Heterogeneity of mast cells and expression of Annexin A1 protein in a second degree burn model with silver sulfadiazine treatment.

Mast cells (MCs) participate in all stages of skin healing and one of their mediators is the Annexin A1 protein (AnxA1), linked to inflammation, proliferation, migration and apoptosis processes, but not studied in thermal burns yet. Therefore, our objectives were to evaluate the behavior of MCs and AnxA1 in a second degree burn model, treated or not with silver sulfadiazine 1% (SDP 1%) and associated to macrophages quantification and cytokines dosages. MCs counts showed few cells in the early stages of repair but increased MCs in the final phases in the untreated group. The normal skin presented numerous tryptase-positive MCs that were reduced after burning in all analyzed periods. Differently, few chymase-positive MCs were observed in the early stages of healing, however, increased chymase-positive MCs were found at the final phase in the untreated group. MCs also showed high immunoreactivity for AnxA1 on day 3 in both groups. In the tissue there was a strong protein expression in the early stages of healing, but in the final phases only in the SDP treated animals. TNF-α, IL-1β, IL-6, IL-10 and MCP-1 levels and macrophages quantification were increased in inflammation and reepithelialization phases. Reduced IL-1β, IL-6 and IL-10 levels and numerous macrophages occurred in the treated animals during tissue repair. Our results indicate modulation in the profile of MCs and AnxA1expression during healing by the treatment with SDP 1%, pointing them as targets for therapeutic interventions on skin burns

Differential expression of AMPA subunits induced by NMDA intrahippocampal injection in rats

Glutamate is involved in excitotoxic mechanisms by interacting with different receptors. Such interactions result in neuronal death associated with several neurodegenerative disorders of the central nervous system (CNS). The aim of this work was to study the time course of changes in the expression of GluR1 and GluR2 subunits of glutamate amino-acid-3-hydroxy-5-methyl-isoxazol-4-propionic acid (AMPA) receptors in rat hippocampus induced by NMDA intrahippocampal injection. Rats were submitted to stereotaxic surgery for NMDA or saline (control) microinjection into dorsal hippocampus and the parameters were evaluated 24 hours, 1, 2 and 4 weeks after injection. The extension and efficacy of the NMDA-induced injury were evaluated by Morris water maze (MWM) behavioral test and Nissl staining. The expression of GluR1 and GluR2 receptors, glial fibrillary acidic protein (GFAP) and neuronal marker (NeuN) was analyzed by immunohistochemistry. It was observed the impairment of learning and memory functions, loss of neuronal cells and glial proliferation in CA1 area of NMDA compared with control groups, confirming the injury efficacy. In addition, NMDA injection induced distinct changes in GluR1 and GluR2 expression over the time. In conclusion, such changes may be related to the complex mechanism triggered in response to NMDA injection resulting in a local injury and in the activation of neuronal plasticity

Ethanolic Extract of Garcinia kola Stem Bark Inhibits LDL-Uptake and LPS-Induced Anxa-1 and ICAM-1 Expression in Human Umbilical Vein Endothelial Cells (HUVECS)

The endothelial cells’ dysfunction linked to the development of atherosclerosis plays an important role in the regulation of inflammatory responses. Previous studies demonstrated the antiatherogenic effects of Garcinia kola seed extracts based on their lipid lowering effects. Our recent studies showed the in vitro antioxidant and anti-inflammatory activities of the ethanolic extract of Garcinia kola stem barks (EE). For more insight on the antiatherogenic effect of EE, we investigated its activity on some key points of the atherosclerosis process. The cytotoxicity of EE as well as its effects on LDL-uptake, LPS-induced InterCellular Adhesion Molecule (ICAM-1) and Annexin-1 (Anxa-1) expression and LPS-induced DNA damage were evaluated in Human Umbilical Vein Endothelial Cells (HUVECs). EE significantly (p<0.0001) increased the cell viability in both naive and LPS-treated HUVECs. At the concentrations of 50 and 100 µg/ml, EE significantly reduced LDL-uptake by endothelial cells stimulated with LPS. The immunohistochemistry results showed a significant (p<0.01) decrease in the ICAM-1 expression at the EE concentration of 250 µg/ml. EE also showed a significant (p<0.0001) concentration-dependent reduction of Annexin-1 expression in LPS-treated HUVECs. Besides, EE exhibited significant inhibition on LPS-induced genotoxicity marked by a decrease in tail DNA expression (p<0.0001) and tail movement expression (p<0.001) for the concentrations of 50 and 100 µg/ml. These findings showed that EE may mitigate the atherogenic process by reducing LDL-uptake and the expression of adhesion molecules. Thus, the EE of Garcinia kola turns out to be a potential candidate for the treatment of atherosclerosis with a lower risk of toxicity. Keywords: Garcinia kola, LDL-uptake, LPS-induced inflammation, comet assay

Cytokines and macrophages in wound healing in a second degree burn.

<p>(a) TNF-α: high dosages in C3 and in both groups after 7 days. (b) IL-1β: high levels in both groups on days 3 and 7 and also in C14. (c) IL-6: overexpression on day 7, especially in the control group. (d) IL-10: Increased levels on days 3 and 7. (e) MCP-1: overexpression on days 3 and 7. (f) Macrophages: numerous macrophages on day 3 and SDP7. (g): Macrophage. Counter-staining: Hematoxilin. Bar 2μm. Values are presented as mean ± S.E.M. (n = 5/group). * <i>p<0</i>.<i>05 vs</i> Normal Skin (N).</p

Morphology of MCs in burn healing process.

<p>(a) MCs (arrows), most intact in Normal Skin (N). (b) C3 and (c) SDP3 with few MCs and mostly degranulated. (d) C14 (e) SDP14, MCs degranulated, mainly in control group. (f) C21, numerous intact MCs. Details of intact (a and f) and degranulated MCs (b and c). Staining: Toluidine Blue. Bars 50 μm. Quantification of MCs: (g) numerous intact MCs in N and C21 and (h) degranulated MCs in SDP7 and in both groups on day 14. (i) Differences between quantification of MCs evidenced by Toluidine blue or Safranin-O, with few MCs S-O+ (MCs histamine storage) in SDP7, C14 and SDP14 compared to the MC TB+ (total MCs in the tissue). (j) MC stained with Safranin-O (MC S-O+); (k) reactive MC for tryptase (MCT) and (l) chymase (MCC); Bars 50 μm. Heterogeneity of MCs, showing many MCTs (m) in N and numerous MCCs (n) in C21. Values are presented as mean ± S.E.M. (n = 5/group). * <i>p<0</i>.<i>05 vs</i> N and γ <i>p<0</i>.<i>001 vs</i> C14.</p

Macroscopic and histopathologic analysis of the healing process in a second degree burn.

<p>(a) C3 and (b) SDP3, inflammation phases, with leukocytes influx and presence of adipocytes in both groups. (c) C7 and (d) SDP7, proliferation phase with re-epithelialization (arrows). Up to 7 days weakly stained collagen fibers (a3, b3, c3 and d3) and low birefringent after polarization (a4, b4, c4 and d4) in the dermis of both groups. (e) C14 and (f) SDP14, complete reepithelialization and fast healing in SDP 1% group (e1 and f1). Dermis and hypodermis (e2, f2—arrows) are better organized and collagen fibers more strongly stained in the group treated with SDP 1% (e3, f3). (g) C21 and (h) SDP21, remodeling phase, the epithelial attachments (g1, h1, g2, h2—arrows) may be observed in larger amount and with increased birefringence under polarized light in the groups treated with SDP 1% (g4 and h4). Normal Skin (i). (a1, b1, c1, d1, e1, f1, g1, h1 and i1) Macroscopic analisys. (a2, b2, c2, d2, e2, f2, g2, h2 and i2) Staining: HE. Bars 500 μm. Picrossirius staining without (a3, b3, c3, d3, e3, f3, g3, h3 and i3) and after (a4, b4, c4, d4, e4, f4, g4, h4 and i4) polarization. Bars 200μm.</p

Anti-inflammatory actions of herbal medicines in a model of chronic obstructive pulmonary disease induced by cigarette smoke

The effects of four medicinal herbs (Arctium lappa, Plantago major, Mikania glomerata Spreng and Equisetum arvense) with anti-inflammatory properties were evaluated in a chronic obstructive pulmonary disease (COPD) model. Wistar rats were exposed to cigarette smoke during 8 weeks and one of the groups was orally given a solution containing 4% of each alcoholic herbal extracts during the exposure period. Control group was not exposed to smoke or treated. Histopathological, immunohistochemical and biochemical analyzes were performed. Normal blood plasma levels of gamma glutamyl transferase indicated no toxicity of the administered herbal extracts. The treatment reduced leukocytes influx in bronchoalveolar lavage, mast cell and macrophages numbers in lungs, as well as prevented pulmonary congestion and tracheal metaplasia. Herbal mixture also decreased plasma inflammatory mediator levels and pulmonary expression of annexin A1 and nuclear factor-k beta. Our data indicate synergistic and protective effects of the used herbal medicines in animals exposed to cigarette smoke as a potential therapeutic strategy.Faculdades Integradas Padre Albino (FIPA), Catanduva, SP, BrazilFundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP), SP, BrazilIntegrated Coll Padre Albino Fdn FIPA, Catanduva, SP, BrazilSao Paulo State Univ UNESP, Inst Biosci Humanities & Exact Sci IBILCE, Dept Biol, Lab Immunomorphol, Sao Jose do Rio Preto Campus, Sao Paulo, SP, BrazilMunicipal Inst Higher Educ Catanduva IMES, Catanduva, SP, BrazilSao Paulo Fed Univ UNIFESP, Sao Paulo, SP, BrazilUniv Aveiro, Aveiro Inst Mat, Dept Biol, Aveiro, PortugalCICECO, Aveiro, PortugalSao Paulo Fed Univ UNIFESP, Sao Paulo, SP, BrazilFAPESP: 2015/033595Web of Scienc