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1.
World J Gastroenterol ; 28(35): 5154-5174, 2022 Sep 21.
Artigo em Inglês | MEDLINE | ID: mdl-36188720

RESUMO

BACKGROUND: Colorectal cancer (CRC) is a common malignant tumor. Alcohol consumption is positively correlated with CRC malignant metastasis; however, the mechanism is unclear. The interaction between laminin-γ2 (LAMC2) and integrin-ß1 (ITGB1) plays a role in premetastatic niche signaling, which may induce epithelial mesenchymal transformation (EMT) and lead to metastasis. AIM: To investigate the effects of alcohol on CRC metastasis from the molecular mechanism of the premetastatic niche. METHODS: The interaction between LAMC2 and ITGB1 was measured by Duolink assay, and the expression levels of LAMC2, ITGB1 and focal adhesion kinase (FAK), snail, fibronectin, N-cadherin and special AT-rich sequence binding protein 1 (SATB1) were measured by quantitative real-time polymerase chain reaction, immunohistochemistry and western blotting. Interleukin-1ß (IL-1ß), tumor necrosis factor-α (TNF-α) and IL-6 levels were measured via enzyme-linked immunosorbent assay, histopathological assessment via hematoxylin eosin staining, and determination of aberrant crypt foci via methylene blue. RESULTS: The lymph node metastasis rate was higher in the alcohol group than non-alcohol group. There was a significant increase in interaction signals between LAMC2 and ITGB1, and an increase in phosphorylate-FAK/FAK, snail, fibronectin, N-cadherin and SATB1, whereas E-cadherin was reduced in the alcohol group compared to the non-alcohol group in both animal and clinical samples. Serum IL-1ß, TNF-α and IL-6 were higher in alcohol group than in non-alcohol group. Alcohol may promote CRC metastasis by influencing the molecular mechanism of the premetastatic niche. CONCLUSION: Our study suggests that alcohol promotes EMT-mediated premetastatic niche formation of CRC by activating the early interaction between LAMC2 and ITGB1 and lead to CRC metastasis.


Assuntos
Neoplasias Colorretais , Proteínas de Ligação à Região de Interação com a Matriz , Animais , Caderinas , Linhagem Celular Tumoral , Movimento Celular , Neoplasias Colorretais/patologia , Amarelo de Eosina-(YS)/farmacologia , Transição Epitelial-Mesenquimal , Fibronectinas/farmacologia , Proteína-Tirosina Quinases de Adesão Focal/metabolismo , Hematoxilina/farmacologia , Integrina beta1/metabolismo , Integrina beta1/farmacologia , Interleucina-1beta , Interleucina-6 , Laminina , Azul de Metileno , Fator de Necrose Tumoral alfa/farmacologia
2.
ACS Appl Mater Interfaces ; 14(42): 48179-48193, 2022 Oct 26.
Artigo em Inglês | MEDLINE | ID: mdl-36251059

RESUMO

The synthesis and study of the tripeptide Arg-Gly-Asp (RGD), the binding site of different extracellular matrix proteins, e.g., fibronectin and vitronectin, has allowed the production of a wide range of cell adhesive surfaces. Although the surface density and spacing of the RGD peptide at the nanoscale have already shown a significant influence on cell adhesion, the impact of its hierarchical nanostructure is still rather unexplored. Accordingly, a versatile colloidal system named quatsomes, based on fluid nanovesicles formed by the self-assembling of cholesterol and surfactant molecules, has been devised as a novel template to achieve hierarchical nanostructures of the RGD peptide. To this end, RGD was anchored on the vesicle's fluid membrane of quatsomes, and the RGD-functionalized nanovesicles were covalently anchored to planar gold surfaces, forming a state of quasi-suspension, through a long poly(ethylene glycol) (PEG) chain with a thiol termination. An underlying self-assembled monolayer (SAM) of a shorter PEG was introduced for vesicle stabilization and to avoid unspecific cell adhesion. In comparison with substrates featuring a homogeneous distribution of RGD peptides, the resulting hierarchical nanoarchitectonic dramatically enhanced cell adhesion, despite lower overall RGD molecules on the surface. The new versatile platform was thoroughly characterized using a multitechnique approach, proving its enhanced performance. These findings open new methods for the hierarchical immobilization of biomolecules on surfaces using quatsomes as a robust and novel tissue engineering strategy.


Assuntos
Fibronectinas , Integrinas , Integrinas/metabolismo , Adesão Celular , Fibronectinas/farmacologia , Fibronectinas/metabolismo , Vitronectina , Oligopeptídeos/farmacologia , Polietilenoglicóis , Tensoativos , Compostos de Sulfidrila , Ouro/farmacologia
3.
J Craniofac Surg ; 33(7): 2272-2275, 2022 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-36201689

RESUMO

This study aimed to investigate the effects of systemic irisin hormone application on new bone formation in peri-implant bone defects. After surgically creating peri-implant bone defects in the metaphyseal part of the tibiae of rats, the rats were randomly divided into 2 equal groups: a control group and an irisin group. In the control group, the rats received no further treatment during the 4-week experimental period after the surgery. The rats in the irisin group, 100 ng/kg irisin was administered intraperitoneally 3 days a week during the 8 weeks experimental period after the surgery. At the end of the experimental period, the rats were euthanized. Implants and surrounding bone tissues were collected for histological new bone formation analysis. The Student t test was used for statistical analysis. There were no significant differences between the groups in the histological analysis, new bone formation and fibrosis (P>0.05). Also, in the irisin group, there was numerically but not statistically more new bone formation detected compared with the controls. Within the limitations of this study, irisin did not affect new bone formation in peri-implant defects, although the numerical values favored the irisin group.


Assuntos
Implantes Dentários , Animais , Regeneração Óssea , Osso e Ossos , Fibronectinas/farmacologia , Hormônios , Osseointegração , Ratos
4.
J Coll Physicians Surg Pak ; 32(9): 1175-1180, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-36089716

RESUMO

OBJECTIVE: To determine the protective role of irisin in attenuating nicotine-induced oxidative stress in vascular tissue in mice. STUDY DESIGN: Experimental study. PLACE AND DURATION OF STUDY: Foundation University, Islamabad, Pakistan, from January 2019 to June 2020. METHODOLOGY: Thirty healthy BALB/c mice were divided into 3 groups. Group 1 was control, group II received nicotine 2 mg/Kg body weight intraperitoneally for 28 days, and group III, in addition, received r-irisin 0.5 µg/g body weight /day via tail vein, for the last 14 days. The tissue anti-oxidant enzymes (SOD, CAT, and GR) and lipid peroxidation marker (TBARS) were estimated. Aortic endothelium was analysed for atherosclerotic changes. The significant difference across groups was calculated using ANOVA. RESULTS: Group II showed statistically significant increase in lipid peroxidation marker (TBARS) levels (1059.04±32.31 ng/ml, p<0.001) and reduction in anti-oxidative enzymes (SOD, CAT and GR) levels (5479.24±25.38 pg/ml, 11.51±0.24 ng/ml and 1924.88±31.23 ng/ml, p<0.001) in aortic tissue homogenate as compared to group I. In Group III, with co- administration of r-irisin, significant improvement in antioxidant enzymes i.e. SOD, CAT, and GR levels (7958.70±110.54 pg/ml, 20.86±0.57 ng/ml, and 2897.18±52.93 ng/ml) and reduction in TBARS levels (239.14±19.90 ng/ml) was observed as compared to Group II (p<0.001). Endothelial damage manifested to type IV on histological examination. Co-administration of r-irisin in group III showed significant improvement in histological grading (only Type I and II lesions were seen). CONCLUSION: Exogenous administration of irisin improves anti-oxidant enzyme levels, ameliorates nicotine-induced oxidative stress, and endothelial dysfunction in the BALB/c mice. KEY WORDS: Irisin/FNDC-5, Oxidative stress, Anti-oxidant enzymes, Endothelial dysfunction, Atherosclerosis.


Assuntos
Aterosclerose , Fibronectinas , Animais , Antioxidantes/farmacologia , Aterosclerose/induzido quimicamente , Peso Corporal , Fibronectinas/farmacologia , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Nicotina/farmacologia , Estresse Oxidativo , Superóxido Dismutase/metabolismo , Substâncias Reativas com Ácido Tiobarbitúrico
5.
Bull Exp Biol Med ; 173(4): 464-467, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-36058964

RESUMO

We studied the effect of conditioned media from limbal epithelial stem cells, fibroblasts, and corneal keratocytes on the functional activity of human limbal mesenchymal stem cells. It was shown that the conditioned media from limbal epithelial stem cells reduced proliferative activity and inhibited migration of limbal mesenchymal stem cells. In the conditioned media of limbal epithelial stem cells, increased concentrations of VEGF and TNFα and reduced concentration of BDNF, vimentin, and fibronectin were found. The conditioned medium from corneal stromal cells did not affect functional activity of mesenchymal stem cells in the limbus. These data contribute to the understanding of the interaction of cells in the limbal niche and with corneal cells essential for the maintenance of the cellular homeostasis in the cornea.


Assuntos
Epitélio Corneano , Limbo da Córnea , Células-Tronco Mesenquimais , Fator Neurotrófico Derivado do Encéfalo/farmacologia , Diferenciação Celular , Córnea , Meios de Cultivo Condicionados/farmacologia , Células Epiteliais , Fibronectinas/farmacologia , Humanos , Células Estromais , Fator de Necrose Tumoral alfa/farmacologia , Fator A de Crescimento do Endotélio Vascular , Vimentina/genética
6.
Turk J Med Sci ; 52(2): 514-521, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36161624

RESUMO

BACKGROUND: Irisin, a newly identified exercise-derived myokine, has been found involved in a peripheral vasodilator effect. However, little is known regarding the potential vascular activity of irisin, and the mechanisms underlying its effects on vascular smooth muscle have not been fully elucidated. This study was aimed to investigate the effects of irisin on vascular smooth muscle contractility in rat thoracic aorta, and the hypothesis that protein kinase C (PKC) may have a role in these effects. METHODS: Isometric contraction-relaxation responses of thoracic aorta rings were measured with an isolated organ bath model. The steady contraction was induced with 10 µM phenylephrine (PHE), and then the concentration-dependent responses of irisin (0.001-1 µM) were examined. The time-matched vehicle control (double distilled water) group was also formed. To evaluate the role of PKC, endothelium-intact thoracic aorta rings were incubated with 150 nM bisindolylmaleimide I (BIM I) for 20 min before the addition of 10 µM PHE and irisin. Also, a vehicle control group was formed for dimethyl sulfoxide (DMSO). RESULTS: Irisin exerted the vasorelaxant effects at concentrations of 0.01, 0.1, and 1 µM compared to the control group (p < 0.001). Besides, PKC inhibitor BIM I incubation significantly inhibited the relaxation responses induced by varying concentrations of irisin (p: 0.000 for 0.01 µM; p: 0.000 for 0.1 µM; p: 0.000 for 1 µM). However, DMSO, a solvent of BIM I, did not modulate the relaxant effects of irisin (p > 0.05). DISCUSSION: In conclusion, physiological findings were obtained regarding the functional relaxing effects of irisin in rat thoracic aorta. The findings demonstrated that irisin induces relaxation responses in endothelium-intact thoracic aorta rings in a concentration-dependent manner. Furthermore, this study is the first to report that irisin-induced relaxation responses are regulated probably via activating signaling pathways implicating PKC.


Assuntos
Aorta Torácica , Fibronectinas , Animais , Endotélio Vascular , Fibronectinas/farmacologia , Fenilefrina/farmacologia , Proteína Quinase C/farmacologia , Ratos , Transdução de Sinais , Vasodilatadores/farmacologia
7.
Microcirculation ; 29(8): e12786, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36151930

RESUMO

PURPOSE: NLRP3 inflammasome mediates myocardial ischemia/reperfusion (MI/R) injury and diabetic vascular endothelia dysfunction. However, the role of NLRP3 inflammasome in MI/R injury with diabetes has not been fully described. Irisin plays an important role in anti-inflammation and improves endothelial function in type 2 diabetes. The current study aimed to investigate the effect of irisin on regulating NLRP3 inflammasome activation in diabetic vascular endothelia dysfunction. METHODS: Cardiac microvascular endothelial cells (CMECs) were cultured and subjected to high glucose/high fat (HG/HF) receiving hypoxia/reoxygenation (H/R) with irisin incubation or not. Then, apoptosis, viability, migration, NO secretion, and inflammasome activation were examined. RESULTS: The hypoxic CMECs exhibited increased apoptosis, impaired viability, and migration, even decreased NO secretion and enhanced inflammasome activation. Moreover, irisin incubation decreased NLRP3 activation and attenuated cell injury in HG/HF cultured CMECs subjected to H/R injury, which was abolished by NLRP3 inflammasome activation. Meanwhile, NLRP3 inflammasome siRNA also attenuated H/R injury in CMECs under HG/HF condition. CONCLUSION: The current study demonstrated for the first time that irisin inhibits NLRP3 inflammasome activation in CMECs as a novel mechanism in myocardial ischemia/reperfusion injury in diabetes.


Assuntos
Diabetes Mellitus Tipo 2 , Traumatismo por Reperfusão Miocárdica , Traumatismo por Reperfusão , Humanos , Inflamassomos/farmacologia , Células Endoteliais , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Fibronectinas/farmacologia
8.
In Vitro Cell Dev Biol Anim ; 58(8): 658-668, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-36125694

RESUMO

Tendinopathy is a common tendon disorder characterized by pain, swelling, and dysfunction. Current evidence has demonstrated that the depletion of stem cell pool and non-tenogenic differentiation of tendon-derived stem/progenitor cells (TSPCs) might account for the pathogenesis of tendinopathy. FNDC5/Irisin, as a novel exercise-induced myokine, is proved to be involved in the exercise-induced protective effects on musculoskeletal disorders. However, whether irisin can affect TSPCs fate is still unknown. To ascertain the roles of irisin on the proliferation and tenogenic differentiation of TSPCs, rat TSPCs were isolated and incubated with irisin. Cell viability, phenotypic changes, and related signaling pathways were evaluated by CCK-8 assay, colony formation assay, real-time PCR, Western blot, immunofluorescence, and proteasome activity assay. We found that irisin treatment increased the proliferative and colony-forming abilities, and promoted the tenogenic differentiation of TSPCs by upregulating the expression of YAP/TAZ. In conclusion, our work showed for the first time that irisin promotes the proliferation and tenogenic differentiation of rat TSPCs in vitro by activating YAP/TAZ, and the process was associated with a ubiquitin-proteasome proteolytic pathway. In conclusion, irisin and agents targeting YAP/TAZ may be promising therapeutic options for tendinopathy.


Assuntos
Doenças dos Roedores , Tendinopatia , Animais , Diferenciação Celular , Proliferação de Células , Fibronectinas/metabolismo , Fibronectinas/farmacologia , Complexo de Endopeptidases do Proteassoma/metabolismo , Complexo de Endopeptidases do Proteassoma/farmacologia , Ratos , Doenças dos Roedores/metabolismo , Doenças dos Roedores/patologia , Sincalida/metabolismo , Sincalida/farmacologia , Células-Tronco , Tendinopatia/metabolismo , Tendinopatia/patologia , Tendões , Ubiquitinas/metabolismo , Ubiquitinas/farmacologia
9.
Eur J Med Chem ; 242: 114685, 2022 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-36037790

RESUMO

Liver fibrosis is characterized by the excessive deposition of extracellular matrix components and results from chronic liver injury. At present, there is no approved drug for the treatment of liver fibrosis by the Food and Drug Administration. Here, we have reported a series of novel compounds with phenacrylanilide scaffolds that potently inhibit the transfer growth factor ß1 (TGF-ß1)-induced activation of LX-2, a hepatic stellate cell (HSC) line. Among them, compound 42 suppressed TGF-ß1-induced upregulation of fibrotic markers (α-SMA and fibronectin) and showed excellent safety in vitro. Furthermore, in a carbon tetrachloride (CCl4) -induced liver fibrosis model, 42 at a dose of 30 mg/kg/day through oral administration for 3 weeks effectively improved liver function, restored damaged liver structures, and reduced collagen deposition, with a greater effect than Tranilast. In addition, epithelial-mesenchymal transition (EMT) is inhibited by compound 42 in the process of fibrosis. Meanwhile, the imbalanced immune microenvironment could also be effectively reversed. More interestingly, compound 42 prolongs the survival of CCl4 mice and ameliorates CCl4-induced injury to spleen, kidney, lung and heart. Altogether, these results suggest that 42 could be a potential drug candidate for the treatment of liver fibrosis.


Assuntos
Tetracloreto de Carbono , Fibronectinas , Animais , Tetracloreto de Carbono/metabolismo , Fibronectinas/metabolismo , Fibronectinas/farmacologia , Fibronectinas/uso terapêutico , Fibrose , Células Estreladas do Fígado , Cirrose Hepática/induzido quimicamente , Cirrose Hepática/tratamento farmacológico , Cirrose Hepática/metabolismo , Camundongos , Fator de Crescimento Transformador beta1/metabolismo
10.
J Bone Miner Metab ; 40(5): 735-747, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-35925402

RESUMO

INTRODUCTION: Irisin is a proteolytic product of fibronectin type II domain-containing 5, which is related to the improvement in glucose metabolism. Numerous studies have suggested that irisin is a crucial myokine linking muscle to bone in physiological and pathophysiological states. MATERIALS AND METHODS: We examined the effects of local irisin administration with gelatin hydrogel sheets and intraperitoneal injection of irisin on the delayed femoral bone repair caused by streptozotocin (STZ)-induced diabetes in female mice. We analyzed the femurs of mice using quantitative computed tomography and histological analyses and then measured the mRNA levels in the damaged mouse tissues. RESULTS: Local irisin administration significantly blunted the delayed bone repair induced by STZ 10 days after a femoral bone defect was generated. Local irisin administration significantly blunted the number of Osterix-positive cells that were suppressed by STZ at the damaged site 4 days after a femoral bone defect was generated, although it did not affect the mRNA levels of chondrogenic and adipogenic genes 4 days after bone injury in the presence or absence of diabetes. On the other hand, intraperitoneal injection of irisin did not affect delayed bone repair induced by STZ 10 days after bone injury. Irisin significantly blunted the decrease in Osterix mRNA levels induced by advanced glycation end products or high-glucose conditions in ST2 cells in the presence of bone morphogenetic protein-2. CONCLUSIONS: We first showed that local irisin administration with gelatin hydrogel sheets improves the delayed bone repair induced by diabetic state partially by enhancing osteoblastic differentiation.


Assuntos
Diabetes Mellitus Experimental , Fibronectinas , Animais , Osso e Ossos , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Experimental/genética , Feminino , Fibronectinas/genética , Fibronectinas/metabolismo , Fibronectinas/farmacologia , Gelatina , Hidrogéis , Camundongos , RNA Mensageiro/genética
11.
Stem Cell Res Ther ; 13(1): 392, 2022 08 03.
Artigo em Inglês | MEDLINE | ID: mdl-35922833

RESUMO

BACKGROUND: Human mesenchymal stem cells (hMSCs) have been proven to have inherent chondrogenic differentiation potential, which appears to be used in cartilage regeneration. Increasing evidence suggests that irisin enhances osteoblast differentiation of MSCs, but little is known about its potential on chondrogenic differentiation. METHODS: In the study, we investigated the effects of irisin on chondrogenic differentiation of hMSCs using a high-density pellet culture system. The cartilage pellets were evaluated by morphology, and the metabolism of cartilage matrix was detected by qPCR, western blot and immunohistochemistry. Next, RNA-seq was performed to explore the underlying mechanism. Furthermore, using the transduction of plasmid, miRNAs mimics and inhibitor, the activation of Rap1/PI3K/AKT axis, the expression level of SIPA1L2, and the functional verification of miR-125b-5p were detected on day 7 of chondrogenic differentiation of hMSCs. RESULTS: Compared with the controls, we found that irisin treatment could significantly enhance the chondrogenic differentiation of hMSCs, enlarge the induced-cartilage tissue and up-regulate the expression levels of cartilage markers. RNA-seq indicated that irisin activated the Rap1 and PI3K/AKT signaling pathway, and the lower expression level of SIPA1L2 and the higher expression level of miR-125b-5p were found in irisin-treated group. Further, we found that irisin treatment could up-regulate the expression level of miR-125b-5p, targeting SIPA1L2 and consequently activating the Rap1/PI3K/AKT axis on the process of chondrogenic differentiation of hMSCs. CONCLUSIONS: Collectively, our study reveals that irisin can enhance chondrogenic differentiation of hMSCs via the Rap1/PI3K/AKT pathway, suggesting that irisin possesses prospects in cartilage regeneration.


Assuntos
Células-Tronco Mesenquimais , MicroRNAs , Células Cultivadas , Fibronectinas/metabolismo , Fibronectinas/farmacologia , Humanos , Células-Tronco Mesenquimais/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas rap1 de Ligação ao GTP/metabolismo
12.
Biomater Adv ; 135: 212741, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35929214

RESUMO

Surface functionalization to improve the blood compatibility is pivotal for the application of biomaterials. In this article, the surface of silicon was first functionalized with chemical groups, such as amino, quinone and phenol groups by the self-polymerization of dopamine, which were used to immobilize anticoagulant drugs hirudin. The detailed analysis and discussion about the grafting groups, morphology, wettability, the dynamic adsorption of proteins, the cytological property and the blood compatibility on the surfaces were carried on by the technology of contact angle, X-ray photoelectron spectroscopy, quartz crystal microbalance, endothelial cells culture and anticoagulant blood test in vivo. The surface with hirudin modification exhibited hydrophilic property and significantly inhibited the nonspecific adsorption of albumin, while it was more approachable to fibronectin. In vitro study displayed that the surface loaded with hirudin could promote the proliferation of endothelial cells. The evaluation of anticoagulant showed good anti-adhesion effect on platelets and the hemolysis rate decreased significantly to less than 0.4%. Activated partial thromboplastin time (APTT) of the silicon wafer loaded with hirudin can exceed 38 s, and the APTT prolongs as the hirudin concentration rises. This study suggested that such simple but effective surface functionalization technique, combining excellent anticoagulant activity together with reendothelialization potential due to the preferable fibronectin adsorption, provide great practical significance to the application of cardiovascular materials.


Assuntos
Fibronectinas , Hirudinas , Adsorção , Anticoagulantes/farmacologia , Células Endoteliais , Fibronectinas/farmacologia , Hirudinas/química , Silício/química
13.
J Dermatol ; 49(11): 1148-1157, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-35983802

RESUMO

Staphylococcus aureus (S. aureus) is frequently detected in the skin of patients with atopic dermatitis (AD). AD skin-derived strains of S. aureus (AD strain) are selectively internalized into keratinocytes (HaCaT cells) compared to standard strains. However, the mechanism of AD strain internalization by keratinocytes and effect of the skin environment on internalization remain unclear. HaCaT cells were exposed to heat-killed AD or standard strains of fluorescently labeled S. aureus, with or without interferon (IFN)-γ, interleukin (IL)-4, and IL-13 cytokines, for 24 h. Filaggrin and fibronectin expression in HaCaT cells was knocked down using small interfering RNA. The amount of internalized S. aureus was evaluated using a cell imaging system. The effects of INF-γ, IL-4, and S. aureus exposure on mRNA expression in HaCaT cells were analyzed using single-cell RNA sequencing. AD strains adhered to HaCaT cells in approximately 15 min and were increasingly internalized for up to 3 h (2361 ± 467 spots/100 cells, mean ± SD), whereas the standard strain was not (991 ± 71 spots/100 cells). In the presence of IFN-γ, both the number of internalized strains and fibronectin expression significantly decreased compared to in the control, whereas Th2 cytokines had no significant effects. The number of internalized AD strains was significantly higher in filaggrin knockdown and lower in fibronectin knockdown HaCaT cells compared to in the control. RNA sequencing revealed that IFN-γ decreased both fibronectin and filaggrin expression. Keratinocyte internalization of the AD strain may be predominantly mediated by the INF-γ-fibronectin pathway and partially regulated by filaggrin expression.


Assuntos
Dermatite Atópica , Infecções Estafilocócicas , Humanos , Staphylococcus aureus/metabolismo , Proteínas Filagrinas , Interferons/metabolismo , Interferons/farmacologia , Fibronectinas/metabolismo , Fibronectinas/farmacologia , Queratinócitos/metabolismo , Dermatite Atópica/metabolismo , Citocinas/metabolismo
14.
Protein Pept Lett ; 29(9): 760-768, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35909268

RESUMO

BACKGROUND: This study aimed to investigate the effects of irisin on rat tracheal smooth muscle contraction-relaxation responses and the roles of voltage-gated potassium (KV) channels, ATP-sensitive potassium (KATP) channels, and large-conductance calcium-activated potassium (BKCa) channels in these effects. METHODS: Isometric contraction and relaxation responses of tracheal segments were measured using the tissue bath method. Submaximal contractions were induced by ACh (10-5 M) or KCl (60 mM), and then concentration-response curves of irisin (10-9 to 10-6 M) were obtained. For the temporal control, a double-distilled water group was formed. ACh and irisin were added to the baths after tracheal segments were incubated with 4-AP (KV channel blocker), glibenclamide (KATP channel blocker), TEA, and iberiotoxin (BKCa channel blockers) to assess the role of K+ channels. In addition, a vehicle group was performed for the solvent dimethyl sulfoxide (DMSO). RESULTS: Irisin exhibited the relaxant effects in tracheal segments precontracted with both ACh and KCl at concentrations of 10-8-10-6 M (p<0.05). Besides, incubations of 4-AP, glibenclamide, TEA, and iberiotoxin significantly inhibited the irisin-mediated relaxation (p<0.05), whereas DMSO incubation did not modulate irisin responses (p>0.05). CONCLUSION: In conclusion, the first physiological results on the relaxant effects of irisin in rat trachea were obtained. Our findings demonstrated that irisin mediates concentration-dependent relaxation in rat tracheas. Moreover, the present study reported for the first time that irisin-induced bronchorelaxation is associated with the activity of the K+ channels.


Assuntos
Glibureto , Traqueia , Ratos , Animais , Glibureto/farmacologia , Fibronectinas/farmacologia , Canais de Potássio/farmacologia , Canais de Potássio/fisiologia , Dimetil Sulfóxido , Potássio/farmacologia , Trifosfato de Adenosina , Canais KATP/farmacologia
15.
Endocrinol Metab (Seoul) ; 37(4): 620-629, 2022 08.
Artigo em Inglês | MEDLINE | ID: mdl-35871605

RESUMO

BACKGRUOUND: Hepatic stellate cells (HSCs) are the central players interacting with multiple cell types in liver fibrosis. The crosstalk between HSCs and macrophages has recently become clearer. Irisin, an exercise-responsive myokine, was known to have a potentially protective role in liver and renal fibrosis, especially in connection with stellate cells. This study investigated the effects of irisin on the interaction between HSCs and macrophages. METHODS: Tamm-Horsfall protein-1 (THP-1) human monocytes were differentiated into macrophages, polarized into the inflammatory M1 phenotype with lipopolysaccharide. Lieming Xu-2 (LX-2) cells, human HSCs, were treated with conditioned media (CM) from M1 macrophages, with or without recombinant irisin. HSCs responses to CM from M1 macrophages were evaluated regarding activation, proliferation, wound healing, trans-well migration, contractility, and related signaling pathway. RESULTS: CM from M1 macrophages significantly promoted HSC proliferation, wound healing, transwell migration, and contractility, but not activation of HSCs. Irisin co-treatment attenuated these responses of HSCs to CM. However, CM and irisin treatment did not induce any changes in HSC activation. Further, irisin co-treatment alleviated CM-induced increase of phopho-protein kinase B (pAKT), matrix metalloproteinase-9 (MMP-9), and tissue inhibitor of metalloproteinases-1 (TIMP-1). CONCLUSION: These findings suggested that irisin may play a protective role in the pathogenesis of liver fibrosis, especially when working in the crosstalk between HSCs and macrophages.


Assuntos
Fibronectinas , Células Estreladas do Fígado , Fibronectinas/metabolismo , Fibronectinas/farmacologia , Humanos , Cirrose Hepática/patologia , Macrófagos/metabolismo , Macrófagos/patologia
16.
PLoS One ; 17(7): e0259482, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35857759

RESUMO

Serum-containing medium is widely used to support cell attachment, stable growth and serial passaging of various cancer cell lines. However, the presence of cholesterols and lipids in serum greatly hinders the analysis of the effects of cholesterol depletion on cells in culture. In this study, we developed a defined serum-free culture condition accessible to a variety of different types of adherent cancer cells. We tested different factors that are considered essential for cell culture and various extracellular matrix for plate coating, and found cells cultured in Dulbecco's Modified Eagle's Medium (DMEM) basal media supplemented with Albumin (BSA) and insulin-transferrin-selenium-ethanolamine (ITS-X) on fibronectin-precoated plate (called as "DA-X condition") showed comparable proliferation and survival to those in a serum-containing medium. Interestingly, we observed that DA-X condition could be adapted to a wide variety of adherent cancer cell lines, which enabled the analysis of how cholesterol depletion affected cancer cells in culture. Mechanistically, we found the beneficial effects of the DA-X condition in part can be attributed to the appropriate level of membrane cholesterol, and fibronectin-mediated signaling plays an important role in the suppression of cholesterol production.


Assuntos
Colesterol , Fibronectinas , Adesão Celular , Proliferação de Células , Células Cultivadas , Colesterol/farmacologia , Meios de Cultura/farmacologia , Fibronectinas/farmacologia
17.
Exp Mol Med ; 54(7): 1038-1048, 2022 07.
Artigo em Inglês | MEDLINE | ID: mdl-35882943

RESUMO

Intervertebral disc degeneration (IVDD) is a major cause of low back pain (LBP), and excessive senescence and apoptosis of nucleus pulposus (NP) cells are major pathological changes in IVDD. Physical exercise could effectively delay the process of intervertebral disc degeneration; however, its mechanism is still largely unknown. Irisin is an exercise-induced myokine released upon cleavage of the membrane-bound precursor protein fibronectin type III domain-containing protein 5 (FNDC5), and its levels increase after physical exercise. Here, we show that after physical exercise, FNDC5/irisin levels increase in the circulation and NP, senescence and apoptosis are reduced, autophagy is activated in NP tissue, and the progression of IVDD is delayed. Conversely, after knocking out FNDC5, the benefits of physical exercise are compromised. Moreover, the overexpression of FNDC5 in NP tissue effectively alleviated the degeneration of the intervertebral disc (IVD) in rats. By showing that FNDC5/irisin is an important mediator of the beneficial effects of physical exercise in the IVDD model, the study proposes FNDC5/irisin as a novel agent capable of activating autophagy and protecting NP from senescence and apoptosis.


Assuntos
Degeneração do Disco Intervertebral , Núcleo Pulposo , Animais , Apoptose , Autofagia , Fibronectinas/metabolismo , Fibronectinas/farmacologia , Degeneração do Disco Intervertebral/metabolismo , Camundongos , Núcleo Pulposo/metabolismo , Núcleo Pulposo/patologia , Ratos , Natação , Fatores de Transcrição/metabolismo
18.
CNS Neurosci Ther ; 28(11): 1883-1894, 2022 11.
Artigo em Inglês | MEDLINE | ID: mdl-35906830

RESUMO

AIM: To investigate the effect of apigenin on fibrous scar formation after mouse spinal cord injury (SCI). METHODS: The pneumatic impactor strike method was used to establish an SCI model. Mice were intraperitoneally injected with 5 mg/kg or 20 mg/kg apigenin daily for 28 days after SCI. The Basso Mouse Scale (BMS) score, hematoxylin-eosin staining, and immunohistochemical staining were used to assess the effect of apigenin on scar formation and motor function recovery. Western blotting and qRT-PCR were used to detect the expression of fibrosis-related parameters in spinal cord tissue homogenates. NIH-3 T3 cells and mouse primary spinal cord fibroblasts, α-Smooth muscle actin (α-SMA), collagen 1, and fibronectin were used to evaluate apigenin's effect in vitro. Western blotting and immunofluorescence techniques were used to study the effect of apigenin on TGFß/SMADs signaling. RESULTS: Apigenin inhibited fibrous scar formation in the mouse spinal cord and promoted the recovery of motor function. It reduced the expression of fibroblast-related parameters and increased the content of nerve growth factor in vivo, decreasing myofibroblast activation and collagen fiber formation by inhibiting TGFß-induced SMAD2/3 phosphorylation and nuclear translocation in vitro. CONCLUSION: Apigenin inhibits fibrous scar formation after SCI by decreasing fibrosis-related factor expression through TGFß/SMADs signaling.


Assuntos
Cicatriz , Traumatismos da Medula Espinal , Actinas/metabolismo , Animais , Apigenina/farmacologia , Apigenina/uso terapêutico , Cicatriz/tratamento farmacológico , Cicatriz/etiologia , Cicatriz/metabolismo , Colágeno/metabolismo , Colágeno/farmacologia , Amarelo de Eosina-(YS)/metabolismo , Amarelo de Eosina-(YS)/farmacologia , Fibronectinas/metabolismo , Fibronectinas/farmacologia , Hematoxilina/metabolismo , Hematoxilina/farmacologia , Camundongos , Fatores de Crescimento Neural/metabolismo , Recuperação de Função Fisiológica , Transdução de Sinais , Medula Espinal/patologia , Traumatismos da Medula Espinal/complicações , Traumatismos da Medula Espinal/tratamento farmacológico , Traumatismos da Medula Espinal/metabolismo , Fator de Crescimento Transformador beta/metabolismo , Fator de Crescimento Transformador beta/farmacologia
19.
Cancer Gene Ther ; 29(11): 1686-1696, 2022 11.
Artigo em Inglês | MEDLINE | ID: mdl-35701616

RESUMO

Acquired resistance to epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs), such as gefitinib, has dramatically impaired the clinical outcomes in non-small cell lung cancer (NSCLC) patients, but the mechanisms are still unclear in substantial cases. In our previous study, we demonstrated that a novel long non-coding RNA (lncRNA), lnc-ABCA12-8, was overexpressed in gefitinib-resistant NSCLC cells, but the exact function is unknown. In this study, we confirmed that lnc-ABCA12-8 was significantly upregulated both in NSCLC cell lines and the plasma samples of NSCLC patients with acquired resistance to gefitinib. Downregulation of lnc-ABCA12-8 could reverse gefitinib resistance both in vitro and in vivo. Mechanistically, lnc-ABCA12-8 interacted with alternative splicing factor/splicing factor 2 (ASF/SF2), promoted the binding of ASF/SF2 to the IIICS exon of fibronectin 1 (FN1) gene and enhanced the IIICS region inclusion during fibronectin 1 (FN1) alternative splicing, resulting in the upregulation of entire IIICS region, and enhanced cell proliferation, migration, invasion, and adhesion. Taken together, our study suggest that lnc-ABCA12-8 is involved in the acquired resistance to gefitinib, and may be a novel biomarker and therapeutic target for monitoring and overcoming gefitinib resistance in NSCLC.


Assuntos
Antineoplásicos , Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , Humanos , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Gefitinibe/farmacologia , Processamento Alternativo , Fibronectinas/genética , Fibronectinas/metabolismo , Fibronectinas/farmacologia , Receptores ErbB/genética , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Resistencia a Medicamentos Antineoplásicos/genética , Linhagem Celular Tumoral , Inibidores de Proteínas Quinases/farmacologia , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Transportadores de Cassetes de Ligação de ATP/genética , Transportadores de Cassetes de Ligação de ATP/metabolismo , Transportadores de Cassetes de Ligação de ATP/farmacologia
20.
Clin Mol Hepatol ; 28(4): 827-840, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35730208

RESUMO

BACKGROUND/AIMS: We aim to evaluate the effects of auranofin, a known antioxidant, on hepatic steatosis, inflammation, and fibrosis, contributing to non-alcoholic steatohepatitis (NASH) development in vivo and in vitro. METHODS: Transcriptome analysis of LX-2 cells was that expression patterns of genes changed by auranofin, and their related pathways were estimated. We used the gene set enrichment analysis (GSEA) program to determine the pathway involved in overall genetic change. In vitro, LX-2 and HepG2 cells were treated with transforming growth factor (TGF)-ß1 and palmitic acid (PA), respectively, and the antifibrotic and antiadipogenic effect function of auranofin was evaluated. RESULTS: Transcriptome analysis revealed that auranofin decreased the expression of 15 genes, including thrombospondin 1, endothelin 1 (ET-1), fibronectin 1, and LOX. The molecular functions of these genes are involved in collagen binding. GSEA of the overall gene expression pattern revealed that many genes increased in the reactive oxygen species pathway and decreased in the inflammatory response. Auranofin decreased nuclear factor kappa B (NF-κB) and IκBα in TGF-ß1-induced LX-2 cells, thereby reducing ET-1 and fibrosis. Furthermore, increased pNRF2 in PA-induced HepG2 cells led to increased antioxidant marker expression and decreased lipid accumulation. In the bile duct ligation model mice, auranofin reduced the fibrosis area and increased the survival rate. Auranofin reduced liver fibrosis and lipid accumulation in NASH model mice fed on a Western diet. CONCLUSION: Auranofin inhibits lipogenesis and fibrosis formation and is a proposed candidate for NASH treatment.


Assuntos
Hepatopatia Gordurosa não Alcoólica , Camundongos , Humanos , Animais , Hepatopatia Gordurosa não Alcoólica/complicações , Hepatopatia Gordurosa não Alcoólica/tratamento farmacológico , Hepatopatia Gordurosa não Alcoólica/metabolismo , NF-kappa B/metabolismo , NF-kappa B/farmacologia , Fator 2 Relacionado a NF-E2/genética , Fator 2 Relacionado a NF-E2/metabolismo , Fator 2 Relacionado a NF-E2/farmacologia , Fator de Crescimento Transformador beta1/genética , Fator de Crescimento Transformador beta1/metabolismo , Fator de Crescimento Transformador beta1/farmacologia , Auranofina/farmacologia , Auranofina/uso terapêutico , Auranofina/metabolismo , Inibidor de NF-kappaB alfa/metabolismo , Trombospondina 1/metabolismo , Trombospondina 1/farmacologia , Fibronectinas/metabolismo , Fibronectinas/farmacologia , Ácido Palmítico/toxicidade , Ácido Palmítico/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Endotelina-1/metabolismo , Endotelina-1/farmacologia , Antioxidantes/farmacologia , Antioxidantes/uso terapêutico , Antioxidantes/metabolismo , Camundongos Endogâmicos C57BL , Transdução de Sinais , Cirrose Hepática/patologia , Fatores de Crescimento Transformadores/metabolismo , Fatores de Crescimento Transformadores/farmacologia , Colágeno/metabolismo , Colágeno/farmacologia , Fígado/patologia
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