RESUMO
OBJECTIVE: High-fat diet (HFD)-induced metabolic disorders can lead to impaired sperm production. We aim to investigate if HFD-induced gut microbiota dysbiosis can functionally influence spermatogenesis and sperm motility. DESIGN: Faecal microbes derived from the HFD-fed or normal diet (ND)-fed male mice were transplanted to the mice maintained on ND. The gut microbes, sperm count and motility were analysed. Human faecal/semen/blood samples were collected to assess microbiota, sperm quality and endotoxin. RESULTS: Transplantation of the HFD gut microbes into the ND-maintained (HFD-FMT) mice resulted in a significant decrease in spermatogenesis and sperm motility, whereas similar transplantation with the microbes from the ND-fed mice failed to do so. Analysis of the microbiota showed a profound increase in genus Bacteroides and Prevotella, both of which likely contributed to the metabolic endotoxaemia in the HFD-FMT mice. Interestingly, the gut microbes from clinical subjects revealed a strong negative correlation between the abundance of Bacteroides-Prevotella and sperm motility, and a positive correlation between blood endotoxin and Bacteroides abundance. Transplantation with HFD microbes also led to intestinal infiltration of T cells and macrophages as well as a significant increase of pro-inflammatory cytokines in the epididymis, suggesting that epididymal inflammation have likely contributed to the impairment of sperm motility. RNA-sequencing revealed significant reduction in the expression of those genes involved in gamete meiosis and testicular mitochondrial functions in the HFD-FMT mice. CONCLUSION: We revealed an intimate linkage between HFD-induced microbiota dysbiosis and defect in spermatogenesis with elevated endotoxin, dysregulation of testicular gene expression and localised epididymal inflammation as the potential causes. TRIAL REGISTRATION NUMBER: NCT03634644.
Assuntos
Bacteroides/isolamento & purificação , Dieta Hiperlipídica/efeitos adversos , Disbiose , Prevotella/isolamento & purificação , Motilidade dos Espermatozoides/imunologia , Espermatogênese/imunologia , Animais , Correlação de Dados , Citocinas/análise , Disbiose/etiologia , Disbiose/microbiologia , Endotoxemia/microbiologia , Epididimo/imunologia , Epididimo/patologia , Fezes/microbiologia , Microbioma Gastrointestinal/imunologia , Humanos , Macrófagos/imunologia , Masculino , Camundongos , Linfócitos T/imunologiaRESUMO
Baicalin has shown multiple neuroprotective biological activities, including antiapoptotic and anti-inflammatory functions in neurodegeneration diseases. However, whether baicalin can regulate Aß-induced microglial activation or inhibit inflammatory cytokine secretion has not been confirmed. We demonstrated that baicalin can inhibit beta amyloid peptides (Aß42)-induced BV2 microglial cell proliferation, reduce the expression of CD11b, decrease chemotactic ability of BV2 cells and significantly inhibit the secretion of IL-6, TNF-α and NO. Moreover, baicalin pretreatment can effectively inhibit Aß-induced phosphorylation of JAK2 and STAT3. Baicalin can inhibit Aß-induced microglial cell activation by regulating the JAK2/STAT3 signaling pathway in AD transgenic mice. The modulation of microglial proliferation, activation and secretion by baicalin could be a promising therapeutic option for the treatment of Alzheimer's disease.
Assuntos
Peptídeos beta-Amiloides/antagonistas & inibidores , Flavonoides/farmacologia , Janus Quinase 2/antagonistas & inibidores , Microglia/efeitos dos fármacos , Fragmentos de Peptídeos/antagonistas & inibidores , Fator de Transcrição STAT3/antagonistas & inibidores , Transdução de Sinais/efeitos dos fármacos , Peptídeos beta-Amiloides/metabolismo , Animais , Anti-Inflamatórios não Esteroides/farmacologia , Humanos , Janus Quinase 2/metabolismo , Camundongos , Camundongos Transgênicos , Microglia/metabolismo , Fragmentos de Peptídeos/metabolismo , Distribuição Aleatória , Fator de Transcrição STAT3/metabolismo , Transdução de Sinais/fisiologiaRESUMO
AIMS: To further investigate the antineuroblastoma effect of rutin which is a type of flavonoid. METHODS: The antiproliferation of rutin in human neuroblastoma cells LAN-5 were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Chemotaxis of LAN-5 cells was assessed using transwell migration chambers and scratch wound migration assay. The cell cycle arrest and apoptosis in a dose-dependent manner was measured by flow cytometric and fluorescent microscopy analyses. The apoptosis-related proteins BAX and BCL2 as well as MYCN mRNA express were determined by RT-PCR analysis. Secreted TNF- α level were determined using specific enzyme-linked immunosorbent assay kits. RESULTS: Rutin significantly inhibited the growth of LAN-5 cells and chemotactic ability. Flow cytometric analysis revealed that rutin induced G2/M arrest in the cell cycle progression and induced cell apoptosis. The RT-PCR showed that rutin could decrease BCL2 expression and BCL2/BAX ratio. In the meantime, the MYCN mRNA level and the secretion of TNF- α were inhibited. CONCLUSION: These results suggest that rutin produces obvious antineuroblastoma effects via induced G2/M arrest in the cell cycle progression and induced cell apoptosis as well as regulating the expression of gene related to apoptosis and so on. It supports the viability of developing rutin as a novel therapeutic prodrug for neuroblastoma treatment, as well as providing a new path on anticancer effect of Chinese traditional drug.
Assuntos
Apoptose/efeitos dos fármacos , Pontos de Checagem da Fase G2 do Ciclo Celular/efeitos dos fármacos , Pontos de Checagem da Fase M do Ciclo Celular/efeitos dos fármacos , Neuroblastoma/tratamento farmacológico , Apoptose/genética , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Movimento Celular/genética , Pontos de Checagem da Fase G2 do Ciclo Celular/genética , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Pontos de Checagem da Fase M do Ciclo Celular/genética , Proteína Proto-Oncogênica N-Myc , Neuroblastoma/genética , Neuroblastoma/metabolismo , Neuroblastoma/patologia , Proteínas Nucleares/biossíntese , Proteínas Nucleares/genética , Proteínas Oncogênicas/biossíntese , Proteínas Oncogênicas/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Rutina , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo , Proteína X Associada a bcl-2/biossíntese , Proteína X Associada a bcl-2/genéticaRESUMO
This study was to investigate antidepressant activities of Shuyusan (a Chinese herb), using a rats model of depression induced by unpredictable chronic mild stress (UCMS). The administration groups were treated with Shuyusan decoction for 3 weeks and compared with fluoxetine treatment. In order to understand the potential antidepressant-like activities of Shuyusan, tail suspension test (TST) and forced swimming test (FST) were used as behavioral despair study. The level of corticotropin-releasing factor (CRH), adrenocorticotropic hormone (ACTH), corticosterone (CORT) and hippocampus glucocorticoid receptor expression were examined. After modeling, there was a significant prolongation of immobility time in administration groups with the TST and FST. High-dose Shuyusan could reduce the immobility time measured with the TST and FST. The immobility time in high-dose herbs group and fluoxetine group was increased significantly compared with the model group. After 3 weeks herbs fed, the serum contents level of CRH, ACTH, and CORT in high-dose herb group was significantly decreased compared to the model group. The result indicated that Shuyusan had antidepressant activity effects on UCMS model rats. The potential antidepressant effect may be related to decreasing glucocorticoid levels activity, regulating the function of HPA axis, and inhibiting glucocorticoid receptor expression in hippocampus.
RESUMO
This study is to investigate the effect of baicalin (BL) against oxidative injury stress of SH-SY5Y cells induced by H2O2 and the possible mechanism. SH-SY5Y cells were pre-incubated with baicalin (25, 50, and 100 micromol x L(-1)) for 12 h prior to exposure to H2O2 (150 micromol x L(-1)) for 24 h. The viability of SH-SY5Y cells was measured by MTT assay. The contents of LDH and NO were determined. The percentage of apoptotic cells was assessed by flow cytometry (FCM). The content of Caspase-3 was tested by immunofluorescence histochemical method. BL at 50 and 100 micromol x L(-1) separately increased the cell viability and up-regulated SIRT1, reduced the contents of LDH, NO, Caspase-3 and the apoptotic percentage of SH-SY5Y cells. This study results suggest that baicalin could inhibit the H2O2-induced neuronal apoptosis. The further mechanism studies show that baicalin inhibit apoptosis via reducing Caspase-3 expression and up-regulating SIRT1.
Assuntos
Apoptose/efeitos dos fármacos , Flavonoides/farmacologia , Neuroblastoma , Estresse Oxidativo/efeitos dos fármacos , Sirtuína 1/metabolismo , Antioxidantes/isolamento & purificação , Antioxidantes/farmacologia , Caspase 3/metabolismo , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Flavonoides/isolamento & purificação , Humanos , Peróxido de Hidrogênio/toxicidade , L-Lactato Desidrogenase/metabolismo , Neuroblastoma/metabolismo , Neuroblastoma/patologia , Óxido Nítrico/metabolismo , Plantas Medicinais/química , Scutellaria/química , Regulação para CimaRESUMO
OBJECTIVE: To explore the pathogenesis of brain damage after chronic cerebral ischemia through analysis of the differences in proteins expression in hippocampus between chronic cerebral ischemia rats and normal rats. METHODS: The chronic cerebral ischemia model was established by ligating the bilateral common carotid arteries.Twenty rats were randomly divided into a model group (n=10)and a sham operation group(n=10). Four weeks later, the differences of proteins expression in hippocampus between model group and sham operation group were analyzed by two dimensional polyacryalmide gel electrophoresis and ultraflex TOF/TOF mass spectrograph. RESULTS: Compared to the sham operation group, the expressions of 4 proteins were up-regulated and that of 2 proteins were down-regulated in the model group. Six proteins were identified by ultraflex TOF/TOF, which were ubiquitin carboxy-terminal hydrolase L1; Dynamin-1; TMF regulated nuclear protein-like, partial; ATP synthase; rCG50513, isoform CRA_a; and expressed sequence AU016693, isoform CRA_b. CONCLUSION: Well-resolved and reproducible 2-DE patterns of chronic cerebral ischemia rats were established. Six proteins that correlate with nerve damage after chronic cerebral ischemia are identified.
Assuntos
Isquemia Encefálica/metabolismo , Hipocampo/metabolismo , Proteômica/métodos , Animais , Doença Crônica , Dinamina I/análise , Eletroforese em Gel Bidimensional , Feminino , Masculino , Proteoma/análise , Ratos , Ratos Wistar , Ubiquitina Tiolesterase/análiseRESUMO
The aim of this study is to investigate the effects and mechanism of extract of Apocynum venetum (AV) on kidneys of streptozotocin-induced diabetic rats. Diabetes mellitus (DM) was induced in rats by intraperitoneal injection of streptozotocin (STZ). The indexes of the blood glucose, renal function and oxidative stress were observed. The DM rats were administrated with the AV for 8 weeks, the above-mentioned indexes were detected. The blood glucose level, BUN, 24 h urine protein excretion, urine volume, renal index, renal cortex's MDA level in model groups all increased significantly. Renal cortex's SOD and GSH activities decreased significantly compared with the normal control group (P < 0.05). The above-mentioned indexes were significantly improved by the AV treatment (P < 0.05). AV have protective effects on renal function of kidneys of streptozotocin-induced diabetic rats, and maybe via inhibition of the renal oxidative stress.
Assuntos
Apocynum/química , Diabetes Mellitus Experimental/tratamento farmacológico , Medicamentos de Ervas Chinesas/farmacologia , Córtex Renal/patologia , Animais , Glicemia/metabolismo , Nitrogênio da Ureia Sanguínea , Creatinina/sangue , Diabetes Mellitus Experimental/sangue , Diabetes Mellitus Experimental/patologia , Medicamentos de Ervas Chinesas/isolamento & purificação , Frutosamina/sangue , Glutationa Peroxidase/metabolismo , Rim/fisiopatologia , Masculino , Malondialdeído/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Ratos , Ratos Wistar , Superóxido Dismutase/metabolismoRESUMO
The aim of this study is to investigate the effect of hyperin on the cccDNA of duck hepatitis B virus and its immunological regulation. Duck hepatitis B virus (DHBV) infection model and normal mouse spleen lymphocyte were used to evaluate the anti-HBV and immunoregulation effects. The DHBV-DNA of serum was detected at different time points by using serum DOT-BLOT hybridization. Polymerase chain reaction (PCR) was used for the determination of nuclear covalent closed circular DNA (cccDNA). Cytokine secretion was determined by ELISA method. DHBV-DNA were inhibited by hyperin (25 or 50 mg x kg(-1)), while cccDNA of liver could be eliminated efficiently by hyperin (25 or 50 mg x kg(-1), P < 0.05, P < 0.01). The T helper 1 effector cytokine was markedly enhanced by hyperin (25 or 50 microg x mL(-1), P < 0.01). In conclusion, hyperin has anti-HBV activity via multiple targets and pathways, and cccDNA may be one of the important targets.
Assuntos
DNA Circular/metabolismo , DNA Viral/metabolismo , Vírus da Hepatite B do Pato/genética , Interferon gama/metabolismo , Interleucina-12/metabolismo , Quercetina/análogos & derivados , Animais , Antivirais/farmacologia , Infecções por Hepadnaviridae/virologia , Hepatite Viral Animal/virologia , Fígado/virologia , Linfócitos/metabolismo , Camundongos , Quercetina/farmacologia , Baço/patologia , Baço/virologiaRESUMO
The active ingredient of ginseng, ginsenosides Rg1, has been shown to scavenge free radicals and improve antioxidant capacity. This study hypothesized that ginsenosides Rg1 has a protective role in human neuroblastoma cells injured by H2O2. Ginsenosides Rg1 at different concentrations (50 and 100 µM) was used to treat H2O2 (150 µM)-injured SH-SY5Y cells. Results demonstrated that ginsenoside Rg1 elevated the survival rate of SH-SY5Y cells injured by H2O2, diminished the amount of leaked lactate dehydrogenase, and increased superoxide dismutase activity. Ginsenoside Rg1 effectively suppressed caspase-3 immunoreactivity, and contributed to heat shock protein 70 gene expression, in a dose-dependent manner. These results indicate that ginsenoside Rg1 has protective effects on SH-SY5Y cells injured by H2O2 and that its mechanism of action is associated with anti-oxidation and the inhibition of apoptosis.
RESUMO
OBJECTIVE: To describe the methods which were used to develop collagen-based materials for wound dressing. METHODS: Fresh frozen bovine tendon was treated with 0.05 mol/L acetic acid at pH 3.2 for 48-72 hours, homogenized, filtered, mixed with 8% chondroitin sulphate, for creating a deaerated 1.5%-2.5% collagen solution. The solution was lyophilized in either a pre-frozen or non-pre-frozen mould. The collagen sponge was then cross-linked with 0.25% glutaraldehyde for 24 hours. Three other types of wound dressings were developed using a similar method: collagen membrane with a polyurethane membrane onlay, polyurethane-coated collagen membrane and collagen membrane on gauze. RESULTS: It was demonstrated that the use of frozen bovine tendon was stable, and that the prepared collagen sponge contained pores of 50-400 microm in diameter. CONCLUSIONS: Collagen could be used as wound dressing.
Assuntos
Curativos Biológicos , Colágeno , Aminoácidos/análise , Animais , Bovinos , Colágeno/análise , Colágeno/química , Colágeno/isolamento & purificação , Liofilização , PoliuretanosRESUMO
In this study the treatment effect of combined implantation of autologous skin on pig dermis in injured rats was observed. Twenty-one Wistar rats were used, and the wounds were formed by excising a piece of full thickness skin on the back. After the pig dermis was implanted, the autologous skin was grafted on the dermis at 0.7 and 10 days. In the group with perforated pig dermis, the autograft skin was implanted on the day when the pig dermis was implanted. The healing effect was evaluated by measuring wound area, and by observing the growth of the autograft skin. Two weeks after the autograft skin was implanted, the skin securely adhered to the dermis, and the edge of autograft skin expanded clearly. The wound of the autograft skin implanted in the perforation of the dermis healed completely after 3 weeks, but the other 3 groups had remnant small wound. The autograft skin merged with the dermis and its surrounding tissue, but a clear dividing line still existed between autograft skin and dermis after implantation. The area of the implanted dermis and autograft skin varied from 51.8% to 65.9% compared to its original size. The results suggested that the time and the way of autologous skin grafting on xenogenous dermis may influence wound contraction and healing time.
Assuntos
Derme/transplante , Transplante de Pele/métodos , Porco Miniatura , Animais , Feminino , Sobrevivência de Enxerto , Masculino , Ratos , Ratos Wistar , Suínos , Transplante Autólogo , Transplante HeterólogoRESUMO
Alzheimer's disease (AD) is the most common form of dementia. Accumulation of amyloid-beta (Aß) peptides is regarded as the critical component associated with AD pathogenesis, which is derived from the amyloid precursor protein (APP) cleavage. Recent studies suggest that synaptic activity is one of the most important factors that regulate Aß levels. It has been found that synaptic activity facilitates APP internalization and influences APP cleavage. Glutamatergic, cholinergic, serotonergic, leptin, adrenergic, orexin, and gamma-amino butyric acid receptors, as well as the activity-regulated cytoskeleton-associated protein (Arc) are all involved in these processes. The present review summarizes the evidence for synaptic activity-modulated Aß levels and the mechanisms underlying this regulation. Interestingly, the immediate early gene product Arc may also be the downstream signaling molecule of several receptors in the synaptic activity-modulated Aß levels. Elucidating how Aß levels are regulated by synaptic activity may provide new insights in both the understanding of the pathogenesis of AD and in the development of therapies to slow down the progression of AD.
Assuntos
Doença de Alzheimer/etiologia , Peptídeos beta-Amiloides/metabolismo , Encéfalo/metabolismo , Sinapses/fisiologia , Doença de Alzheimer/tratamento farmacológico , Doença de Alzheimer/genética , Doença de Alzheimer/metabolismo , Secretases da Proteína Precursora do Amiloide/fisiologia , Precursor de Proteína beta-Amiloide/metabolismo , Animais , Proteínas do Citoesqueleto/fisiologia , Descoberta de Drogas , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/fisiologia , Terapia de Alvo Molecular , Proteínas do Tecido Nervoso/fisiologia , Receptores Colinérgicos/fisiologia , Receptores de Glutamato/fisiologia , Receptores de N-Metil-D-Aspartato/fisiologiaRESUMO
The Chinese herb Shuyusan, whose main constituent is jasminoidin, has been shown to protect SH-SY5Y cells against corticosterone-induced damage. SH-SY5Y cells injured by 400 µmol/L corticosterone were treated with 5 and 30 µg/mL Shuyusan-containing serum. Results revealed that Shuyusan-containing serum elevated the survival rate of SH-SY5Y cells, reduced Bax expression, increased Bcl-2 expression, markedly elevated brain-derived neurotrophic factor mRNA expression, and blocked cell apoptosis. Moreover, the effect of high-dose (30 µg/mL) Shuyusan-containing serum was more remarkable. Therefore, Shuyusan-containing serum appears to protect SH-SY5Y cells against corticosterone-induced impairment by adjusting the expression of apoptosis-associated proteins and brain-derived neurotrophic factor. Moreover, high-dose Shuyusan-containing serum has a protective effect on high-dose corticosterone-induced impairment.
RESUMO
The transcription factor NRF2 (NFE2L2) is a pivotal activator of genes encoding cytoprotective and detoxifying enzymes that limit the action of cytotoxic therapies in cancer. NRF2 acts by binding antioxidant response elements (ARE) in its target genes, but there is relatively limited knowledge about how it is negatively controlled. Here, we report that retinoic X receptor alpha (RXRα) is a hitherto unrecognized repressor of NRF2. RNAi-mediated knockdown of RXRα increased basal ARE-driven gene expression and induction of ARE-driven genes by the NRF2 activator tert-butylhydroquinone (tBHQ). Conversely, overexpression of RXRα decreased ARE-driven gene expression. Biochemical investigations showed that RXRα interacts physically with NRF2 in cancer cells and in murine small intestine and liver tissues. Furthermore, RXRα bound to ARE sequences in the promoters of NRF2-regulated genes. RXRα loading onto AREs was concomitant with the presence of NRF2, supporting the hypothesis that a direct interaction between the two proteins on gene promoters accounts for the antagonism of ARE-driven gene expression. Mutation analyses revealed that interaction between the two transcription factors involves the DNA-binding domain of RXRα and a region comprising amino acids 209-316 in human NRF2 that had not been defined functionally, but that we now designate as the NRF2-ECH homology (Neh) 7 domain. In non-small cell lung cancer cells where NRF2 levels are elevated, RXRα expression downregulated NRF2 and sensitized cells to the cytotoxic effects of therapeutic drugs. In summary, our findings show that RXRα diminishes cytoprotection by NRF2 by binding directly to the newly defined Neh7 domain in NRF2.
Assuntos
Elementos de Resposta Antioxidante/fisiologia , Fator 2 Relacionado a NF-E2/antagonistas & inibidores , Receptor X Retinoide alfa/fisiologia , Transdução de Sinais/fisiologia , Animais , Linhagem Celular Tumoral , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/fisiologia , Proteína 1 Associada a ECH Semelhante a Kelch , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Fator 2 Relacionado a NF-E2/química , Fator 2 Relacionado a NF-E2/fisiologia , Estrutura Terciária de ProteínaRESUMO
OBJECTIVE: To investigate the effect of intensive rosuvastatin therapy on adhesion molecules in patients with peripheral atherosclerosis and explore the possible upstream mechanism. METHODS: Twenty asymptomatic patients with peripheral atherosclerosis were enrolled and given 5-20 mg/day rosuvastatin for 3 months. Before and after the treatment, the lipid profile and plasma vascular cell adhesion molecule-1 (VCAM-1) levels were examined. The expression of intercellular adhesion molecule-1 (ICAM-1) in the mononuclear cells was measured using flow cytometry, and the mRNA and protein expressions of peroxisome proliferator-activated receptor γ (PPARγ) were detected using RT-PCR and Western blotting, respectively. RESULTS: Compared with the baseline levels, ICAM-1 expression decreased and PPARγ protein expression increased in the lymphocytes. Rosuvastatin therapy did not produce obvious effects on plasma VCAM-1 level or ICAM-1 expression in the monocytes in these patients. CONCLUSION: Rosuvastatin produces anti-inflammatory effects by decreasing the expression of ICAM-1 in mononuclear cells, and its upstream mechanism may involve the PPARγ pathway.
Assuntos
Aterosclerose/tratamento farmacológico , Aterosclerose/metabolismo , Moléculas de Adesão Celular/metabolismo , Fluorbenzenos/uso terapêutico , Pirimidinas/uso terapêutico , Sulfonamidas/uso terapêutico , Feminino , Fluorbenzenos/administração & dosagem , Humanos , Molécula 1 de Adesão Intercelular/metabolismo , Masculino , Pessoa de Meia-Idade , Monócitos/efeitos dos fármacos , Monócitos/metabolismo , PPAR gama/metabolismo , Pirimidinas/administração & dosagem , Rosuvastatina Cálcica , Sulfonamidas/administração & dosagem , Molécula 1 de Adesão de Célula Vascular/metabolismoRESUMO
OBJECTIVE: To get a better understanding of the mechanisms underlying type 1 diabetes mellitus, the differentially expressed pancreatic proteins from multiple low-dose streptozotocin (MLD-SIZ) mouse and normal mouse were analyzed and compared. METHODS: 20 male rats were separated into 2 groups (n=10): model mice treated with MLD-STZ and normal mice,differences of pancreatic proteome among in the two groups of mice, were analyzed by two dimensional polyacryamide gel electrophoresis (2DE). Protein quantification was analyzed and the differentially expressed spots were identified using mass spectrometry and MASCOT database searching. RESULTS: Compared with control group, 23 proteins had changed significantly in the model group, 8 proteins expression were up-regulated, 15 proteins expressions down-regulated significantly. Using MALDI-TOF-MS, 15 proteins with significant change were identified by peptide fingerprinting map and the results were searched in MASCOT database. The function analyzed showed that proteins with change were associated with metabolic, anti-oxidant, structural, catalytic enzymes and chaperone, et al. CONCLUSION: Type 1 diabetes is probably exerted via multi-target and multi-path mechanism. The proteins with significant change are newly target for type 1 diabetes early diagnosis and treatment.
Assuntos
Diabetes Mellitus Tipo 1/metabolismo , Pâncreas/metabolismo , Proteínas/metabolismo , Proteômica/métodos , Animais , Diabetes Mellitus Experimental/fisiopatologia , Diabetes Mellitus Tipo 1/induzido quimicamente , Diabetes Mellitus Tipo 1/fisiopatologia , Masculino , Camundongos , EstreptozocinaRESUMO
Plasmid DNA vaccine is an appealing cancer immunotherapy. However, it is a weak immunogen and immunization with plasmid DNA encoding self-antigens, such as melanoma-associated antigens, could not induce antitumor immunity because of tolerance. In this study, we investigated the feasibility of using a plasmid DNA encoding Xenopus laevis transforming growth factor-beta 5 (aTGF-beta5) as an immunogen to induce neutralizing antibodies against murine TGF-beta1 (mTGF-beta1) and thus enhance the efficacy of plasmid DNA vaccine encoding murine tyrosinase-related protein 2 (mTRP-2) through neutralization of TGF-beta. The results showed that immunization with aTGF-beta5 resulted in the generation of mTGF-beta1-neutralizing antibodies, and immunization with a combination of aTGF-beta5 and mTRP-2 induced specific cytotoxic T lymphocytes (CTLs). On the contrary, immunization with mTRP-2 alone could not elicit the CTL response. Moreover, immunization of C57BL/6 wild-type mice with a combination of aTGF-beta5 and mTRP-2 induced the protective and therapeutic antitumor immunity to B16F10 melanoma, whereas the antitumor activity was abrogated in both CD4-deficient mice and CD8-deficient mice on the C57BL/6 background. Our results indicate that immunization with aTGF-beta5 is capable of breaking immune tolerance and induces mTGF-beta1-neutralizing antibodies. Neutralization of TGF-beta can enhance the efficacy of DNA vaccine encoding mTRP-2 and the induction of antitumor immunity by this immunization strategy is associated with CD4+ and CD8+ T cells.
Assuntos
Vacinas Anticâncer/uso terapêutico , Melanoma Experimental/tratamento farmacológico , Proteínas de Membrana/imunologia , Fragmentos de Peptídeos/imunologia , Fator de Crescimento Transformador beta/genética , Fator de Crescimento Transformador beta/imunologia , Animais , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Vacinas Anticâncer/genética , Vacinas Anticâncer/imunologia , Feminino , Tolerância Imunológica/imunologia , Oxirredutases Intramoleculares/imunologia , Melanoma Experimental/imunologia , Melanoma Experimental/prevenção & controle , Camundongos , Camundongos Endogâmicos C57BL , Plasmídeos/genética , Linfócitos T Citotóxicos/imunologia , Fator de Crescimento Transformador beta1 , Vacinas de DNA/genética , Vacinas de DNA/imunologia , Vacinas de DNA/uso terapêutico , Proteínas de XenopusRESUMO
Vaccination with antigenic peptide-pulsed antigen-presenting cells (APC) represents an attractive approach for therapy for cancer and diseases caused by intracellular infections. It has been suggested that sufficient stable MHC/peptide complexes on the surface of APC might play an important role in the generation of antitumor and antiviral immunity in vivo. In this study, we observed that exogenous peptides that were artificially fused with an endoplasmic reticulum (ER) retrieval signal, a C-terminal Lys-Asp-Glu-Leu sequence, could be efficiently presented by intracellular MHC class I molecules in a TAP- and proteasome-independent, but brefeldin A-sensitive manner. The APC retained the capacity to display surface MHC/peptide complexes for a prolonged period. In addition, our results show that vaccination with DC bearing our fusion peptides induced greatly enhanced specific CTL response, and resulted in significant inhibition of tumor growth. Thus, the ER retrieval signal modification can be regarded as a novel method for targeting exogenous peptides into the intracellular MHC class I presentation pathway, and may improve the clinical utility of vaccines based on synthetic peptide pulsed DC.
Assuntos
Apresentação de Antígeno/imunologia , Retículo Endoplasmático/imunologia , Antígenos de Histocompatibilidade Classe I/imunologia , Oligopeptídeos/imunologia , Peptídeos/imunologia , Animais , Apresentação de Antígeno/efeitos dos fármacos , Apresentação de Antígeno/fisiologia , Brefeldina A/farmacologia , Células Dendríticas/imunologia , Cinética , Camundongos , Microscopia Confocal , Neoplasias/imunologia , Neoplasias/prevenção & controle , Complexo de Endopeptidases do Proteassoma/fisiologia , Sinais Direcionadores de Proteínas/fisiologia , Baço/imunologiaRESUMO
Rotaviruses (RV) are a common cause of severe diarrhea in young children, resulting in nearly one million deaths worldwide annually. Rotavirus VP7 was the rotavirus neutralizing protein. Previous study reported that VP7 DNA vaccine can induce high levels of IgG in mice but cannot protect mice against challenge (Choi, A.H., Basu, M., Rae, M.N., McNeal, M.M., Ward, R.L., 1998. Virology 250, 230-240). We found that rotavirus VP7 could maintain its neutralizing immunity when it was transformed into the potato genome. Mice immunized with the transformed tubers successfully elicited serum IgG and mucosal IgA specific for VP7. The mucosal IgA titer was as high as 1000, while serum IgG titer was only 600. Neutralizing assays indicated that IgA could neutralize rotavirus. These results indicate the potential usefulness of plants for production and delivery of edible rotavirus vaccines.