RESUMO
Nonalcoholic fatty liver disease and diabetes always coexist. The relationship of fatty liver and hyperglycemia is not clear. We studied the influence of hyperglycemia on triglyceride (TG) accumulation in the liver and explored its possible mechanisms. SD rats were divided into three groups: Group A (sham operation control), Group B (partially pancreatectomized rats), and Group C (partially pancreatectomized rats treated with insulin). At 4 weeks after surgery, pancreatic weights and liver TG contents were measured. Serum biochemical parameters were determined, and oral glucose tolerance tests (OGTT) were performed. The gene expression of sterol regulatory element-binding protein1c (SREBP-1c), carbohydrate regulatory element-binding protein (ChREBP), fatty acid synthase(FAS), carnitine palmitoyltransferase 1 (CPT-1), and fibroblast growth factor 21 (FGF21) was determined by real-time PCR. Compared with Group A, postprandial glucose increased significantly; the concentrations of insulin and C-peptides, pancreatic weights and serum FGF21 levels were decreased, liver TG was increased significantly in Group B, and insulin treatment improved these changes. Compared with Group A, the gene expressions of FGF21, CPT-1 and FAS in the liver were decreased in Group B (all p<0.05). Compared with Group B, the gene expressions of FGF21, FAS, ChREBP, SREBP-1c and CPT-1 in the liver in Group C were all increased significantly (p<0.05, respectively). Hyperglycemia induced by partial pancreatectomy could lead to increased liver TG. Insulin treatment could decrease glucose levels and improve fatty liver, and genes related to lipid metabolism may play a role in this process.
Assuntos
Hiperglicemia , Hepatopatia Gordurosa não Alcoólica , Ratos , Animais , Triglicerídeos/metabolismo , Proteína de Ligação a Elemento Regulador de Esterol 1/genética , Proteína de Ligação a Elemento Regulador de Esterol 1/metabolismo , Ratos Sprague-Dawley , Fígado/metabolismo , Hepatopatia Gordurosa não Alcoólica/genética , Metabolismo dos Lipídeos/genética , Insulina/metabolismo , Glucose/metabolismoRESUMO
BACKGROUND: Elevation of exogenous free fatty acid (FFA) level leads to insulin resistance (IR) in liver, IR is manifested by elevated hepatic glucose production. We aim to study whether inhibition of endogenous fatty acid synthesis could decrease hepatic glucose production. METHODS: Low-passage HepG2 cells derived from human liver tissue were cultured in medium supplemented with FFA to induce IR, the influences of sterol regulatory element binding protein-1c (SREBP-1c) silencing on glucose production of HepG2 cells were investigated, and genes responsible for fatty acid and glucose metabolism were detected by real-time PCR. RESULTS: Compared with HepG2 cells cultured in normal growth medium, glucose production of HepG2 cells treated by FFA was significantly increased {[(0.28 ± 0.01) vs (0.83 ± 0.02)] umol.ug- 1 protein, n = 6 wells, P < 0.01}; the mRNA expression of phosphoenolpyruvate carboxylase kinase (PEPCK) and glucose-6-phosphatase (G6PC) in HepG2 cells increased by more than 5-fold and 3-fold, respectively; the mRNA expression of fatty acid synthase (FAS) and stearoyl-CoA desaturase-1 (SCD1) increased by approximately 4-fold and 1.1-fold, respectively; the mRNA expression of carnitine palmitoyltransferase-1 (CPT-1) changed slightly. Compared with the scrambled siRNA control, glucose production of HepG2 cells treated by FFA significantly increased after SREBP-1c silencing {[(0.018 ± 0.001) vs (0.028 ± 0.002)] umol.ug- 1 protein, n = 6 wells, P < 0.01}; the mRNA expression of PEPCK and G6PC increased by approximately 1.5-fold and 5-fold, respectively, but the mRNA expression of FAS, SCD1 and CPT-1 changed slightly. CONCLUSIONS: SREBP-1c silencing further augmented glucose production of HepG2 cells treated by FFA significantly, genes responsible for fatty acid synthesis and gluconeogenesis played an important role in this process. SREBP-1c functions not only as a lipid regulator but also plays an important role in regulation of glucose metabolism.
Assuntos
Meios de Cultura/farmacologia , Ácidos Graxos não Esterificados/farmacologia , Glucose/metabolismo , Proteína de Ligação a Elemento Regulador de Esterol 1/genética , Carnitina O-Palmitoiltransferase/genética , Ácido Graxo Sintases/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Inativação Gênica/efeitos dos fármacos , Células Hep G2 , Humanos , Resistência à Insulina/genética , Metabolismo dos Lipídeos/efeitos dos fármacos , Lipídeos/genética , Proteína de Ligação a Elemento Regulador de Esterol 1/antagonistas & inibidores , Triglicerídeos/metabolismoRESUMO
BACKGROUND: The prevalence of impaired glucose tolerance and diabetes is much higher in people with cirrhosis than that in the general population. However, there are inadequate concrete guidelines for the management of diabetes in these patients, particularly in the early stage. Bile aids (BAs) have been found to exert hormone-like functions in the control of lipid and glucose metabolism. We studied the effect of ursodeoxycholic acid (UDCA) on glucose levels in rats with cirrhosis induced by bile duct ligation (BDL). METHODS: SD rats were divided into three groups: sham operation (Group A); BDL (Group B), and UDCA plus BDL (Group C). After 4 weeks, oral glucose tolerance tests were performed. Serum biochemical parameters and the levels of glucose, insulin, and glucagon-like peptide 1 (GLP-1) were measured. Histopathology of the liver and islet was observed. The gene expression of cholesterol 7α-hydroylase (CYP7A1), microsomal oxysterol 7a-hydroxylase (CYP7B1) in the liver, and Takeda G-protein-coupled receptor-5 (TGR5) in the intestine was determined by real-time PCR. RESULTS: Compared with Group A, fasting glucose and 1-h and 2-h postprandial glucose levels increased slightly (all P > 0.05), 2-h postprandial insulin levels increased significantly (P < 0.05), 15 min postprandial GLP-1 levels decreased (P < 0.05) in Group B. Compared with Group B, fasting glucose and 1-h postprandial glucose levels decreased (all P < 0.05), 2-h postprandial insulin levels decreased (P < 0.01), and 15 min postprandial GLP-1 levels increased (P < 0.05) in Group C. After UDCA intervention, liver fibrosis induced by BDL was alleviated, and the islet areas were increased (P < 0.05). Compared with Group A, the mRNA expression of CYP7A1 and CYP7B1 in the liver increased, and the mRNA expression of TGR5 in the intestine decreased in Group B (all P < 0.05). Compared with Group B, the mRNA expression of CYP7A1 and CYP7B1 in the liver decreased, and TGR5 in the intestine increased in Group C (P < 0.05). CONCLUSIONS: After 4 weeks of BDL, the rats developed liver fibrosis and abnormal glucose metabolism. UDCA administration improved liver fibrosis, increased islet area, decreased glucose levels, inhibited genes in BA synthesis, enhanced TGR5 gene expression in the intestine, and further improved islet function.
RESUMO
BACKGROUND: The relationship between cirrhosis and diabetes is controversial. We studied the influence of cirrhosis on glucose levels and islet function and explored its possible mechanisms. MATERIALS AND METHODS: Cirrhosis was induced in male C57BL/6 mice by bile duct ligation (BDL). Serum biochemical parameters were determined, and oral glucose tolerance tests (OGTT) were performed at 4 and 8 weeks after BDL. Histopathology and phospho-NF-κB-p65/I-kappa B α immunohistochemical staining of the liver and islet were observed. The protein levels of the insulin signaling system and the gene expression of insulin-degrading enzyme (IDE) in the liver and muscle were determined. The activity of glucokinase (GCK) and glucose 6-phosphatase (G6P) and glycogen levels in liver homogenates were measured. RESULTS: After BDL, the mice developed cirrhosis, and fasting glucose decreased significantly, but 2 h postprandial glucose increased, and the insulin areas under the curves increased. At 4 weeks of BDL, the ratios of phospho-NF-κB-p65/I-kappa B α accumulation in the liver and islet increased, the activity of G6P and the glycogen content in liver homogenates decreased, the insulin signaling system and the gene expression of IDE in the liver was downregulated, and the islet areas were decreased. After 8 weeks, these changes were more severe. CONCLUSIONS: In different periods of cirrhosis, the levels of fasting glucose and 2 h postprandial glucose changed in different amplitudes. Glycogen concentrations and the activity of G6P in the liver were decreased. The mice developed insulin resistance and the islet areas were decreased. The NF-κB pathway may play a role in the process.
Assuntos
Glicemia/análise , Resistência à Insulina/fisiologia , Ilhotas Pancreáticas/fisiopatologia , Cirrose Hepática/fisiopatologia , Animais , Ilhotas Pancreáticas/fisiologia , Masculino , Camundongos , Camundongos Endogâmicos C57BLRESUMO
OBJECTIVE: To observe the effects of fenofibrate on the expression of peroxisome proliferator-activated (PPAR)-gamma coactivator-1α (PGC-1α) in skeletal muscle of rats with insulin resistance (IR) induced by elevated plasma free fatty acid (FFA) levels. METHODS: Male Sprague-Dawley (SD) rats were randomly divided into three groups: control group (Con, infused with saline), lipid infusion group (FFA) and fenofibrate treatment plus lipid infusion group (F-FFA). Plasma glucose, insulin and FFA were measured. Glucose infusion rate (GIR) was used to evaluate the insulin sensitivity by euglycemic-hyperinsulinemic clamp. The gene expression of PGC-1α in skeletal muscle was determined by real-time polymerase chain reaction (PCR). RESULTS: Compared with the control group, the levels of plasma FFA and insulin were elevated significantly in rats infused with lipid, but they decreased significantly in rats pretreated with fenofibrate then infused with lipid [FFA, Con: 0.46 (0.08 - 0.72) mmol/L, FFA: 1.45(0.87-1.70) mmol/L, F-FFA: 0.54 (0.06 - 0.84) mmol/L, all P < 0.01; Insulin, Con: (6.56 ± 0.78) µIU/ml, FFA: (10.54 ± 0.86) µIU/ml, F-FFA: (6.69 ± 0.84) µIU/ml, all P < 0.01]. In addition, the plasma glucose levels did not change markedly after lipid infusion; GIR was significantly reduced by 55.6% in lipid infusion group, but fenofibrate-pretreatment increased glucose infusion rate (GIR) [Con: (25.13 ± 2.10) mg×kg(-1)×min(-1), FFA: (10.16 ± 0.75) mg×kg(-1)×min(-1), F-FFA: (21.72 ± 2.89) mg×kg(-1)×min(-1), all P < 0.01]; the mRNA expression of PGC-1α decreased by about 71% in lipid infusion group but fenofibrate increased the gene expression by about 150% as compared with FFA group (all P < 0.01). CONCLUSION: The elevation of plasma FFA levels may induce IR, and it also decreases the mRNA expression of PGC-1α in skeletal muscle. And fenofibrate pretreatment reverses these changes.
Assuntos
Fenofibrato/farmacologia , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/metabolismo , Proteínas de Ligação a RNA/metabolismo , Fatores de Transcrição/metabolismo , Animais , Ácidos Graxos/efeitos adversos , Expressão Gênica/efeitos dos fármacos , Insulina/metabolismo , Resistência à Insulina , Masculino , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo , Proteínas de Ligação a RNA/genética , Ratos , Ratos Sprague-Dawley , Fatores de Transcrição/genéticaRESUMO
BACKGROUND: Panax ginseng is a well-known medicinal herb that is widely used in traditional Chinese medicine for treating various diseases. Ginsenoside Rg3 (Rg3) is thought to be one of the most important active ingredients of Panax ginseng. However, the molecular mechanism underlying the beneficial effects of Rg3 has been elusive. METHODS: In the mouse heart injury model induced by isoproterenol (ISO), we used brain natriuretic peptide (BNP), lactate dehydrogenase (LDH) and caspase-3 ELISA kits to test myocardium injury. To test whether Rg3 protects myocardial injury through AMPK mediated autophagy, we used specific AMPK inhibitor in combination with Rg3. NLRP3 inflammasome related molecules such as NLRP3, ASC and caspase-1 were measured by western-blot following Rg3 treatment. RESULTS: We found that Rg3 significantly reduced ISO induced myocardial injury indicated by the downregulation of serum BNP and LDH. In addition, we showed that the improvement of myocardial injury by Rg3 was associated with enhanced expression of autophagy related protein and activation of AMPK downstream signaling pathway. CONCLUSIONS: We observed that inhibition of AMPK significantly reversed the myocardial protective effect of Rg3, which is associated with a decrease of Rg3 induced autophagy. These together suggested that Rg3 may improve myocardial injury during MI through AMPK mediated autophagy. Our study also provides important translational evidence for using Rg3 in treating myocardial infarction (MI).
RESUMO
OBJECTIVE: To investigate the effect of beta cell lipoapoptosis after long term high-fat feeding in rats, and to investigate the relationship between oxidative stress, gene expression and beta cell lipoapoptosis. METHODS: Forty-one SD male rats were randomly divided into 2 groups: high-fat diet group (HF group) and control group (NC group). At the end of 28 weeks, the levels of malondialdehyde (MDA) and glutamylcysteinylglycine (GSH) in plasma and pancreatic tissue,the early-phase insulin secretion in beta cells, the beta cell apoptosis (TUNEL technology) and the uncoupling protein 2 (UCP2) gene expression in islets were measured. RESULT: The concentrations of MDA both in plasma and pancreatic tissue were higher in HF group than those in NC group.In contrast, The contents of GSH both in plasma and pancreatic tissue were lower in HF group. Insulin secretion response to glucose load was significantly decreased in HF group (3.0 fold Compared with 5.7 fold, P<0.01). Blood glucose levels at 3 min, 5 min and 10 min during IVGTT were significantly higher in HF group than those in NC group (P<0.05). The frequency of beta cell apoptosis was increased by 40.0% in HF group (P<0.01). The gene expression of UCP2 in islets was increased by 22.4% in HF group (P<0.01). CONCLUSION: The frequency of beta cell apoptosis in high-fat feeding rats is affected by oxidative stress, which results in increasing UCP2 gene expression.
Assuntos
Apoptose/fisiologia , Gorduras na Dieta/administração & dosagem , Células Secretoras de Insulina/metabolismo , Malondialdeído/metabolismo , Estresse Oxidativo/fisiologia , Animais , Células Secretoras de Insulina/patologia , Canais Iônicos/genética , Canais Iônicos/metabolismo , Masculino , Proteínas Mitocondriais/genética , Proteínas Mitocondriais/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Proteína Desacopladora 2RESUMO
OBJECTIVE: To study the effects of fenofibrate (FF), a peroxisome proliferator activated receptor (PPAR) alpha activator, on the expression of carnitine palmitoyltransferase 1 (CPT-1) mRNA in liver and muscle and its influence on insulin sensitivity. METHODS: Thirty-two normal 8 week-old male SD rats were randomly divided into 3 groups: normal control group, fed with normal food for 3 weeks (NC group, n = 10), high fat diet group, fed with high fat food (HF group, n = 10), and high fat diet supplemented with FF group, fed with high fat food and given with gastric perfusion of FF (50 mg x kg(-1) x d(-1)) (FF group, n = 12). Fast serum triglyceride (TG) level was tested by automatic biochemical analyzer after 8-10 h fasting. Euglycemic-hyperinsulinemic clamp method was used to calculate the glucose infusion rate (GIR) so as to evaluate the insulin sensitivity. By the end of experiment the rats were killed with their internal organs taken out. The triglyceride (TG) contents of liver and skeletal muscle were measured using Folch method. Real-time PCR was used to detect the mRNA expression. of CPT-1 in the liver and skeletal muscles. RESULTS: As compared with the NC group, the TG levels of serum, liver, and skeletal muscle of the HF group were higher than those of the NC group by 0.45-fold, 2.14-fold, and 10.64-fold respectively. The GIR of the HF group was (6.2 +/- 0.8) mg x kg(-1) x min(-1), significantly lower than that of the NC group (15.8 +/- 2.1) mg x kg(-1) x min(-1), (P < 0.01), and that of the P < 0.01. The CPT-1 mRNA expression in liver of the HF group was not significantly different from that of the NC group (P > 0.05); the expression of CPT-1 mRNA in skeletal muscle of the HF group was lower than that of the NC group by 71% (P < 0.01). The CPT-1 mRNA expression in liver and skeletal muscle of the FF group were significantly higher than those of the HF group by 1.00 and 1.05 times respectively (both P < 05). The GIR was negatively correlated with the levels in the liver (r = -0.87, P < 0.01) and in the skeletal muscle (r = -0.78, P < 0.01). CONCLUSION: Fenofibrate promotes the oxygenation of fatty acids by up-regulating the CPT-1 mRNA expression in the liver and skeletal muscles, thus improving the insulin sensitivity.
Assuntos
Carnitina O-Palmitoiltransferase/genética , Fenofibrato/farmacologia , Resistência à Insulina , Fígado/efeitos dos fármacos , Músculo Esquelético/efeitos dos fármacos , Animais , Glicemia/metabolismo , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Hipoglicemiantes/farmacologia , Hipolipemiantes/farmacologia , Insulina/farmacologia , Fígado/metabolismo , Masculino , Músculo Esquelético/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
OBJECTIVE: To study the effects of high fat diet on the functions of islet beta cells and the role of uncoupling protein-2 (UCP2) therein and possible mechanism. METHODS: Forty SD rats were randomly divided into two equal groups: high-fat-(HF) diet group, fed with HF diet for 20 weeks, and normal diet control (NC) group, fed with normal diet. At the end of the twentieth week blood samples were collected from the heart to determine the serum fasting blood glucose (FBG) and fasting insulin (FINS), and plasma nitrotyrosine, malondialdehyde (MDA), and glutamylcysteinylglycine (GSH), indicators of oxidative stress. Glucose infusion rate (GIR) was measured using euglycemic hyperinsulinemic clamp test to evaluate the peripheral insulin resistance. Pancreatic islets were isolated and collected. Islet perfusion was conducted to evaluate the insulin secretion in the islet beta cells. Real-time PCR was used to detect the expression of insulin receptor substrate-1 (IRS-1), IRS-2, and uncoupling protein 2 (UCP2) genes in the islet. Immunohistochemistry was used to detect the protein expression of IRS-1 and IRS-2. RESULTS: (1) The concentrations of plasma nitrotyrosine and MDA of the HF group were both significantly higher than those of the NC group (both P < 0.05). However, the plasma GSH of the HF group was significantly lower than that of the NC group (P < 0.01). (2) The blood glucose of both groups became stable since 60 min after the experiment and the GIR of the HF group was (5.25 +/- 1.2) mg x min(-1) x kg(-1), significantly lower r than that of the NC group [(13.6 +/- 1l.7) mg x min(-1) x kg(-1), P < 0.01). (3) The peak of glucose-stimulated insulin secretion (GSIS) of the HF group was significantly lower than that of the NC group; and the GSIS peak increase In comparison with the NC group. (4) In comparison with the NC group, the mRNA expression levels of IRS-1 and IRS-2 genes of the HF group were significantly lower, by 42.3% and 28.1% respectively (both P < 0.05), and the expression of UCP2 was significantly higher, by 32.5% (P < 0.05). (5) Compared with the NC group, the protein expression levels of IRS-1 and IRS-2 in the islets of the HF group were lower, by 26.3% and 11.2% respectively, however not significantly (both P > 0.05). (6) There was a significantly negative correlation between the UCP2 and IRS-1/IRS-2 gene expression in islet beta cells in the HF group (r = -0.621 and r = -0.436, both P < 0.05). CONCLUSION: High-fat-diet impairs the expression of insulin signal transduction molecules and the function of islet beta cells that may be correlated with overexpression of UCP2. The basic insulin secretion of HF group was significantly higher than that of the NC group; but the glucose-stimulated insulin secretion (GSIS) peak decreased in comparison with the NC group. Compared with the NC group, the protein expression levels of IRS-1 and IRS-2 in the islets of the HF group were lower, by 26.3% (P < 0.05) and 11.2% (P > 0.05) respectively.
Assuntos
Insulina/metabolismo , Canais Iônicos/genética , Ilhotas Pancreáticas/metabolismo , Proteínas Mitocondriais/genética , Animais , Células Cultivadas , Gorduras na Dieta/administração & dosagem , Expressão Gênica , Glutationa/sangue , Imuno-Histoquímica , Proteínas Substratos do Receptor de Insulina/genética , Proteínas Substratos do Receptor de Insulina/metabolismo , Ilhotas Pancreáticas/citologia , Masculino , Malondialdeído/sangue , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Tirosina/análogos & derivados , Tirosina/sangue , Proteína Desacopladora 2RESUMO
OBJECTIVE: To observe the relationship between changes of genes expression related to lipid metabolism and insulin resistance induced by high fat diet in SD rats. METHODS: Normal 8-week old male SD rats were randomly divided into 3 groups. They were fed with normal chow (NC, n = 10), high fat diet (HF, n = 10) and high fat diet supplemented with pioglitazone 15 mg x kg(-1) x d(-1) (HP, n = 12). The TG content of liver and skeletal muscle of the rats was measured. Glucose infusion rate (GIR) was used to evaluate the insulin sensitivity by using euglycemic-hyperinsulinemic clamp method. Genes expression was investigated using real-time PCR method. RESULTS: After high fat feeding for 20 weeks, the serum TG, the content of TG in the liver and skeletal muscle of the rats in the HF group increased 0.45, 2.28 and 9.31 fold respectively as compared with those in the NC group. The change of TG content in the liver and skeletal muscle was associated with the reduction of GIR [(6.16 +/- 0.75) mg x kg(-1) x min(-1) vs (15.82 +/- 2.10) mg x kg(-1).min(-1) (P < 0.01)]. As compared with the NC group, the expression of hormone-sensitive lipase (HSL) and fatty-acid synthase (FAS) gene in the HF group was enhanced by 28.2% and 21.3%, respectively (P < 0.05). In addition, the expression of acetyl-CoA carboxylase1 (ACC1) mRNA in the liver increased 48.3% (P < 0.05), increased 101.1% (P < 0.01) and carnitine palmitoyltransferase 1 (CPT-1) decreased 71.0% (P < 0.01) in the skeletal muscle in the rats of HF group. As compared with those in the HF group, GIR increased 1.54 fold in the HP group and on the contrary, serum TG, liver TG and muscle TG decreased about 66%, 64.5% and 59.6% respectively in the HP group (P < 0.05). Accordingly, the expression of FAS and HSL in the adipose tissue and the expression of ACC1 in the liver were reduced (P < 0.05) and the expression of CPT-1 was enhanced and ACC2 was reduced in the muscle (P < 0.01) in the HP group. CONCLUSIONS: The changing expression of genes related to lipid metabolism may play a role in the accumulation of lipids in non-adipose tissue and the induction of insulin resistance in rats fed with high fat diet.
Assuntos
Perfilação da Expressão Gênica , Resistência à Insulina , Metabolismo dos Lipídeos/genética , Acetiltransferases/genética , Animais , Glicemia/metabolismo , Carnitina O-Palmitoiltransferase/genética , Gorduras na Dieta/administração & dosagem , Regulação da Expressão Gênica/efeitos dos fármacos , Hipoglicemiantes/farmacologia , Fígado/efeitos dos fármacos , Fígado/metabolismo , Masculino , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/metabolismo , Pioglitazona , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Tiazolidinedionas/farmacologiaRESUMO
OBJECTIVE: To study the changes of inflammatory path molecules in the islet alpha cells in high-fat-diet fed plus beta cell-deleted rat models and the effects of pioglitazone intervention. METHODS: Forty five normal male SD rats, 8 week old, were randomly divided into 3 groups, i.e., a normal diet group (NC), a high fat diet fed group (HF), and a high fat diet fed and pioglitazone treated group (HP, pioglitazone 15 mg kg(-1) d(-1)). At the end of twenty weeks of feeding, fasting serum insulin (FIns), glucagon, free fatty acid (FFA) and high sensitive C reactive protein (hsCRP) were determined. Glucose infusion rate (GIR) was measured by using euglycemic hyperinsulinemia clamp to evaluate the peripheral insulin resistance. The contents of glucagon in perfusion medium during islet cell perfusion was measured with RIA. At the same time, beta cell-deleted rat models were established by injecting large dose streptozocin (100 mg/kg) in 8 rats in each of the three groups, i.e., HF-B group, P-B group and NC-B group. Five days later, the rats were sacrificed and the pancreatic islets were isolated and collected. The expression of NF-kappaB and inhibitor kappaBalpha (IkappaBalpha) gene in the islets was detected with real-time PCR. RESULTS: (1) GIR was decreased significantly in HF group as compared with NC group (P < 0.01). The concentrations of serum FIns, glucagon, FFA and hsCRP in HF group were higher than those in NC group. Pioglitazone intervention could reverse these effects. (2) 16.7 mmol/L glucose could inhibit the glucagon secretion by the islet alpha cells of the NC group rats, but not of the HF group rats. Pioglitazone intervention could reverse these effects. (3) The gene expression of NF-kappaB was significantly increased by 20.5% in the HF-B group than in the NC-B group (P < 0.01). In contrast, the expression of IkappaBalpha was significantly decreased by 24.3% (P < 0.01). The expression of NF-kappaB and IkappaBalpha mRNAs in HP-B group, when compared with that of HF-B group, was improved 78.3% and 58.8%, respectively. CONCLUSIONS: High-fat-diet feeding induces islet alpha cell insulin resistance and activates the mRNA expression of inflammatory path molecules in beta cell-deleted rat models and it may relate with the increased plasma FFA concentration. Pioglitazone intervention can reverse these effects.
Assuntos
Inflamação/fisiopatologia , Resistência à Insulina , Ilhotas Pancreáticas/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Tiazolidinedionas/farmacologia , Animais , Ácidos Graxos não Esterificados/sangue , Glucagon/sangue , Hipoglicemiantes/farmacologia , Inflamação/sangue , Insulina/sangue , Ilhotas Pancreáticas/patologia , Ilhotas Pancreáticas/fisiopatologia , Masculino , Pioglitazona , Distribuição Aleatória , Ratos , Ratos Sprague-DawleyRESUMO
OBJECTIVE: To study the changes and mechanism of the function of islet beta cells and insulin signal transduction molecules after lipid infusion. METHODS: Twenty five SD rats were randomly divided into 2 groups, FFA group and NS group. Catheters were implanted under pentobarbital anesthesia in the right atrium via the jugular vein and the left carotid artery. A technique for a 48 h infusion in unrestrained rats was used for triglyceride and heparin or saline infusion. The infusion period started on day 2 after surgery. After 48 h infusion, we determined fasting serum insulin (Ins), free fat acid (FFA) in the blood. The glucose infusion rate (GIR) was measured by hyperinsulinemia euglycemic clamp to evaluate the peripheral insulin resistance. The ivgtt and islet cell perifusion was conducted to evaluate the function of islet beta cells. The rats in the two groups were sacrificed, and the pancreatic islets were isolated and collected. The expressions of insulin receptor substrate-1(IRS-1), insulin receptor substrate-2 (IRS-2) glucose transporter-2 (Glut-2) gene in islets and IRS-1, IRS-2 in muscle were detected by real-time PCR. RESULTS: (1) The serum FFA and insulin concentrations of blood in FFA group were higher than in NS group (P<0.05). (2) The GIR was decreased significantly in FFA group compared with that in NS group (P<0.05). (3) The glucose stimulated insulin secretion increased in the FFA group. (4) The gene expression of IRS-1 in muscle was significantly decreased by 87.7% in FFA group, and the expression of IRS-2 was decreased by 50.7% (all P<0.05). The gene expression of IRS-1 in islets was significantly increased by 29.3% (P<0.05), and the expressions of IRS-2, Glut-2 were increased by 345.1% and 536.4% respectively, in FFA group (all P<0.01). CONCLUSION: Lipid infusion in short time increased the secretion of insulin and impaired expression of insulin signal transduction molecules in muscle but it also increased the expression of insulin signal transduction molecules in islet beta cells.
Assuntos
Ácidos Graxos não Esterificados/farmacologia , Insulina/sangue , Ilhotas Pancreáticas/efeitos dos fármacos , Ilhotas Pancreáticas/metabolismo , Transdução de Sinais/efeitos dos fármacos , Animais , Ácidos Graxos não Esterificados/sangue , Resistência à Insulina , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
OBJECTIVE: To study the changes of insulin signal transduction molecules in islet alpha cells in high-fat-diet plus beta cell-deleting rat models and its underlying mechanism. METHODS: Thirty SD rats were randomly divided into 2 equal groups and fed with high-fat-diet (HF group) or normal diet (normal control group, NC group) respectively. At the end of twenty-week feeding, the fasting serum insulin (Ins), glucagon (Glc), free fatty acid (FFA), and triglyceride (TG were measured. The glucose infusion rate (GIR) was measured by using euglycemic hyperinsulinemia clamp to evaluate the peripheral insulin resistance. At the same time, large dose streptozocin (100 mg/kg) was injected so as to establish beta cell-deleting rat models, i.e., HF-B group (n = 8) and NC-B group (n = 8). Five days later, the rats of the HF-B and NC-B subgroups were sacrificed, and the pancreatic islets were isolated and collected. The expression of Glc, insulin receptor substrate-1 (IRS-1), IRS-2, and phosphatidylinositol-3-kinase (PI3K) gene in the islets were detected by RT-PCR. RESULTS: (1) The serum FFA, insulin and Glc concentrations of the HF group were 508 (394 - 622) micromol/L, 23.7 (14.0 - 33.4) mIU/L, and 345 (298.6 - 391.4) pg/ml respectively, all significantly higher than those of the NC group [325 (240 - 410) micromol/L, 11.5 (3.6 - 19.4) mIU/L, 256 (226.4 - 285.6) pg/ml; respectively, all P < 0.05]. The GIR of the HF group was 5.25 mgxmin(-1)xkg(-1) +/- 1.2 mgxmin(-1)xkg(-1), significantly lower than that of the NC group (13.6 mgxmin(-1)xkg(-1) +/- 1.7 mgxmin(-1)xkg(-1), P < 0.01)). (2) The gene expression of Glc of the HF-B subgroup was significantly higher than that of the NC-B subgroup by 34.2% +/- 2.1%. In contrast, the expression of IRS-2, and PI3K of the HF-B subgroup was significantly lower than that of the NC-B subgroup by 28.5% +/- 1.8% and 21.3% +/- 1.6% respectively (both P < 0.01). (3) The plasma FFA concentration was asignificantly negatively correlated with GIR (r = -0.675, P < 0.05) and IRS-2 gene expression in islet alpha cells (r = -0.458, P < 0.05) in the HF-B group. CONCLUSION: High-fat-diet feeding plus beta cell-deleting rat model shows an impaired expression of insulin signal transduction molecules in islet alpha cells which may relate with the increased plasma FFA concentration.
Assuntos
Resistência à Insulina/genética , Insulina/metabolismo , Ilhotas Pancreáticas/metabolismo , Transdução de Sinais/genética , Proteínas Adaptadoras de Transdução de Sinal/genética , Animais , Glicemia/metabolismo , Ácidos Graxos não Esterificados/administração & dosagem , Ácidos Graxos não Esterificados/sangue , Expressão Gênica , Glucagon/sangue , Técnica Clamp de Glucose , Insulina/sangue , Proteínas Substratos do Receptor de Insulina , Masculino , Fosfatidilinositol 3-Quinases/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Receptor de Insulina/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Triglicerídeos/sangueRESUMO
OBJECTIVES: CST has been recently identified as a mediator of various beneficial effects in animal models of sepsis. At present, no data are available concerning the levels of CST in sepsis patients. In sepsis the plasma amino acid pattern is characterized by decreased branced chain amino acids (BCAAs). We investigated the levels of plasma CST or branched-chain α-ketoacid dehydrogenase kinase (BCKDK) and their relationship to component traits in patients with sepsis. DESIGN AND METHODS: We studied 228 patients and divided them into two groups based on severity of infection. Blood samples were taken at study entry, and CST, BCKDK were measured. RESULTS: CST and BCKDK levels were significantly higher in patients with sepsis than in controls: the median plasma CST concentration was 103.1ng/ml (range, <83.13-189.7ng/ml) in patients with sepsis and 49.69ng/ml (range, <19.38-100.8ng/ml) in controls (p=0.0022); the median plasma BCKDK concentration was 801.7ng/ml in sepsis group and 745ng/ml in controls (p=0.0292). Additionally, there was correlation between the plasma concentrations of CST and BCKDK in sepsis patients (r2=0.6357, p<0.01). CONCLUSIONS: We conclude that the plasma levels of CST in sepsis patients were higher than in controls, and there is a relationship between CST and BCKDK in sepsis patients. Future experimental and clinical studies are needed to evaluate CST as a novel prognostic tool in sepsis patients and its potential therapeutic use in sepsis.
Assuntos
Neuropeptídeos/sangue , Proteínas Quinases/sangue , Sepse/sangue , Adulto , Biomarcadores/sangue , Feminino , Humanos , Ácido Láctico/sangue , Masculino , Pessoa de Meia-IdadeRESUMO
BACKGROUND: Currently it is unclear whether lipid accumulation occurs in a particular sequence and its relationship with whole body insulin resistance (IR). This study aimed to answer this question. METHODS: Male Sprague-Dawley (SD) rats were fed on a normal or a high-fat diet for 20 weeks. Serum triglycerides (TG), serum free fatty acids (FFA), fasting plasma glucose (FPG), and liver and skeletal muscle TG were measured. The glucose infusion rate (GIR) and mRNA levels of acetyl-CoA carboxylase (ACC) and carnitine palmitoyltransferase-1 (CPT-1) in the liver and skeletal muscle were determined at different stages. RESULTS: Compared with rats fed on the normal diet, serum FFA was not significantly increased in rats fed on the high-fat diet until 20 weeks. In contrast, liver TG was significantly increased by the high-fat diet by four weeks (20-fold; P < 0.01), and remained elevated until the end of the study. However, skeletal muscle TG was not significantly increased by the high-fat diet until 20 weeks (10.6-fold; P < 0.01), and neither was the FPG. The GIR was significantly reduced (1.6-fold; P < 0.01) by the high-fat diet after 8 weeks. The mRNA levels of ACC gradually increased over time and CPT-1 decreased over time, in both the liver and skeletal muscle in rats fed the high-fat diet. CONCLUSIONS: Lipid accumulation in the liver occurs earlier than lipid accumulation in the skeletal muscle. Fatty liver may be one of the early markers of whole body IR. Changes in the gene expression levels of ACC and CPT-1 may have important roles in the process of IR development.