GPR40/120-NF-κB signaling pathway, leading to inflammation and reduced glucose consumption (p < 0.05 for both). Luciferase reporter gene assay and ChIP assay showed that p65 could transcriptionally up-regulates the expression of KLF7. In high-fat diet (HFD) mice, after intraperitoneal injection of GPR40 or GPR120 blocker, the levels of p-p65 and KLF7 in epididymal white adipose tissue and liver were significantly decreased (p < 0.05 for both). Pharmacological inhibition of p-p65 significantly attenuated KLF7 expression and improved glucose tolerant and insulin sensitive (p < 0.05 for both). CONCLUSIONS: Our results indicate that obesity-induced elevated palmitic acid promotes inflammation and glucose metabolism disorders through GPRs/NF-κB/KLF7 signaling pathway.
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Trastornos del Metabolismo de la Glucosa , FN-kappa B , Animales , Glucosa , Trastornos del Metabolismo de la Glucosa/complicaciones , Inflamación/complicaciones , Factores de Transcripción de Tipo Kruppel/genética , Factores de Transcripción de Tipo Kruppel/metabolismo , Ratones , FN-kappa B/metabolismo , Obesidad/metabolismo , Ácido Palmítico/farmacologíaRESUMEN
AIM/INTRODUCTION: Obesity is considered an important risk factor for many metabolic disorders, especially type 2 diabetes mellitus, and microRNAs (miRNAs) play a vital role in the development of type 2 diabetes mellitus. Therefore, we conducted this study to investigate the role of miR-4431 in the obesity-associated pathobiology of type 2 diabetes mellitus. MATERIALS AND METHODS: Subjects were divided into normal control (n = 36), obese (n = 36), and type 2 diabetes mellitus (n = 12) groups, and serum miR-4431 levels were analyzed. Adenovirus-vectored miR-4431 mimic or sponge was intraperitoneally injected into the normal diet group and the high-fat diet group (HFD) mice to investigate glucose tolerance, insulin sensitivity, and lipid levels. The downstream target genes of miR-4431 were predicted using bioinformatics, and they were verified in vitro. RESULTS: Serum miR-4431 levels were significantly high in obese and type 2 diabetes mellitus individuals, and positively correlated with the body mass index and fasting plasma glucose levels. In HFD mice, miR-4431 levels in the serum, white adipose tissue, and liver were significantly increased. Moreover, miR-4431 impaired glucose tolerance, insulin sensitivity, and lipid metabolism in mice. Bioinformatic prediction suggested that TRIP10 and PRKD1 could be the downstream target genes of miR-4431. The HFD mice showed a remarkable reduction in the mRNA levels of TRIP10 and PRKD1 in the liver, which were countered by blocking miR-4431. In HepG2 and L02 cells, miR-4431 could downregulate TRIP10 and PRKD1 while blocking glucose uptake. The luciferase reporter assay showed that miR-4431 could bind TRIP10 and PRKD1 3'-UTR. CONCLUSION: miR-4431 targets TRIP10/PRKD1 and impairs glucose metabolism.
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Diabetes Mellitus Tipo 2 , Resistencia a la Insulina , MicroARNs , Animales , Diabetes Mellitus Tipo 2/genética , Diabetes Mellitus Tipo 2/metabolismo , Dieta Alta en Grasa/efectos adversos , Glucosa/metabolismo , Humanos , Resistencia a la Insulina/genética , Metabolismo de los Lípidos , Ratones , Ratones Endogámicos C57BL , MicroARNs/genética , MicroARNs/metabolismo , Obesidad/complicaciones , Obesidad/genética , Obesidad/metabolismoRESUMEN
BACKGROUND: Obesity-induced elevated serum free fatty acids (FFAs) levels result in the occurrence of type 2 diabetes mellitus (T2DM). However, the molecular mechanism remains largely enigmatic. This study was to explore the effect and mechanism of KLF15 on FFAs-induced abnormal glucose metabolism. METHODS: Levels of TG, TC, HDL-C, LDL-C, and glucose were measured by different assay kits. qRT-PCR and Western Blot were used to detect the levels of GPR120, GPR40, phosphorylation of p38 MAPK, KLF15, and downstream factors. RESULTS: KLF15 was decreased in visceral adipose tissue of obesity subjects and high-fat diet (HFD) mice. In HFD mice, GPR120 antagonist significantly promoted KLF15 protein expression level and phosphorylation of p38 MAPK, meanwhile reduced the blood glucose levels. While, blocking GPR40 inhibited the KLF15 expression. In 3T3-L1 adipocytes, 1500 µM PA inhibited KLF15 through a GPR120/P-p38 MAPK signal pathway, and 750 µM OA inhibited KLF15 mainly through GPR120 while not dependent on P-p38 MAPK, ultimately resulting in abnormal glucose metabolism. Unfortunately, GPR40 didn't contribute to PA or OA-induced KLF15 reduction. CONCLUSIONS: Both PA and OA inhibit KLF15 expression through GPR120, leading to abnormal glucose metabolism in adipocytes. Notably, the inhibition of KLF15 expression by PA depends on phosphorylation of p38 MAPK.
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AIMS/INTRODUCTION: Microribonucleic acid-155 (microRNA155) and microRNA29 are reported to inhibit glucose metabolism in some cell and animal models, but no evidence from susceptible populations that examines the relationship between microRNA155 or microRNA29 and type 2 diabetes mellitus currently exists. Furthermore, target genes regulated by microRNA155 and microRNA29 that affect glucose and lipid metabolism remain unknown. MATERIALS AND METHODS: Human participants were divided into normal weight (n = 72), obesity (n = 120) and type 2 diabetes (n = 59) groups. The contents of microRNA155 and microRNA29 abundance in serum were measured, and candidate genes potentially related to glucose and lipid metabolism targeted by either microRNA155 or microRNA29 were screened. Overexpression of microRNA155 and microRNA29 in HepG2 cells was used to verify candidate gene expression, and measure the effects on glucose and lipid metabolism. RESULTS: Serum levels of microRNA155 and microRNA29 show a significant increase in individuals with obesity and type 2 diabetes compared with normal weight individuals. Identified target genes for microRNA155 were MAPK14, MAP3K10, DUSP14 and PRKAR2B. Identified target genes for microRNA29 were PEX11A and FADS1. Overexpression of microRNA155 or microRNA29 in HepG2 cells was found to downregulate the expression of identified target genes, and result in inhibition of triglyceride synthesis and glucose incorporation. CONCLUSIONS: MicroRNA155 and microRNA29 were significantly higher in type 2 diabetes patients compared with the control patients, their levels were also positively correlated with fasting plasma glucose levels, and over-expression of microRNA155 or microRNA29 were found to downregulate glucose and lipid metabolism target genes, and reduce lipid synthesis and glucose incorporation in HepG2 cells.
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Biomarcadores/análisis , Diabetes Mellitus Tipo 2/patología , MicroARNs/genética , Glucemia/análisis , Estudios de Casos y Controles , delta-5 Desaturasa de Ácido Graso , Diabetes Mellitus Tipo 2/epidemiología , Diabetes Mellitus Tipo 2/genética , Estudios de Seguimiento , Células Hep G2 , Humanos , PronósticoRESUMEN
AIMS/INTRODUCTION: Type 2 diabetes mellitus is closely linked to increased levels of free fatty acids (FFAs) in obese individuals, although which FFA is most associated with type 2 diabetes mellitus is unclear. This study aimed to identify the specific FFAs that best predict the occurrence of type 2 diabetes mellitus in obese individuals, and assess their potential application value. MATERIALS AND METHODS: Participants were divided into three groups: a normal weight group (n = 20), an obese group (n = 10) and a type 2 diabetes mellitus group (n = 10). FFAs in serum samples were determined by ultra-high-pressure liquid chromatography-mass spectrometry, and orthogonal partial least squares discriminant analysis models were used to study the FFA profile among the three groups. RESULTS: Compared with the normal weight group, 14 FFAs (C8:0/10:0/14:0/16:1/18:1/20:2/ 20:3 /20:4/ 20:5/ 22:6/7:0/9:0/11:0 and C13:0) were significantly increased in the obese group, and nine FFAs (C14:0, C18:1, C20:1, C 18:2, C20:2, C20:3, C18:3, C20:5 and C22:6) were significantly increased in the type 2 diabetes mellitus group. Subsequently, the Venn diagram results showed that six FFAs (C14:0, C18:1, C20:2, C20:3, C20:5 and C22:6) were significantly increased in both the obese and type 2 diabetes mellitus groups. Among these six, C22:6 was finally identified as an independent risk factor for type 2 diabetes mellitus, and had a great potential to predict the susceptibility to type 2 diabetes mellitus (area under the curve 0.803). CONCLUSIONS: C22:6 can be an independent risk factor for type 2 diabetes mellitus, and it has a great potential to predict the susceptibility to type 2 diabetes mellitus.
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Diabetes Mellitus Tipo 2/etiología , Susceptibilidad a Enfermedades/sangre , Ácidos Grasos no Esterificados/sangre , Obesidad/sangre , Adulto , Biomarcadores/sangre , Femenino , Cromatografía de Gases y Espectrometría de Masas , Humanos , Masculino , Persona de Mediana Edad , Obesidad/complicaciones , Valor Predictivo de las Pruebas , Factores de RiesgoRESUMEN
The morphology of nanomaterials significantly affects their physical, chemical, and biological properties. In the present study, nano-hydroxyapatite coatings with different morphologies were produced on the surface of a titanium-niobium shape memory alloy via a hydrothermal process. The effect of the nano-hydroxyapatite coatings on the in vitro proliferation of SaOS-2 osteoblast-like cells was investigated. Factors including crystallinity, surface micro-roughness, and surface energy of the nano-hydroxyapatite coatings were discussed. Results show that in vitro proliferation of the osteoblast-like cells was significantly enhanced on the nano-hydroxyapatite-coated titanium-niobium alloy compared to the titanium-niobium alloy without coating. The cell numbers on the nano-hydroxyapatite-coated titanium-niobium alloy changed consistently with the surface energy of the hydroxyapatite coatings. This study suggests that surface energy as a characteristic parameter influencing the in vitro proliferation of osteoblast-like cells was predominant over the crystallinity and surface micro-roughness of the nano-hydroxyapatite coatings.
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Aleaciones/química , Proliferación Celular , Durapatita/química , Nanoestructuras/química , Niobio/química , Osteoblastos/fisiología , Titanio/química , Animales , Línea Celular , Materiales Biocompatibles Revestidos/química , Materiales Biocompatibles Revestidos/metabolismo , Durapatita/metabolismo , Glicol de Etileno/química , Humanos , Ensayo de Materiales , Microscopía Electrónica de Rastreo , Osteoblastos/citología , Propiedades de Superficie , Agua/química , Difracción de Rayos XRESUMEN
A novel one-step hydrothermal coating process was used to produce nanohydroxyapatite (nano-HA) coating on a titanium-niobium (TiNb) alloy substrate in a newly designed solution containing calcium and phosphate ions. The morphology of the coating was studied using scanning electron microscopy. The phase identification of the coating was carried out using X-ray diffraction, attenuated total reflectance Fourier transform infrared spectroscopy and transmission electron microscopy. The reaction between the surface of TiNb alloy and the solution during the hydrothermal process was studied by X-ray photoelectron spectroscopy. Results show that the coating formed on the surface of TiNb alloy was composed of nano-HA particles. During the hydrothermal process, TiO(2) and Nb(2)O(5) formed on the TiNb alloy surface and hydrated to Ti(OH)(4) and Nb(OH)(5), respectively. Calcium phosphate nucleated and grew into a layer of nano-HA particles on the surface of TiNb alloy under the hydrothermal conditions. The crystallinity of the nano-HA coating was improved with the increase in hydrothermal treatment temperature and time duration. Nano-HA coating with good crystallinity was produced on the TiNb alloy via the hydrothermal process at a temperature of 200 degrees C for 12 h.
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Aleaciones/química , Materiales Biocompatibles Revestidos/química , Durapatita/química , Nanoestructuras/química , Nanotecnología/métodos , Niobio/química , Titanio/química , Elementos Químicos , Nanoestructuras/ultraestructura , Espectroscopía de Fotoelectrones , Espectroscopía Infrarroja por Transformada de Fourier , Propiedades de Superficie , Temperatura , Agua , Difracción de Rayos XRESUMEN
In the present study, porous Ti-10Nb-10Zr alloy scaffolds with different porosities were successfully fabricated by a "space-holder" sintering method. By the addition of biocompatible alloying elements the porous TiNbZr scaffolds achieved significantly higher strength than unalloyed Ti scaffolds of the same porosity. In particular, the porous TiNbZr alloy with 59% porosity exhibited an elastic modulus and plateau stress of 5.6 GPa and 137 MPa, respectively. The porous alloys exhibited excellent ductility during compression tests and the deformation mechanism is mainly governed by bending and buckling of the struts. Cell cultures revealed that SaOS2 osteoblast-like cells grew on the surface and inside the pores and showed good spreading. Cell viability for the porous scaffold was three times higher than the solid counterpart. The present study has demonstrated that the porous TiNbZr alloy scaffolds are promising scaffold biomaterials for bone tissue engineering by virtue of their appropriate mechanical properties, highly porous structure and excellent biocompatibility.
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Aleaciones/química , Materiales Biocompatibles/química , Niobio/química , Andamios del Tejido/química , Titanio/química , Circonio/química , Línea Celular , Fuerza Compresiva , Módulo de Elasticidad , Humanos , Ensayo de Materiales , Osteoblastos/citología , Osteoblastos/metabolismo , Porosidad , Polvos/química , Estrés Mecánico , Resistencia a la TracciónRESUMEN
Porous titanium (Ti) and titanium alloys are promising scaffold biomaterials for bone tissue engineering, because they have the potential to provide new bone tissue ingrowth abilities and low elastic modulus to match that of natural bone. In the present study, a new highly porous Ti6Ta4Sn alloy scaffold with the addition of biocompatible alloying elements (tantalum (Ta) and tin (Sn)) was prepared using a space-holder sintering method. The strength of the Ti6Ta4Sn scaffold with a porosity of 75% was found to be significantly higher than that of a pure Ti scaffold with the same porosity. The elastic modulus of the porous alloy can be customized to match that of human bone by adjusting its porosity. In addition, the porous Ti6Ta4Sn alloy exhibited an interconnected porous structure, which enabled the ingrowth of new bone tissues. Cell culture results revealed that human SaOS(2) osteoblast-like cells grew and spread well on the surfaces of the solid alloy, and throughout the porous scaffold. The surface roughness of the alloy showed a significant effect on the cell behavior, and the optimum surface roughness range for the adhesion of the SaOS(2) cell on the alloy was 0.15 to 0.35 mum. The present study illustrated the feasibility of using the porous Ti6Ta4Sn alloy scaffold as an orthopedic implant material with a special emphasis on its excellent biomechanical properties and in vitro biocompatibility with a high preference by osteoblast-like cells.
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Aleaciones/química , Huesos/citología , Tantalio/química , Ingeniería de Tejidos/métodos , Andamios del Tejido/química , Titanio/química , Animales , Materiales Biocompatibles/química , Fenómenos Biomecánicos , Huesos/ultraestructura , Proliferación Celular , Supervivencia Celular/fisiología , Células Cultivadas , Humanos , Microscopía Electrónica de Rastreo , Porosidad , RatasRESUMEN
A porous Ti-18 at.%Nb-4 at.%Sn (hereafter, Ti-18Nb-4Sn) alloy was prepared by powder metallurgy. The porous structures were examined by scanning electron microscopy and the phase constituents were analysed by X-ray diffraction. Mechanical properties of the porous alloy were investigated using a compressive test. To enhance the bioactivity of the alloy surface, alkali-heat treatment was used to modify the surface. The bioactivity of the pre-treated alloy sample was investigated using a biomimetic process by soaking the sample into simulated body fluid (SBF). Results indicate that the elastic modulus and plateau stress of the porous Ti-18Nb-4Sn alloy decrease with decreasing relative density. The mechanical properties of the porous alloy can be tailored to match those of human bone. After soaking in SBF for 7 days, a hydroxyapatite layer formed on the surface of the pre-treated porous Ti-18Nb-4Sn alloy. The pre-treated porous Ti-18Nb-4Sn alloy therefore has the potential to be a bioactive implant material.