RESUMO
OBJECTIVE: To observe the effects of different doses of propofol on the growth of transplanted liver tumor in BALB/C mice and check the expression of PCNA, CD34 and pAKT proteins to clarify the mechanism on molecule level. METHOD: Human primary liver cancer cells SMMC-7721 were subcutaneously cultured in BALB/C mice, and the transplanted tumor model of BALB/C mice was constructed. Forty mice successfully modeled were randomly divided into 5 groups (n = 8): the blank control group (group C), low-fat milk group (group I), low-dose (50 mg/kg) propofol group (P1), middle-dose (100 mg/kg) propofol group (P2) and high dose (150 mg/kg) propofol group (P3). Tumor volume changes were observed at 3, 6, 9, 12, 15 and 18 days (T1, T2, T3, T4, T5, T6 and T7) before and after administration of the drug, and tumor growth curves were plotted. After 19 days of administration, all mice were killed for tumor collection, tumor weight was measured, and the tumor inhibition rate of propofol was calculated. The protein expression of cluster of differentiation 34 (CD34) in transplanted tumor was detected by immunohistochemistry, and the protein expression of proliferating cell nuclear antigen (PCNA) and phospho-Akt (pAKT) was detected by immunofluorescence. RESULTS: Compared with group C, there was no significant difference in tumor volume in group I. At T2 ~ 7, the tumor volume of group P1, P2 and P3 decreased successively (P < 0.05). There was no significant difference in the inhibitory rate of tumor in group I, and the inhibitory rate of tumor in group P1, P2 and P3 increased successively (P < 0.05). There was no significant difference in PCNA, CD34, and pAKT protein expression in group I, while PCNA, CD34, and pAKT protein content in P1, P2, P3 groups were successively decreased (P < 0.05). CONCLUSION: Propofol had a dose-dependent effect on the growth of liver cancer xenografts in mice, inhibiting the expression of PCNA, CD34 and pAKT proteins, and the effect was most obvious in the 150 mg/kg propofol group.
Assuntos
Antígenos CD34/análise , Neoplasias Hepáticas Experimentais/tratamento farmacológico , Antígeno Nuclear de Célula em Proliferação/análise , Propofol/uso terapêutico , Proteínas Proto-Oncogênicas c-akt/análise , Animais , Linhagem Celular Tumoral , Relação Dose-Resposta a Droga , Humanos , Neoplasias Hepáticas Experimentais/patologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Propofol/farmacologia , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
BACKGROUND/AIMS: Skeletal mass loss is reported in several catabolic conditions and it has been associated with a reduced intracellular L-glutamine content. We investigated the association of intracellular L-glutamine concentration with the protein content in skeletal muscle cells. METHODS: We cultivated C2C12 myotubes in the absence or presence of 2 (reference condition), 8 or 16 mM L-glutamine for 48 hours, and the variations in the contents of amino acids and proteins measured. We used an inhibitor of L-glutamine synthesis (L-methionine sulfoximine - MSO) to promote a further reduction in intracellular L-glutamine levels. Amino acids contents in cells and media were measured using LC-MS/MS. We measured changes in phosphorylated Akt, RP-S6, and 4E-BP1contents in the absence or presence of insulin by western blotting. RESULTS: Reduced intracellular L-glutamine concentration was associated with decreased protein content and increased protein breakdown. Low intracellular glutamine levels were also associated with decreased p-Akt contents in the presence of insulin. A further decrease in intracellular L-glutamine caused by glutamine synthetase inhibitor reduced protein content and levels of amino acids generated from glutamine metabolism and increased bAib still further. Cells exposed to high medium glutamine levels did not have any change in protein content but exhibited increased contents of the amino acids derived from L-glutamine metabolism. CONCLUSION: Intracellular L-glutamine levels per se play a role in the control of protein content in skeletal muscle myotubes.
Assuntos
Proteínas de Transporte/metabolismo , Glutamina/metabolismo , Insulina/metabolismo , Fibras Musculares Esqueléticas/metabolismo , Fosfoproteínas/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteína S6 Ribossômica/metabolismo , Proteínas Adaptadoras de Transdução de Sinal , Animais , Proteínas de Transporte/análise , Proteínas de Ciclo Celular , Linhagem Celular , Cromatografia Líquida , Fatores de Iniciação em Eucariotos , Glutamina/análise , Insulina/análise , Camundongos , Fibras Musculares Esqueléticas/química , Fosfoproteínas/análise , Fosforilação , Proteínas Proto-Oncogênicas c-akt/análise , Proteína S6 Ribossômica/análise , Espectrometria de Massas em TandemRESUMO
Pituitary adenoma is one of the most common tumors in the neuroendocrine system. This study investigated the effects of long non-coding RNAs (lncRNAs) highly up-regulated in liver cancer (HULC) on rat secreting pituitary adenoma GH3 cell viability, migration, invasion, apoptosis, and hormone secretion, as well as the underlying potential mechanisms. Cell transfection and qRT-PCR were used to change and measure the expression levels of HULC, miR-130b, and FOXM1. Cell viability, migration, invasion, and apoptosis were assessed using trypan blue staining assay, MTT assay, two-chamber transwell assay, Guava Nexin assay, and western blotting. The concentrations of prolactin (PRL) and growth hormone (GH) in culture supernatant of GH3 cells were assessed using ELISA. The targeting relationship between miR-130b and FOXM1 was verified using dual luciferase activity. Finally, the expression levels of key factors involved in PI3K/AKT/mTOR and JAK1/STAT3 pathways were evaluated using western blotting. We found that HULC was highly expressed in GH3 cells. Overexpression of HULC promoted GH3 cell viability, migration, invasion, PRL and GH secretion, as well as activated PI3K/AKT/mTOR and JAK1/STAT3 pathways. Knockdown of HULC had opposite effects and induced cell apoptosis. HULC negatively regulated the expression of miR-130b, and miR-130b participated in the effects of HULC on GH3 cells. FOXM1 was a target gene of miR-130b, which was involved in the regulation of GH3 cell viability, migration, invasion, and apoptosis, as well as PI3K/AKT/mTOR and JAK1/STAT3 pathways. In conclusion, HULC tumor-promoting roles in secreting pituitary adenoma might be via down-regulating miR-130b, up-regulating FOXM1, and activating PI3K/AKT/mTOR and JAK1/STAT3 pathways.
Assuntos
Adenoma/patologia , Neoplasias Hipofisárias/patologia , RNA Longo não Codificante/fisiologia , Adenoma/genética , Adenoma/metabolismo , Animais , Apoptose/fisiologia , Western Blotting , Linhagem Celular Tumoral , Ensaios de Migração Celular , Movimento Celular/fisiologia , Sobrevivência Celular/fisiologia , Ensaio de Imunoadsorção Enzimática , Proteína Forkhead Box M1/análise , Proteína Forkhead Box M1/metabolismo , Humanos , Janus Quinase 1/análise , Janus Quinase 1/metabolismo , Luciferases , MicroRNAs/análise , MicroRNAs/metabolismo , Fosfatidilinositol 3-Quinases/análise , Fosfatidilinositol 3-Quinases/metabolismo , Neoplasias Hipofisárias/genética , Neoplasias Hipofisárias/metabolismo , Proteínas Proto-Oncogênicas c-akt/análise , Proteínas Proto-Oncogênicas c-akt/metabolismo , RNA Longo não Codificante/análise , Ratos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fator de Transcrição STAT3/análise , Fator de Transcrição STAT3/metabolismo , Serina-Treonina Quinases TOR/análise , Serina-Treonina Quinases TOR/metabolismo , TransfecçãoRESUMO
Ginsenoside Rg1, one of the most notable active components of Panax ginseng, has been widely reported to exert anti-inflammatory actions. This study aimed to reveal whether ginsenoside Rg1 also exhibits beneficial roles against lipopolysaccharide (LPS)-induced apoptosis and inflammation in human renal tubular epithelial cells, and to evaluate the potential role of the component on tubulointerstitial nephritis treatment. HK-2 cells were treated with various doses of ginsenoside Rg1 (0, 50, 100, 150, and 200 µM) in the absence or presence of 5 µg/mL LPS. Thereafter, CCK-8 assay, flow cytometry, western blot, migration assay, reactive oxygen species (ROS) assay, and ELISA were carried out to respectively assess cell viability, apoptosis, migration, ROS activity, and the release of inflammatory cytokines. As a result, ginsenoside Rg1 protected HK-2 cells from LPS-induced injury, as cell viability was increased, cell apoptosis was decreased, and the release of MCP-1, IL-1ß, IL-6, and TNF-α was reduced. Ginsenoside Rg1 functioned to HK-2 cells in a dose-dependent manner, and the 150 µM dose exhibited the most protective functions. Ginsenoside Rg1 had no significant impact on cell migration and ROS activity, while it alleviated LPS-induced ROS release and migration impairment. Furthermore, the down-regulations of p-PI3K, p-AKT, and up-regulations of PTEN, p-IκBα, p-p65, Bcl-3 induced by LPS were recovered to some extent after ginsenoside Rg1 treatment. In conclusion, ginsenoside Rg1 protects HK-2 cells against LPS-induced inflammation and apoptosis via activation of the PI3K/AKT pathway and suppression of NF-κB pathway.
Assuntos
Anti-Inflamatórios/farmacologia , Apoptose/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Ginsenosídeos/farmacologia , Túbulos Renais/citologia , Lipopolissacarídeos , Nefrite/prevenção & controle , Análise de Variância , Western Blotting , Linhagem Celular , Ensaios de Migração Celular , Sobrevivência Celular/efeitos dos fármacos , Citocinas/análise , Citocinas/efeitos dos fármacos , Ensaio de Imunoadsorção Enzimática , Humanos , Túbulos Renais/efeitos dos fármacos , Fosfatidilinositol 3-Quinases/análise , Fosfatidilinositol 3-Quinases/efeitos dos fármacos , Substâncias Protetoras/farmacologia , Proteínas Proto-Oncogênicas c-akt/análise , Proteínas Proto-Oncogênicas c-akt/efeitos dos fármacos , Espécies Reativas de Oxigênio/análise , Reprodutibilidade dos TestesRESUMO
Cetuximab is widely used in patients with metastatic colon cancer expressing wildtype KRAS. However, acquired drug resistance limits its clinical efficacy. Exosomes are nanosized vesicles secreted by various cell types. Tumor cell-derived exosomes participate in many biological processes, including tumor invasion, metastasis, and drug resistance. In this study, exosomes derived from cetuximab-resistant RKO colon cancer cells induced cetuximab resistance in cetuximab-sensitive Caco-2 cells. Meanwhile, exosomes from RKO and Caco-2 cells showed different levels of phosphatase and tensin homolog (PTEN) and phosphor-Akt. Furthermore, reduced PTEN and increased phosphorylated Akt levels were found in Caco-2 cells after exposure to RKO cell-derived exosomes. Moreover, an Akt inhibitor prevented RKO cell-derived exosome-induced drug resistance in Caco-2 cells. These findings provide novel evidence that exosomes derived from cetuximab-resistant cells could induce cetuximab resistance in cetuximab-sensitive cells, by downregulating PTEN and increasing phosphorylated Akt levels.
Assuntos
Antineoplásicos Imunológicos/farmacologia , Cetuximab/farmacologia , Neoplasias do Colo/tratamento farmacológico , Exossomos/efeitos dos fármacos , PTEN Fosfo-Hidrolase/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-akt/efeitos dos fármacos , Análise de Variância , Western Blotting , Células CACO-2 , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Neoplasias do Colo/metabolismo , Resistência a Medicamentos , Exossomos/metabolismo , Formazans , Humanos , PTEN Fosfo-Hidrolase/análise , PTEN Fosfo-Hidrolase/metabolismo , Proteínas Proto-Oncogênicas c-akt/análise , Proteínas Proto-Oncogênicas c-akt/metabolismo , Sais de Tetrazólio , Fatores de TempoRESUMO
OBJECTIVES: To evaluate the expression of p-AKT, p-JNK, FoxO3a, and Ki-67 in samples of oral squamous cell carcinoma (OSCC) and oral epithelial dysplasias (OEDs) to understand their possible involvement in the malignant transformation process of oral lesions. MATERIALS AND METHODS: Tissue samples of 20 cases of OSCCs, 20 OEDs, and normal oral mucosa were subjected to immunohistochemistry reactions for anti-p-AKT, anti-p-JNK, anti-FoxO3a, and anti-Ki-67 antibodies. It was analyzed using quantitative (number of immunostained cells) and qualitative (immunostaining intensity) parameters in different cell immunostaining sublocations. RESULTS: Nuclear p-AKT was observed significantly greater immunostaining in OSCC (21.2 ± 19.0) than in dysplasias (7.9 ± 8.1) and controls (1.8 ± 4.7) (P = 0.002). Immunostaining of strong nuclear p-JNK was greater in controls (48.3 ± 13.7) than in OEDs (11.0 ± 10.3) and OSCCs (1.1 ± 1.3) (P < 0.001). Strong nuclear immunostaining of FoxO3a proved to be absent in OSCCs (0.0 ± 0.1) with little staining on dysplasias (3.2 ± 5.4) and increased expression in controls (13.5 ± 4.8) (P < 0.001). Immunostaining of strong nuclear Ki-67 was grater in OSCCs (48.1 ± 49.6) than in OED (11.8 ± 10.6) and controls (1.9 ± 2.0) (P < 0.001). CONCLUSIONS: Malignant process of OEDs in this research may involve the same mechanisms of established malignant lesions.
Assuntos
Carcinoma de Células Escamosas/química , Proteína Forkhead Box O3/análise , Proteínas Quinases JNK Ativadas por Mitógeno/análise , Mucosa Bucal/química , Neoplasias Bucais/química , Proteínas Proto-Oncogênicas c-akt/análise , Estudos Transversais , Humanos , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Antígeno Ki-67/análise , Mucosa Bucal/patologia , Fosforilação , Proteínas Proto-Oncogênicas c-akt/metabolismoRESUMO
ABSTRACT The present study describes the histopathological and molecular effects of P-MAPA (Protein aggregate magnesium-ammonium phospholinoleate-palmitoleate anhydride) intravesical immunotherapy combined with systemic doxorubicin or cisplatin for treatment of non-muscle invasive bladder cancer (NMIBC) in an appropriate animal model. Our results showed an undifferentiated tumor, characterizing a tumor invading mucosa or submucosa of the bladder wall (pT1) and papillary carcinoma in situ (pTa) in the Cancer group. The histopathological changes were similar between the combined treatment with intravesical P-MAPA plus systemic Cisplatin and P-MAPA immunotherapy alone, showing decrease of urothelial neoplastic lesions progression and histopathological recovery in 80% of the animals. The animals treated systemically with cisplatin or doxorubicin singly, showed 100% of malignant lesions in the urinary bladder. Furthemore, the combined treatment with P-MAPA and Doxorubicin showed no decrease of urothelial neoplastic lesions progression and histopathological recovery. Furthermore, Akt, PI3K, NF-kB and VEGF protein levels were significantly lower in intravesical P-MAPA plus systemic cisplatin and in intravesical P-MAPA alone treatments than other groups. In contrast, PTEN protein levels were significantly higher in intravesical P-MAPA plus systemic cisplatin and in intravesical P-MAPA alone treatments. Thus, it could be concluded that combination of intravesical P-MAPA immunotherapy and systemic cisplatin in the NMIBC animal model was effective, well tolerated and showed no apparent signs of antagonism between the drugs. In addition, intravesical P-MAPA immunotherapy may be considered as a valuable option for treatment of BCG unresponsive patients that unmet the criteria for early cystectomy.
Assuntos
Animais , Feminino , Neoplasias da Bexiga Urinária/terapia , Carcinoma/terapia , Doxorrubicina/uso terapêutico , Cisplatino/uso terapêutico , Imunoterapia/métodos , Proteínas de Membrana/uso terapêutico , Antineoplásicos/uso terapêutico , Ratos Endogâmicos F344 , Neoplasias da Bexiga Urinária/patologia , Administração Intravesical , Vacina BCG , Carcinoma/patologia , Western Blotting , Reprodutibilidade dos Testes , NF-kappa B/análise , Resultado do Tratamento , Terapia Combinada , Progressão da Doença , Fosfatidilinositol 3-Quinases/análise , Modelos Animais , Fator A de Crescimento do Endotélio Vascular/análise , PTEN Fosfo-Hidrolase/análise , Proteínas Proto-Oncogênicas c-akt/análiseRESUMO
Quercetin shows protective effects against hepatopulmonary syndrome (HPS), as demonstrated in a rat model. However, whether these effects involve pulmonary vascular angiogenesis in HPS remains unclear. Therefore, this study aimed to assess the effect of quercetin on pulmonary vascular angiogenesis and explore the underlying mechanisms. Male Sprague-Dawley rats weighing 200-250 g underwent sham operation or common bile duct ligation (CBDL). Two weeks after surgery, HIF-1α and NFκB levels were assessed in rat lung tissue by immunohistochemistry and western blot. Then, CBDL and sham-operated rats were further divided into 2 subgroups each to receive intraperitoneal administration of quercetin (50 mg/kg daily) or 0.2% Tween for two weeks: Sham (Sham+Tween; n=8), CBDL (CBDL+Tween; n=8), Q (Sham+quercetin; n=8), and CBDL+Q (CBDL+quercetin; n=8). After treatment, lung tissue specimens were assessed for protein (immunohistochemistry and western blot) and/or gene expression (quantitative real-time PCR) levels of relevant disease markers, including VEGFA, VEGFR2, Akt/p-Akt, HIF-1α, vWf, and IκB/p-IκB. Finally, arterial blood was analyzed for alveolar arterial oxygen pressure gradient (AaPO2). Two weeks after CBDL, HIF-1α expression in the lung decreased, but was gradually restored at four weeks. Treatment with quercetin did not significantly alter HIF-1α levels, but did reduce AaPO2 as well as lung tissue NF-κB activity, VEGFA gene and protein levels, Akt activity, and angiogenesis. Although hypoxia is an important feature in HPS, our findings suggest that HIF-1α was not the main cause for the VEGFA increase. Interestingly, quercetin inhibited pulmonary vascular angiogenesis in rats with HPS, with involvement of Akt/NF-κB and VEGFA/VEGFR-2 pathways.
Assuntos
Antioxidantes/farmacologia , Síndrome Hepatopulmonar/tratamento farmacológico , Pulmão/irrigação sanguínea , Neovascularização Patológica/tratamento farmacológico , Quercetina/farmacologia , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos/análise , Western Blotting , Ducto Colédoco/cirurgia , Modelos Animais de Doenças , Síndrome Hepatopulmonar/patologia , Imuno-Histoquímica , Ligadura , Pulmão/patologia , Masculino , NF-kappa B/análise , Neovascularização Patológica/patologia , Proteínas Proto-Oncogênicas c-akt/análise , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real , Reprodutibilidade dos Testes , Fatores de Tempo , Resultado do Tratamento , Fator A de Crescimento do Endotélio Vascular/análise , Fator A de Crescimento do Endotélio Vascular/efeitos dos fármacos , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/análise , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/efeitos dos fármacosRESUMO
BACKGROUND: Impaired angiogenesis in cardiac tissue is a major complication of diabetes. Protein kinase B (AKT) and extracellular signal regulated kinase (ERK) signaling pathways play important role during capillary-like network formation in angiogenesis process. OBJECTIVES: To determine the effects of testosterone and voluntary exercise on levels of vascularity, phosphorylated Akt (P- AKT) and phosphorylated ERK (P-ERK) in heart tissue of diabetic and castrated diabetic rats. METHODS: Type I diabetes was induced by i.p injection of 50 mg/kg of streptozotocin in animals. After 42 days of treatment with testosterone (2mg/kg/day) or voluntary exercise alone or in combination, heart tissue samples were collected and used for histological evaluation and determination of P-AKT and P-ERK levels by ELISA method. RESULTS: Our results showed that either testosterone or exercise increased capillarity, P-AKT, and P-ERK levels in the heart of diabetic rats. Treatment of diabetic rats with testosterone and exercise had a synergistic effect on capillarity, P-AKT, and P-ERK levels in heart. Furthermore, in the castrated diabetes group, capillarity, P-AKT, and P-ERK levels significantly decreased in the heart, whereas either testosterone treatment or exercise training reversed these effects. Also, simultaneous treatment of castrated diabetic rats with testosterone and exercise had an additive effect on P-AKT and P-ERK levels. CONCLUSION: Our findings suggest that testosterone and exercise alone or together can increase angiogenesis in the heart of diabetic and castrated diabetic rats. The proangiogenesis effects of testosterone and exercise are associated with the enhanced activation of AKT and ERK1/2 in heart tissue.
Assuntos
Androgênios/farmacologia , Diabetes Mellitus Experimental/metabolismo , MAP Quinases Reguladas por Sinal Extracelular/análise , Coração/efeitos dos fármacos , Condicionamento Físico Animal/fisiologia , Proteínas Proto-Oncogênicas c-akt/análise , Testosterona/farmacologia , Androgênios/uso terapêutico , Animais , Diabetes Mellitus Experimental/fisiopatologia , Diabetes Mellitus Tipo 1/metabolismo , Diabetes Mellitus Tipo 1/fisiopatologia , Ensaio de Imunoadsorção Enzimática , MAP Quinases Reguladas por Sinal Extracelular/efeitos dos fármacos , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Coração/fisiopatologia , Terapia de Reposição Hormonal/métodos , Masculino , Miocárdio/química , Miocárdio/metabolismo , Miócitos Cardíacos/efeitos dos fármacos , Neovascularização Fisiológica/efeitos dos fármacos , Neovascularização Fisiológica/fisiologia , Proteínas Proto-Oncogênicas c-akt/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-akt/metabolismo , Ratos Wistar , Reprodutibilidade dos Testes , Transdução de Sinais/efeitos dos fármacos , Testosterona/uso terapêutico , Fatores de TempoRESUMO
The present study describes the histopathological and molecular effects of P-MAPA (Protein aggregate magnesium-ammonium phospholinoleate-palmitoleate anhydride) intravesical immunotherapy combined with systemic doxorubicin or cisplatin for treatment of non-muscle invasive bladder cancer (NMIBC) in an appropriate animal model. Our results showed an undifferentiated tumor, characterizing a tumor invading mucosa or submucosa of the bladder wall (pT1) and papillary carcinoma in situ (pTa) in the Cancer group. The histopathological changes were similar between the combined treatment with intravesical P-MAPA plus systemic Cisplatin and P-MAPA immunotherapy alone, showing decrease of urothelial neoplastic lesions progression and histopathological recovery in 80% of the animals. The animals treated systemically with cisplatin or doxorubicin singly, showed 100% of malignant lesions in the urinary bladder. Furthemore, the combined treatment with P-MAPA and Doxorubicin showed no decrease of urothelial neoplastic lesions progression and histopathological recovery. Furthermore, Akt, PI3K, NF-kB and VEGF protein levels were significantly lower in intravesical P-MAPA plus systemic cisplatin and in intravesical P-MAPA alone treatments than other groups. In contrast, PTEN protein levels were significantly higher in intravesical P-MAPA plus systemic cisplatin and in intravesical P-MAPA alone treatments. Thus, it could be concluded that combination of intravesical P-MAPA immunotherapy and systemic cisplatin in the NMIBC animal model was effective, well tolerated and showed no apparent signs of antagonism between the drugs. In addition, intravesical P-MAPA immunotherapy may be considered as a valuable option for treatment of BCG unresponsive patients that unmet the criteria for early cystectomy.
Assuntos
Antineoplásicos/uso terapêutico , Carcinoma/terapia , Cisplatino/uso terapêutico , Doxorrubicina/uso terapêutico , Imunoterapia/métodos , Proteínas de Membrana/uso terapêutico , Neoplasias da Bexiga Urinária/terapia , Administração Intravesical , Animais , Vacina BCG , Western Blotting , Carcinoma/patologia , Terapia Combinada , Progressão da Doença , Feminino , Modelos Animais , NF-kappa B/análise , PTEN Fosfo-Hidrolase/análise , Fosfatidilinositol 3-Quinases/análise , Proteínas Proto-Oncogênicas c-akt/análise , Ratos Endogâmicos F344 , Reprodutibilidade dos Testes , Resultado do Tratamento , Neoplasias da Bexiga Urinária/patologia , Fator A de Crescimento do Endotélio Vascular/análiseRESUMO
Skin-wound healing is a complex and dynamic biological process involving inflammation, proliferation, and remodeling. Recent studies have shown that statins are new therapeutical options because of their actions, such as anti-inflammatory and antioxidant activity, on vasodilation, endothelial dysfunction and neoangiogenesis, which are independent of their lipid-lowering action. Our aim was to investigate the effect of atorvastatin on tissue repair after acute injury in healthy animals. Rats were divided into four groups: placebo-treated (P), topical atorvastatin-treated (AT), oral atorvastatin-treated (AO), topical and oral atorvastatin-treated (ATO). Under anesthesia, rats were wounded with an 8-mm punch in the dorsal region. Lesions were photographed on Days 0, 1, 3, 7, 10, 12, and 14 post-injury and samples taken on Days 1, 3, 7, and 14 for protein-expression analysis of insulin receptor substrate (IRS)-1, phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt), glycogen synthase kinase (GSK)-3, endothelial nitric oxide synthase (eNOS), vascular endothelial growth factor (VEGF), extracellular signal-regulated kinase (ERK), interleukin (IL)-10, IL-1ß, IL-6, and tumor necrosis factor (TNF)-α. Upon macroscopic examination, we observed significant reductions of lesion areas in groups AT, AO, and ATO compared to the P group. Additionally, AT and AO groups showed increased expression of IRS-1, PI3K, Akt, GSK-3, and IL-10 on Days 1 and 3 when compared with the P group. All atorvastatin-treated groups showed higher expression of IRS-1, PI3K, Akt, GSK-3, IL-10, eNOS, VEGF, and ERK on Day 7. On Days 1, 3, and 7, all atorvastatin-treated groups showed lower expression of IL-6 and TNF-α when compared with the P group. We conclude that atorvastatin accelerated tissue repair of acute lesions in rats and modulated expressions of proteins and cytokines associated with cell-growth pathways.
Assuntos
Atorvastatina/farmacologia , Inibidores de Hidroximetilglutaril-CoA Redutases/farmacologia , Cicatrização/efeitos dos fármacos , Administração Oral , Administração Tópica , Animais , Atorvastatina/administração & dosagem , MAP Quinases Reguladas por Sinal Extracelular/análise , Quinase 3 da Glicogênio Sintase/análise , Inibidores de Hidroximetilglutaril-CoA Redutases/administração & dosagem , Immunoblotting , Interleucina-10/análise , Interleucina-1beta/análise , Interleucina-6/análise , Masculino , Óxido Nítrico Sintase Tipo III/análise , Fragmentos de Peptídeos/análise , Fosfatidilinositol 3-Quinase/análise , Proteínas Proto-Oncogênicas c-akt/análise , Ratos , Receptor de Insulina/análise , Fator de Necrose Tumoral alfa/análise , Fator A de Crescimento do Endotélio Vascular/análiseRESUMO
OBJECTIVE: Endometriosis is linked to altered cell proliferation and stem cell markers c-kit/stem cell factor (SCF) in ectopic endometrium. Our aim was to investigate whether c-kit/SCF also plays a role in eutopic endometrium. DESIGN: Eutopic endometrium obtained from 35 women with endometriosis and 25 fertile eumenorrheic women was analyzed for in situ expression of SCF/c-kit, Ki67, RAC-alpha serine/threonine-protein kinase (Akt), phosphorylated RAC-alpha serine/threonin-protein kinase (pAkt), Glycogen synthase kinase 3 beta (GSK3ß), and phosphorylated glycogen synthase kinase 3 beta (pGSK3ß), throughout the menstrual cycle. RESULTS: Expression of Ki67 and SCF was higher in endometriosis than in control tissue (P < .05) and greater in secretory rather than proliferative (P < .01) endometrium in endometriosis. Expression of c-kit was also higher in endometriosis although similar in both phases. Expression of Akt and GSK3ß was identical in all samples and cycle phases, whereas pAkt and pGSK3ß, opposed to control tissue, remained overexpressed in the secretory phase in endometriosis. CONCLUSION: Unceasing cell proliferation in the secretory phase of eutopic endometriosis is linked to deregulation of c-kit/SCF-associated signaling pathways.
Assuntos
Proliferação de Células , Endometriose/enzimologia , Endométrio/enzimologia , Quinase 3 da Glicogênio Sintase/análise , Proteínas Proto-Oncogênicas c-akt/análise , Adulto , Biópsia , Estudos de Casos e Controles , Endometriose/patologia , Endometriose/fisiopatologia , Endométrio/metabolismo , Endométrio/patologia , Feminino , Glicogênio Sintase Quinase 3 beta , Humanos , Imuno-Histoquímica , Antígeno Ki-67/metabolismo , Fosforilação , Proteínas Proto-Oncogênicas c-kit/metabolismo , Transdução de Sinais , Adulto JovemRESUMO
Ginkgo biloba extract (GbE) has been indicated as an efficient medicine for the treatment of diabetes mellitus type 2. It remains unclear if its effects are due to an improvement of the insulin signaling cascade, especially in obese subjects. The aim of the present study was to evaluate the effect of GbE on insulin tolerance, food intake, body adiposity, lipid profile, fasting insulin, and muscle levels of insulin receptor substrate 1 (IRS-1), protein tyrosine phosphatase 1B (PTP-1B), and protein kinase B (Akt), as well as Akt phosphorylation, in diet-induced obese rats. Rats were fed with a high-fat diet (HFD) or a normal fat diet (NFD) for 8 weeks. After that, the HFD group was divided into two groups: rats gavaged with a saline vehicle (HFD+V), and rats gavaged with 500 mg/kg of GbE diluted in the saline vehicle (HFD+Gb). NFD rats were gavaged with the saline vehicle only. At the end of the treatment, the rats were anesthetized, insulin was injected into the portal vein, and after 90s, the gastrocnemius muscle was removed. The quantification of IRS-1, Akt, and Akt phosphorylation was performed using Western blotting. Serum levels of fasting insulin and glucose, triacylglycerols and total cholesterol, and LDL and HDL fractions were measured. An insulin tolerance test was also performed. Ingestion of a hyperlipidic diet promoted loss of insulin sensitivity and also resulted in a significant increase in body adiposity, plasma triacylglycerol, and glucose levels. In addition, GbE treatment significantly reduced food intake and body adiposity while it protected against hyperglycemia and dyslipidemia in diet-induced obesity rats. It also enhanced insulin sensitivity in comparison to HFD+V rats, while it restored insulin-induced Akt phosphorylation, increased IRS-1, and reduced PTP-1B levels in gastrocnemius muscle. The present findings suggest that G. biloba might be efficient in preventing and treating obesity-induced insulin signaling impairment.
Assuntos
Animais , Masculino , Adiposidade/efeitos dos fármacos , Dislipidemias/tratamento farmacológico , Ginkgo biloba/química , Obesidade/tratamento farmacológico , Fitoterapia , Glicemia/análise , HDL-Colesterol/sangue , LDL-Colesterol/sangue , Dieta Hiperlipídica/efeitos adversos , Dislipidemias/metabolismo , Ingestão de Alimentos/efeitos dos fármacos , Teste de Tolerância a Glucose , Hipoglicemia/sangue , Proteínas Substratos do Receptor de Insulina/análise , Resistência à Insulina/fisiologia , Insulina/metabolismo , Músculo Esquelético/química , Obesidade/etiologia , Extratos Vegetais/uso terapêutico , Proteína Tirosina Fosfatase não Receptora Tipo 1/análise , Proteínas Proto-Oncogênicas c-akt/análise , Ratos Wistar , Transdução de Sinais/efeitos dos fármacos , Triglicerídeos/sangueRESUMO
Ginkgo biloba extract (GbE) has been indicated as an efficient medicine for the treatment of diabetes mellitus type 2. It remains unclear if its effects are due to an improvement of the insulin signaling cascade, especially in obese subjects. The aim of the present study was to evaluate the effect of GbE on insulin tolerance, food intake, body adiposity, lipid profile, fasting insulin, and muscle levels of insulin receptor substrate 1 (IRS-1), protein tyrosine phosphatase 1B (PTP-1B), and protein kinase B (Akt), as well as Akt phosphorylation, in diet-induced obese rats. Rats were fed with a high-fat diet (HFD) or a normal fat diet (NFD) for 8 weeks. After that, the HFD group was divided into two groups: rats gavaged with a saline vehicle (HFD+V), and rats gavaged with 500 mg/kg of GbE diluted in the saline vehicle (HFD+Gb). NFD rats were gavaged with the saline vehicle only. At the end of the treatment, the rats were anesthetized, insulin was injected into the portal vein, and after 90s, the gastrocnemius muscle was removed. The quantification of IRS-1, Akt, and Akt phosphorylation was performed using Western blotting. Serum levels of fasting insulin and glucose, triacylglycerols and total cholesterol, and LDL and HDL fractions were measured. An insulin tolerance test was also performed. Ingestion of a hyperlipidic diet promoted loss of insulin sensitivity and also resulted in a significant increase in body adiposity, plasma triacylglycerol, and glucose levels. In addition, GbE treatment significantly reduced food intake and body adiposity while it protected against hyperglycemia and dyslipidemia in diet-induced obesity rats. It also enhanced insulin sensitivity in comparison to HFD+V rats, while it restored insulin-induced Akt phosphorylation, increased IRS-1, and reduced PTP-1B levels in gastrocnemius muscle. The present findings suggest that G. biloba might be efficient in preventing and treating obesity-induced insulin signaling impairment.
Assuntos
Adiposidade/efeitos dos fármacos , Dislipidemias/tratamento farmacológico , Ginkgo biloba/química , Obesidade/tratamento farmacológico , Fitoterapia , Animais , Glicemia/análise , HDL-Colesterol/sangue , LDL-Colesterol/sangue , Dieta Hiperlipídica/efeitos adversos , Dislipidemias/metabolismo , Ingestão de Alimentos/efeitos dos fármacos , Teste de Tolerância a Glucose , Hipoglicemia/sangue , Insulina/metabolismo , Proteínas Substratos do Receptor de Insulina/análise , Resistência à Insulina/fisiologia , Masculino , Músculo Esquelético/química , Obesidade/etiologia , Extratos Vegetais/uso terapêutico , Proteína Tirosina Fosfatase não Receptora Tipo 1/análise , Proteínas Proto-Oncogênicas c-akt/análise , Ratos Wistar , Transdução de Sinais/efeitos dos fármacos , Triglicerídeos/sangueRESUMO
OBJECTIVE: To investigate the effects of different intensities of acute exercise on insulin sensitivity and protein kinase B/Akt activity in skeletal muscle of obese mice. METHODS: Swiss mice were randomly divided into four groups, and fed either a standard diet (control group) or high fat diet (obese sedentary group and obese exercise group 1 and 2) for 12 weeks. Two different exercise protocols were used: swimming for 1 hour with or without an overload of 5% body weight. The insulin tolerance test was performed to estimate whole-body sensitivity. Western blot technique was used to determine protein levels of protein kinase B/Akt and phosphorylation by protein Kinase B/Akt in mice skeletal muscle. RESULTS: A single bout of exercise inhibited the high fat diet-induced insulin resistance. There was increase in phosphorylation by protein kinase B/Akt serine, improve in insulin signaling and reduce of fasting glucose in mice that swam for 1 hour without overload and mice that swan for 1 hour with overload of 5%. However, no significant differences were seen between exercised groups. CONCLUSION: Regardless of intensity, aerobic exercise was able to improve insulin sensitivity and phosphorylation by protein kinase B/Ak, and proved to be a good form of treatment and prevention of type 2 diabetes.
Assuntos
Resistência à Insulina/fisiologia , Músculo Esquelético/metabolismo , Obesidade/metabolismo , Condicionamento Físico Animal/fisiologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Animais , Glicemia/análise , Western Blotting , Diabetes Mellitus Tipo 2/prevenção & controle , Dieta Hiperlipídica , Ensaio de Imunoadsorção Enzimática , Insulina/sangue , Masculino , Camundongos , Camundongos Obesos , Obesidade/fisiopatologia , Fosforilação/fisiologia , Proteínas Proto-Oncogênicas c-akt/análise , Distribuição Aleatória , Fatores de TempoRESUMO
Objective : To investigate the effects of different intensities of acute exercise on insulin sensitivity and protein kinase B/Akt activity in skeletal muscle of obese mice. Methods : Swiss mice were randomly divided into four groups, and fed either a standard diet (control group) or high fat diet (obese sedentary group and obese exercise group 1 and 2) for 12 weeks. Two different exercise protocols were used: swimming for 1 hour with or without an overload of 5% body weight. The insulin tolerance test was performed to estimate whole-body sensitivity. Western blot technique was used to determine protein levels of protein kinase B/Akt and phosphorylation by protein Kinase B/Akt in mice skeletal muscle. Results : A single bout of exercise inhibited the high fat diet-induced insulin resistance. There was increase in phosphorylation by protein kinase B/Akt serine, improve in insulin signaling and reduce of fasting glucose in mice that swam for 1 hour without overload and mice that swan for 1 hour with overload of 5%. However, no significant differences were seen between exercised groups. Conclusion : Regardless of intensity, aerobic exercise was able to improve insulin sensitivity and phosphorylation by protein kinase B/Ak, and proved to be a good form of treatment and prevention of type 2 diabetes. .
Objetivo : Investigar os efeitos do exercício físico agudo com diferentes intensidades sobre a sensibilidade à insulina e a atividade da proteína quinase B/Akt no músculo esquelético de camundongos obesos. Métodos : Foram utilizados camundongos Swiss, divididos aleatoriamente em quatro grupos, que receberam dieta padrão (grupo controle) ou dieta hiperlipídica (grupos obeso sedentário e grupos obesos exercitados 1 e 2), por período de 12 semanas. Dois diferentes protocolos de exercício foram utilizados: natação durante 1 hora com ou sem sobrecarga de 5% da massa corporal. O teste de tolerância à insulina foi realizado para estimar a sensibilidade à insulina. E os níveis protéicos da proteína quinase B/Akt e de sua fosforilação foram determinados no músculo esquelético dos camundongos, através da técnica de Western blot. Resultados : Uma sessão de exercício físico foi capaz de inibir a resistência à insulina em decorrência de uma dieta hiperlipídica. Foi possível demonstrar um aumento na fosforilação da proteína quinase B/Akt, melhora da sinalização da insulina e redução da glicemia de jejum nos camundongos que realizaram 1 hora de natação sem sobrecarga adicional e nos camundongos que realizaram 1 hora de natação com sobrecarga adicional de 5% de sua massa corporal. Entretanto, não houve diferença significativa entre os grupos que realizaram o exercício em diferentes intensidades. Conclusão : Independente da intensidade, o exercício físico aeróbio conseguiu aumentar a sensibilidade à insulina e a fosforilação da proteína quinase B/Akt, revelando ser uma boa forma de tratamento e prevenção do diabetes tipo 2. .
Assuntos
Animais , Masculino , Camundongos , Resistência à Insulina/fisiologia , Músculo Esquelético/metabolismo , Obesidade/metabolismo , Condicionamento Físico Animal/fisiologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Western Blotting , Glicemia/análise , Dieta Hiperlipídica , /prevenção & controle , Ensaio de Imunoadsorção Enzimática , Insulina/sangue , Camundongos Obesos , Obesidade/fisiopatologia , Fosforilação/fisiologia , Proteínas Proto-Oncogênicas c-akt/análise , Distribuição Aleatória , Fatores de TempoRESUMO
BACKGROUND: Ameloblastoma is an odontogenic neoplasm with local invasiveness and high recurrence. We previously suggested that growth factors, matrix metalloproteinases (MMPs), and TIMPs influence ameloblastoma invasiveness (Pathol. Res. Pract., 208, 2012, 225; Oral. Surg. Oral. Med. Oral. Pathol. Oral Radiol. Endod., 111, 2011, 474). Signals generated by this molecular network would be transduced by ERK 1/2 pathway (Oral. Surg. Oral. Med. Oral. Pathol. Oral Radiol. Endod., 111, 2011, 474). Others signaling pathways may influence ameloblastoma biology. Here, we studied expression of AKT and related molecules in ameloblastoma. METHODS: Fourteen cases of solid/multicystic ameloblastomas were examined. Immunohistochemistry was carried out to detected AKT (phospho-AKT), NF-ÒB (phospho-NF-ÒB), ß-catenin, cyclin-D1, and COX-2 in ameloblastoma samples. These molecules were evaluated in neoplastic cells and stroma. RESULTS: All proteins were detected in ameloblastoma. Expression of these markers was quantified and compared. Spearman's rank test was carried out to address positive correlations between proteins (P < 0.05). Ameloblastoma had a significant positive correlation of AKT (phospho-AKT) with ß-catenin. ß-catenin correlated with Cyclin-D1 and COX-2 in neoplastic cells. AKT (phospho-AKT) correlated with ß-catenin; ß-catenin with Cyclin-D1; AKT (phospho-AKT) with NF-ÒB (phospho-NF-ÒB); and NF-ÒB (phospho-NF-ÒB) with COX-2 in stromal cells. CONCLUSIONS: Results suggest that proteins studied are present and probably involved in a functional pathway in neoplastic cells and stroma and may therefore influence the local invasiveness of ameloblastoma.
Assuntos
Ameloblastoma/patologia , Proteínas Proto-Oncogênicas c-akt/análise , Núcleo Celular/patologia , Ciclina D1/análise , Ciclo-Oxigenase 2/análise , Citoplasma/patologia , Humanos , Imuno-Histoquímica , NF-kappa B/análise , Invasividade Neoplásica , Transdução de Sinais/fisiologia , Células Estromais/patologia , beta Catenina/análiseRESUMO
Tissue microarray technology enables us to evaluate the pattern of protein expression in large numbers of samples. However, manual data acquisition and analysis still represent a challenge because they are subjective and time-consuming. Automated analysis may thus increase the speed and reproducibility of evaluation. However, the reliability of automated analysis systems should be independently evaluated. Herein, the expression of phosphorylated AKT and mTOR was determined by ScanScope XT (Aperio; Vista, CA) and ACIS III (Dako; Glostrup, Denmark) and compared with the manual analysis by two observers. The percentage of labeled pixels or nuclei analysis had a good correlation between human observers and automated systems (κ = 0.855 and 0.879 for ScanScope vs. observers and κ = 0.765 and 0.793 for ACIS III vs. observers). The intensity of labeling determined by ScanScope was also correlated with that found by the human observers (correlation index of 0.946 and 0.851 for pAKT and 0.851 and 0.875 for pmTOR). However, the correlation between ACIS III and human observation varied for labeling intensity and was considered poor in some cases (correlation index of 0.718 and 0.680 for pAKT and 0.223 and 0.225 for pmTOR). Thus, the percentage of positive pixels or nuclei determination was satisfactorily performed by both systems; however, labeling intensity was better identified by ScanScope XT.
Assuntos
Automação , Proteínas Proto-Oncogênicas c-akt/análise , Serina-Treonina Quinases TOR/análise , Análise Serial de Tecidos , Humanos , Imuno-Histoquímica , Fosforilação , Proteínas Proto-Oncogênicas c-akt/biossíntese , Proteínas Proto-Oncogênicas c-akt/metabolismo , Serina-Treonina Quinases TOR/biossíntese , Serina-Treonina Quinases TOR/metabolismoRESUMO
AIMS/HYPOTHESIS: Ciliary neurotrophic factor (CNTF) improves metabolic variables of obese animals with characteristics of type 2 diabetes, mainly by reducing insulin resistance. We evaluated whether CNTF was able to improve other metabolic variables in mouse models of type 2 diabetes, such as beta cell mass and insulin clearance, and whether CNTF has any effect on non-obese mice with characteristics of type 2 diabetes. METHODS: Neonatal mice were treated with 0.1 mg/kg CNTF or citrate buffer via intraperitoneal injections, before injection of 250 mg/kg alloxan. HEPG2 cells were cultured for 3 days in the presence of citrate buffer, 1 nmol/l CNTF or 50 mmol/l alloxan or a combination of CNTF and alloxan. Twenty-one days after treatment, we determined body weight, epididymal fat weight, blood glucose, plasma insulin, NEFA, glucose tolerance, insulin resistance, insulin clearance and beta cell mass. Finally, we assessed insulin receptor and protein kinase B phosphorylation in peripheral organs, as well as insulin-degrading enzyme (IDE) protein production and alternative splicing in the liver and HEPG2 cells. RESULTS: CNTF improved insulin sensitivity and beta cell mass, while reducing glucose-stimulated insulin secretion and insulin clearance in Swiss mice, improving glucose handling in a non-obese type 2 diabetes model. This effect was associated with lower IDE production and activity in liver cells. All these effects were observed even at 21 days after CNTF treatment. CONCLUSIONS/INTERPRETATION: CNTF protection against type 2 diabetes is partially independent of the anti-obesity actions of CNTF, requiring a reduction in insulin clearance and increased beta cell mass, besides increased insulin sensitivity. Furthermore, knowledge of the long-term effects of CNTF expands its pharmacological relevance.
Assuntos
Fator Neurotrófico Ciliar/farmacologia , Diabetes Mellitus Experimental/prevenção & controle , Diabetes Mellitus Tipo 2/prevenção & controle , Células Secretoras de Insulina/efeitos dos fármacos , Insulina/metabolismo , Animais , Glicemia/efeitos dos fármacos , Peso Corporal/efeitos dos fármacos , Diabetes Mellitus Tipo 2/metabolismo , Epididimo/efeitos dos fármacos , Ácidos Graxos não Esterificados/sangue , Células Hep G2 , Humanos , Insulina/sangue , Resistência à Insulina , Secreção de Insulina , Insulisina/biossíntese , Masculino , Camundongos , Proteínas Proto-Oncogênicas c-akt/análise , Receptor de Insulina/análiseRESUMO
El Síndrome Metabólico corresponde a una serie de trastornos relacionados con obesidad e inactividad física. Poco se conoce respecto de la falta de ejercicio, en estadios tempranos del desarrollo, en la susceptibilidad a un fenotipo insulinoresistente inducido por una dieta alta en grasas. Akt juega un rol clave en la síntesis de proteínas y el transporte de glucosa en el músculo esquelético y ha mostrado ser regulada por la actividad muscular. El objetivo del presente estudio fue determinar el efecto de la inactividad física temprana sobre el crecimiento muscular y la susceptibilidad de adquirir un fenotipo diabético y evaluar su relación con la expresión de Akt. Cuarenta ratas Wistar fueron distribuidas en 2 grupos (Grupos Control, Std) y Restricción de movimiento (RM). Entre los días postnatal 23 y 70 los animales del grupo RM fueron alojados en pequeñas jaulas que no permitían una actividad motora relevante. A partir del día postnatal 71 y hasta el día 102, 10 ratas de cada grupo fueron alimentadas con una Dieta Alta en Grasas (RM-DAG y Std-DAG). No se observaron diferencias en el peso corporal total pero DAG generó un significativo incremento en la grasa epididimal. RM generó una disminución significativa en el peso de los músculos sóleo. La captación de glucosa estimulada por insulina fue menor en el grupo RM-DAG. Los niveles de proteína Akt fueron menores en los grupos RM. El análisis de PCR a tiempo real mostró que la restricción de movimiento disminuyó los niveles de ARNm de AKT1 en el músculo sóleo, independiente de la dieta administrada. Estos hallazgos sugieren que la inactividad física temprana limita el crecimiento muscular y contribuye en la instauración un fenotipo insulino resistente, lo cual puede ser en parte explicado por una desregulación en la expresión de Akt.
Metabolic Syndrome is a group of conditions related to obesity and physical inactivity. Little is known about the role of physical inactivity, in early stages of development, in the susceptibility to insulin resistant phenotype induced by high fat diet. Akt plays a key role in protein synthesis and glucose transport in skeletal muscle and has been regulated by muscle activity. The objective of present study was to determine the effect of early physical inactivity on muscle growth and susceptibility to acquire a diabetic phenotype and to assess its relationship with Akt expression. Forty Wistar male rats were distributed in two groups (standard group, Std) and movement restriction (RM). Between days 23 and 70 after birth, RM group was kept in small cages that did not allow them to perform relevant motor activity. From day 71 to 102 after birth, 10 rats of each group were fed with hyperlipidic diet (groups Std-DAG and RM-DAG). No differences were observed in total body weight although DAG increased epididymal fat pad weight. RM decreased significantly the soleus weight. Insulin-mediated glucose uptake was lower in RM-DAG group. Akt protein levels were lower in RM groups. Real time RT-PCR analysis showed that movement restriction decreased mRNA levels of AKT1 in soleus muscle, regardless of supplied diet. These findings suggest that early physical inactivity limits muscle's growth and contributes to instauration of insulin resistant phenotype, which can be partly explained by dysregulation of Akt expression.