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1.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 36(10): 890-896, 2020 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-33148383

RESUMO

Objective To investigate the reducing effects of shikimic acid from the total extract of Chaenomeles speciose on the differentiation of chondrocytes into hypertrophic chondrocytes by inhibiting RBL-2H3 cell degranulation. Methods The chondrocytes were identified by toluidine blue staining and tryptase immunohistochemical staining. The chondrocytes were divided into normal chondrocytes control group, C48/80 activated RBL-2H3 cell culture supernatant treatment group, 3, 10 and 30 µg/mL SA activated RBL-2H3 cell culture supernatant treatment groups. The toxicity of SA and RBL-2H3 cell supernatant were detected by MTT assay. Western blotting was used to detect the expression of collagen type II (Col2) and collagen type X (Col10) in chondrocytes. The levels of matrix metalloproteinase 13 (MMP13), soluble nuclear factor B receptor activated protein ligand (sRANKL) and bone protective factor (OPG) were determined by ELISA, and glycosaminoglycan polysaccharide (GAG) were tested by dimethylmethylene blue (DMB) colorimetry. Results (0~30) µg/mL SA had no significant effects on the growth of chondrocytes. Compared with the C48/80 activated RBI-2H3 cell supernatant treatment group, the expression of Col2 and GAG proteins increased significantly, while the expression of Col10 and MMP13 and the ratio of sRANKL/OPG decreased significantly in the SA treatment groups in a dose-dependent manner. Conclusion SA can effectively reduce the differentiation of chondrocytes into hypertrophic chondrocytes by inhibiting RBL-2H3 cell degranulation.


Assuntos
Degranulação Celular , Diferenciação Celular , Condrócitos/efeitos dos fármacos , Mastócitos/efeitos dos fármacos , Rosaceae/química , Ácido Chiquímico/farmacologia , Animais , Linhagem Celular Tumoral , Condrócitos/citologia , Mastócitos/citologia , Ratos
2.
J Pharmacol Exp Ther ; 374(1): 104-112, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32434944

RESUMO

Statin drugs are widely employed in the clinic to reduce serum cholesterol. Because of their hydroxymethylglutaryl coenzyme A reductase antagonism, statins also reduce isoprenyl lipids necessary for the membrane anchorage and signaling of small G-proteins in the Ras superfamily. We previously found that statins suppress immunoglobulin E (IgE)-mediated mast cell activation, suggesting these drugs might be useful in treating allergic disease. Although IgE-induced function is critical to allergic inflammation, mast cell proliferation and survival also impact atopic disease and mast cell neoplasia. In this study, we describe fluvastatin-mediated apoptosis in primary and transformed mast cells. An IC50 was achieved between 0.8 and 3.5 µM in both cell types, concentrations similar to the reported fluvastatin serum Cmax value. Apoptosis was correlated with reduced stem cell factor (SCF)-mediated signal transduction, mitochondrial dysfunction, and caspase activation. Complementing these data, we found that p53 deficiency or Bcl-2 overexpression reduced fluvastatin-induced apoptosis. We also noted evidence of cytoprotective autophagy in primary mast cells treated with fluvastatin. Finally, we found that intraperitoneal fluvastatin treatment reduced peritoneal mast cell numbers in vivo These findings offer insight into the mechanisms of mast cell survival and support the possible utility of statins in mast cell-associated allergic and neoplastic diseases. SIGNIFICANCE STATEMENT: Fluvastatin, a statin drug used to lower cholesterol, induces apoptosis in primary and transformed mast cells by antagonizing protein isoprenylation, effectively inhibiting stem cell factor (SCF)-induced survival signals. This drug may be an effective means of suppressing mast cell survival.


Assuntos
Apoptose/efeitos dos fármacos , Fluvastatina/farmacologia , Mastócitos/citologia , Mastócitos/efeitos dos fármacos , Animais , Células da Medula Óssea/citologia , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Humanos , Metabolismo dos Lipídeos/efeitos dos fármacos , Mastócitos/metabolismo , Camundongos
3.
PLoS One ; 15(3): e0230633, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32208440

RESUMO

Mast cells are connective tissue resident cells with morphological and functional characteristics that contribute to their role in allergic and inflammatory processes, host defense and maintenance of tissue homeostasis. Mast cell activation results in the release of pro-inflammatory mediators which are largely responsible for the physiological functions of mast cells. The lectin ArtinM, extracted from Artocarpus heterophyllus (jackfruit), binds to D-manose, thus inducing degranulation of mast cells. ArtinM has several immunomodulatory properties including acceleration of wound healing, and induction of cytokine release. The aim of the present study was to investigate the role of ArtinM in the activation and proliferation of mast cells. The rat mast cell line RBL-2H3 was used throughout this study. At a low concentration (0.25µg/mL), ArtinM induced mast cell activation and the release of IL-6 without stimulating the release of pre-formed or newly formed mediators. Additionally, when the cells were activated by ArtinM protein tyrosine phosphorylation was stimulated. The low concentration of ArtinM also activated the transcription factor NFkB, but not NFAT. ArtinM also affected the cell cycle and stimulated cell proliferation. Therefore, ArtinM may have therapeutic applications by modulating immune responses due to its ability to activate mast cells and promote the release of newly synthesized mediators. Additionally, ArtinM could have beneficial effects at low concentrations without degranulating mast cells and inducing allergic reactions.


Assuntos
Degranulação Celular/efeitos dos fármacos , Lectinas/farmacologia , Proteínas de Plantas/farmacologia , Animais , Artocarpus/metabolismo , Linhagem Celular , Membrana Celular/efeitos dos fármacos , Membrana Celular/ultraestrutura , Proliferação de Células/efeitos dos fármacos , Interleucina-6/metabolismo , Mastócitos/citologia , Mastócitos/metabolismo , Mitose/efeitos dos fármacos , NF-kappa B/metabolismo , Fosforilação/efeitos dos fármacos , Ratos
4.
Life Sci ; 250: 117570, 2020 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-32205088

RESUMO

Accidental exposure to ionizing radiation is a serious concern to human life. Studies on the mitigation of side effects following exposure to accidental radiation events are ongoing. Recent studies have shown that radiation can activate several signaling pathways, leading to changes in the metabolism of free radicals including reactive oxygen species (ROS) and nitric oxide (NO). Cellular and molecular mechanisms show that radiation can cause disruption of normal reduction/oxidation (redox) system. Mitochondria malfunction following exposure to radiation and mutations in mitochondria DNA (mtDNA) have a key role in chronic oxidative stress. Furthermore, exposure to radiation leads to infiltration of inflammatory cells such as macrophages, lymphocytes and mast cells, which are important sources of ROS and NO. These cells generate free radicals via upregulation of some pro-oxidant enzymes such as NADPH oxidases, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). Epigenetic changes also have a key role in a similar way. Other mediators such as mammalian target of rapamycin (mTOR) and peroxisome proliferator-activated receptor (PPAR), which are involved in the normal metabolism of cells have also been shown to regulate cell death following exposure to radiation. These mechanisms are tissue specific. Inhibition or activation of each of these targets can be suggested for mitigation of radiation injury in a specific tissue. In the current paper, we review the cellular and molecular changes in the metabolism of cells and ROS/NO following exposure to radiation. Furthermore, the possible strategies for mitigation of radiation injury through modulation of cellular metabolism in irradiated organs will be discussed.


Assuntos
Estresse Oxidativo/efeitos da radiação , Lesões por Radiação/metabolismo , Animais , Ciclo-Oxigenase 2/metabolismo , DNA Mitocondrial/genética , Epigênese Genética , Humanos , Inflamação , Linfócitos/citologia , Mastócitos/citologia , Camundongos , Mitocôndrias/efeitos da radiação , Mutação , NADPH Oxidases/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Oxirredução , Radiação Ionizante , Espécies Reativas de Oxigênio/metabolismo
5.
Sci Rep ; 10(1): 1771, 2020 02 04.
Artigo em Inglês | MEDLINE | ID: mdl-32019985

RESUMO

Immune cells resident in adipose tissue have important functions in local and systemic metabolic homeostasis. Nevertheless, these immune cell populations remain poorly characterized in bovines. Recently, we described diverse lymphocyte subpopulations in adipose tissue of Holstein-Friesian cows. Here, we aimed at characterising myeloid cell populations present in bovine adipose tissue using multicolour flow cytometry, cell sorting and histochemistry/immunohistochemistry. Macrophages, CD14+CD11b+MHC-II+CD45+ cells, were identified in mesenteric and subcutaneous adipose tissue, though at higher proportions in the latter. Mast cells, identified as SSC-AhighCD11b-/+CD14-MHC-II-CH138A-CD45+ cells, were also observed in adipose tissue and found at higher proportions than macrophages in mesenteric adipose tissue. Neutrophils, presenting a CH138A+CD11b+ phenotype, were also detected in mesenteric and subcutaneous adipose tissue, however, at much lower frequencies than in the blood. Our gating strategy allowed identification of eosinophils in blood but not in adipose tissue although being detected by morphological analysis at low frequencies in some animals. A population not expressing CD45 and with the CH138A+ CD11b-MHC-II- phenotype, was found abundant and present at higher proportions in mesenteric than subcutaneous adipose tissue. The work reported here may be useful for further studies addressing the function of the described cells.


Assuntos
Antígenos CD/metabolismo , Mesentério/metabolismo , Células Mieloides/metabolismo , Gordura Subcutânea/metabolismo , Animais , Bovinos , Feminino , Citometria de Fluxo , Imuno-Histoquímica , Mastócitos/citologia , Mastócitos/metabolismo , Mesentério/citologia , Células Mieloides/citologia , Células Progenitoras Mieloides/citologia , Células Progenitoras Mieloides/metabolismo , Gordura Subcutânea/citologia
6.
Immunity ; 52(2): 404-416.e5, 2020 02 18.
Artigo em Inglês | MEDLINE | ID: mdl-32049054

RESUMO

Mast cells are rare tissue-resident cells of importance to human allergies. To understand the structural basis of principle mast cell functions, we analyzed the proteome of primary human and mouse mast cells by quantitative mass spectrometry. We identified a mast-cell-specific proteome signature, indicative of a unique lineage, only distantly related to other immune cell types, including innate immune cells. Proteome comparison between human and mouse suggested evolutionary conservation of core mast cell functions. In addition to specific proteases and proteins associated with degranulation and proteoglycan biosynthesis, mast cells expressed proteins potentially involved in interactions with neurons and neurotransmitter metabolism, including cell adhesion molecules, ion channels, and G protein coupled receptors. Toward targeted cell ablation in severe allergic diseases, we used MRGPRX2 for mast cell depletion in human skin biopsies. These proteome analyses suggest a unique role of mast cells in the immune system, probably intertwined with the nervous system.


Assuntos
Mastócitos/citologia , Mastócitos/imunologia , Animais , Biomarcadores/metabolismo , Degranulação Celular , Linhagem da Célula , Células Cultivadas , Tecido Conjuntivo/imunologia , Humanos , Imunoterapia , Mastócitos/metabolismo , Proteínas de Membrana/imunologia , Proteínas de Membrana/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Proteínas do Tecido Nervoso/imunologia , Proteínas do Tecido Nervoso/metabolismo , Neuroimunomodulação , Proteoglicanas/biossíntese , Proteoma , Receptores Acoplados a Proteínas-G/imunologia , Receptores Acoplados a Proteínas-G/metabolismo , Receptores de Neuropeptídeos/imunologia , Receptores de Neuropeptídeos/metabolismo , Pele/imunologia
7.
Cell Biol Int ; 44(4): 1068-1075, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31889352

RESUMO

Cationic liposomes are commonly used as vectors to effectively introduce foreign genes into target cells. In another function, we recently showed that cationic liposomes bound to the mast cell surface suppress the degranulation induced by the cross-linking of high-affinity immunoglobulin E receptor in a time- and dose-dependent manner. This suppression is mediated by the impairment of the sustained level of intracellular Ca2+ concentration ([Ca2+ ]i ) via the inhibition of store-operated Ca2+ entry. Further, we revealed that the mechanism underlying an impaired [Ca2+ ]i increase is the inhibition of the activation of the phosphatidylinositol 3-kinase (PI3K)-Akt pathway. Yet, how cationic liposomes inhibit the PI3K-Akt pathway is still unclear. Here, we focused on caveolin-1, a major component of caveolae, which is reported to be involved in the activation of the PI3K-Akt pathway in various cell lines. In this study, we showed that caveolin-1 translocated from the cytoplasm to the plasma membrane after the activation of mast cells and colocalized with the p85 subunit of PI3K, which seemed to be essential for PI3K activity. Meanwhile, cationic liposomes suppressed the translocation of caveolin-1 to the plasma membrane and the colocalization of caveolin-1 with PI3K p85 also at the plasma membrane. This finding provides new information for the development of therapies using cationic liposomes against allergies.


Assuntos
Cálcio/metabolismo , Cavéolas/metabolismo , Caveolina 1/metabolismo , Lipossomos/metabolismo , Fosfatidilinositol 3-Quinase/metabolismo , Animais , Linhagem Celular Tumoral , Membrana Celular/metabolismo , Mastócitos/citologia , Mastócitos/metabolismo , Ratos
8.
Bull Exp Biol Med ; 168(3): 381-384, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31938914

RESUMO

Serotonin-containing cytostructures of mouse bone marrow after autotransplantation were studied using Falk-Hillarp luminescent histochemical method, and expression of antiapoptotic marker was determined by the immunohistochemical reaction. Autotransplantation of the bone marrow in mice led to an increase in the number of mast cells; in 40 and 120 min after autotransplantation, serotonin content in mast cells increased by 12 and 23%, respectively, and in hemopoietic cells of the bone marrow - by 24 and 78%, respectively, which affected the early stages of proliferation and differentiation of hemopoietic cells. The mitotic index in bone marrow smears at these terms increased by 3.8 and 4.5 times. According to myelogram data, the fraction of early undifferentiated forms (lymphoblasts, erythroblasts) in the primary organ of hemopoiesis (bone marrow) increased.


Assuntos
Transplante de Medula Óssea/métodos , Hematopoese/fisiologia , Transplante Autólogo/métodos , Animais , Células da Medula Óssea/citologia , Células da Medula Óssea/metabolismo , Diferenciação Celular/genética , Diferenciação Celular/fisiologia , Hematopoese/genética , Masculino , Mastócitos/citologia , Mastócitos/metabolismo , Camundongos
9.
Immunology ; 159(4): 441-449, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-31957000

RESUMO

Signaling by Kit has been extensively studied in hematopoietic cells and is essential for the survival, proliferation and maintenance of hematopoietic stem and progenitor cells. In addition to the activation of intrinsic signaling pathways, Kit has been shown to interact with lineage-restricted type I cytokine receptors and produce cross signals, e.g. erythropoietin receptor, interleukin-7 receptor (IL-7R), IL-3R. Based on the earlier studies, we hypothesize that Kit activate other type I cytokine receptors in a cell-specific manner and execute cell-specific function. To investigate other Kit-activated receptors, we tested Kit and IL-4R cross-receptor activation in murine bone-marrow-derived mast cells, which express both Kit and IL-4R at the surface level. Kit upon activation by Kit ligand (KL), activated IL-4Rα, γC , and signal transducer and activator of transcription 6 independent of its cognate ligand IL-4. Though KL and IL-4 are individually mitogenic, combinations of KL and IL-4 synergistically promoted mast cell proliferation. Furthermore, inhibition of lipid raft formation by methyl-ß-cyclodextrin resulted in loss of synergistic proliferation. Together the data suggest IL-4R as a novel Kit-activated receptor. Such cross-receptor activations are likely to be a universal mechanism of Kit signaling in hematopoiesis.


Assuntos
Interleucina-4/farmacologia , Mastócitos/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-kit/genética , Receptores de Interleucina-4/genética , Fator de Transcrição STAT6/genética , Fator de Células-Tronco/farmacologia , Animais , Células da Medula Óssea/citologia , Células da Medula Óssea/efeitos dos fármacos , Células da Medula Óssea/imunologia , Proliferação de Células/efeitos dos fármacos , Regulação da Expressão Gênica , Hematopoese/efeitos dos fármacos , Hematopoese/genética , Hematopoese/imunologia , Interleucina-4/genética , Interleucina-4/imunologia , Mastócitos/citologia , Mastócitos/imunologia , Microdomínios da Membrana/efeitos dos fármacos , Microdomínios da Membrana/imunologia , Microdomínios da Membrana/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Cultura Primária de Células , Proteínas Proto-Oncogênicas c-kit/imunologia , Receptores da Eritropoetina/genética , Receptores da Eritropoetina/imunologia , Receptores de Interleucina-3/genética , Receptores de Interleucina-3/imunologia , Receptores de Interleucina-4/imunologia , Receptores de Interleucina-7/genética , Receptores de Interleucina-7/imunologia , Fator de Transcrição STAT6/imunologia , Transdução de Sinais , Fator de Células-Tronco/genética , Fator de Células-Tronco/imunologia , beta-Ciclodextrinas/farmacologia
10.
Ann Lab Med ; 40(3): 193-200, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-31858758

RESUMO

BACKGROUND: Waldenström macroglobulinemia (WM) is a subset of lymphoplasmacytic lymphoma (LPL) with bone marrow (BM) involvement and an IgM monoclonal gammopathy of any level. We aimed to identify the clinical, laboratory, and BM findings of patients with WM and to evaluate the usefulness of CD154 for the diagnosis and prognosis of WM. METHODS: We reviewed the medical records and BM studies and/or flow cytometric immunotyping of 31 patients with untreated WM. Semiquantitative immunohistochemistry (CD20, CD138, tryptase, and CD154) of BM was performed. RESULTS: Only six patients presented with symptoms of hyperviscosity syndrome. Eleven patients had solid cancer and/or another hematologic malignancy. Mast cells (MC) increased in all samples, with some in close contact with tumor cells. Tryptase-positive MC (17.1/ high-power fields [HPF], 1.2-72.0/HPF) and CD154-positive MC (8.6/HPF, 0.1-31.1/HPF) were observed. The high CD154-positive MC (≥8.6/HPF) group showed a lower overall five-year survival rate than the low CD154-positive MC (<8.6/HPF) group (71.9% vs. 100.0%; P=0.012). Flow cytometric immunophenotyping of BM aspirates showed increased B lymphocytes and plasma cells with a normal phenotype (CD138⁺/CD38⁺/CD19⁺/CD45⁺/CD56⁻). CONCLUSIONS: Approximately one third of WM patients showed other malignancies and all patients had increased MC. Immunohistochemistry and flow cytometric immunophenotyping are useful for diagnosing WM, and increased CD154-positive MC can indicate poor prognosis.


Assuntos
Medula Óssea/patologia , Macroglobulinemia de Waldenstrom/diagnóstico , Idoso , Idoso de 80 Anos ou mais , Antígenos CD20/metabolismo , Linfócitos B/citologia , Linfócitos B/metabolismo , Medula Óssea/metabolismo , Ligante de CD40/metabolismo , Diagnóstico Diferencial , Citometria de Fluxo/métodos , Humanos , Imunofenotipagem , Estimativa de Kaplan-Meier , Mastócitos/citologia , Mastócitos/metabolismo , Pessoa de Meia-Idade , Paraproteinemias/complicações , Paraproteinemias/diagnóstico , Plasmócitos/citologia , Plasmócitos/metabolismo , Estudos Retrospectivos , Macroglobulinemia de Waldenstrom/complicações , Macroglobulinemia de Waldenstrom/mortalidade
11.
Int J Mol Sci ; 20(24)2019 Dec 10.
Artigo em Inglês | MEDLINE | ID: mdl-31835486

RESUMO

Mitochondrial aldehyde dehydrogenase (ALDH2) metabolizes endogenous and exogenous aldehydes and protects cells against oxidative injury. Inactivating genetic polymorphisms in humans are common and associate with alcohol flush reactions. However, whether mast cell Aldh2 activity impacts normal mast cell responses is unknown. Using bone marrow-derived mast cells from Aldh2 knockout mice, we found evidence for a role of mast cell Aldh2 in Kit-mediated responses. Aldh2-deficient mast cells showed enhanced Kit tyrosine kinase phosphorylation and activity after stimulation with its ligand (stem cell factor) and augmentation of downstream signaling pathways, including Stat4, MAPKs, and Akt. The activity of the phosphatase Shp-1, which attenuates Kit activity, was reduced in Aldh2-/- mast cells, along with an increase in reactive oxygen species, known to regulate Shp-1. Reduced Shp-1 activity concomitant with sustained Kit signaling resulted in greater proliferation following Kit engagement, and increased mediator and cytokine release when Aldh2-/- mast cells were co-stimulated via Kit and FcεRI. However, FcεRI-mediated signaling and responses were unaffected. Therefore, our findings reveal a functional role for mast cell intrinsic Aldh2 in the control of Kit activation and Kit-mediated responses, which may lead to a better understanding of mast cell reactivity in conditions related to ALDH2 polymorphisms.


Assuntos
Aldeído-Desidrogenase Mitocondrial/genética , Mastócitos/citologia , Proteínas Proto-Oncogênicas c-kit/metabolismo , Fator de Células-Tronco/metabolismo , Animais , Linhagem Celular , Proliferação de Células , Técnicas de Inativação de Genes , Mastócitos/metabolismo , Fosforilação , Proteína Tirosina Fosfatase não Receptora Tipo 6 , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais
12.
J Photochem Photobiol B ; 201: 111634, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31715551

RESUMO

Skin Flap is used in reconstructive plastic surgery. However, complications such as ischemia followed by local necrosis may occur, requiring a new surgical procedure. It is well known that photobiomodulation therapy (PBMT) is an effective technique for improving microcirculation and neoangiogenesis, which contributes positively to the blood supply in the pre and post surgical period. Thus, the objective of the present study was to investigate the effects of preemptive treatment with laser PBMT with different energies on the viability in skin flaps in rats. Sixty-three Wistar rats, male, were randomized into five groups: Control Group (CG) (n = 15): PBMT simulation; Preemptive group 1.1 J laser (GP1) (n = 15): preemptive laser PBMT with 1.1 J of energy per point; Preemptive group 4 J laser (GP4) (n = 15): preemptive PBMT with 4 J of energy per point; Laser group 11 J (G1) (n = 9): PBMT immediately after surgery with 1.1 J of energy per point; Laser group 4 J (G4) (n = 9): TFMB immediately after surgery with 4 J of energy per point. The CG, GP1 and GP4 groups started treatment 72 h prior to surgery and were subdivided into two experimental periods, one of them on the day of the flap and the other along with the other groups on the seventh postoperative day. Three days after the randomization, the animals underwent random skin flap surgery. PBMT was performed with a 660 nm laser at three points. In the first experimental period, a greater number of vessels were found, as well as mast cells in GP1 compared to the CG and greater expression of fibroblast growth factor and vascular endothelial growth factor in the GP1 and GP4 groups compared to the CG. In the second experimental period, GP1 presented a lower percentage of necrotic tissue, a higher number of vessels and a percentage of cells labeled with both VEGF and hypoxia indicible factor alpha (HIF-1α) compared to the CG, FGF in GP1, GP4 and G4 when compared to the CG. Thus, it was concluded that preemptive treatment with PBMT with the application of 1.1 J of energy per point is effective in improving the viability of the skin flap.


Assuntos
Lasers Semicondutores , Retalhos Cirúrgicos/patologia , Animais , Vasos Sanguíneos/patologia , Vasos Sanguíneos/efeitos da radiação , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Masculino , Mastócitos/citologia , Mastócitos/metabolismo , Mastócitos/efeitos da radiação , Necrose , Distribuição Aleatória , Ratos , Ratos Wistar , Retalhos Cirúrgicos/irrigação sanguínea , Fator A de Crescimento do Endotélio Vascular/metabolismo
13.
Int J Mol Sci ; 20(22)2019 11 06.
Artigo em Inglês | MEDLINE | ID: mdl-31698677

RESUMO

BACKGROUND: Laboratory of allergic diseases 2 (LAD2) human mast cells were developed over 15 years ago and have been distributed worldwide for studying mast cell proliferation, receptor expression, mediator release/inhibition, and signaling. LAD2 cells were derived from CD34+ cells following marrow aspiration of a patient with aggressive mastocytosis with no identified mutations in KIT. Another aspiration gave rise to a second cell line which has recently been re-established (LADR). We queried whether LADR had unique properties for the preclinical study of human mast cell biology. METHODS: LADR and LAD2 cells were cultured under identical conditions. Experiments examined proliferation, beta-hexosaminidase (ß-hex) release, surface receptor and granular protease expression, infectivity with HIV, and gene expression. RESULTS: LADR cells were larger and more granulated as seen with Wright-Giemsa staining and flow cytometry, with cell numbers doubling in 4 weeks, in contrast to LAD2 cells, which doubled every 2 weeks. Both LADR and LAD2 cells released granular contents following aggregation of FcεRI. LADR cells showed log-fold increases in FcεRI/CD117 and expressed CD13, CD33, CD34, CD63, CD117, CD123, CD133, CD184, CD193, and CD195, while LAD2 cells expressed CD33, CD34, CD63, CD117, CD133, CD193 but not CD13, CD123, CD184, or CD195. LADR tryptase expression was one-log-fold increased. LADR cell and LAD2 cell chymase expression were similar. Both cell lines could be infected with T-tropic, M-tropic, and dual tropic HIV. Following monomeric human IgE stimulation, LADR cells showed greater surface receptor and mRNA expression for CD184 and CD195. Expression arrays revealed differences in gene upregulation, especially for the suppressor of cytokine signaling (SOCS) family of genes with their role in JAK2/STAT3 signaling and cellular myelocytomatosis oncogene (c-MYC) in cell growth and regulation. CONCLUSIONS: LADR cells are thus unique in that they exhibit a slower proliferation rate, are more advanced in development, have increased FcεRI/CD117 and tryptase expression, have a different profile of gene expression, and show earlier infectivity with HIV-BAL, LAV, and TYBE when compared to LAD2 cells. This new cell line is thus a valuable addition to the few FcεRI+ human mast cell lines previously described and available for scientific inquiry.


Assuntos
Linhagem Celular/citologia , Mastócitos/citologia , Antígenos CD/metabolismo , Degranulação Celular , Proliferação de Células , Quimases/metabolismo , Regulação da Expressão Gênica , Infecções por HIV/patologia , Humanos , Mastócitos/fisiologia , Transdução de Sinais , Triptases/metabolismo , beta-N-Acetil-Hexosaminidases/metabolismo
14.
Eur J Pharmacol ; 864: 172713, 2019 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-31586631

RESUMO

The role of mast cells during inflammatory bowel diseases (IBD) is discussed controversially. Whereas several studies report an increase in mast cell density during IBD, others found a decrease. Recently, we observed a reduced response to mast cell degranulation induced by antigen contact in a colitis model. As the effects of mast cell mediators on epithelial ion transport are mediated indirectly via stimulation of secretomotor neurons, we investigated in vitro whether proinflammatory cytokines change the response to mast cell degranulation. Tumor necrosis factor α (TNFα) and a mix of proinflammatory cytokines caused an increase of short-circuit current (Isc) and tissue conductance in rat colon. Anion secretion induced by histamine was downregulated in the presence of interleukin-1ß (IL-1ß) and the cytokine mix, whereas the response to the mast cell stimulator compound 48/80 was not changed significantly. In a coculture of rat submucosal ganglionic cells with a mast cell line (RBL-2H3), TNFα preincubation for 1 d increased the percentage of neurons responding to mast cell degranulation with an increase of the cytosolic Ca2+ concentration and enhanced the amplitude of this response. Consequently, the downregulation of epithelial secretion is compensated by an increased sensitivity of secretomotor neurons leading to a constant response of the epithelium to compound 48/80. Furthermore, enteric neurons can modify mast cell functions as nicotine inhibited the increase in cytosolic Ca2+ concentration of RBL-2H3 cells and the Isc evoked by compound 48/80. Consequently, these in vitro models deliver new insights into cellular interactions in the gut wall under inflammatory conditions.


Assuntos
Comunicação Celular/efeitos dos fármacos , Citocinas/farmacologia , Mucosa Intestinal/citologia , Mastócitos/citologia , Neurônios/citologia , Neurotransmissores/farmacologia , Animais , Cálcio/metabolismo , Citosol/efeitos dos fármacos , Citosol/metabolismo , Feminino , Mucosa Intestinal/metabolismo , Masculino , Mastócitos/efeitos dos fármacos , Mastócitos/metabolismo , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Permeabilidade/efeitos dos fármacos , Ratos , Ratos Wistar , p-Metoxi-N-metilfenetilamina/farmacologia
15.
Bull Exp Biol Med ; 167(5): 689-693, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31630303

RESUMO

Morphological structures involved in local regulation of tissue processes and the neurotransmitters they produce are assumed to play an important role in the pathogenesis of oncological diseases. We studied the dynamics of neurotransmitter cell counts and distribution of catecholamines and serotonin in them and analyzed proliferative activity of hemopoietic cells in the bone marrow of mice after bone marrow autotransplantation. The bone marrow of male mice was studied by fluorescent and immunohistochemical methods after injection of the bone marrow from the same mouse into the caudal vein. The count of neurotransmitter cells and the cell proliferation index increased after bone marrow autotransplantation.


Assuntos
Células da Medula Óssea/metabolismo , Transplante de Medula Óssea , Catecolaminas/metabolismo , Mastócitos/metabolismo , Neurotransmissores/metabolismo , Serotonina/metabolismo , Animais , Medula Óssea/imunologia , Medula Óssea/metabolismo , Células da Medula Óssea/citologia , Células da Medula Óssea/imunologia , Contagem de Células , Proliferação de Células , Imuno-Histoquímica , Masculino , Mastócitos/citologia , Mastócitos/imunologia , Camundongos , Microscopia de Fluorescência , Transplante Autólogo
16.
Histochem Cell Biol ; 152(6): 439-451, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31549232

RESUMO

Superficial fascia has abundant preadipocytes capable of spontaneous and induced differentiation and is thought to be a novel origin of adipocytes. This study aimed to quantitatively evaluate the spatial distribution and correlation of adipocytes and mast cells in rat superficial fascia. Panoramic images were obtained from whole-mounted fascia stained by toluidine blue. Adipocytes increased gradually in superficial fascia of growing rats. Abundant mast cells, with the degranulation and exocytosis of abundant secretory granules, appeared in fascia where partially differentiating adipocytes and mature adipocytes occurred. Quantitative histological analysis by variance-mean ratio and Morisita index of dispersion indicated that both mast cells and adipocytes in fascia were distributed individually in cluster, but not random or uniform. Spearman's correlation coefficient revealed that the spatial cluster distributions of mast cells and adipocytes positively correlated with each other and correlated with increased number and size of adipocytes and adipogenic areas in fascia. Morphometry analysis indicated the strong correlation between fascial adipocytes and mast cells during the periods of rat growth. The correlation coefficient increased significantly at 8 weeks compared to 4 weeks, consistent with the increasing trend of the number and size of fascia adipocytes in growing rats. This finding provided the first quantitative histological analysis for the spatial distribution and correlation of fascia adipocytes and mast cells, which could be the histocytological basis for further exploring spatial and functional interactions between fascial mast cells and adipocytes. Also, the present data were informative for the research on physiologies and pathologies of fascia and fascia-related connective tissues.


Assuntos
Adipócitos/citologia , Fáscia/citologia , Mastócitos/citologia , Análise Espacial , Animais , Masculino , Ratos , Ratos Sprague-Dawley
17.
Int J Mol Sci ; 20(18)2019 Sep 06.
Artigo em Inglês | MEDLINE | ID: mdl-31500217

RESUMO

The pathophysiological roles of mast cells are still not fully understood, over 140 years since their description by Paul Ehrlich in 1878. Initial studies have attempted to identify distinct "subpopulations" of mast cells based on a relatively small number of biochemical characteristics. More recently, "subtypes" of mast cells have been described based on the analysis of transcriptomes of anatomically distinct mouse mast cell populations. Although mast cells can potently alter homeostasis, in certain circumstances, these cells can also contribute to the restoration of homeostasis. Both solid and hematologic tumors are associated with the accumulation of peritumoral and/or intratumoral mast cells, suggesting that these cells can help to promote and/or limit tumorigenesis. We suggest that at least two major subsets of mast cells, MC1 (meaning anti-tumorigenic) and MC2 (meaning pro-tumorigenic), and/or different mast cell mediators derived from otherwise similar cells, could play distinct or even opposite roles in tumorigenesis. Mast cells are also strategically located in the human myocardium, in atherosclerotic plaques, in close proximity to nerves and in the aortic valve. Recent studies have revealed evidence that cardiac mast cells can participate both in physiological and pathological processes in the heart. It seems likely that different subsets of mast cells, like those of cardiac macrophages, can exert distinct, even opposite, effects in different pathophysiological processes in the heart. In this chapter, we have commented on possible future needs of the ongoing efforts to identify the diverse functions of mast cells in health and disease.


Assuntos
Fenômenos Fisiológicos Celulares , Mastócitos/fisiologia , Animais , Biomarcadores , Movimento Celular , Suscetibilidade a Doenças , Homeostase , Humanos , Mastócitos/citologia
18.
Proc Natl Acad Sci U S A ; 116(41): 20500-20504, 2019 10 08.
Artigo em Inglês | MEDLINE | ID: mdl-31548430

RESUMO

Skin wound infections are a significant health problem, and antibiotic resistance is on the rise. Mast cells (MCs) have been shown to contribute to host-defense responses in certain bacterial infections, but their role in skin wound superinfection is unknown. We subjected 2 MC-deficient mouse strains to Pseudomonas aeruginosa skin wound infection and found significantly delayed wound closure in infected skin wounds. This delay was associated with impaired bacterial clearance in the absence of MCs. Engraftment of MCs restored both bacterial clearance and wound closure. Bacterial killing was dependent on IL-6 released from MCs, and engraftment with IL-6-deficient MCs failed to control wound infection. Treatment with recombinant IL-6 enhanced bacterial killing and resulted in the control of wound infection and normal wound healing in vivo. Taken together, our results demonstrate a defense mechanism for boosting host innate immune responses, namely effects of MC-derived IL-6 on antimicrobial functions of keratinocytes.


Assuntos
Queratinócitos/imunologia , Mastócitos/imunologia , Infecções por Pseudomonas/prevenção & controle , Pseudomonas aeruginosa/imunologia , Pele/imunologia , Cicatrização/imunologia , Infecção dos Ferimentos/prevenção & controle , Animais , Antibacterianos/farmacologia , Células Cultivadas , Humanos , Interleucina-6/farmacologia , Queratinócitos/efeitos dos fármacos , Mastócitos/citologia , Camundongos , Infecções por Pseudomonas/imunologia , Infecções por Pseudomonas/microbiologia , Pele/efeitos dos fármacos , Cicatrização/efeitos dos fármacos , Infecção dos Ferimentos/imunologia , Infecção dos Ferimentos/microbiologia
19.
Psychiatr Danub ; 31(Suppl 3): 357-370, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31488753

RESUMO

Efforts to disclose the mechanisms of transcranial therapeutic electro-magnetic fields (EMF) acting on the brain's cells (Marino, Kibleur) and recently immune cells (Gülöksüz) meet unsolved physiological details of blood vessels, exclusively arterial vasomotion or the non-glial-related former g(lia)-lymphatic flow (Iliff; Liu DX) - now replaced by an astrocytic AQP4-pipeline cooling the brain (Nakada 2014). Here within the convergent dyn4TAM-framework, which had suggested the first mast cell behavioral experiment (Fitzpatrick & Morrow 2017), three intertwined physiological concepts are contributed: A) "autocrinicity" - how flushed, thus absent, autocrine signals integrate external fluidics into cellular computations e.g. on motility: EMFs could increase such absences by targeting e.g. dipole-cytokines; B) a new concept of the arterial wall based on a tangible interpretation of the coronal histology of all arteries as a co-axial pulse-dampening engine (Treviranus 2012). In the brain this engine might provide the quickest cerebral outflow via the Cerebral IntraMUral Reverse Arterial Flow (Treviranus 2018b), while transmitting further forces acting upstream to the paravascular spaces; C) some key roles for mast cells in neuro-psychiatry (Silver & Curley 2013) and their interactive lymphatic and non-luminal vascular routes to the brain dictated by peripheral imprinting as to destiny (Csaba 1987) and destination (Treviranus 2013). Within the skull they might advance against para-arterial upstream currents. Some known causal mediators of the effects of transcranially applied EFMs and puzzling results are then put tentatively in perspective with the above "tangible" models, e.g. by aligning probable induced currents with arterial segments or the new direct meningeal-calvario-myeloid channels. RESULTS: The case for a role of mast cells and diverse flows in transcranial electromagnetic brain therapy seems promising.


Assuntos
Artérias/citologia , Comunicação Autócrina , Encéfalo/irrigação sanguínea , Campos Eletromagnéticos , Sistema Glinfático/fisiologia , Mastócitos/citologia , Humanos
20.
Acupunct Med ; 37(6): 348-355, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31429590

RESUMO

OBJECTIVE: Cannabinoid CB2 receptors (CB2Rs) are mainly present on immune cells including mast cells, which participate in 2,4-dinitrofluorobenzene (DNFB)-induced allergic contact dermatitis (ACD). In this study, we aimed to investigate whether inhibition of mast cell degranulation was involved in the anti-ACD effect of electroacupuncture (EA) at ST36 via CB2R. METHODS: Sprague-Dawley rats were sensitised and challenged with DNFB following EA stimulation for 1 week. Ear swelling, serum IgE levels, local cytokine production and mast cell infiltration were evaluated. Additionally, rat peritoneal mast cells (RPMCs) were isolated and cultured for detection of CB2R expression, mitogen-activated protein kinase (MAPK) signalling activation and mast cell degranulation (including ß-hexosaminidase and histamine release) in the presence or absence of CB2R antagonists. RESULTS: EA treatment inhibited ear swelling, suppressed IgE and cytokine production, decreased the number of mast cells and curbed mast cell degranulation, which was associated with the inhibition of p38 phosphorylation in DNFB-induced ACD. Importantly, EA enhanced the expression of CB2R mRNA and protein in the RPMCs. CB2R antagonist AM630 but not CB1R antagonist AM251 effectively reversed the suppressive effect of EA on p38 activation, mast cell infiltration and degranulation. CONCLUSION: These findings provide more evidence to support the hypothesis that EA promotes CB2R expression in mast cells, which is followed by inhibition of the p38 MAPK pathway, potentially resulting in the anti-ACD effect of EA. This suggests that EA at ST36 may be an effective candidate therapy for treating inflammatory skin diseases such as ACD.


Assuntos
Dermatite Alérgica de Contato/terapia , Eletroacupuntura , Mastócitos/imunologia , Receptor CB2 de Canabinoide/imunologia , Pontos de Acupuntura , Animais , Degranulação Celular , Citocinas/genética , Citocinas/imunologia , Dermatite Alérgica de Contato/genética , Dermatite Alérgica de Contato/imunologia , Humanos , Imunoglobulina E/imunologia , Masculino , Mastócitos/citologia , Ratos , Ratos Sprague-Dawley , Receptor CB2 de Canabinoide/genética , Proteínas Quinases p38 Ativadas por Mitógeno/genética , Proteínas Quinases p38 Ativadas por Mitógeno/imunologia
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