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1.
Mol Cell Endocrinol ; 559: 111780, 2023 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-36179941

RESUMO

Molecular pathways that contribute to orbital fibroblast activation during thyroid-eye disease (TED) may promote TED progression. Non-coding RNAs, especially miRNAs, play a critical role in the pathogenesis of TED. In the present study, miR-103a-3p was dramatically upregulated and TGFBR3 was downregulated within TED orbital tissue samples and TGF-ß-stimulated TED orbital fibroblasts. miR-103a-3p inhibition in TGF-ß-stimulated TED orbital fibroblasts partially abolished TGF-ß-induced fibrotic alterations, as manifested by the impaired fibroblast cell viability and decreased vimentin and fibronectin levels. miR-103a-3p directly targeted TGFBR3 in TED orbital samples and TGF-ß-stimulated TED orbital fibroblasts. In TGF-ß-stimulated TED orbital fibroblasts, TGFBR3 overexpression inhibited fibroblast cell viability and decreased vimentin and fibronectin levels. TGFBR3 overexpression partially attenuated the inhibitory effects of miR-103a-3p overexpression on TGFBR3 expression and the promotive effects of miR-103a-3p overexpression on TGF-ß-induced fibrotic alterations. Under TGF-ß stimulation, miR-103a-3p overexpression significantly promoted, whereas TGFBR3 overexpression inhibited the phosphorylation of Erk1/2, JNK, Smad2, and Smad3. TGFBR3 overexpression also partially abolished the effects of miR-103a-3p overexpression on Erk1/2, JNK, Smad2, and Smad3 phosphorylation. In conclusion, the miR-103a-3p/TGFBR3 axis regulated TGF-ß-induced TED orbital fibroblast activation and fibrosis in TED, with the possible involvement of the Erk/JNK and TGF-ß/Smad signaling pathways.


Assuntos
Oftalmopatia de Graves , MicroRNAs , Humanos , Fator de Crescimento Transformador beta/metabolismo , Fibronectinas/metabolismo , Vimentina/metabolismo , Fibroblastos/metabolismo , Fibrose , Oftalmopatia de Graves/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Proliferação de Células
2.
J Cell Biol ; 222(2)2023 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-36416725

RESUMO

Fibronectin (FN) is an essential structural and regulatory component of the extracellular matrix (ECM), and its binding to integrin receptors supports cell adhesion, migration, and signaling. Here, using live-cell microscopy of fibroblasts expressing FN tagged with a pH-sensitive fluorophore, we show that FN is secreted predominantly at the ventral surface of cells in an integrin-independent manner. Locally secreted FN then undergoes ß1 integrin-dependent fibrillogenesis. We find that the site of FN secretion is regulated by cell polarization, which occurs in bursts under stabilized lamellipodia at the leading edge. Moreover, analysis of FN secretion and focal adhesion dynamics suggest that focal adhesion formation precedes FN deposition and that deposition continues during focal adhesion disassembly. Lastly, we show that the polarized FN deposition in spreading and migrating cells requires both intact microtubules and myosin II-mediated contractility. Thus, while FN secretion does not require integrin binding, the site of exocytosis is regulated by membrane and cytoskeletal dynamics with secretion occurring after new adhesion formation.


Assuntos
Fibronectinas , Microtúbulos , Miosina Tipo II , Pseudópodes , Proteínas do Citoesqueleto/metabolismo , Fibroblastos/metabolismo , Fibronectinas/genética , Fibronectinas/metabolismo , Integrinas/metabolismo , Microtúbulos/genética , Microtúbulos/metabolismo , Miosina Tipo II/genética , Miosina Tipo II/metabolismo , Pseudópodes/genética , Pseudópodes/metabolismo , Matriz Extracelular/metabolismo , Exocitose
3.
J Pharm Biomed Anal ; 222: 115103, 2023 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-36272276

RESUMO

The urothelial basement membrane (UBM) contains type IV collagen, laminin-5, and fibronectin. Urothelial neoplasm can break through the UBM underlying the urothelium to invade the lamina propria. Conceptually, all bladder cancer staging over Ta (T1-T4) may demonstrate disturbances in the UBM structure, as well as alterations in the serum concentrations of the studied components. The aim of this study was to determine the blood serum concentration of collagen IV, laminin-5 and fibronectin in bladder cancer patients. Quantification of their concentrations and correlation with various clinicopathological parameters may be useful for making more accurate predictions and identifying high-risk patients. The study included 96 patients with bladder cancer confirmed by transurethral resection or cystectomy and 26 patients with diagnosed cystitis chronica or BPH (benign prostate hyperplasia). Collagen IV, laminin-5 and fibronectin were detected using Surface Plasmon Resonance Imaging biosensors. Significant differences in blood serum concentrations of the studied biomarkers were observed between the control group and bladder cancer patients, as well as between nonmuscle-invasive and muscle-invasive groups. ROC analysis gave satisfactory results for differentiation between the control group and bladder cancer group (AUC 0.92-0.99), with lower values only for collagen IV between nonmuscle-invasive and muscle-invasive patients (AUC 0.71), and a statistically insignificant difference for laminin-5. Laminin-5 concentration was more closely correlated to tumour grade, size and recurrence rate; fibronectin to tumour stage, size and morphology; and collagen IV to tumour stage, grade and recurrence rate. The relations between serum concentrations of the presented biomarkers of the urothelial basement membrane may be useful for bladder cancer detection and for determination of the tumour stage, hence simplifying the making of therapeutic decisions.


Assuntos
Técnicas Biossensoriais , Neoplasias da Bexiga Urinária , Masculino , Humanos , Neoplasias da Bexiga Urinária/cirurgia , Colágeno Tipo IV , Fibronectinas , Ressonância de Plasmônio de Superfície , Soro , Biomarcadores
4.
Dev Comp Immunol ; 138: 104541, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36108933

RESUMO

As the most typical example of mRNA variable splicing, Down Syndrome Cell Adhesion Molecule (Dscam) can produce a large number of mRNA isomers. It plays an important role not only in the nervous system, but also in the immune system. In Eriocheir sinensis, the extracellular region of Dscam has three variable domains, which can produce 25, 34 and 18 exons and encode the N-terminal region of immunoglobulin (Ig) 2 and Ig3 domains, and the entire Ig7 domain, respectively. In addition to three variable domains, the extracellular non-variable region of Dscam also includes many Ig domains and fibronectin type III (FNIII) domains. However, the role of the extracellular non-variable region function of Dscam in the immune defense of E. sinensis is unclear. In this study, we focused on the role of the extracellular non-variable region of Dscam in crab immune defense. The results indicate that the extracellular non-variable region of Dscam can bind bacteria and has bacteriostatic function. At the same time, the extracellular non-variable region of Dscam can also directly promote bacterial clearance by promoting phagocytosis of hemocytes.


Assuntos
Braquiúros , Sequência de Aminoácidos , Animais , Bactérias , Moléculas de Adesão Celular/metabolismo , Fibronectinas/genética , Fibronectinas/metabolismo , Hemócitos/metabolismo , Imunoglobulinas/genética , Fagocitose , Filogenia , RNA Mensageiro/genética , Alinhamento de Sequência
5.
Phys Chem Chem Phys ; 24(45): 27989-28002, 2022 Nov 23.
Artigo em Inglês | MEDLINE | ID: mdl-36373734

RESUMO

Protein adsorption is the first key step in cell-material interactions. The initial phase of such an adsorption process can only be probed using modelling approaches like molecular dynamics (MD) simulations. Despite a large number of studies on the adsorption behaviour of proteins on different biomaterials including calcium phosphates (CaP), little attention has been paid towards the quantitative assessment of the effects of various physicochemical influencers like surface modification, pH, and ionic strength. In the case of doped CaPs, surface modification through isomorphic substitution of foreign ions inside the apatite structure is of particular interest in the context of protein-HA interactions, as it is widely used to tailor the biological response of HA. Given this background, we present here the molecular-level understanding of the fibronectin (FN) adsorption mechanism and kinetics on a Sr2+-doped hydroxyapatite, HA, (001) surface at 300 K by means of all-atom molecular dynamics simulations. Electrostatic interactions involved in the adsorption of FN on HA were found to be significantly modified due to Sr2+ doping into the apatite lattice. In harmony with the published experimental observations, the Sr-doped surfaces were found to better support FN adhesion compared to pure HA, with 10 mol% Sr-doped HA exhibiting the best FN adsorption. The observed altered adsorption behaviour of FN on Sr-doped HA was correlated with the Hofmeister effect. Moreover, the non-monotonous trend of the FN-material interaction energy can be attributed to the spatial rearrangement of the functional groups (PO43-, OH-) in the apatite crystal. Sr2+ ions also influence the stability of the secondary structure of FN, as observed from the root mean square deviation (RMSD) and root mean square fluctuation (RMSF) analysis. The presence of Sr2+ enhances the flexibility of specific residues (residue nos. 20-44, 74-88) of the FN module. Rupture forces to disentangle FN from the biomaterial surface, obtained from steered molecular dynamics (SMD) simulations, were found to corroborate well with the results of equilibrium MD simulations. One particular observation is that the availability of an RGD motif (Arginine-Glycine-aspartate sequence, which interacts with cell surface receptor integrin to form a focal adhesion complex) for the interaction with cell surface receptor integrin is not significantly influenced by Sr2+ substitution.


Assuntos
Durapatita , Estrôncio , Durapatita/química , Estrôncio/química , Fibronectinas/química , Íons , Adsorção , Apatitas , Materiais Biocompatíveis , Integrinas
6.
Diabetes ; 71(12): 2486-2489, 2022 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-36409786
7.
BMC Endocr Disord ; 22(1): 281, 2022 Nov 18.
Artigo em Inglês | MEDLINE | ID: mdl-36401269

RESUMO

BACKGROUND: Irisin is a novel myokine both in mice and humans, and it can also be secreted by adipose tissue and the liver in a small amounts. There are few studies on irisin and bone metabolism. The aim of this study was to assess the relationship between serum irisin levels and bone metabolism and analyze its related factors in Han young male with pre-diabetic individuals. METHODS: This cross-sectional study included 41 pre-diabetes and 45 normal glucose tolerance (NGT). Anthropometric measurements, including height, weight, waist circumference (WC), and bone mineral content (BMC), were performed. All patients underwent an oral glucose tolerance test (OGTT) after 8 h of fasting, and the levels of glucose, insulin, lipids, serum irisin and bone turnover markers were measured. RESULTS: The levels of serum irisin (4.4 ± 1.4 vs. 6.3 ± 1.5 µg/mL), P1NP and OC were significantly lower and CTX was significantly higher in the pre-diabetes group (P < 0.05). BMC did not differ in the two groups (P > 0.05). Serum irisin levels negatively correlated with BMI (r =-0.325), FPG (r =-0.329), TG (r =-0.339) (P < 0.05) in NGT individuals. Serum irisin levels positively correlated with P1NP (r = 0.398), OC (r = 0.351), HDL-C (r = 0.432) and negatively correlated with FPG (r = -0.725), 2 h-PG (r = -0.360) (P < 0.05) in pre-diabetic individuals. Multiple regression analysis revealed that Serum irisin (ß = 9.768, P = 0.025) and WC (ß = -2.355, P = 0.002) were significant independent predictors for P1NP. CONCLUSION: Bone turnover markers were changed rather than bone mineral content in young men with pre-diabetes. In pre-diabetes individuals, serum irisin levels were reduced and close relationship with P1NP. Falling irisin levels may be a predictor of decreased bone formation in Han young men with pre-diabetes individuals.


Assuntos
Estado Pré-Diabético , Humanos , Masculino , Camundongos , Animais , Estado Pré-Diabético/diagnóstico , Fibronectinas , Estudos Transversais , Glucose , China/epidemiologia
8.
J Biomed Sci ; 29(1): 98, 2022 Nov 18.
Artigo em Inglês | MEDLINE | ID: mdl-36401329

RESUMO

BACKGROUND: Tumor vascular mimicry is an emerging issue that affects patient survival while having no treatment at the current moment. Despite several factors implicated in vascular mimicry, little is known about stromal factors that modulate tumor microenvironment and shape malignant transformation. CD248, a type-I transmembrane protein dominantly expressed in stromal cells, mediates the interaction between cells and extracellular matrix proteins. CD248 protein expression is associated with the metastatic melanoma phenotype and promotes tumor progression in the stromal cells. This study aimed to explore the cell-autonomous effects of CD248 in melanoma vascular mimicry to aid cancer therapy development. METHODS: Loss-of-function approaches in B16F10 melanoma cells were used to study the cell-autonomous effects of CD248 on cell adhesion, migration, proliferation, and vascular mimicry. A solid-phase binding assay was performed to identify the interaction between CD248 and fibronectin. Horizontal and vertical cell migration assays were performed to analyze cell migration activity, and cell-patterned network formation on Matrigel was used to evaluate vascular mimicry activity. Recombinant CD248 (rCD248) proteins were generated, and whether rCD248 interfered with melanoma CD248 functions was evaluated in vitro. An experimental lung metastasis mouse model was used to investigate the effect of rCD248 treatment in vivo. RESULTS: CD248 protein expression in melanoma cells was increased by a fibroblast-conditioned medium. Knockdown of CD248 expression significantly decreased cell adhesion to fibronectin, cell migration, and vascular mimicry in melanoma cells. The lectin domain of CD248 was directly involved in the interaction between CD248 and fibronectin. Furthermore, rCD248 proteins containing its lectin domain inhibited cell adhesion to fibronectin and slowed down cell migration and vascular mimicry. Treatment with rCD248 protein could reduce pulmonary tumor burden, accompanied by a reduction in vascular mimicry in mice with melanoma lung metastasis. CONCLUSION: CD248 expression in melanoma cells promotes malignant transformation by increasing the activity of cell adhesion, migration, and vascular mimicry, whereas rCD248 protein functions as a molecular decoy interfering with tumor-promoting effects of CD248 in melanoma cells.


Assuntos
Neoplasias Pulmonares , Melanoma , Camundongos , Animais , Fibronectinas , Melanoma/genética , Adesão Celular , Neoplasias Pulmonares/genética , Lectinas/farmacologia , Microambiente Tumoral , Antígenos de Neoplasias/metabolismo , Antígenos CD/genética , Antígenos CD/metabolismo , Antígenos CD/farmacologia
9.
Cells ; 11(21)2022 Nov 02.
Artigo em Inglês | MEDLINE | ID: mdl-36359868

RESUMO

Cisplatin is a potent chemotherapeutic used for the treatment of many types of cancer, but it has nephrotoxic side effects leading to acute kidney injury and subsequently chronic kidney disease (CKD). Previous work has focused on acute kidney tubular injury induced by cisplatin, whereas the chronic sequelae post-injury has not been well-explored. In the present study, we established a kidney fibroblast model of CKD induced by repeated administration of cisplatin (RAC) as a clinically relevant model. In NRK-49F rat kidney fibroblasts, RAC upregulated α-smooth muscle actin (α-SMA) and fibronectin proteins, suggesting that RAC induces kidney fibroblast-to-myofibroblast transformation. RAC also enhanced cell size, including the cell attachment surface area, nuclear area, and cell volume. Furthermore, RAC induced p21 expression and senescence-associated ß-galactosidase activity, suggesting that kidney fibroblasts exposed to RAC develop a senescent phenotype. Inhibition of p21 reduced cellular senescence, hypertrophy, and myofibroblast transformation induced by RAC. Intriguingly, after RAC, kidney fibroblasts were arrested at the G2/M phase. Repeated treatment with paclitaxel as an inducer of G2/M arrest upregulated p21, α-SMA, and fibronectin in the kidney fibroblasts. Taken together, these data suggest that RAC transforms kidney fibroblasts into myofibroblasts through G2/M arrest and cellular senescence.


Assuntos
Cisplatino , Insuficiência Renal Crônica , Ratos , Animais , Cisplatino/farmacologia , Cisplatino/metabolismo , Fibronectinas/metabolismo , Apoptose , Linhagem Celular Tumoral , Pontos de Checagem da Fase G2 do Ciclo Celular , Senescência Celular , Fibroblastos/metabolismo , Rim/metabolismo , Insuficiência Renal Crônica/metabolismo
10.
PLoS One ; 17(11): e0277561, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36355857

RESUMO

Acute myocardial infarction (AMI) results in weakening of the heart muscle and an increased risk for chronic heart failure. Therapeutic stem cells have been shown to reduce inflammatory signaling and scar tissue expansion, despite most of these studies being limited by poor retention of cells. Gelatin methacrylate (GelMA) coatings have been shown to increase the retention of these therapeutic cells near the infarct. In this work, we evaluate two different potential binding partners for GelMA-coated bone marrow cells (BMCs) and myocardial tissue: the extracellular matrix (ECM) and interstitial non-cardiomyocytes. While cells containing ß1 integrins mediate cell-ECM adhesion in vivo, these cells do not promote binding to our collagen-degraded, GelMA coating. Specifically, microscopic imagining shows that even with high integrin expression, GelMA-coated BMCs do not bind to cells within the myocardium. Alternatively, BMC incubation with decellularized heart tissue results in higher adhesion of coated cells versus uncoated cells supporting our GelMA-ECM binding mode. To further evaluate the ECM binding mode, cells were incubated on slides modified with one of three different major heart ECM components: collagen, laminin, or fibronectin. While all three components promoted higher adhesion than unmodified glass, collagen-coated slides resulted in a significantly higher adhesion of GelMA-coated BMCs over laminin and fibronectin. Incubation with unmodified BMCs confirmed that without a GelMA coating minimal adhesion of BMCs occurred. We conclude that GelMA cellular coatings significantly increase the binding of cells to collagen within the ECM. Our results provide progress towards a biocompatible and easily translatable method to enhance the retention of transplanted cells in human studies.


Assuntos
Gelatina , Infarto do Miocárdio , Humanos , Gelatina/farmacologia , Gelatina/metabolismo , Adesão Celular , Fibronectinas/metabolismo , Laminina/metabolismo , Miocárdio , Matriz Extracelular/metabolismo , Colágeno/metabolismo , Metacrilatos , Infarto do Miocárdio/terapia , Infarto do Miocárdio/metabolismo
11.
Cell Mol Life Sci ; 79(12): 602, 2022 Nov 24.
Artigo em Inglês | MEDLINE | ID: mdl-36434305

RESUMO

Experimental autoimmune-orchitis (EAO), a rodent model of chronic testicular inflammation and fibrosis, replicates pathogenic changes seen in some cases of human spermatogenic disturbances. During EAO, increased levels of pro-inflammatory and pro-fibrotic mediators such as TNF, CCL2, and activin A are accompanied by infiltration of leukocytes into the testicular parenchyma. Activin A levels correlate with EAO severity, while elevated CCL2 acting through its receptor CCR2 mediates leukocyte trafficking and recruits macrophages. CCR2 + CXCR4 + macrophages producing extracellular matrix proteins contribute widely to fibrogenesis. Furthermore, testicular macrophages (TMs) play a critical role in organ homeostasis. Therefore, we aimed to investigate the role of the activin A/CCL2-CCR2/macrophage axis in the development of testicular fibrosis. Following EAO induction, we observed lower levels of organ damage, collagen deposition, and leukocyte infiltration (including fibronectin+, collagen I+ and CXCR4+ TMs) in Ccr2-/- mice than in WT mice. Furthermore, levels of Il-10, Ccl2, and the activin A subunit Inhba mRNAs were lower in Ccr2-/- EAO testes. Notably, fibronectin+ TMs were also present in biopsies from patients with impaired spermatogenesis and fibrotic alterations. Overexpression of the activin A antagonist follistatin reduced tissue damage and collagen I+ TM accumulation in WT EAO testes, while treating macrophages with activin A in vitro increased the expression of Ccr2, Fn1, Cxcr4, and Mmp2 and enhanced migration along a CCL2 gradient; these effects were abolished by follistatin. Taken together, our data indicate that CCR2 and activin A promote fibrosis during testicular inflammation by regulating macrophage function. Inhibition of CCR2 or activin A protects against damage progression, offering a promising avenue for therapeutic intervention.


Assuntos
Orquite , Masculino , Humanos , Camundongos , Animais , Folistatina , Fibronectinas , Macrófagos , Fibrose , Inflamação , Receptores CCR2/genética
12.
Artigo em Inglês | MEDLINE | ID: mdl-36429644

RESUMO

BACKGROUND: Exercise training programs have the potential to improve cognitive function in older subjects. However, the majority of training programs are based on aerobic modality. In the current study, the influence of 3 months programs of sitting callisthenic balance (SCB) and resistance training (RT) on cognitive functioning and the mediating role that a change in the level of neurotrophic factors and strength in older, healthy participants plays were examined. MATERIAL AND METHODS: Global cognitive function was examined using MoCA, short-term memory using Digit Span and Delayed Matching to Sample, set shifting using Trial Making Test Part B, speed of processing simple visual stimuli using Simple Reaction Time, decision making using Choice Reaction Time, visual attention with Visual Attention Test (VAT), tests. Strength of lower and upper limbs, neurotrophin level (irisin, brain-derived neurotrophic factor (BDNF), neurotrophin 3 (NT-3), neurotrophin 4/5 (NT 4/5) were examined. RESULTS: Improved scores in RT vs. SCB were noted in MoCA (p = 0.02), reaction time in SRT (p = 0.02), TMT B (p = 0.03), errors committed in CRT (p = 0.04) and VAT (p = 0.02) were observed. No significant changes in the level of neurotrophic factors were observed. Changes in upper limb strength were related to changes in the number of errors committed in the SRT (p = 0.03). Lower limb strength changes explained the dynamics of the number of correct answers (p = 0.002) and errors committed (p = 0.006) in VAT. CONCLUSIONS: Both SCB and RT influenced multiple cognitive domains. The RT program improved global cognitive functioning, while no improvement was noticed in the SCB group. Decision making, visual attention and global cognitive function were improved after the RT program. Set-shifting, short-term visual memory processing speed of simple visual stimuli were improved after the SCB program, while a decrease in the processing speed of simple visual stimuli was noted in the RT group. Changes in irisin were related to set-shifting and short-term memory, while in BDNF to an improvement in the processing speed of simple visual stimuli. Resistance exercise training programs could be applied to prevent age related declines of cognitive function in healthy older subjects.


Assuntos
Treinamento de Força , Humanos , Idoso , Fator Neurotrófico Derivado do Encéfalo , Ginástica , Fibronectinas , Cognição , Exercício Físico
13.
Int J Mol Sci ; 23(22)2022 Nov 12.
Artigo em Inglês | MEDLINE | ID: mdl-36430467

RESUMO

Chronic obstructive pulmonary disease (COPD) is characterized by irreversible deterioration of the airway wall. Cigarette smoking is the major trigger, and in vitro studies showed that cigarette smoke extract (CSE) induced mitophagy in airway epithelial cells via oxidative stress, but this mechanism was not studied in airway smooth muscle cells (ASMCs). Primary ASMCs isolated from COPD patients or non-disease donors were investigated for CSE-induced remodeling and mitochondria structure. Proteins were assessed by Western blots for remodeling: collagen type-I, α-smooth muscle actin (α-SMA) and fibronectin; autophagy: beclin-1, protein62 (p62), light chain (LC)3A/B; mitochondria activity: mitochondrially encoded cytochrome c oxidase II & -IV (MTCO2, MTCO4), peroxisome proliferator activated receptor gamma coactivator 1α (PGC-1α); lysosomes: early endosome antigen 1, lysosome activated membrane protein 1; and cell signaling: extracellular signal regulated kinase (ERK1/2). Lysotracker and Mitotracker were used to monitor mitochondria morphology and organelle co-localization. Compared with controls, untreated COPD ASMCs showed lower collagen type-I and α-SMA expressions, but increased fibronectin levels. CSE further downregulated collagen type-I and α-SMA expression, but upregulated fibronectin. CSE decreased PGC-1α, MTCO2, and MTCO4, but increased beclin-1, p62, and LC3. CSE upregulated mitophagy and lysosomes activity via ERK1/2 phosphorylation. In vitro, cigarette smoke induced the deterioration of ASMCs, which might explain the tissue loss and structural remodeling in COPD bronchi. The results suggest that preventing exceeded mitophagy in ASMCs might present a novel therapeutic target for COPD.


Assuntos
Mitofagia , Doença Pulmonar Obstrutiva Crônica , Humanos , Fibronectinas/metabolismo , Sistema de Sinalização das MAP Quinases , Proteína Beclina-1/metabolismo , Doença Pulmonar Obstrutiva Crônica/metabolismo , Miócitos de Músculo Liso/metabolismo , Brônquios/metabolismo , Mitocôndrias/metabolismo , Colágeno Tipo I/metabolismo , Tabaco/metabolismo
14.
Int J Mol Sci ; 23(22)2022 Nov 14.
Artigo em Inglês | MEDLINE | ID: mdl-36430504

RESUMO

The immunolocalization of the cytoskeletal and the extracellular matrix proteins was investigated in the testicular excurrent duct system of healthy Japanese quail at 4, 6-7, 12 and 52 weeks of age. TdT dUTP Nick End Labeling (TUNEL) assay was used to assess apoptotic cell formation. The epithelia of the testicular excurrent duct system in birds of all age groups displayed various immunolabeling intensities and localization of cytokeratin 5 and beta-tubulin, while α-SMA was observed in epithelia only of 4-week-old birds. In all age groups, vimentin immunostaining was observed in the rete testes and efferent ductular epithelia, but not in the epididymal duct unit. The periductal smooth muscle cells of the excurrent duct system displayed variably intense immunopositivity with cytokeratin 5, desmin, fibronectin, α-SMA, and beta-tubulin. Furthermore, beta-tubulin and vimentin immunolabeled endothelial cells and fibroblasts with various intensities, while fibronectin immunostained extracellular matrices surrounding these cells. TUNEL-positive apoptotic cells were observed in the rete testes and efferent ductular epithelia, with increased frequency (p < 0.001) in 52-week-old birds. The study serves as a baseline normal for this region in healthy birds at 4, 6-7, 12, and 52 weeks of age, for comparison in future similar immunohistochemical studies involving environmental toxins affecting this region.


Assuntos
Coturnix , Testículo , Animais , Masculino , Testículo/metabolismo , Vimentina/metabolismo , Queratina-5 , Fibronectinas/metabolismo , Tubulina (Proteína)/metabolismo , Células Endoteliais/metabolismo
15.
Int J Mol Sci ; 23(22)2022 Nov 17.
Artigo em Inglês | MEDLINE | ID: mdl-36430689

RESUMO

The rapid growth and division of cancer cells are associated with mitochondrial biogenesis or switching to glycolysis. ERRα, PGC-1α and irisin/FNDC5 are some of the proteins that can influence these processes. The aim of this study was to determine the correlation of these proteins in non-small cell lung cancer (NSCLC) and to investigate their association with clinicopathological parameters. Immunohistochemistry reactions were performed on tissue microarrays (860 NSCLC, 140 non-malignant lung tissue). The normal fibroblast cell line (IMR-90) and lung cancer cell lines (NCI-H1703 and NCI-H522) were used as co-cultures. The mRNA levels of FNDC5 and ESRRA (encoding ERRα) were assessed in IMR-90 cells after co-culture with lung cancer cells. We observed a decreased level of ERRα with an increase in tumor size (T), stages of the disease, and lymph node metastases (N). In the adenocarcinoma (AC) subtype, patients with a higher ERRα expression had significantly longer overall survival. A moderate positive correlation was observed between FNDC5 mRNA and ESRRA mRNA in NSCLCs. The expression of FNDC5 mRNA in IMR-90 cells increased after 24 h, and ESRRA gene expression increased after 48 h of co-culture. The ERRα receptor with PGC-1α participates in the control of FNDC5/irisin expression. Normal fibroblasts revealed an upregulation of the FNDC5 and ESRRA genes under the influence of lung cancer cells.


Assuntos
Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , Humanos , Carcinoma Pulmonar de Células não Pequenas/genética , Fibronectinas/genética , Neoplasias Pulmonares/genética , Receptores de Estrogênio/genética , Fatores de Transcrição/genética , RNA Mensageiro/genética
16.
Int J Mol Sci ; 23(21)2022 Oct 31.
Artigo em Inglês | MEDLINE | ID: mdl-36362050

RESUMO

Diabetic kidney disease (DKD) frequently leads to end-stage renal disease and other life-threatening illnesses. The dysregulation of glomerular cell types, including mesangial cells, endothelial cells, and podocytes, appears to play a vital role in the development of DKD. Myeloid-derived suppressor cells (MDSCs) exhibit immunoregulatory and anti-inflammatory properties through the depletion of L-arginine that is required by T cells, through generation of oxidative stress, interference with T-cell recruitment and viability, proliferation of regulatory T cells, and through the promotion of pro-tumorigenic functions. Under hyperglycemic conditions, mouse mesangial cells reportedly produce higher levels of fibronectin and pro-inflammatory cytokines. Moreover, the number of MDSCs is noticeably decreased, weakening inhibitory immune activities, and creating an inflammatory environment. In diabetic mice, immunotherapy with MDSCs that were induced by a combination of granulocyte-macrophage colony-stimulating factor, interleukin (IL)-1ß, and IL-6, reduced kidney to body weight ratio, fibronectin expression, and fibronectin accumulation in renal glomeruli, thus ameliorating DKD. In conclusion, MDSCs exhibit anti-inflammatory activities that help improve renal fibrosis in diabetic mice. The therapeutic targeting of the proliferative or immunomodulatory pathways of MDSCs may represent an alternative immunotherapeutic strategy for DKD.


Assuntos
Diabetes Mellitus Experimental , Nefropatias Diabéticas , Células Supressoras Mieloides , Animais , Camundongos , Diabetes Mellitus Experimental/imunologia , Diabetes Mellitus Experimental/metabolismo , Nefropatias Diabéticas/imunologia , Nefropatias Diabéticas/metabolismo , Células Endoteliais/metabolismo , Fibronectinas/metabolismo , Camundongos Endogâmicos C57BL , Células Supressoras Mieloides/imunologia
17.
Lipids Health Dis ; 21(1): 115, 2022 Nov 05.
Artigo em Inglês | MEDLINE | ID: mdl-36335399

RESUMO

BACKGROUND: Compared with typical visceral fat deposits in obesity and metabolic syndrome, perirenal adipose tissue (PRAT) dysfunction is more closely linked to obesity-related chronic kidney disease (OB-CKD). The myokine irisin reportedly promotes positive outcomes in metabolic disease. This study investigated whether irisin could reduce urinary albumin excretion and demonstrate renoprotective effects through the regulation of PRAT function in obese mice. METHODS: C57BL/6 J mice received a high-fat diet (HFD) with or without concurrent administration of irisin. Glucose tolerance, plasma levels of free fatty acids, and urinary albumin excretion were assessed, along with renal morphology. The vascular endothelial growth factor and nitric oxide in glomeruli were also analyzed, in addition to PRAT function-associated proteins. RESULTS: Irisin administration significantly reduced the final body weight, fat mass, and free fatty acids, without reducing PRAT mass, in HFD mice. Furthermore, irisin decreased urinary albumin excretion and attenuated both renal fibrosis and lipid accumulation. Irisin administration led to increases in PRAT function-associated proteins, including sirtuin1, uncoupling protein-1, and heme-oxygenase-1. Ex vivo treatment of PRAT and glomeruli with irisin also restored PRAT function. Finally, irisin treatment restored the vascular endothelial growth factor-nitric oxide axis. CONCLUSIONS: Irisin attenuated metabolic disorders and protected against OB-CKD by normalizing the PRAT-kidney axis. These results suggest that agents targeting PRAT activation might be useful for treatment of OB-CKD.


Assuntos
Fibronectinas , Insuficiência Renal Crônica , Camundongos , Animais , Camundongos Obesos , Óxido Nítrico/metabolismo , Ácidos Graxos não Esterificados/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Camundongos Endogâmicos C57BL , Obesidade/metabolismo , Tecido Adiposo/metabolismo , Dieta Hiperlipídica/efeitos adversos , Rim/metabolismo , Insuficiência Renal Crônica/tratamento farmacológico , Insuficiência Renal Crônica/prevenção & controle , Insuficiência Renal Crônica/metabolismo , Albuminas/metabolismo
18.
Mol Med ; 28(1): 134, 2022 11 18.
Artigo em Inglês | MEDLINE | ID: mdl-36401176

RESUMO

BACKGROUND: Circular RNA (circ) AFF4 was documented to regulate osteogenesis but the underlying mechanism remains to be elucidated. The preliminary study showed that circ_AFF4 may promote osteogenesis via FNDC5/Irisin. Furthermore, the online prediction tool indicated the interaction of circ_AFF4, insulin-like growth factor-2 mRNA-binding protein 3 (IGF2BP3), FNDC5 and lysine (K)-specific demethylase 1 A (KDM1A). Therefore, this study aims to elucidate the relationships of KDM1A, circ_AFF4, IGF2BP3 and FNDC5/Irisin during osteogenesis. METHODS: The alkaline phosphatase (ALP) activities and osteogenic-related factors were determined using ALP and alizarin red S (ARS) staining, real-time quantitative PCR(RT-qPCR) and western blot. Immunoprecipitation (RIP), pull-down assay and fluorescence in situ hybridization (FISH) were used to examine the interactions among circ_AFF4/IGF2BP3/FNDC5. A mouse in vivo model was utilized to further confirm the regulatory effect on bone formation. RESULTS: Circ_AFF4 and KDM1A expression levels were increased during osteoinduction of BM-MSCs. Knockdown of circ_AFF4 and KDM1A significantly suppressed BM-MSC osteogenesis. We also proved that KDM1A directly bound to circ_AFF4 and FNDC5 promoter and induced circ_AFF4 and FNDC5 expression. Furthermore, circ_AFF4 enhanced the stability of FNDC5 by generating a circ_AFF4, IGF2BP3 and FNDC5 RNA-protein complex, and thereby induced Irisin and osteogenesis. The in vitro data was confirmed with in vivo model. CONCLUSION: These findings elucidate that KDM1A induces circ_AFF4, which promotes promote osteogenesis via IGF2BP3. This study indicates that circ_AFF4 may potentially represent a critical therapeutic target for the diseases.


Assuntos
Osteogênese , RNA Circular , Camundongos , Animais , RNA Circular/genética , Osteogênese/genética , Fibronectinas/genética , Hibridização in Situ Fluorescente , Fatores de Transcrição/genética
19.
Zhongguo Ying Yong Sheng Li Xue Za Zhi ; 38(4): 356-360, 2022 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-36414561

RESUMO

Objective: To investigate the effects of aerobic versus resistance exercise on soleus muscle contractile properties and the expressions of MuRF1, PGC-1α and FNDC5 in amyotrophic rats after unloading, and the possible molecular biological mechanisms. Methods: Male Wistar rats were randomly divided into recovery group (CT), aerobic exercise group (A), resistance exercise group (R) and control group (C), with 6 rats in each group. The control group did not receive any treatment. The other three groups underwent tail suspension for 2 weeks, and then the recovery group recovered quietly. The aerobic group and the resistance group underwent a 2-week exercise intervention. Exercise plan: the aerobic group rats were treated with treadmill speed corresponding to 65% maximum oxygen uptake (VO2max), 60 min/d, 5 d/w; the rats in the resistance group were allowed to climb the ladder with 65% of the maximum voluntary weight-bearing (MVCC) for 3 times, with a total of 5 sets. Each time had a rest of 1 min, with an interval of 2 min among sets, and 5 d/w. Fasting for 24 hours after the last exercise, the soleus muscle samples were collected to observe the histological changes, test the contractility, and detect MuRF1 and PGC-1α and FNDC5 expressions. Results: compared with the control group, the body weight, soleus muscle wet weight, average cross-sectional area of muscle fibers and muscle contractility of the recovery group were decreased significantly(P<0.01), and the expression of MuRF1 was increased significantly(P<0.01). Compared with the recovery group, the body weight, wet weight of soleus muscle, the average cross-sectional area of muscle fiber and muscle contractility of rats in aerobic group and resistance group were increased (P<0.01), the expression of PGC-1α/FNDC5 was increased (P<0.01) and the expression of MuRF1 was decreased significantly (P<0.01). Compared with the aerobic group, the expression of PGC-1α in soleus muscle of rats in the resistance group was increased (P<0.05), while the expression of MuRF1 was decreased (P<0.05). Conclusion: Aerobic and resistance exercise can significantly improve muscle contractility, upregulate the expression of PGC-1α/FNDC5, and inhibit the expression of MuRF1, indicating that the molecular mechanisms of aerobic and resistance exercise to improve unloaded muscular atrophy may be related to PGC-1α and MuRF1.


Assuntos
Consumo de Oxigênio , Oxigênio , Animais , Masculino , Ratos , Peso Corporal , Fibronectinas , Fibras Musculares Esqueléticas , Atrofia Muscular , Ratos Wistar , Fatores de Transcrição
20.
Hum Exp Toxicol ; 41: 9603271221143040, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36444838

RESUMO

Clerodane diterpene, a class of bicyclic diterpenoids, is found in hundreds of plant species. 16-hydroxycleroda-3,13-dien-15,16-olide (CD) can be isolated from the plant Polyalthia longifolia and has been applied against oral cancer and glioma by xenograft model. In this study, we aim to explore its antitumour action by examining its histone deacetylase (HDAC) activity and integrin-associated intracellular signalling pathway on T24 human bladder cancer (BC) cells. Our results revealed that CD-inhibited colony formation, HDAC activity, HDAC (1, 2 and 3) mRNA and cell spreading on fibronectin-coated surfaces in a concentration-dependent manner. Furthermore, decreased cFLIP and increased caspase-8 cleavage accompanied CD-induced cell death. At non-toxic concentrations, CD blocked the migration and invasion of T24 cells. CD hindered migration and invasion by the downregulation of fibronectin, integrin α5ß1, ß-catenin, FAK, vinculin and Rho A, as well as by reduction of phosphorylated glycogen synthase kinase 3ß (pGSK3ß), pSrc, pstat3 and pNFκB. We observed that the MMP9 gene was closely linked with prognosis of patients with bladder cancer. MMP9 protein levels and activity were largely attenuated by CD in a concentration-dependent manner. In conclusion, CD-induced caspase-8-dependent apoptosis and suppressed migration and invasion by blocking several intracellular signalling pathways, including downregulation of HDAC activity and integrin-FAK and MMP9 pathways.


Assuntos
Diterpenos Clerodânicos , Neoplasias da Bexiga Urinária , Humanos , Proteína-Tirosina Quinases de Adesão Focal , Anoikis , Metaloproteinase 9 da Matriz/genética , Integrinas , Neoplasias da Bexiga Urinária/tratamento farmacológico , Histona Desacetilases/genética , Caspase 8/genética , Fibronectinas
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