RESUMO
Our prior investigations have evidenced that bone marrow mesenchymal stem cell (BMSC) therapy can significantly improve the outcomes of rheumatoid arthritis (RA). This study aims to conduct a comprehensive analysis of the proteomics between BMSCs and BMSCs-Exos, and to further elucidate the potential therapeutic effect of BMSCs-Exos on RA, so as to establish a theoretical framework for the prevention and therapy of BMSCs-Exos on RA. The 4D label-free LC-MS/MS technique was used for comparative proteomic analysis of BMSCs and BMSCs-Exos. Collagen-induced arthritis (CIA) rat model was used to investigate the therapeutic effect of BMSCs-Exos on RA. Our results showed that some homology and differences were observed between BMSCs and BMSCs-Exos proteins, among which proteins highly enriched in BMSCs-Exos were related to extracellular matrix and extracellular adhesion. BMSCs-Exos can be taken up by chondrocytes, promoting cell proliferation and migration. In vivo results revealed that BMSCs-Exos significantly improved the clinical symptoms of RA, showing a certain repair effect on the injury of articular cartilage. In short, our study revealed, for the first time, that BMSCs-Exos possess remarkable efficacy in alleviating RA symptoms, probably through shuttling proteins related to cell adhesion and tissue repair ability in CIA rats, suggesting that BMSCs-Exos carrying expressed proteins may become a useful biomaterial for RA treatment.
Assuntos
Artrite Reumatoide , Exossomos , Células-Tronco Mesenquimais , Ratos , Animais , Exossomos/metabolismo , Cromatografia Líquida , Proteômica , Espectrometria de Massas em Tandem , Células-Tronco Mesenquimais/metabolismo , Artrite Reumatoide/terapia , Artrite Reumatoide/metabolismoRESUMO
Curcumin (CUR) is a kind of natural orange-yellow phenolic compound mainly extracted from the stems and roots of turmeric plants and other species in the genus Curcuma, furthermore, it is also the most important active ingredient exerting pharmacological functions in turmeric. In recent years, CUR has been frequently reported and has attracted widespread attention from scholars all over the world due to its numerous biological functions and good application prospects, such as anti-inflammatory, anticancer, antioxidant and providing lipid-lowering effects, etc. In addition, adding a certain dose of CUR to livestock and poultry feed is important for animal growth and development, which plays a key role in animal metabolism, reproduction, immunity and clinical health care. This review aims to summarize, based on the published papers and our own observations, the physical and chemical properties and the biological functions of the plant-derived bioactive ingredient CUR, especially regarding the latest research progress in regulating intestinal health as well as its current development and future application prospects in livestock and poultry as a novel feed additive, so as to provide theoretical and practical references for the further study of the application of CUR as a novel feed additive and a potential new antibiotic substitute, thereby improving the research field of plant-derived bioactive ingredients and promoting the healthy development of livestock and poultry.
Assuntos
Antineoplásicos , Curcumina , Animais , Antibacterianos , Anti-Inflamatórios/farmacologia , Antineoplásicos/farmacologia , Antioxidantes/química , Antioxidantes/farmacologia , Curcumina/química , Curcumina/farmacologia , Lipídeos , Gado , Polifenóis/farmacologia , Aves DomésticasRESUMO
Lead (Pb) is a toxic heavy metal pollutants and can damage male reproductive function. Selenium (Se) possesses an ability of antagonizing Pb toxicity. However, biological events in the process of Pb toxicity and mitigative effect of Se are not well understood. The aim of present research was to investigate potential mechanism of Se against Pb toxicity from the perspective of oxidative stress, heat shock response and autophagy in the spermatogonia and Leydig cell of chicken. The cells from one-day-old male Hyline chickens were treated with Se (0.5 µmol/L) and/or Pb (20 µmol/L) for 24 h, respectively. Cell viability, cell ultrastucture, Pb and Se concentrations, testosterone level, oxidative stress indicators and relative expression of heat shock proteins (HSPs) and autophagy-related genes were measured. The results showed that spermatogonia was more tolerant to Pb than Leydig cell; cell injury was confirmed via histological assessment, cell viability and testosterone level; oxidative stress was further indicated by the decrease of catalase, glutathione peroxidase, glutathione-s-transferase and superoxide dismutase activities and the increase of malondialdehyde and reactive oxygen species contents. Pb increased expression of HSPs (27, 40, 60, 70 and 90). Meanwhile Pb induced autophagy through up-regulation of autophagy-related proteins 5, Beclin 1, Dynein, light chain 3 (LC3)-I and LC3-II and down-regulation of mammalian target of rapamycin in two type cells of chicken. However, Se intervention mitigated the aforementioned alterations caused by Pb. In conclusion, Pb led to oxidative stress, which triggered heat shock response and autophagy; Se administration mitigated reproductive toxicity of Pb through strengthening antioxidant defense in the spermatogonia and Leydig cell of chicken.
Assuntos
Antioxidantes/farmacologia , Chumbo/toxicidade , Células Intersticiais do Testículo/efeitos dos fármacos , Selênio/farmacologia , Espermatogônias/fisiologia , Animais , Antioxidantes/metabolismo , Autofagia/efeitos dos fármacos , Catalase/metabolismo , Galinhas/metabolismo , Poluentes Ambientais/metabolismo , Glutationa Peroxidase/metabolismo , Proteínas de Choque Térmico/metabolismo , Chumbo/metabolismo , Células Intersticiais do Testículo/metabolismo , Masculino , Malondialdeído/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Espermatogônias/metabolismoRESUMO
PURPOSE: The study aimed to determine the effects of maternal low-protein (LP) diet on subcutaneous fat deposition of weaning piglets and the potential mechanism. METHODS: Sows were fed either a standard protein (SP, 15 and 18% crude protein) or a LP diet (50% protein levels of SP) throughout pregnancy and lactation. Subcutaneous fat and blood were sampled from male piglets at 28 days of age. Serum biochemical metabolites and hormone concentrations were detected with the enzymatic colorimetric methods. Serum-free amino acid (FAA) levels were measured by amino acid auto-analyzer. The mRNA and protein were measured by qRT-PCR and Western blot. RESULTS: Body weight, back fat thickness, triglycerides concentrations in subcutaneous fat tissue, and serum, as well as FFA concentrations were significantly reduced in LP piglets when compared with SP piglets. Further studies showed that mRNA and protein expression of acetyl-CoA carboxylase and fatty acid synthetase, two key enzymes of de novo lipogenesis, were significantly reduced in LP piglets, while mRNA expression and the lipolytic enzymes activities of lipolysis genes, adipose triglyceride lipase and hormone-sensitive lipase, were significantly increased. Furthermore, expression of autophagy-related gene 7 and autophagy maker gene microtubule-associated protein 1A/1B-light chain 3 (LC 3) as well as the conversion of LC3I to LC3II were significantly elevated, along with the expression of activating transcription factor-4 and eukaryotic translation initiation factor-2a. CONCLUSION: These results indicate that amino acid starvation-induced autophagy is involved in reduced subcutaneous fat deposition in maternal LP weaning piglets, demonstrating links between maternal protein restriction and offspring fat deposition.
Assuntos
Autofagia , Desenvolvimento Fetal , Lactação , Fenômenos Fisiológicos da Nutrição Materna , Deficiência de Proteína/fisiopatologia , Gordura Subcutânea/patologia , Magreza/etiologia , Adiposidade , Animais , Biomarcadores/sangue , Biomarcadores/metabolismo , China , Cruzamentos Genéticos , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Regulação Enzimológica da Expressão Gênica , Metabolismo dos Lipídeos , Masculino , Gravidez , Distribuição Aleatória , Gordura Subcutânea/enzimologia , Gordura Subcutânea/metabolismo , Sus scrofa , Magreza/sangue , Magreza/metabolismo , Magreza/patologia , Desmame , Aumento de PesoRESUMO
OBJECTIVES: Wnt signalling has been implicated in activating a fibrogenic programme in fibroblasts in systemic sclerosis (SSc). Porcupine is an O-acyltransferase required for secretion of Wnt proteins in mammals. Here, we aimed to evaluate the antifibrotic effects of pharmacological inhibition of porcupine in preclinical models of SSc. METHODS: The porcupine inhibitor GNF6231 was evaluated in the mouse models of bleomycin-induced skin fibrosis, in tight-skin-1 mice, in murine sclerodermatous chronic-graft-versus-host disease (cGvHD) and in fibrosis induced by a constitutively active transforming growth factor-ß-receptor I. RESULTS: Treatment with pharmacologically relevant and well-tolerated doses of GNF6231 inhibited the activation of Wnt signalling in fibrotic murine skin. GNF6231 ameliorated skin fibrosis in all four models. Treatment with GNF6231 also reduced pulmonary fibrosis associated with murine cGvHD. Most importantly, GNF6231 prevented progression of fibrosis and showed evidence of reversal of established fibrosis. CONCLUSIONS: These data suggest that targeting the Wnt pathway through inhibition of porcupine provides a potential therapeutic approach to fibrosis in SSc. This is of particular interest, as a close analogue of GNF6231 has already demonstrated robust pathway inhibition in humans and could be available for clinical trials.
Assuntos
Aminopiridinas/uso terapêutico , Proteínas de Membrana/antagonistas & inibidores , Piperazinas/uso terapêutico , Esclerodermia Localizada/prevenção & controle , Escleroderma Sistêmico/prevenção & controle , Pele/patologia , Via de Sinalização Wnt/efeitos dos fármacos , Aciltransferases , Aminopiridinas/farmacologia , Animais , Bleomicina , Modelos Animais de Doenças , Progressão da Doença , Feminino , Fibrose , Doença Enxerto-Hospedeiro/complicações , Camundongos Endogâmicos BALB C , Piperazinas/farmacologia , Proteínas Serina-Treonina Quinases/genética , Fibrose Pulmonar/etiologia , Fibrose Pulmonar/prevenção & controle , Receptor do Fator de Crescimento Transformador beta Tipo I , Receptores de Fatores de Crescimento Transformadores beta/genética , Esclerodermia Localizada/etiologia , Esclerodermia Localizada/metabolismo , Escleroderma Sistêmico/induzido quimicamente , Escleroderma Sistêmico/metabolismo , Escleroderma Sistêmico/patologia , Pele/metabolismo , Fator de Crescimento Transformador beta/metabolismoRESUMO
To investigate the effects of maternal dietary protein restriction on offspring Fe metabolism, twenty-four second-parity Landrace×Yorkshire sows were randomly allocated to standard-protein (SP) and low-protein (LP) groups. The SP sows were fed diets containing 15 and 18 % crude protein throughout pregnancy and lactation, respectively, whereas the LP sows were subjected to 50 % dietary protein restriction. Offspring birth weight was not affected, but the body weight at weaning (P=0·06) and average daily gain (P=0·01) of the female piglets were significantly decreased. Serum Fe level in the LP piglets was markedly decreased at weaning, especially in males (P=0·03). Serum ferritin level (P=0·08) tended to be lower, yet serum transferrin was greatly higher (P=0·01) in male weaning piglets of the LP group. Duodenal expression of the divalent metal transporter 1 (DMT1) and ferroportin (FPN) was surprisingly reduced (P<0·05) at the level of protein, but not at the mRNA level, in male weaning piglets of the LP group. Male weaning piglets born to the LP sows exhibited higher hepatic hepcidin levels (P=0·09), lower hepatic expression of transferrin (P<0·01) and transferrin receptor 1 (P<0·05) at the level of mRNA. However, no significant differences were observed for hepatic Fe storage, ferritin, transferrin and transferrin receptor 1 protein expression in male weaning piglets of the two groups. These results indicate that maternal protein restriction during pregnancy and lactation influences growth of female offspring at weaning, reduces duodenal expression of Fe transporters (DMT1 and FPN) and decreases serum Fe level in male weaning piglets.
Assuntos
Anemia Ferropriva/veterinária , Proteínas de Transporte de Cátions/metabolismo , Dieta com Restrição de Proteínas/veterinária , Duodeno/metabolismo , Mucosa Intestinal/metabolismo , Fenômenos Fisiológicos da Nutrição Materna , Anemia Ferropriva/sangue , Anemia Ferropriva/etiologia , Anemia Ferropriva/metabolismo , Animais , Proteínas de Transporte de Cátions/genética , Dieta com Restrição de Proteínas/efeitos adversos , Feminino , Hepcidinas/genética , Hepcidinas/metabolismo , Ferro/sangue , Ferro/metabolismo , Lactação , Fígado/metabolismo , Masculino , Gravidez , Receptores da Transferrina/genética , Receptores da Transferrina/metabolismo , Caracteres Sexuais , Suínos , Doenças dos Suínos/sangue , Doenças dos Suínos/etiologia , Doenças dos Suínos/metabolismo , Transferrina/análise , Transferrina/genética , Transferrina/metabolismo , Desmame , Aumento de PesoRESUMO
PURPOSE: In this study, we sought to investigate the effects of maternal betaine supplementation on the expression and regulation of GALK1 gene in the liver of neonatal piglets. METHODS: Sixteen sows of two groups were fed control or betaine-supplemented diets (3 g/kg), respectively, throughout the pregnancy. Newborn piglets were individually weighed immediately after birth, and one male piglet close to mean body weight from the same litter was selected and killed before suckling. Serum samples of newborn piglets were analyzed for biochemical indexes, hormone and amino acid levels. Liver samples were analyzed for GALK1 expression by real-time PCR and western blotting, while GALK1 regulational mechanism was analyzed by methylated DNA immunoprecipitation, chromatin immunoprecipitation and microRNAs expression. RESULTS: Betaine-exposed neonatal piglets had lower serum concentration of galactose, which was associated with significantly down-regulated hepatic GALK1 expression. The repression of GALK1 mRNA expression was associated with DNA hypermethylation and more enriched repression histone mark H3K27me3 on its promoter. Binding sites of SP1, GR and STAT3 were predicted on GALK1 promoter, and decreased SP1 protein content and lower SP1 binding to GALK1 promoter were detected in the liver of betaine-exposed piglets. Furthermore, the expression of miRNA-149 targeting GALK1 was up-regulated in the liver of betaine-exposed piglets, along with elevated miRNAs-processing enzymes Dicer and Ago2. CONCLUSIONS: Our results suggest that maternal dietary betaine supplementation during gestation suppresses GALK1 expression in the liver of neonatal piglets, which involves complex gene regulation mechanisms including DNA methylation, histone modification, miRNAs expression and SP1-mediated transcriptional modulation.
Assuntos
Betaína/administração & dosagem , Repressão Epigenética , Galactoquinase/genética , Fator de Transcrição Sp1/metabolismo , Aminoácidos/sangue , Animais , Animais Recém-Nascidos , Betaína/sangue , Biomarcadores/sangue , Imunoprecipitação da Cromatina , Metilação de DNA/efeitos dos fármacos , Dieta , Suplementos Nutricionais , Feminino , Galactoquinase/metabolismo , Galactose/metabolismo , Regulação da Expressão Gênica , Insulina/sangue , Fígado/metabolismo , Masculino , MicroRNAs/genética , MicroRNAs/metabolismo , Tamanho do Órgão/efeitos dos fármacos , Regiões Promotoras Genéticas , Reprodutibilidade dos Testes , Fator de Transcrição Sp1/genética , SuínosRESUMO
A growing number of agents targeting ligand-induced Wnt/ß-catenin signaling are being developed for cancer therapy. However, clinical development of these molecules is challenging because of the lack of a genetic strategy to identify human tumors dependent on ligand-induced Wnt/ß-catenin signaling. Ubiquitin E3 ligase ring finger 43 (RNF43) has been suggested as a negative regulator of Wnt signaling, and mutations of RNF43 have been identified in various tumors, including cystic pancreatic tumors. However, loss of function study of RNF43 in cell culture has not been conducted, and the functional significance of RNF43 mutations in cancer is unknown. Here, we show that RNF43 inhibits Wnt/ß-catenin signaling by reducing the membrane level of Frizzled in pancreatic cancer cells, serving as a negative feedback mechanism. Inhibition of endogenous Wnt/ß-catenin signaling increased the cell surface level of Frizzled. A panel of 39 pancreatic cancer cell lines was tested for Wnt dependency using LGK974, a selective Porcupine inhibitor being examined in a phase 1 clinical trial. Strikingly, all LGK974-sensitive lines carried inactivating mutations of RNF43. Inhibition of Wnt secretion, depletion of ß-catenin, or expression of wild-type RNF43 blocked proliferation of RNF43 mutant but not RNF43-wild-type pancreatic cancer cells. LGK974 inhibited proliferation and induced differentiation of RNF43-mutant pancreatic adenocarcinoma xenograft models. Our data suggest that mutational inactivation of RNF43 in pancreatic adenocarcinoma confers Wnt dependency, and the presence of RNF43 mutations could be used as a predictive biomarker for patient selection supporting the clinical development of Wnt inhibitors in subtypes of cancer.
Assuntos
Carcinoma Ductal Pancreático/metabolismo , Proteínas de Ligação a DNA/metabolismo , Mutação , Proteínas Oncogênicas/metabolismo , Neoplasias Pancreáticas/metabolismo , Proteínas Wnt/metabolismo , beta Catenina , Aciltransferases , Antineoplásicos/farmacologia , Carcinoma Ductal Pancreático/tratamento farmacológico , Carcinoma Ductal Pancreático/genética , Carcinoma Ductal Pancreático/patologia , Linhagem Celular Tumoral , Ensaios Clínicos Fase I como Assunto , Proteínas de Ligação a DNA/genética , Receptores Frizzled/genética , Receptores Frizzled/metabolismo , Células HEK293 , Humanos , Proteínas de Membrana/antagonistas & inibidores , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Proteínas Oncogênicas/genética , Neoplasias Pancreáticas/tratamento farmacológico , Neoplasias Pancreáticas/genética , Ubiquitina-Proteína Ligases , Proteínas Wnt/genética , Via de Sinalização WntRESUMO
Wnt signaling is one of the key oncogenic pathways in multiple cancers, and targeting this pathway is an attractive therapeutic approach. However, therapeutic success has been limited because of the lack of therapeutic agents for targets in the Wnt pathway and the lack of a defined patient population that would be sensitive to a Wnt inhibitor. We developed a screen for small molecules that block Wnt secretion. This effort led to the discovery of LGK974, a potent and specific small-molecule Porcupine (PORCN) inhibitor. PORCN is a membrane-bound O-acyltransferase that is required for and dedicated to palmitoylation of Wnt ligands, a necessary step in the processing of Wnt ligand secretion. We show that LGK974 potently inhibits Wnt signaling in vitro and in vivo, including reduction of the Wnt-dependent LRP6 phosphorylation and the expression of Wnt target genes, such as AXIN2. LGK974 is potent and efficacious in multiple tumor models at well-tolerated doses in vivo, including murine and rat mechanistic breast cancer models driven by MMTV-Wnt1 and a human head and neck squamous cell carcinoma model (HN30). We also show that head and neck cancer cell lines with loss-of-function mutations in the Notch signaling pathway have a high response rate to LGK974. Together, these findings provide both a strategy and tools for targeting Wnt-driven cancers through the inhibition of PORCN.
Assuntos
Proteínas de Membrana/antagonistas & inibidores , Neoplasias/tratamento farmacológico , Pirazinas/farmacologia , Piridinas/farmacologia , Via de Sinalização Wnt/efeitos dos fármacos , Aciltransferases , Animais , Proteína Axina/antagonistas & inibidores , Western Blotting , Linhagem Celular Tumoral , Clonagem Molecular , Ensaios de Triagem em Larga Escala , Humanos , Camundongos , Mutagênese , Fosforilação/efeitos dos fármacos , Pirazinas/uso terapêutico , Piridinas/uso terapêutico , Ensaio Radioligante , Ratos , Receptores Notch/genética , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
BACKGROUND: We have shown previously that microvesicle (MV)-delivered miR-130b (miR-130b-MV) is able to target PPAR-γ and subsequently reduce the lipid accumulation in vitro. However, the in vivo effect of miR-130b on fat deposition and glucose homeostasis remains unknown. RESULTS: Three-week-old C57BL/6 mice were fed a high-fat diet for 8 weeks and then intravenously injected with MV-packaged scrambled control microRNA (miRNA) or miR-130b every other day for 10 days. Glucose tolerance test was performed and body weight, epididymal fat weight, as well as the expression of lipid metabolic genes were determined. We showed that mice fed on high-fat diet for 8 weeks demonstrated significantly higher body weight, elevated blood glucose and impaired glucose tolerance. miR-130b-MV injection significantly reduced body weight and epididymal fat weight and partly restored glucose tolerance. miR-130b expression was significantly increased in the epididymal fat after miR-130b-MV injection while the protein content of its target gene PPAR-γ was significantly suppressed, together with a significant up-regulation of the lipolysis genes, hormone sensitive lipase, monoglyceride lipase and leptin. Moreover, miR-130b-MV injection increased the expression of miR-378a and miR-378-3p that are reported to participate in the regulation of fat deposition. CONCLUSION: Our results indicate that miR-130b-MV is able to reduce the epididymal fat deposition and partly restore glucose tolerance, through translational repression of PPAR-γ in a high-fat diet-induced obese mouse model.
Assuntos
Gorduras na Dieta/efeitos adversos , Portadores de Fármacos/farmacologia , MicroRNAs/farmacologia , Obesidade/tratamento farmacológico , PPAR gama/biossíntese , Biossíntese de Proteínas/efeitos dos fármacos , Animais , Gorduras na Dieta/farmacologia , Portadores de Fármacos/química , Lipólise/efeitos dos fármacos , Masculino , Camundongos , MicroRNAs/química , Obesidade/induzido quimicamente , Obesidade/metabolismoRESUMO
PURPOSE: We tested the hypothesis that maternal low-protein (LP) diet during gestation and lactation can program myostatin (MSTN) signaling and protein synthesis in skeletal muscle of offspring at weaning stage (35 days). METHODS: Fourteen Meishan sows were fed either LP or standard-protein diets throughout gestation and lactation, male offspring piglets were killed at weaning stage and longissimus dorsi (LD) muscles were taken. The cross-sectional areas (CSA) of LD muscles were measured by hematoxylin and eosin staining. The levels of free amino acids in plasma were measured by amino acid auto-analyzer. Proteins and mRNA were determined by Western blot and RT-qPCR, respectively. RESULTS: Body weight, LD muscle weight and the myofiber CSA were significantly decreased (P < 0.05) in LP piglets; meanwhile, the concentration of branched-chain amino acids was also significantly decreased (P < 0.001). MSTN protein content tended to be higher (P = 0.098) in LP piglets, while the expression of MSTN receptors, activin type II receptor-beta and transforming growth factor type-beta type I receptor kinase, was significantly up-regulated (P < 0.05). Furthermore, p38 mitogen-activated protein kinase, the downstream signaling factor of MSTN, was also enhanced significantly (P < 0.05). In addition, key factors of translation initiation, phosphorylated eukaryotic initiation factor 4E and the 70 kDa ribosomal protein S6 kinase, were significantly decreased (P < 0.05) in LP piglets. CONCLUSIONS: Our results suggest that maternal LP diet during gestation and lactation affects MSTN signaling and protein synthesis in skeletal muscle of offspring at weaning stage.
Assuntos
Dieta com Restrição de Proteínas/efeitos adversos , Fenômenos Fisiológicos da Nutrição Materna , Proteínas Musculares/biossíntese , Miostatina/metabolismo , Efeitos Tardios da Exposição Pré-Natal , Sus scrofa , Aminoácidos/sangue , Aminoácidos de Cadeia Ramificada/sangue , Fenômenos Fisiológicos da Nutrição Animal , Animais , Fator de Iniciação 4E em Eucariotos/análise , Feminino , Lactação , Masculino , Músculo Esquelético/crescimento & desenvolvimento , Tamanho do Órgão , Gravidez , Proteínas Quinases S6 Ribossômicas 70-kDa/análise , Transdução de Sinais , DesmameRESUMO
Obesity is a worldwide epidemic, and a risk factor for cardiovascular disease and type 2 diabetes. Consequently, the development of safe and effective anti-obesity drugs is an area of ongoing clinical interest. MicroRNAs play a vital role in anti-obesity by inhibiting the expression of genes involved in adipogenesis and lipogenesis. However, the clinical application of miRNAs has been limited by a lack of appropriate delivery systems. The discovery of microvesicles (MVs) has shed new light on the search for more efficient drug transport tools. In a previous study, we demonstrated that miRNA-130b suppressed fat deposition by inhibiting PPAR-g expression. In order to demonstrate whether miRNA-130b can be packaged into MVs and function as an endogenous form of miRNA-130b in recipient cells, we transfected HeLa-229 cells with plasmid to overexpress miRNA-130b. This enabled HeLa-229 cells to selectively package miRNA-130b into MVs and actively secrete the miRNA-130b enriched MVs into the culture media. We further verified that MVs enriched with miRNA-130b contain elevated concentrations of Argonaute 2 and heat shock protein 90a which are known to protect the circulating miRNAs from degradation. Exposure of primary cultured porcine adipocytes to purified, miRNA-130b-enriched MVs resulted in a significant down-regulation of PPAR-g expression which was associated with reduced adipogenesis and lipogenesis. Taken together, our results suggest that MVs may provide an effective transport systems for the deliver of miRNAs for therapeutic use. We also showed that MV-shuttled miRNA-130b inhibited adipogenesis and lipogenesis, and reduced fat deposition in recipient adipocytes by targeting PPAR-g.
Assuntos
Adipócitos/metabolismo , Gorduras/metabolismo , MicroRNAs/metabolismo , PPAR gama/metabolismo , Animais , Proteínas Argonautas/genética , Proteínas Argonautas/metabolismo , Regulação da Expressão Gênica , Proteínas de Choque Térmico HSP90/genética , Proteínas de Choque Térmico HSP90/metabolismo , Células HeLa , Humanos , MicroRNAs/genética , PPAR gama/antagonistas & inibidores , PPAR gama/genética , Plasmídeos , Reação em Cadeia da Polimerase em Tempo Real , SuínosRESUMO
This study aimed to explore alterations in growth performance, glycolipid metabolism disorders, intestinal mucosal barrier, cecal microbiota community, and metabolites in a chronic corticosterone (CORT)-induced stress (CCIS) broiler model. Results showed that compared with control (CON) broilers, in CCIS broilers: (i) the final body weight (BW), BW gain, and average daily gain were significantly reduced. (ii) The glycolipid metabolism disorder and impairement of intestinal immune barrier and physical barrier function were observed. (iii) Diversity and richness of cecal microbiota were obviously increased. From phylum to genus level, the abundances of Firmicutes and Faecalibacterium were significantly decreased, while the abundances of Proteobacteria, RuminococcaceaeUCG-005, and Escherichia coli (Shigella) were significantly increased. Microbial network analysis and function pathways prediction showed that cecal microbiota was mainly concentrated in translation, metabolism, nucleotide metabolism, and endocrine system. (iv) The main differential metabolites identified include steroids and their derivatives, amino acids, fatty acids, and carbohydrates; among which 37 metabolites were significantly upregulated, while 27 metabolites were significantly downregulated. These differential metabolites were mainly enriched in pathways related to steroid hormone biosynthesis and tyrosine metabolism. (v) Correlation between cecal microbiota and glycolipid metabolism indexes showed that BW and total cholesterol (TC) were positively correlated with Christensenellaceae_R.7_group and Escherichia_Shigella, respectively. Furthermore, the downregulated Faecalibacterium and Christensenellaceae were negatively correlated with the upregulated differentially expressed metabolites. These findings suggested that CCIS altered cecal microbiota composition and metabolites, which led to glycolipid metabolism disorder and impaired the nutritional metabolism and immune homeostasis, providing a theoretical basis for efforts to eliminate the harm of chronic stress to human health and animal production. IMPORTANCE: The study aimed to determine the influence of altered intestinal mucosal barrier, cecum flora community, and metabolites on anti-growth performance, glycolipid metabolism disorders of chronic corticosterone (CORT)-induced stress (CCIS) broilers. Compared with control (CON) broilers, in CCIS broilers: (i) anti-growth performance, glycolipid metabolism disorder, and impaired intestinal immune barrier and physical barrier function were observed. (ii) From phylum to genus level, the abundances of Firmicutes and Faecalibacterium were decreased; whereas, the abundances of Proteobacteria, RuminococcaceaeUCG-005, and Escherichia coli (Shigella) were increased. (iii) Differential metabolites in cecum were mainly enriched in steroid hormone biosynthesis and tyrosine metabolism. (iv) Body weight (BW) and total cholesterol (TC) were positively correlated with Christensenellaceae_R.7_group and Escherichia_Shigella, respectively, while downregulated Faecalibacterium and Christensenellaceae were negatively correlated with upregulated metabolites. Our findings suggest that CCIS induces anti-growth performance and glycolipid metabolism disorder by altering cecum flora and metabolites, providing a theoretical basis for efforts to eliminate the effect of chronic stress on human health and animal production.
Assuntos
Bactérias , Ceco , Galinhas , Corticosterona , Microbioma Gastrointestinal , Glicolipídeos , Mucosa Intestinal , Estresse Fisiológico , Animais , Galinhas/microbiologia , Galinhas/crescimento & desenvolvimento , Corticosterona/metabolismo , Glicolipídeos/metabolismo , Mucosa Intestinal/metabolismo , Mucosa Intestinal/microbiologia , Ceco/microbiologia , Ceco/metabolismo , Bactérias/classificação , Bactérias/metabolismo , Bactérias/genética , Bactérias/isolamento & purificaçãoRESUMO
Chronic stress refers to the activation of the hypothalamic-pituitary-adrenal (HPA) axis and elevated blood contents of ACTH and corticosterone (CORT), exhibiting significant adverse effects on health outcomes. Currently, natural polyphenol compounds are increasingly being explored as potential therapeutic agents and have been considered as a treatment option for a variety of stress-induced diseases. Curcumin (CUR) is the main substance in Curcuma longa (Zingiberacea) rhizome that has strong health-beneficial properties. The study aimed to assess the potential protective effects of CUR on hepatic oxidative stress damage and abnormal lipid deposition in a chronic CORT-induced stress (CCIS) model in broilers. One hundred and twenty experimental broilers were randomly divided into 1) control group (CON), 2) CUR group (200â¯mg/kg feed), 3) CORT group (4â¯mg/kg BW CORT) and 4) CORT+CUR group (200â¯mg/kg feed plus 4â¯mg/kg BW CORT). The liver histology, glycolipid metabolism and oxidative stress were determined. In addition, qPCR was performed to identify shifts in genes expression. Compared with CON group, broilers under CCIS showed a decreased body weight, body weight gain and average daily gain, while dietary CUR significantly reversed these adverse effects. Furthermore, the plasma contents of TCH, TG, HDL-C, LDL-C, TP, GLB and AST were all significantly increased in CCIS broilers, while dietary CUR obviously alleviated the increase of TCH, HDL-C, LDL-C and AST, and relieved the hepatic lipid deposition disorder and liver injury. Moreover, CCIS significantly increased the contents of MDA in both liver and plasma, and decreased the content of plasma SOD, while CUR obviously reversed these changes, showing reduced oxidative stress damage. Finally, the mRNA expressions of FAS, ACC, SCD and the protein level of PPAR-γ were significantly increased, meanwhile the mRNA expression of lipolytic genes ACOX1, ATGL and CPT as well as two major intracellular antioxidant enzymes SOD1 and GPX1 were obviously decreased, while CUR effectively reversed these effects. These results showed that dietary CUR effectively alleviated CCIS-induced body weight loss, hepatic oxidative damage and lipid deposition disorder, suggesting the possible therapeutic effectiveness of CUR against hepatic damage and function abnormality caused by CCIS.
RESUMO
To investigate whether the effect of maternal dietary protein on offspring lipid metabolism is mediated by microRNA (miRNA), fourteen Meishan sows were fed either low-protein (LP, half of standard protein (SP) level, n 7) or SP (n 7) diets throughout gestation and lactation periods. PPAR-γ and CCAAT/enhancer-binding protein-ß (C/EBP-ß) protein expression was evaluated. The expression of miRNA predicted to directly target PPAR-γ and C/EBP-ß in the subcutaneous fat of offspring at weaning age was determined, and the functions of these potential miRNA were verified. The results showed that piglet body weight and back fat thickness were significantly decreased in the LP group compared with the SP group (P<0·05). The protein level of PPAR-γ was significantly decreased and C/EBP-ß protein expression was also decreased, though not significantly (P=0·056), in the subcutaneous fat of the LP group. Furthermore, miRNA expression analysis showed that miR-130b, targeting the PPAR-γ 3'-untranslated region (UTR), and miR-374b, targeting the C/EBP-ß 3'-UTR, were significantly increased in the LP group compared with the SP group; other candidate regulatory miRNA were expressed similarly in both groups. Dual luciferase activity assay results indicated that miR-130b directly recognised and bound to the 3'-UTR of PPAR-γ and thereby suppressed PPAR-γ gene expression. Similar results were found for miR-374b and the 3'-UTR of C/EBP-ß. The present study showed that miR-130b and miR-374b are involved in the effect of maternal dietary protein on offspring lipid metabolism in pigs. These results shed new light on our understanding of the maternal effect on offspring lipid deposition.
Assuntos
Tecido Adiposo/efeitos dos fármacos , Peso Corporal/efeitos dos fármacos , Dieta/veterinária , Proteínas Alimentares/farmacologia , Metabolismo dos Lipídeos/efeitos dos fármacos , MicroRNAs/metabolismo , Fenômenos Fisiológicos da Nutrição Pré-Natal , Fenômenos Fisiológicos da Nutrição Animal , Animais , Peso Corporal/genética , Proteína beta Intensificadora de Ligação a CCAAT/genética , Proteína beta Intensificadora de Ligação a CCAAT/metabolismo , Dieta com Restrição de Proteínas/veterinária , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Lactação , Metabolismo dos Lipídeos/genética , Mães , PPAR gama/genética , PPAR gama/metabolismo , Gravidez , Efeitos Tardios da Exposição Pré-Natal , SuínosRESUMO
The study aimed to investigate bone marrow mesenchymal stem cells (BMSCs) and extracellular signal-regulated kinase (ERK) gene-modified BMSCs (ERK-BMSCs) transplantation in ameliorating cognitive deficits in Parkinson's disease (PD). The PD rat model was built by 6-hydroxydopamine (6-OHDA) injection into the right striatum for 8 weeks, then successful PD rats were randomly divided into three groups and respectively transplanted in the same position of striatum as modeling with PBS, BMSCs and ERK-BMSCs for another 8 weeks. The 6-OHDA-induced PD rat model was successfully established, as demonstrated by reduced active avoidance response (AAR) times, percentage of time exploring in the light area (Ltime%) and platform quadrant time (PQT), as well as p-ERK expression. Compared with PBS rats, both BMSCs and ERK-BMSCs transplantation significantly reduced the left turn number, while increased AAR, Ltime%, PQT and p-ERK expression, suggesting improved cognitive abilities through restoring p-ERK expression. In addition, ERK-BMSCs injection exhibited higher therapeutic efficacy against cognitive deficits compared with BMSCs injection. These results demonstrated that BMSCs transplantation ameliorated cognitive deficits, and ERK-BMSCs exerted synergistic effects, which may prove beneficial against cognitive impairments in PD.
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Disfunção Cognitiva , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais , Doença de Parkinson , Ratos , Animais , Doença de Parkinson/terapia , Doença de Parkinson/metabolismo , Oxidopamina/toxicidade , Oxidopamina/metabolismo , Ratos Sprague-Dawley , Transplante de Células-Tronco Mesenquimais/métodos , Células-Tronco Mesenquimais/metabolismo , Disfunção Cognitiva/terapia , Disfunção Cognitiva/metabolismo , Cognição , Células da Medula Óssea , Transplante de Medula ÓsseaRESUMO
Through scaffold morphing of a known Smoothened antagonist Antag691, a series of novel phenyl imidazole derivatives were developed. Structure-activity-relationship studies and lead optimization led to the discovery of potent, selective and orally bioavailable Smoothened antagonist 19 that is suitable for in vivo studies.
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Desenho de Fármacos , Imidazóis/síntese química , Imidazóis/farmacocinética , Receptores Acoplados a Proteínas G/antagonistas & inibidores , Administração Oral , Animais , Antineoplásicos/síntese química , Antineoplásicos/química , Antineoplásicos/farmacocinética , Antineoplásicos/farmacologia , Área Sob a Curva , Humanos , Imidazóis/química , Imidazóis/farmacologia , Concentração Inibidora 50 , Camundongos , Ligação Proteica/efeitos dos fármacos , Ratos , Receptor Smoothened , Relação Estrutura-AtividadeRESUMO
BACKGROUND: Rheumatoid arthritis (RA) is an autoimmune disease. Bone marrow mesenchymal stem cells (BMSCs) have multilineage differentiation and anti-inflammatory potential, and small interfering RNAs (siRNAs) can inhibit the target gene expression, which make them suitable for ameliorating RA. The current study was aimed to explore the effect and potential mechanisms of siRNAs targeting IL-1ß/TNF-α combined with BMSCs transplantation in ameliorating RA in rats. METHODS: Collagen-induced arthritis (CIA) model rats were randomly divided into five groups: PBS (Model control group), methotrexate (Positive drug treatment group), BMSCs (BMSCs transplantation group), siRNA (IL-1ß/TNF-α siRNAs injection group), siRNA + BMSCs (Both IL-1ß/TNF-α siRNAs injection and BMSCs transplantation group). After treatment for 0, 7, 14, 21, 28 days, the ameliorating effect was comprehensively assessed through results of the body weight, toe swelling value, the immobility time of forced swimming, the serum concentrations of IL-1ß and TNF-α, knee joint DR-X imaging and pathological analysis as well as of IL-1ß, TNF-α and NF-κB mRNA expression in spleen tissue. Furthermore, the potential underlying mechanism involving the NF-κB signaling pathways was also explored. RESULTS: Compared with the PBS group, BMSCs, siRNA, siRNA + BMSCs treatment groups showed significant lower toe swelling value, immobility time, spleen index, serum contents of IL-1ß and TNF-α. In addition, the DR-X results showed that the knee carton surface tended to smoothing without bone hyperplasia, suggesting that these three treatments were all able to successfully ameliorate RA symptoms. In addition, compared with the PBS group, the protein expression of p-NF-κB-p65 was significantly reduced in the knees of siRNA + BMSCs rats. BMSCs labeled with BrdU were also found in the knees of rats. Moreover, the mRNA expression of IL-1ß, TNF-α and NF-κB-P65 in spleen tissue of siRNA + BMSCs rats were all significantly inhibited. CONCLUSIONS: Our results demonstrated for the first time that siRNA + BMSCs was able to ameliorate RA inflammation by inhibiting the activation of NF-κB signaling pathways and reducing the erosion of articular cartilage, and siRNA + BMSCs treatment showed synergism effects in helping ameliorating the inflammation and cartilage repair of RA rats. Therefore, the results of our present study provide a new idea for gene and stem cell therapy for RA.
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The misuse and abuse of antibiotics in livestock and poultry seriously endanger both human health and the continuously healthy development of the livestock and poultry breeding industry. Plant-derived bioactive compounds (curcumin, capsaicin, quercetin, resveratrol, catechin, lignans, etc.) have been widely studied in recent years, due to their extensive pharmacological functions and biological activities, such as anti-inflammatory, antioxidant, antistress, antitumor, antiviral, lowering blood glucose and lipids, and improving insulin sensitivity. Numerous studies have demonstrated that plant-derived bioactive compounds are able to enhance the host's ability to resist or diminish diseases by regulating the abundance of its gut microbiota, achieving great potential as a substitute for antibiotics. Recent developments in both humans and animals have also highlighted the major contribution of gut microbiota to the host's nutrition, metabolism, immunity, and neurological functions. Changes in gut microbiota composition are closely related to the development of obesity and can lead to numerous metabolic diseases. Mounting evidence has also demonstrated that plant-derived bioactive compounds, especially curcumin, can improve intestinal barrier function by regulating intestinal flora. Furthermore, bioactive constituents can be also directly metabolized by intestinal flora and further produce bioactive metabolites by the interaction between the host and intestinal flora. This largely enhances the protective effect of bioactive compounds on the host intestinal and whole body health, indicating that the bidirectional regulation between bioactive compounds and intestinal flora has great application potential in maintaining the host's intestinal health and preventing or treating various diseases. This review mainly summarizes the latest research progress in the bioregulation between gut microbiota and plant-derived bioactive compounds, together with its application potential in humans and animals, so as to provide theoretical support for the application of plant-derived bioactive compounds as new feed additives and potential substitutes for antibiotics in the livestock and poultry breeding industry. Overall, based on this review, it can be concluded that plant-derived bioactive compounds, by modulating gut microbiota, hold great promise toward the healthy development of both humans and animal husbandry.
Assuntos
Curcumina , Microbioma Gastrointestinal , Resistência à Insulina , Animais , Humanos , Curcumina/farmacologia , Intestinos , ObesidadeRESUMO
Exosomes (EXs) are the major types of extracellular vesicles (EVs) of 30-100 nm diameter that can be secreted by most cells to the extracellular environment. EXs transport endogenous cargoes (proteins, lipids, RNAs, etc.) to target cells and thereby triggers the release of these bioactive components, which then play important roles in regulating numerous biological processes under both physiological and pathological conditions. Throughout the studies in recent years, growing evidences have shown that EXs-derived non-coding RNAs (EXs-ncRNAs) are emerging as key players in cell-to-cell communication between adipose tissue and other related tissues in obesity and metabolic diseases. In this review, we will summarize the recent findings about EXs-ncRNAs, especially focus on the following aspects: 1) the biogenesis of EXs and emerging roles of EXs-ncRNAs, 2) the role of EXs-ncRNAs (EXs-miRNAs, EXs-lncRNAs, EXs-circRNAs, etc.) that were secreted by adipose-related tissues in promoting the differentiation of preadipocytes into mature and fully functional adipocytes, and 3) the crosstalk between the adipose tissue derived EXs-ncRNAs and the development of insulin resistance, obesity and various cancers. This review aims to reveal the emerging roles and mechanisms of EXs-ncRNAs in the mutual regulation of adipose tissue and its related tissues in obesity and metabolic diseases, so as to provide references for elucidating the etiology of obesity and related metabolic diseases and screening novel therapeutic targets.