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1.
Biomed Pharmacother ; 118: 109363, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31545277

RESUMO

OBJECTIVE: Alveolar epithelial barrier dysfunction in response to inflammatory reaction contributes to pulmonary edema in acute lung injury(ALI).Irisin,a newly-found myokine,exerts the anti-inflammatory effects. This study aims to investigate the protective effects of irisin on lipopolysaccharide (LPS)-induced ALIin vivo and in vitro, and to explore its underlying mechanism. METHODS: Male SD rats and A549 cells were divided into 4 groups: control group, LPS group, Irisin pretreated group, and Irisin/Compound C(a special inhibitor of AMPK)-treated group. The ALI model was established by intravenous injection of LPS in rats, and LPS challenge in A549 cells. Pulmonary specimens were harvested for microscopic examination of the pathological changes, and the expression of AMPK,SIRT1,NF-κB, p66Shc and caspase-3 in lung tissues. The pulmonary permeability were examined by wet/dry lung weight ratio(W/D) and lung permeability index(LPI). The apoptotic index, and the expression of tumor necrosis factor-α(TNF-α), interleukin-1ß(IL-1ß), monocyte chemoattractant activating protein-1 (MCP-1), tight junctions (occludin,ZO-1) were determined both in lung tissue and A549 cells. RESULTS: Irisin alleviated lung histological changes and decreased pulmonary microvascular permeability in LPS-induced rats. Irisin up-regulated the expression of occludin, ZO-1,AMPK,SIRT1, down-regulated the expression of TNF-α,IL-1ß,MCP-1,NF-κB, p66Shc caspase-3, and decreased the apoptotic index in LPS-induced rats and A549 cells. All these protective effects of irisin could be reversed by Compound C. CONCLUSION: Irisin improved LPS-induced alveolar epithelial barrier dysfunction via suppressing inflammation and apoptosis, and this protective effect might be mediated by activating AMPK/SIRT1 pathways.


Assuntos
Proteínas Quinases Ativadas por AMP/metabolismo , Lesão Pulmonar Aguda/fisiopatologia , Epitélio/fisiopatologia , Fibronectinas/uso terapêutico , Pulmão/fisiopatologia , Transdução de Sinais , Sirtuína 1/metabolismo , Células A549 , Lesão Pulmonar Aguda/patologia , Animais , Apoptose/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/patologia , Células Epiteliais/ultraestrutura , Epitélio/ultraestrutura , Fibronectinas/farmacologia , Humanos , Inflamação/patologia , Pulmão/patologia , Pulmão/ultraestrutura , Masculino , Permeabilidade , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos , Junções Íntimas/efeitos dos fármacos , Junções Íntimas/metabolismo , Regulação para Cima/efeitos dos fármacos
2.
Lipids Health Dis ; 18(1): 166, 2019 Aug 31.
Artigo em Inglês | MEDLINE | ID: mdl-31470857

RESUMO

BACKGROUND: Atherosclerosis is an inflammatory process involving activation of monocytes recruited by various chemoattractant factors, among which lipoprotein(a) and its specific apolipoprotein apo(a). Lp(a) contains a specific apolipoprotein apo(a) which size is determined by a variable number of repeats of a specific structural domain, the kringle IV type 2 (IV-2). Lp(a) plasma concentration and apo(a) size is inversely correlated, and smaller apo(a) are major risk factors for coronary heart disease. DESIGN AND METHODS: The aim of this study was to evaluate the effect of recombinant apo(a) isoforms (containing 10, 18 or 34 kringles) on monocytes interacting with type I collagen. RESULTS: Apo(a) isoforms stimulated reactive oxygen species (ROS) and matrix metalloproteinase-9 (MMP-9) production by monocytes, and not modified monocytes adhesion on type I collagen. This effect was specific of apo(a) since no effect was observed in the presence of plasminogen and was inversely related to apo(a) size. The lysine analogue 6-aminohexanoic acid which blocks the lysine binding sites (LBS), and carboxypeptidase B (CpB) which cleaves carboxy-terminal lysine residues, abolished apo(a)-induced ROS and MMP-9 production, highlighting an effect mediated by apo(a) lysing-binding sites. CONCLUSIONS: These results indicate that activation of collagen-primed monocytes stimulated with apo(a) is a Kringle number-dependent effect and reinforce the hypothesis of a role for small size apo(a) isoforms in atherothrombosis.


Assuntos
Apolipoproteínas A/farmacologia , Colágeno Tipo I/farmacologia , Monócitos/efeitos dos fármacos , Proteínas Recombinantes/farmacologia , Ácido Aminocaproico/farmacologia , Animais , Apolipoproteínas A/biossíntese , Apolipoproteínas A/química , Fibronectinas/farmacologia , Células HEK293 , Humanos , Metaloproteinase 9 da Matriz/biossíntese , Peso Molecular , Monócitos/citologia , Monócitos/metabolismo , Plasminogênio/farmacologia , Cultura Primária de Células , Ligação Proteica , Isoformas de Proteínas/biossíntese , Isoformas de Proteínas/química , Isoformas de Proteínas/farmacologia , Proteólise , Ratos , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismo , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química
3.
Res Vet Sci ; 125: 351-359, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31374445

RESUMO

Fibronectin type III domain-containing protein 5 (FNDC5) plays an important role in white-to-brown adipose tissue conversion in mice. However, there is no report on the role of this protein in Arbas Cashmere goat adipose tissue. We investigated the effect of FNDC5 on the proliferation and differentiation of goat adipose-derived stem cells (gADSCs). We found that FNDC5 promotes the proliferation of gADSCs and increases the levels of p-ERK and p-p38, while it has no effect on the levels of ERK, p38, AKT and p-AKT. What's more, FNDC5 promotes the differentiation of gADSCs into lipid droplets. Overexpression of FNDC5 increased the protein levels of ASC1, UCP1, PPARγ and SREBP1. Additionally, mRNA levels of PPARγ, SREBP1, ACC, and FABP4 increased significantly. FNDC5 knockdown was associated with opposite effects. These results suggest that FNDC5 promotes the proliferation of gADSCs via MAPK signaling pathway and also induces the differentiation of these cells.


Assuntos
Adipócitos/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Fibronectinas/farmacologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Adipócitos/citologia , Animais , Células-Tronco Mesenquimais/citologia , Camundongos
4.
Med Sci Monit ; 25: 6085-6096, 2019 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-31412018

RESUMO

BACKGROUND Irisin, a myokine released from skeletal muscle following exercise, has been shown to affect the proliferation of some cancer cells and chemosensitivity of anticancer drugs like doxorubicin (DOX). However, the effects of irisin on chemosensitivity in pancreatic cancer (PC) cells have not been studied. MATERIAL AND METHODS In this study, the effects of irisin co-treatment with DOX or gemcitabine (GEM) on MIA PaCa-2, BxPC-3 PC cells, and H9c2 cardiomyocytes were investigated. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, flow cytometry, and TUNEL (TdT-mediated dUTP nick-end labeling) assays were conducted to evaluate cytotoxicity induced by DOX or GEM. Fluorescence microscopy and flow cytometry experiments were performed to assess the intracellular accumulation of DOX. Cellular levels of apoptosis-related protein expression and protein phosphorylation were determined by Western blot analyses. RESULTS The results showed that irisin can increase the chemosensitivity of PC cells to DOX or GEM. The analyses of apoptosis indicated that irisin enhances DOX-induced cellular apoptosis by increasing the expression of cleaved PARP (poly ADP-ribose polymerase) and cleaved caspase-3, and reducing the expression of B cell lymphoma/lewkmia-2 (BCL-2) and B cell lymphoma-extra large (BCL-xL) in PC cells but not in H9c2 cells. Irisin attenuated serine/threonine kinase AKT (protein kinase B/PKB) phosphorylation and inhibited the activation of nuclear factor kappaB (NF-kappaB) signaling in PC cells. CONCLUSIONS Irisin can potentiate the cytotoxicity of doxorubicin in PC cells without increasing cardiotoxicity, possibly through inactivating the PI3K/AKT/NF-kappaB signaling pathway.


Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Doxorrubicina/farmacologia , Fibronectinas/farmacologia , Neoplasias Pancreáticas/tratamento farmacológico , Neoplasias Pancreáticas/metabolismo , Transdução de Sinais/efeitos dos fármacos , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Desoxicitidina/administração & dosagem , Desoxicitidina/análogos & derivados , Desoxicitidina/farmacologia , Doxorrubicina/administração & dosagem , Sinergismo Farmacológico , Fibronectinas/administração & dosagem , Humanos , Miócitos Cardíacos/efeitos dos fármacos , NF-kappa B/antagonistas & inibidores , NF-kappa B/metabolismo , Neoplasias Pancreáticas/patologia , Fosfatidilinositol 3-Quinases/metabolismo , Fosforilação/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-akt/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-akt/metabolismo
5.
In Vitro Cell Dev Biol Anim ; 55(8): 656-664, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31297698

RESUMO

The migration of lung fibroblasts plays a pivotal role in wound repair and fibrotic processes in the lung. Although the receptor for advanced glycation end products (RAGE) has been implicated in the pathogenesis of lung diseases, its role in lung fibroblast migration is unclear. The current study examined the effect of three different RAGE ligands, namely, high mobility group box 1 (HMGB1), S100A12, and N-epsilon-(carboxymethyl) lysine (CML), on human fibronectin-directed human fetal lung fibroblast (HFL-1) migration. HMGB1 augmented, whereas S100A12 inhibited, HFL-1 migration in a concentration-dependent manner. CML did not affect HFL-1 migration. The effect of HMGB1 was not through RAGE. However, the effect of S100A12 was mediated by RAGE, but not Toll-like receptor 4. S100A12 did not exert a chemoattractant effect, but inhibited HFL-1 chemotaxis and/or chemokinesis. Moreover, S100A12 mediated HFL-1 migration through p38 mitogen-activated protein kinase (MAPK) but not through nuclear factor-kappa B, protein kinase A, phosphatase and tensin homolog deleted on chromosome 10, or cyclooxygenase. In addition, western blot analysis showed that S100A12 augmented p38 MAPK activity in the presence of human fibronectin. In conclusion, S100A12 inhibits lung fibroblast migration via RAGE-p38 MAPK signaling. This pathway could represent a therapeutic target for pulmonary conditions characterized by abnormal tissue repair and remodeling.


Assuntos
Movimento Celular/efeitos dos fármacos , Fibroblastos/citologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Receptor para Produtos Finais de Glicação Avançada/metabolismo , Proteína S100A12/farmacologia , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Fibronectinas/farmacologia , Proteína HMGB1/metabolismo , Humanos , Ligantes , Pulmão/citologia , Receptor 4 Toll-Like/metabolismo
6.
J Coll Physicians Surg Pak ; 29(8): 736-740, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31358094

RESUMO

OBJECTIVE: To determine the effects of recombinant irisin on body mass index (BMI), serum insulin, luteinizing hormone (LH) and testosterone levels, and to correlate the serum insulin levels with serum luteinizing hormone and testosterone levels and to correlate the body mass index with serum insulin levels in obese female BALB/c mice. STUDY DESIGN: Laboratory-based experimental study. PLACE AND DURATION OF STUDY: Department of Physiology, Shifa College of Medicine, Islamabad in collaboration with Research Laboratory of Shifa College of Medicine, National Institute of Health (NIH) and Reproductive Physiology Laboratory, Quaid-e-Azam University, Islamabad, from March 2015 to September 2016. METHODOLOGY: Ninety female BALB/c mice were divided into three equal groups. Group A which was the control group was fed with normal chow diet. Group B and Group C were fed with high fat-high sucrose (HF-HS) diet for five weeks to induce obesity. After four weeks group C was divided into two subgroups. Group C-low dose (LD) was injected with low dose irisin and group C-High dose (HD) was injected with high dose irisin for one week. After five weeks, the BMI, serum insulin, LH and testosterone levels were measured in all the groups. Data was analysed by SPSS version 21. P-value of <0.05 was considered significant. RESULTS: Group B showed statistically significant elevation in BMI, serum insulin, LH and testosterone levels as compared to Group A (p <0.001, <0.001, 0.007 and 0.014, respectively). Group C-HD showed statistically significant decrease in BMI, serum insulin, and LH as compared to Group B (p <0.001, 0.013 and 0.028, respectively). Serum testosterone level was also decreased in group C-HD as compared to Group B, however the difference was not significant. CONCLUSION: Obesity increases the serum insulin, LH and testosterone and irisin significantly lowers the elevated BMI, serum insulin and LH levels in female BALB/c mice. It also lowers the elevated testosterone levels, but not significantly.


Assuntos
Biomarcadores/sangue , Fibronectinas/farmacologia , Obesidade/sangue , Animais , Índice de Massa Corporal , Feminino , Insulina/sangue , Hormônio Luteinizante/sangue , Camundongos , Camundongos Endogâmicos BALB C , Testosterona/sangue
7.
Int J Mol Sci ; 20(13)2019 Jun 27.
Artigo em Inglês | MEDLINE | ID: mdl-31252620

RESUMO

Psoriasis is a chronic inflammatory skin disease characterized by excessive growth of keratinocytes and hyperkeratosis in the epidermis. An abnormality of the non-lesional epidermis at an early stage of psoriasis is involved in triggering inflammatory cell infiltration into the dermis. Integrin α5ß1 acts as a receptor for fibronectin and has been found to be overexpressed in non-lesional psoriatic epidermis. To investigate whether α5ß1 integrin has a potential as a drug target for psoriasis treatment, the α5ß1 integrin-binding peptide, C16, was used to obstruct the HaCat keratinocyte cellular responses induced by fibronectin (Fn) in culture and psoriasis-like skin inflammation induced in mice by imiquimod (IMQ). The C16 exhibited antagonistic activity against α5ß1 integrin in HaCat cells, with evidence of suppression of the Fn-mediated proliferative, cytoskeletal, and inflammatory responses. Topical treatment with C16 greatly reduced the IMQ-induced epidermal hyperplasia, infiltration of neutrophils/macrophages, and expression of pro-inflammatory mediators in mouse skin. The C16SP (C16-derived short peptide; DITYVRLKF) also exhibited antagonistic activity, suppressing α5ß1 integrin activity in culture, and reducing IMQ-induced skin inflammation. Taken together, this study provides the first evidence that α5ß1 integrin may be a potential drug target for psoriasis. The synthetic C16 peptide may serve as an agent for psoriasis therapy.


Assuntos
Anti-Inflamatórios/uso terapêutico , Laminina/química , Fragmentos de Peptídeos/uso terapêutico , Psoríase/tratamento farmacológico , Animais , Anti-Inflamatórios/química , Anti-Inflamatórios/farmacologia , Linhagem Celular , Feminino , Fibronectinas/farmacologia , Humanos , Imiquimode/toxicidade , Integrina alfa5beta1/antagonistas & inibidores , Integrina alfa5beta1/metabolismo , Queratinócitos/efeitos dos fármacos , Queratinócitos/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/farmacologia , Ligação Proteica , Psoríase/etiologia
8.
Exp Eye Res ; 185: 107681, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31150636

RESUMO

Microenvironmental factors regulate stem cell fate. Fibronectin (FN), a key extracellular matrix component of the microenvironment, has been linked to various stem cell behaviors. However, how FN controls self-renewal, proliferation, and homeostasis of limbal stem cells remains unclear. Our study investigated the roles of FN in the self-renewal of rabbit limbal epithelial stem cells (rLESCs) by assessing rLESC proliferation and stemness in the presence and absence of FN. We further examined the effect of FN on non-canonical Wnt signaling during rLESC proliferation by evaluating the expression of cell cycle regulators. We found that rLESC proliferation increased after FN treatment and that 12.5 µg/cm2 FN maintained rLESC stemness. FN facilitated rLESC self-renewal by promoting Wnt11 and Fzd7 interaction. Furthermore, FN modulated cell cycle regulators to enhance rLESC proliferation via the upregulation of ROCK1 and ROCK2. Our study provides new insights into the mechanism through which FN regulates the self-renewal of rLESCs; specifically, this occurs via stimulation of the Wnt11/Fzd7/ROCK non-canonical Wnt pathway. The roles of FN in the self-renewal of limbal epithelial stem cells should be further investigated for the potential treatment of limbal deficiency.


Assuntos
Epitélio Anterior/efeitos dos fármacos , Fibronectinas/farmacologia , Receptores Frizzled/metabolismo , Limbo da Córnea/citologia , Células-Tronco/efeitos dos fármacos , Proteínas Wnt/metabolismo , Quinases Associadas a rho/metabolismo , Animais , Western Blotting , Diferenciação Celular/fisiologia , Proliferação de Células/fisiologia , Células Cultivadas , Epitélio Anterior/metabolismo , Citometria de Fluxo , Técnica Indireta de Fluorescência para Anticorpo , Técnicas de Silenciamento de Genes , Masculino , RNA Interferente Pequeno/genética , Coelhos , Reação em Cadeia da Polimerase em Tempo Real , Células-Tronco/metabolismo , Via de Sinalização Wnt/fisiologia
9.
Int Immunopharmacol ; 73: 225-235, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31108387

RESUMO

Intestinal ischemia/reperfusion (I/R) injury is a serious clinical event that may induce intestinal mucosal injury, whose major underlying mechanisms include reactive oxygen species (ROS) generation, release of inflammatory mediators and induction of apoptosis. Irisin is considered an agent with potent protection against many pathological injures. The aim of this study was to investigate the protective effect of irisin pretreatment on intestinal injury and explore its underlying mechanisms in a mouse model of intestinal I/R injury as well as a cell model (IEC-6 cell) of hypoxia/reoxygenation (H/R). The results showed that irisin pretreatment ameliorated I/R and H/R-induced injury in vivo and in vitro. In addition, irisin reduced the levels of tumor necrosis factor (TNF)-α, interleukin(IL)-1ß and interleukin(IL)-6 in the intestine. Compared with the I/R group, irisin pretreatment effectively reduced malondialdehyde (MDA) and myeloperoxidase (MPO) levels, but increased superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities in the intestine, and significantly reduced oxidative stress. Furthermore, irisin pretreatment downregulated Bax and cleaved Caspase-3 at the protein level, and increased Bcl-2 protein amounts, significantly reducing apoptosis in the intestine of I/R mice. Moreover, both in vivo and in vitro results showed that irisin pretreatment significantly upregulated nuclear factor (erythroid-derived 2)-like 2 (Nrf2) protein. Meanwhile, Nrf2 siRNA treatment partially abrogated the protective effects of irisin pretreatment on H/R induced cellular damage, inflammatory response, oxidative stress, and apoptosis in IEC-6 cells. These findings suggest that irisin pretreatment improves I/R-induced intestinal inflammatory response, reduces oxidative stress and inhibits apoptosis, which could be, at least partially, associated with Nrf2 pathway activation.


Assuntos
Fibronectinas/farmacologia , Fibronectinas/uso terapêutico , Enteropatias/tratamento farmacológico , Fator 2 Relacionado a NF-E2/metabolismo , Substâncias Protetoras/farmacologia , Substâncias Protetoras/uso terapêutico , Traumatismo por Reperfusão/tratamento farmacológico , Animais , Apoptose/efeitos dos fármacos , Linhagem Celular , Citocinas/metabolismo , Glutationa Peroxidase/metabolismo , Enteropatias/metabolismo , Enteropatias/patologia , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patologia , Masculino , Malondialdeído/metabolismo , Camundongos Endogâmicos C57BL , Peroxidase/metabolismo , Ratos , Traumatismo por Reperfusão/metabolismo , Traumatismo por Reperfusão/patologia , Transdução de Sinais/efeitos dos fármacos , Superóxido Dismutase/metabolismo
10.
Colloids Surf B Biointerfaces ; 180: 334-343, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31075687

RESUMO

Breast cancer cell lines lose the inherent gene expression profiles of their source tumor and when cultured as monolayers (two-dimensional) are unable to represent patient tumors. Thus, we engineered a biochemico- and mechano-mimetic three-dimensional (3D) culture platform for primary breast cancer cells by decellularizing cancer-associated fibroblasts (CAFs) cultured on 3D macroporous polymer scaffolds to recapitulate tumor behavior and drug response more realistically. The presence of the CAF-derived extracellular matrix deposited on the polycaprolactone scaffold promoted cell attachment and viability, which is ascribed to higher levels of phosphorylated Focal Adhesion Kinase that mediates cell attachment via integrins. Single cells from primary breast cancers self-organized into tumoroids on prolonged culture. Response of the tumoroids to two chemotherapeutic drugs, doxorubicin and mitoxanthrone, varied significantly across patient samples. This model could be used as an ex vivo platform to culture primary cells toward developing effective and personalized chemotherapy regimens.


Assuntos
Materiais Biomiméticos/química , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/patologia , Organoides/patologia , Medicina de Precisão , Técnicas de Cultura de Tecidos , Tecidos Suporte/química , Fibroblastos Associados a Câncer/patologia , Proliferação de Células , Sobrevivência Celular/efeitos dos fármacos , Junções Célula-Matriz/efeitos dos fármacos , Junções Célula-Matriz/metabolismo , Colágeno Tipo I/farmacologia , Matriz Extracelular/metabolismo , Feminino , Fibronectinas/farmacologia , Proteína-Tirosina Quinases de Adesão Focal/metabolismo , Humanos , Pessoa de Meia-Idade , Poliésteres , Porosidade , Esferoides Celulares/efeitos dos fármacos , Esferoides Celulares/patologia , Células Tumorais Cultivadas
11.
Nature ; 571(7763): 117-121, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31142833

RESUMO

Multipotent self-renewing haematopoietic stem cells (HSCs) regenerate the adult blood system after transplantation1, which is a curative therapy for numerous diseases including immunodeficiencies and leukaemias2. Although substantial effort has been applied to identifying HSC maintenance factors through the characterization of the in vivo bone-marrow HSC microenvironment or niche3-5, stable ex vivo HSC expansion has previously been unattainable6,7. Here we describe the development of a defined, albumin-free culture system that supports the long-term ex vivo expansion of functional mouse HSCs. We used a systematic optimization approach, and found that high levels of thrombopoietin synergize with low levels of stem-cell factor and fibronectin to sustain HSC self-renewal. Serum albumin has long been recognized as a major source of biological contaminants in HSC cultures8; we identify polyvinyl alcohol as a functionally superior replacement for serum albumin that is compatible with good manufacturing practice. These conditions afford between 236- and 899-fold expansions of functional HSCs over 1 month, although analysis of clonally derived cultures suggests that there is considerable heterogeneity in the self-renewal capacity of HSCs ex vivo. Using this system, HSC cultures that are derived from only 50 cells robustly engraft in recipient mice without the normal requirement for toxic pre-conditioning (for example, radiation), which may be relevant for HSC transplantation in humans. These findings therefore have important implications for both basic HSC research and clinical haematology.


Assuntos
Técnicas de Cultura de Células/métodos , Autorrenovação Celular/efeitos dos fármacos , Transplante de Células-Tronco Hematopoéticas/métodos , Células-Tronco Hematopoéticas/citologia , Animais , Proliferação de Células/efeitos dos fármacos , Células Clonais/citologia , Células Clonais/efeitos dos fármacos , Meios de Cultura/química , Meios de Cultura/farmacologia , Feminino , Fibronectinas/farmacologia , Células-Tronco Hematopoéticas/efeitos dos fármacos , Masculino , Camundongos , Álcool de Polivinil/farmacologia , Albumina Sérica , Fator de Células-Tronco/farmacologia , Trombopoetina/farmacologia , Fatores de Tempo , Condicionamento Pré-Transplante
12.
Cell Stress Chaperones ; 24(3): 595-608, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30993599

RESUMO

Irisin plays a protective effect in acute and chronic myocardial damage, but its role in septic cardiomyopathy is unclear. The aim of our study was to explore the in vivo and in vitro effects of irisin using an LPS-induced septic cardiomyopathy model. Our results demonstrated that irisin treatment attenuated LPS-mediated cardiomyocyte death and myocardial dysfunction. At the molecular level, LPS application was associated with mitochondrial oxidative injury, cardiomyocyte ATP depletion and caspase-related apoptosis activation. In contrast, the irisin treatment sustained mitochondrial function by inhibiting DRP1-related mitochondrial fission and the reactivation of mitochondrial fission impaired the protective action of irisin on inflammation-attacked mitochondria and cardiomyocytes. Additionally, we found that irisin modulated DRP1-related mitochondrial fission through the JNK-LATS2 signaling pathway. JNK activation and/or LATS2 overexpression abolished the beneficial effects of irisin on LPS-mediated mitochondrial stress and cardiomyocyte death. Altogether, our results illustrate that LPS-mediated activation of DRP1-related mitochondrial fission through the JNK-LATS2 pathway participates in the pathogenesis of septic cardiomyopathy. Irisin could be used in the future as an effective therapy for sepsis-induced myocardial depression because it corrects DRP1-related mitochondrial fission and normalizes the JNK-LATS2 signaling pathway.


Assuntos
Cardiomiopatias/tratamento farmacológico , Dinaminas/metabolismo , Fibronectinas/farmacologia , Mitocôndrias/efeitos dos fármacos , Dinâmica Mitocondrial/efeitos dos fármacos , Miócitos Cardíacos/efeitos dos fármacos , Animais , Cardiomiopatias/induzido quimicamente , Lipopolissacarídeos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Camundongos Endogâmicos C57BL , Mitocôndrias/patologia , Miócitos Cardíacos/patologia
13.
Int J Nanomedicine ; 14: 1451-1467, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30863071

RESUMO

Background: Irisin is a cytokine produced by skeletal muscle and usually plays a pivotal role in inducing fat browning and regulating energy expenditure. In recent years, it was found that irisin might be the molecular entity responsible for muscle-bone connectivity and is useful in osteogenesis induction. Materials and methods: To study its effect on bone regeneration, we developed silk/calcium silicate/sodium alginate (SCS) composite scaffold based on an interpenetrating network hydrogel containing silk fibroin, calcium silicate, sodium alginate. Then we loaded irisin on the SCS before coating it with polyvinyl alcohol (PVA). The SCS/P scaffold was physically characterized and some in vitro and in vivo experiments were carried out to evaluate the scaffold effect on bone regeneration. Results: The SCS/P scaffold was showed a porous sponge structure pursuant to scanning electron microscopy analysis. The release kinetics assay demonstrated that irisin was stably released from the irisin-loaded hybrid system (i/SCS/P system) to 50% within 7 days. Moreover, osteoinductive studies using bone marrow stem cells (BMSCs) in vitro exhibited the i/SCS/P system improved the activity of alkaline phosphatase (ALP) and enhanced the expression levels of a series of osteogenic markers containing Runx-2, ALP, BMP2, Osterix, OCN, and OPN. Alizarin red staining also demonstrated the promotion of osteogenesis induced by i/SCS/P scaffolds. In addition, in vivo studies showed that increased bone regeneration with better mineralization and higher quality was found during the repair of rat calvarial defects through utilizing the i/SCS/P system. Conclusion: These data provided strong evidence that the composite i/SCS/P would be a promising substitute for bone tissue engineering.


Assuntos
Alginatos/farmacologia , Regeneração Óssea/efeitos dos fármacos , Compostos de Cálcio/farmacologia , Sistemas de Liberação de Medicamentos , Fibronectinas/farmacologia , Músculos/metabolismo , Silicatos/farmacologia , Seda/farmacologia , Tecidos Suporte/química , Animais , Células Cultivadas , Liberação Controlada de Fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Cinética , Masculino , Osteogênese/efeitos dos fármacos , Osteogênese/genética , Ratos Sprague-Dawley , Crânio/diagnóstico por imagem , Crânio/efeitos dos fármacos
14.
Chem Biol Interact ; 302: 11-21, 2019 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-30703374

RESUMO

Angiotensin II-related cardiac fibrosis is one of the key pathological changes of the hypertrophied left ventricle in various heart disease. Irisin was recently reported to confer cardio-protective and anti-oxidative effects, while whether it can reverse the renin-angiotensin-aldosterone system(RAAS) activation related(angiotensin II-induced) cardiac fibrosis is unknown. In this study, we found that angiotensin II-induced cardiac dysfunction and fibrotic responses were dampened by irisin treatment in mice. Mechanistically, angiotensin II induced robust ROS generation, which in turn triggered activation of pro-fibrotic TGFß1-Smad2/3 signaling and subsequent collagen synthesis and fibroblast-myofibroblast transformation in cardiac fibroblasts. In contrast, Irisin treatment suppressed angiotensin II-induced ROS generation, TGFß1 activation, collagen synthesis and fibroblast-myofibroblast transformation, the effects of which was accompanied by Nrf2 activation and also abolished by a Nrf2 targeted siRNA. Taken together, we here identified irisin as a promising anti-fibrotic therapeutic for angiotensin II-related cardiac fibrosis.


Assuntos
Angiotensina II/farmacologia , Fibronectinas/farmacologia , Cardiopatias/patologia , Fator 2 Relacionado a NF-E2/metabolismo , Substâncias Protetoras/farmacologia , Transdução de Sinais/efeitos dos fármacos , Animais , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Fibrose , Cardiopatias/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Miocárdio/citologia , Fator 2 Relacionado a NF-E2/antagonistas & inibidores , Fator 2 Relacionado a NF-E2/genética , Estresse Oxidativo/efeitos dos fármacos , Interferência de RNA , RNA Interferente Pequeno/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Proteína Smad2/metabolismo , Proteína Smad3/metabolismo , Fator de Crescimento Transformador beta1/metabolismo
15.
Gynecol Endocrinol ; 35(5): 395-400, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30626233

RESUMO

To investigate the influence of irisin on endometrial receptivity of rats with polycystic ovarian syndrome (PCOS). PCOS rats were randomly divided into PCOS group and irisin group, and normal rats were used as control group. The PCOS group and control group were injected intraperitoneally with normal saline while the irisin group with recombinant irisin. The serum and uterus were obtained. Detect serum sex hormones, including Testosterone (T), Estradiol (E2), Progesterone (P), and glucose, insulin levels. Observe endometrial morphology by hematoxylin-eosin staining. Then evaluate the expression of leukemia inhibitory factor (LIF) and integrin αvß3 in endometrium using ELISA, immunohistochemistry and Real-time PCR. (1) Levels of serum T, glucose and insulin in PCOS group were significantly higher than those in control and irisin group. (2) For the endometrial morphology, levels of equivalent diameter, area of uterine glands and gland cavity and endometrial average thickness were lower in PCOS group than those in control and irisin group. (3) LIF and integrin αvß3 mRNA were basically consistent with protein expression. Levels of LIF and integrin αvß3 were decreased in PCOS group when compared with control and irisin group. Irisin may improve endometrial receptivity by promoting expression of LIF and integrin αvß3.


Assuntos
Implantação do Embrião/efeitos dos fármacos , Endométrio/efeitos dos fármacos , Fibronectinas/farmacologia , Síndrome do Ovário Policístico/metabolismo , Proteínas Recombinantes/farmacologia , Animais , Glicemia , Gonadotropina Coriônica , Implantação do Embrião/fisiologia , Endométrio/metabolismo , Estradiol/sangue , Feminino , Insulina/sangue , Integrina alfaVbeta3/metabolismo , Fator Inibidor de Leucemia/metabolismo , Síndrome do Ovário Policístico/sangue , Síndrome do Ovário Policístico/induzido quimicamente , Progesterona/sangue , Ratos , Ratos Sprague-Dawley , Testosterona/sangue
16.
J Cell Physiol ; 234(2): 1671-1681, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30171682

RESUMO

Irisin, a newly identified hormone and cardiokine, is critical for modulating body metabolism. New evidence indicates that irisin protects the heart against myocardial ischemic injury. However, whether irisin enhances cardiac progenitor cell (CPC)-induced cardiac repair remains unknown. This study examines the effect of irisin on CPC-induced cardiac repair when these cells are introduced into the infarcted myocardium. Nkx2.5+ CPC stable cells were isolated from mouse embryonic stem cells. Nkx2.5 + CPCs (0.5 × 10 6 ) were reintroduced into the infarcted myocardium using PEGlylated fibrin delivery. The mouse myocardial infarction model was created by permanent ligation of the left anterior descending (LAD) artery. Nkx2.5 + CPCs were pretreated with irisin at a concentration of 5 ng/ml in vitro for 24 hr before transplantation. Myocardial functions were evaluated by echocardiographic measurement. Eight weeks after engraftment, Nkx2.5 + CPCs improved ventricular function as evident by an increase in ejection fraction and fractional shortening. These findings are concomitant with the suppression of cardiac hypertrophy and attenuation of myocardial interstitial fibrosis. Transplantation of Nkx2.5 + CPCs promoted cardiac regeneration and neovascularization, which were increased with the pretreatment of Nkx2.5 + CPCs with irisin. Furthermore, irisin treatment promoted myocyte proliferation as indicated by proliferative markers Ki67 and phosphorylated histone 3 and decreased apoptosis. Additionally, irisin resulted in a marked reduction of histone deacetylase 4 and increased p38 acetylation in cultured CPCs. These results indicate that irisin promoted Nkx2.5 + CPC-induced cardiac regeneration and functional improvement and that irisin serves as a novel therapeutic approach for stem cells in cardiac repair.


Assuntos
Fibronectinas/farmacologia , Células-Tronco Embrionárias Murinas/efeitos dos fármacos , Células-Tronco Embrionárias Murinas/transplante , Infarto do Miocárdio/cirurgia , Miócitos Cardíacos/efeitos dos fármacos , Miócitos Cardíacos/transplante , Regeneração , Transplante de Células-Tronco/métodos , Função Ventricular Esquerda , Animais , Apoptose , Proliferação de Células , Células Cultivadas , Modelos Animais de Doenças , Fibrose , Proteína Homeobox Nkx-2.5/genética , Proteína Homeobox Nkx-2.5/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos ICR , Células-Tronco Embrionárias Murinas/metabolismo , Infarto do Miocárdio/metabolismo , Infarto do Miocárdio/patologia , Infarto do Miocárdio/fisiopatologia , Miócitos Cardíacos/metabolismo , Miócitos Cardíacos/patologia , Neovascularização Fisiológica , Recuperação de Função Fisiológica , Volume Sistólico , Remodelação Ventricular
17.
Redox Biol ; 20: 296-306, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30388684

RESUMO

Current management of liver ischemia-reperfusion (I/R) injury is mainly based on supportive care and no specific treatment is available. Irisin, a recently identified hormone, plays pivotal roles in energy expenditure and oxidative metabolism; however, it remains unknown whether irisin has any protective effects on hepatic I/R injury. In this study, we found that serum and liver irisin levels were markedly decreased at 24 h after hepatic I/R. Treatment with exogenous irisin improved liver function, reduced liver necrosis and cell apoptosis, and relieved inflammatory response after hepatic I/R. Meanwhile, exogenous irisin markedly inhibited mitochondrial fission related protein dynamin related protein 1 (drp-1) and fission 1 (Fis-1) expression in hepatic I/R. Additionally, treatment with exogenous irisin increased mitochondrial content and increased mitochondrial biogenesis related peroxisome proliferative activated receptor-γ (PPARγ) co-activator 1α (PGC-1α) and mitochondrial transcription factor (TFAM) expression. Furthermore, irisin decreased oxidative stress by upregulating uncoupling proteins (UCP) 2 expression in hepatic I/R. The results reveal that treatment with exogenous irisin alleviated hepatic I/R injury by restraining mitochondrial fission, promoting mitochondrial biogenesis and relieving oxidative stress. Irisin treatment appears to be a novel and promising therapeutic approach for hepatic I/R injury.


Assuntos
Fibronectinas/genética , Hepatopatias/genética , Hepatopatias/metabolismo , Dinâmica Mitocondrial , Biogênese de Organelas , Estresse Oxidativo , Traumatismo por Reperfusão/genética , Traumatismo por Reperfusão/metabolismo , Animais , Apoptose/genética , Fibronectinas/metabolismo , Fibronectinas/farmacologia , Hepatopatias/patologia , Testes de Função Hepática , Masculino , Camundongos , Mitocôndrias/genética , Mitocôndrias/metabolismo , Dinâmica Mitocondrial/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Proteínas Recombinantes , Traumatismo por Reperfusão/patologia
18.
Mol Reprod Dev ; 86(2): 224-238, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30582781

RESUMO

Fibronectin (Fn) enhances human sperm capacitation via the cAMP/PKA pathway, and the endocannabinoid system participates in this process. Moreover, Fn has been linked to endocannabinoid system components in different cellular models, even though no evidence of such interactions in human sperm is available. Normal semen samples were evaluated over a 4-year period. Our findings suggest that (a) the capacitating effects of Fn were reversed by preincubating the sperm with a cannabinoid receptor 1 (CB1) or transient receptor potential cation channel subfamily V member 1 (TRPV1) antagonist ( p < 0.001 and p < 0.05, respectively); (b) cooperation between CB1 and TRPV1 may exist ( p < 0.01); (c) the activity of specific fatty acid amide hydroxylase (FAAH) decreased after 1 min ( p < 0.01) and increased after 60 min ( p < 0.01) of capacitation in the presence of Fn; (d) the effects of Fn on FAAH activity were prevented by preincubating spermatozoa with a protein kinase A (PKA) inhibitor ( p < 0.01); (e) Fn modulated both the cyclic adenosine monophosphate concentration and PKA activity ( p < 0.05) during early capacitation; and (f) FAAH was a PKA substrate modulated by phosphorylation. These findings indicate that Fn stimulates human sperm capacitation via the cAMP/PKA pathway through modulation of the endocannabinoid system. Understanding the functional competence of human spermatozoa is essential for facilitating clinical advances in infertility treatment and for developing novel contraceptive strategies.


Assuntos
Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , AMP Cíclico/metabolismo , Endocanabinoides/farmacologia , Fibronectinas/farmacologia , Sistemas do Segundo Mensageiro/efeitos dos fármacos , Capacitação Espermática/efeitos dos fármacos , Espermatozoides/metabolismo , Humanos , Masculino , Espermatozoides/citologia
19.
Life Sci ; 214: 22-33, 2018 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-30449450

RESUMO

Matrikines, peptides originating from the fragmentation of extracellular matrix proteins are identified to play important role in both health and disease. They possess biological activities, much different from their parent protein. Identification of such bioactive cryptic regions in the extracellular matrix proteins has attracted the researchers all over the world in the recent decade. These bioactive peptides could find use in preparation of biomaterials and tissue engineering applications. Matrikines identified in major extracellular matrix (ECM) proteins like collagen, elastin, fibronectin, and laminin are being extensively studied for use in tissue engineering and regenerative medicine. They are identified to modulate cellular activity like cell growth, proliferation, migration and may induce apoptosis. RGD, a well-known peptide identified in fibronectin with cell adhesive property is being investigated in designing biomaterials. Collagen hexapeptide GFOGER was found to promote cell adhesion and differentiation. Laminin also possesses regions with strong cell adhesion property. Recently, cell-penetrating peptides from elastin are used as a targeted delivery system for therapeutic drugs. The continued search for cryptic sequences in the extracellular matrix proteins along with advanced peptide coupling chemistries would lead to biomaterials with improved surface properties. This review article outlines the peptides derived from extracellular matrix and some of the possible applications of these peptides in therapeutics and tissue engineering applications.


Assuntos
Proteínas da Matriz Extracelular/uso terapêutico , Fragmentos de Peptídeos/farmacologia , Adesão Celular/efeitos dos fármacos , Colágeno/química , Colágeno/uso terapêutico , Sistemas de Liberação de Medicamentos/métodos , Elastina/química , Elastina/farmacologia , Proteínas da Matriz Extracelular/metabolismo , Proteínas da Matriz Extracelular/farmacologia , Fibronectinas/química , Fibronectinas/farmacologia , Humanos , Laminina/química , Laminina/farmacologia , Oligopeptídeos/farmacologia , Fragmentos de Peptídeos/química
20.
Cell Physiol Biochem ; 51(2): 924-937, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30466091

RESUMO

BACKGROUND/AIMS: Islet metabolic disorder and inflammation contribute to the pathogenesis and progression of type 2 diabetes mellitus (T2DM). Irisin is a recently identified adipomyokine with protective effects on metabolic homeostasis and inflammation-suppressing effects in hepatic and vascular cells. The present study examined the effects of irisin on lipid metabolism and inflammation in ß cells under glucolipotoxic conditions. METHODS: Rat INS-1E ß cells and islets isolated from C57BL/6 mice were incubated in glucolipotoxic conditions with or without irisin. Intracellular lipid contents and lipogenic gene expression were determined by enzymatic colorimetric assays and real-time PCR, respectively. Inflammatory status was evidenced by Western blot analysis for the phosphorylation of nuclear factor-κB (NF-κB) p65 and real-time PCR analysis for the expression of pro-inflammatory genes. RESULTS: Irisin reversed glucolipotoxicity-induced intracellular non-esterified fatty acid (NEFA) and triglyceride accumulation, suppressed associated elevations in lipogenic gene expression, and phosphorylated acetyl-CoA-carboxylase (ACC) in INS-1E cells. These demonstrated effects were dependent on irisin-activated adenosine monophosphate-activated protein kinase (AMPK). Meanwhile, AMPK signaling mediated the protective effects of irisin on INS-1E cell insulin secretory ability and survival as well. Additionally, irisin inhibited phosphorylation of NF-κB p65 while decreasing the expression of pro-inflammatory genes in INS-1E cells under glucolipotoxic conditions. Moreover, irisin also improved insulin secretion, inhibited apoptosis, and restored ß-cell function-related gene expression in isolated mouse islets under glucolipotoxic conditions. CONCLUSION: Irisin attenuated excessive lipogenesis in INS-1E cells under glucolipotoxic state through activation of AMPK. Irisin also suppressed overnutrition-induced inflammation in INS-1E cells. Our findings implicate irisin as a promising therapeutic target for the treatment of islet lipid metabolic disorder and islet inflammation in T2DM.


Assuntos
Anti-Inflamatórios/farmacologia , Apoptose/efeitos dos fármacos , Fibronectinas/farmacologia , Transdução de Sinais/efeitos dos fármacos , Proteínas Quinases Ativadas por AMP/metabolismo , Acetil-CoA Carboxilase/metabolismo , Animais , Ácidos Graxos não Esterificados/metabolismo , Glucose/farmacologia , Transportador de Glucose Tipo 2/genética , Transportador de Glucose Tipo 2/metabolismo , Secreção de Insulina/efeitos dos fármacos , Células Secretoras de Insulina/citologia , Células Secretoras de Insulina/efeitos dos fármacos , Células Secretoras de Insulina/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Ácido Palmítico/farmacologia , Fosforilação/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Ratos , Fator de Transcrição RelA/metabolismo , Triglicerídeos/metabolismo
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