ABSTRACT
This study aimed to investigate the effect of Wenyang Zhenshuai Granules(WYZSG) on autophagy and apoptosis of myocardial cells in rats with sepsis via regulating the expression of microRNA-132-3p(miR-132-3p)/uncoupling protein 2(UCP2). Sixty SD rats were randomly divided into modeling group(n=50) and sham operation group(n=10). The sepsis rat model was constructed by cecal ligation and perforation in the modeling group. The successfully modeled rats were randomly divided into WYZSG low-, medium-and high-dose groups, model group and positive control group. Rats in the sham operation group underwent opening and cecum division but without perforation and ligation. Hematoxylin-eosin(HE) staining was used to observe the pathological changes of rat myocardial tissue. Myocardial cell apoptosis was detected by TdT-mediated dUTP nick end labeling(TUNEL) assay. Real-time quantitative polymerase chain reaction(RT-qPCR) was performed to detect the expression of miR-132-3p and the mRNA expressions of UCP2, microtubule-associated protein light chain 3(LC3-Ⅱ/LC3-Ⅰ), Beclin-1 and caspase-3 in rat myocardial tissue. The protein expressions of UCP2, LC3-Ⅱ/LC3-Ⅰ, Beclin-1 and caspase-3 in myocardial tissue were detected by Western blot. Dual luciferase reporter assay was used to verify the regulatory relationship between miR-132-3p and UCP2. The myocardial fibers of sepsis model rats were disordered, and there were obvious inflammatory cell infiltration as well as myocardial cell edema and necrosis. With the increase of the WYZSG dose, the histopathological changes of myocardium were improved to varying degrees. Compared with the conditions in the sham operation group, the survival rate and left ventricular ejection fraction(LVEF) of rats in the model group, positive control group and WYZSG low-, medium-and high-dose groups were decreased, and the myocardial injury score and apoptosis rate were increased. Compared with the model group, the positive control group and WYZSG low-, medium-and high-dose groups had elevated survival rate and LVEF, and lowered myocardial injury score and apoptosis rate. The expression of miR-132-3p and the mRNA and protein expressions of UCP2 in myocardial tissue in the model group, positive control group and WYZSG low-, medium-and high-dose groups were lower, while the mRNA and protein expressions of LC3-Ⅱ/LC3-Ⅰ, Beclin-1 and caspase-3 were higher than those in the sham operation group. Compared with model group, the positive control group and the WYZSG low-, medium-and high-dose groups had an up-regulation in the expression of miR-132-3p and the mRNA and protein expressions of UCP2, while a down-regulation in the mRNA and protein expressions of LC3-Ⅱ/LC3-Ⅰ, Beclin-1 and caspase-3. WYZSG inhibited excessive autophagy and apoptosis of myocardial cells in septic rats and improved myocardial injury, possibly by regulating the expression of miR-132-3p/UCP2.
Subject(s)
Animals , Rats , Rats, Sprague-Dawley , Caspase 3 , Beclin-1/genetics , Stroke Volume , Ventricular Function, Left , Apoptosis/genetics , Autophagy/genetics , Heart Injuries , MicroRNAs/geneticsABSTRACT
El ambiente obesogénico promueve la obesidad al facilitar el acceso y consumo de una amplia variedad de alimentos palatables altos en calorías. La activación del receptor de GLP1 (GLP1R) reduce la ingesta de alimentos, enlentece el vaciamiento gástrico y promueve un balance energético negativo a través de su acción en distintos órganos como el músculo esquelético, disminuyendo así el peso corporal. La obesidad inducida por dieta alta en grasa disminuye el efecto anorexigénico de la administración sistémica vía intra-peritoneal de EX4 (agonista de GLP1R). Sin embargo, se desconoce si la exposición a un ambiente obesogénico previo a la manifestación de obesidad disminuye los efectos anorexigénicos de EX4 o un posible efecto de EX4 sobre marcadores de oxidación de ácidos grasos y termogénesis en músculo esquelético. El objetivo de esta investigación fue determinar el efecto a corto plazo de la dieta CAF, un modelo del ambiente obesogénico humano, sobre la capacidad de EX4 de reducir la ingesta y modular la expresión de marcadores proteicos de oxidación de ácidos grasos y termogénesis (CPT1 y UCP2) en músculo de ratones. Nuestros datos muestran que una inyección intraperitoneal de EX4 a ratones C57BL/6J alimentados con dieta CAF o dieta control durante 10 días no altera la ingesta calórica total, peso corporal, o la expresión de proteínas marcadoras de los procesos de beta-oxidación y de termogénesis (CPT1 y UCP2). Estos datos sugieren que protocolos alternativos de administración de EX4 son necesarios para observar los efectos fisiológicos de la activación de GLP1R.
The obesogenic environment promotes obesity by facilitating access to and consumption of a wide variety of palatable, high-calorie foods. Activation of the GLP1 receptor (GLP1R) reduces food intake, slows gastric emptying, and promotes a negative energy balance by acting on organs such as skeletal muscle, thus decreasing body weight. Obesity induced by a high-fat diet decreased the anorexigenic effect of intraperitoneal systemic administration of EX4 (GLP1R agonist). However, it is unknown whether exposure to an obesogenic environment before the manifestation of obesity diminishes the anorexigenic effects of EX4 or a possible effect of EX4 on markers of fatty acid oxidation and thermogenesis in skeletal muscle. This investigation aimed to determine the short-term effect of the CAF diet, a model of the human obesogenic environment, on the ability of EX4 to reduce intake and modulate the expression of protein markers of fatty acid oxidation and thermogenesis (CPT1 and UCP2) in mouse muscle. Our data show that intraperitoneal injection of EX4 to C57BL/6J mice fed CAF diet or control diet for ten days does not alter total caloric intake, body weight, or expression of proteins markers of beta-oxidation and thermogenesis processes (CPT1 and UCP2). These data suggest that alternative EX4 administration protocols are necessary to observe the physiological effects of GLP1R activation.
Subject(s)
Animals , Male , Mice , Diet/adverse effects , Exenatide/administration & dosage , Obesity/etiology , Obesity/metabolism , Oxidation-Reduction , Blotting, Western , Muscle, Skeletal/metabolism , Thermogenesis , Fatty Acids/metabolism , Glucagon-Like Peptide-1 Receptor/metabolism , Uncoupling Protein 2 , Irinotecan , Injections, Intraperitoneal , Mice, Inbred C57BLABSTRACT
HM(hibernating myocardium)is an adaptive phenome-non of myocardium against sustained ischemia,which maintains its tissue vitality through balancing energy supply and demand.It widely exists in patients suffering from coronary heart disease. HMhas its own metabolic pattern,instead of regular FAO(fatty acid β-oxidation)-based metabolism,glycolysis became main pro-cedure.Reduction of FAO,TCA (tricarboxylic cacidcycle),ETC (electron transport chain)enzyme has been observed,ROS(reac-tive oxygen species)and UCP2(uncoupling protein 2)have been up-regulated.UCP2,which promotes proton leak across innermembrane of mitochondrial and leads to ATP reduction,has e-merged as an important regulator of the energy production.It is regulated by up-stream proteins such as AMPK,PPARs,PGC-1α,and other factors like FFA(free fat acid),ROS and purine nucleotide.HM has potential function of ischemic myocardium, which can improve cardiac function through reasonable interven-tion.Modulation of UCP2 can optimize energy production,and is essential to HM metabolism.
ABSTRACT
Objective To research the protective effect of insulin(IN)on lipopolysaccharide(LPS)- induced impairments of rat cardiomyocytes H9c2,and the role of uncoupling protein 2(UCP2)in this process. Methods Using randomized controlled grouping,after cultured for 24 h,H9c2 cells were randomly divided into 5 groups as follows:con-trol group,LPS stimulation group(LPS group),LPS + 70 IU/ L IN group(IN 70 IU/ L group),LPS + 350 IU/ L IN group(IN 350 IU/ L group),and LPS + 700 IU/ L IN group(IN 700 IU/ L group). H9c2 cells in IN group were treated with 70 IU/ L,350 IU/ L or 700 IU/ L IN 15 min before LPS stimulation,and H9c2 cells in control group were treated with an equal volume of saline. After that,cells in group LPS and IN were treated with LPS for 24 h. Lactate dehydro-genase(LDH)in the culture was determined with LDH detecting assay kit. The activity of reactive oxygen species (ROS)and superoxide dismutase(SOD),and content of malonaldehyde(MDA)were determined by colorimetric detec-tion. Cell viability was evaluated by cell count kit - 8. The expressions of UCP2 in transcription and translation levels were detected through transcription polymerase chain reaction and Western blot respectively. Results The levels of LDH,MDA,and intracellular ROS in LPS group significantly increased compared with control group[LDH:(829. 3 ± 75. 3)U/ L vs(223. 5 ± 23. 6)U/ L,MDA:(60. 90 ± 5. 73)nmol/ mgprot vs(19. 70 ± 1. 99)nmol/ mgprot,ROS:(410. 2 ± 81. 6)U/ well vs(94. 3 ± 18. 5)U/ well,all P ﹤ 0. 05)],while the cell viability and SOD activity significantly decreased[cell viability:0. 822 ± 0. 058 vs 1. 012 ± 0. 023,SOD:(49. 20 ± 5. 81)U/ mgprot vs(89. 80 ± 2. 57)U/ mg-prot,all P ﹤ 0. 05]. And the mRNA and protein expressions of UCP2 in LPS stimulation group were up - regulated (1. 867 ± 0. 130 vs 1. 028 ± 0. 097,0. 288 ± 0. 018 vs 0. 180 ± 0. 008,all P ﹤ 0. 05). 350 IU/ L and 700 IU/ L IN inter-vention significantly decreased the levels of LDH,MDA and intracellular ROS[LDH:(568. 2 ± 35. 7)U/ L,(622. 8 ± 27. 6)U/ L vs(829. 3 ± 75. 3)U/ L,MDA:(29. 20 ± 4. 20)nmol/ mgprot,(42. 10 ± 2. 32)nmol/ mgprot vs(60. 90 ± 5. 73)nmol/ mgprot,ROS:(270. 3 ± 46. 8)U/ well,(301. 5 ± 16. 9)U/ well vs(410. 2 ± 81. 6)U/ well,all P ﹤ 0. 05], increased the cell survival and the levels of SOD activity[cell viability:0. 960 ± 0. 029,0. 906 ± 0. 039 vs 0. 822 ± 0. 058,SOD:(75. 20 ± 2. 21)U/ mgprot,(61. 20 ± 3. 38)U/ mgprot vs(49. 20 ± 5. 81)U/ mgprot,all P ﹤ 0. 05]. And IN with 350 IU/ L and 700 IU/ L increased the mRNA and protein expression of UCP2(3. 830 ± 0. 265,2. 855 ± 0. 215 vs 1. 867 ± 0. 130,0. 464 ± 0. 215,0. 355 ± 0. 006 vs 0. 288 ± 0. 018,all P ﹤ 0. 05). Compared with 70 IU/ L and 700 IU/ L IN group,350 IU/ L IN group had better results. Conclusions IN attenuates LPS - induced oxidative injury in H9c2 cells,which is probably mediated through up - regulating the expression of UCP2.
ABSTRACT
The altered maternal/fetal metabolism appears to be associated with a diabetogenic effect in the adult offspring even in the absence of genetic predisposition. Aim: The study aimed to investigate the effect of maternal obesity and malnutrition on the peripheral glucose sensing and mitochondria biogenesis in F1 offspring. Effect of postnatal diet was also evaluated. Methods: Three groups of female Wistar rats were used (control, obese and malnourished). After the pregnancy and delivery the offspring were weaned to control diet or high-caloric (HCD) diet and followed up for 30 weeks. Every 5 weeks OGTT was constructed and serum and tissues were obtained for assessment of glucose homeostasis parameters, mTFA, mtDNA, UCP2, insulin receptor (IR), phospho-insulin receptor (Phosho-IR) and GLUT4. Results: The results indicated that maternal obesity impair glucose tolerance and sensing in the offspring from the 15th week of age even under control diet and the situation is worse under HCD these defects were preceded by significant decline in mtDNA copy number in muscle and adipose tissues as early as 5th week of age. The offspring of malnourished mothers show normal and even better glucose tolerance and sensing and normal mtDNA copy number under control diet, while those offspring under HCD show impaired glucose sensing and tolerance only at older age than obese group. Conclusion: maternal obesity and malnutrition differentially affect glucose sensing and tolerance, mtDNA copy number and the expression of genes involved in the mitochondrial biogenesis and function in the muscles and adipose tissues in the F1 offspring with the postnatal feeding appearing to play a central role in these differential effects. The male F1 offspring appear to be more sensitive for fetal diabetogenic programming than female offspring.
ABSTRACT
La obesidad es una enfermedad multifactorial que se relaciona con estilos de vida y factores medioambientales y genéticos. Uno de los genes candidatos de la obesidad es el UCP2. Su polimorfismo -866G/A se ha asociado con obesidad en algunas poblaciones. Sin embargo, se han reportado resultados contradictorios alrededor del mundo, lo cual indica la necesidad de nuevas investigaciones al respecto. Objetivo: Analizar el polimorfismo -866G/A del gen UCP2 asociado con obesidad en adultos de la ciudad de Valledupar. Materiales y métodos: Se estudiaron 103 individuos con sobrepeso u obesidad y 100 con normopeso. El polimorfismo de UCP2 -866G/A fue determinado por PCR-RFLP. Se evaluaron también medidas antropométricas, perfil de lipoproteínas y glucemia basal. Resultados: Se observó que el alelo mutado y su genotipo homocigoto fueron significativamente más frecuentes en pacientes con IMC > a 25 kg/m². [A: OR= 2,9 (IC 95%= 1,765-4,751) y AA: OR=5,8 (IC 95%= 1,264-2,745)]. No se encontraron diferencias significativas entre UCP2 -866G/A y las variables clínicas estudiadas en individuos obesos. Sin embargo, se observa que los sujetos con alelos y genotipos mutados presentaron cifras más elevadas de triglicéridos, glucemia e ICC y menor promedio de cHDL. Conclusiones: la mutación -866G/A del gen UCP2 se asocia a obesidad en la población estudiada y aunque no parece influir en las medidas antropométricas y bioquímicas en sujetos obesos, podría estar relacionado con aumento de ICC, glucosa y triglicéridos y disminución de cHDL.
Obesity is a multifactorial disease that is related to lifestyles, and environmental and genetic factors. One of the candidate genes for obesity is the UCP2. Its polymorphism -866G/A was associated with obesity in some populations. However, conflicting results have been reported around the world, indicating the need for further investigations. Objective: To analyze the polymorphism -866G/A UCP2 gene associated with obesity in adults in the city of Valledupar. Materials and methods: We studied 103 overweight or obese individuals and 100 normal weight. The polymorphism of UCP2 -866G/A was determined by PCR-RFLP. Anthropometric measures were also evaluated, lipoprotein profile and fasting glucose. Results: We found that the mutated allele and homozygous genotype were significantly more frequent in patients with BMI > 25 kg/m². [A: OR = 2.9 (95% CI= 1,765 to 4,751 ) and AA: OR= 5.8 (95% CI = 1,264 to 2,745)]. No significant differences were found between UCP2 -866G/A and the clinical variables studied in obese individuals. However it is observed that subjects with mutated alleles and genotypes had higher triglycerides, glucose and ICC and lower average HDL cholesterol. Conclusions: mutation -866G/A UCP2 gene is associated with obesity in the population studied, and although it seems to influence the anthropometric and biochemical measures in obese subjects could be related to increased ICC, glucose and triglycerides and decreased HDL.
A obesidade é uma doença multifatorial que se relaciona com estilos de vida, e fatores meio ambientais e genéticos. Um dos genes candidatos da obesidade é o UCP2. Seu polimorfismo -866G/A tem se associado com obesidade em algumas populações. No entanto, tem se reportado resultados contraditórios ao redor do mundo, o qual indica a necessidade de novas pesquisas a respeito. Objetivo: analisar o polimorfismo -866G/A do gene UCP2 associado com obesidade em adultos da cidade de Valledupar. Materiais e métodos: estudaram-se 103 indivíduos com excesso de peso ou obesidade e 100 com normopeso. O polimorfismo UCP2 -866G/A foi determinado por PCR-RFLP. Avaliaram-se também medidas antropométricas, perfil de lipoproteínas e glicemia basal. Resultados: observou-se que o alelo mudado e seu genotipo homozigoto foram significativamente mais frequentes em pacientes com IMC > a 25 Kg/m2. (A: OR= 2,9 (IC 95%= 1,765-4,751) y AA: OR=5,8 (IC 95%= 1,264-2,745)). Não se encontraram diferenças significativas entre UCP2 -866G/A e as variáveis clínicas estudadas em indivíduos obesos. No entanto, observa-se que os sujeitos com alelos e genótipos mudados apresentaram cifras mais elevadas de triglicerídeos, glicemia, ICC e menor média de cHDL. Conclusões: a mutação -866G/A do gene UCP2 associa-se a obesidade na população estudada, e ainda que não parecesse influir nas medidas antropométricas e bioquímicas em sujeitos obesos, poderia estar relacionado com aumento de ICC, glicose, triglicerídeos e diminuição de cHDL.
Subject(s)
Humans , Polymorphism, Genetic , Data Interpretation, Statistical , Colombia , Uncoupling Protein 2 , ObesityABSTRACT
Objective:To observe the effect of metformin on the expression of SIRT1 and UCP2 in rat liver of type 2 diabetes mellitus (T2DM) with nonalcoholic fatty liver disease (NAFLD), and discuss the pathogenesis of T2DM with NAFLD, and the treatment with and possible mechanism of metformin.Methods:Thirty-six male SD rats were randomly divided into a normal control group (group NC, n=12), a T2DM with NAFLD group (group MC, n=12), and a metformin group (group A, n=12). We established the model of T2DM with NAFLD rats by feeding high-fat and high-sugar diet and injecting STZ. After the success establishment of the model, the metformin group was given metformin 300 mg/(kg.d) for 8 weeks. At the end of the experiment, we measured FBG, ALT, AST, TC, TG, HDL-C, LDL-C, VLDL, FFAs, FINs and HOMA-IR respectively in group NC, MC and A. We observed the change of liver tissue pathology by HE, determined the expression of SIRT1 and UCP2 in rat liver by immunohistochemical method and real-time quantitative method. Results:FBG, ALT, AST, TC, TG, LDL-C, VLDL, FFAs, FINs and HOMA-IR were higher in group MC than in group NC (P Conclusion:The expression of SIRT1 is low and the expression of UCP2 is high in rat liver of T2DM with NAFLD. Metformin can increase the expression of SIRT1 and reduce the expression of UCP2, with negative correlation between the expression of SIRT1 and UCP2.
ABSTRACT
It is well established that genetic factors play an important role in the development of type 2 diabetes mellitus (DM2) and its chronic complications, and that genetically susceptible subjects can develop the disease after being exposed to environmental risk factors. Therefore, great efforts have been made to identify genes associated with DM2. Uncoupling protein 2 (UCP2) is expressed in several tissues, and acts in the protection against oxidative stress; in the negative regulation of insulin secretion by beta cells, and in fatty acid metabolism. All these mechanisms are associated with DM2 pathogenesis and its chronic complications. Therefore, UCP2 is a candidate gene for the development of these disorders. Indeed, several studies have reported that three common polymorphisms in UCP2 gene are possibly associated with DM2 and/or obesity. Only a few studies investigated these polymorphisms in relation to chronic complications of diabetes, with inconclusive results.
Está bem estabelecido que fatores genéticos têm papel importante no desenvolvimento do diabetes melito tipo 2 (DM2) bem como de suas complicações crônicas e que indivíduos geneticamente suscetíveis podem desenvolver essa doença após exposição a fatores de risco ambientais. Assim, grandes esforços têm sido feitos para a identificação de genes associados ao DM2. A proteína desacopladora 2 (UCP2) é expressa em diversos tecidos e atua na proteção contra o estresse oxidativo, na regulação negativa da secreção de insulina pelas células-beta e no metabolismo dos ácidos graxos, mecanismos associados tanto à patogênese do DM2 como a suas complicações crônicas. Portanto, o gene UCP2 é um gene candidato para o desenvolvimento dessas doenças. De fato, diversos estudos têm relatado que três polimorfismos comuns no gene UCP2 estão possivelmente associados ao DM2 e/ou à obesidade. Apenas poucos estudos investigaram esses polimorfismos em relação às complicações crônicas do diabetes, obtendo resultados pouco conclusivos.