ABSTRACT
Although bone marrow-derived mononuclear cells (BMNC) have been extensively used in cell therapy for cardiac diseases, little mechanistic information is available to support reports of their efficacy. To address this shortcoming, we compared structural and functional recovery and associated global gene expression profiles in post-ischaemic myocardium treated with BMNC transplantation. BMNC suspensions were injected into cardiac scar tissue 10 days after experimental myocardial infarction. Six weeks later, mice undergoing BMNC therapy were found to have normalized antibody repertoire and improved cardiac performance measured by ECG, treadmill exercise time and echocardiography. After functional testing, gene expression profiles in cardiac tissue were evaluated using high-density oligonucleotide arrays. Expression of more than 18% of the 11981 quantified unigenes was significantly altered in the infarcted hearts. BMNC therapy restored expression of 2099 (96.2%) of the genes that were altered by infarction but led to altered expression of 286 other genes, considered to be a side effect of the treatment. Transcriptional therapeutic efficacy, a metric calculated using a formula that incorporates both recovery and side effect of treatment, was 73%. In conclusion, our results confirm a beneficial role for bone marrow-derived cell therapy and provide new information on molecular mechanisms operating after BMNC transplantation on post ischemic heart failure in mice.
Subject(s)
Bone Marrow Transplantation , Heart/physiopathology , Myocardial Infarction/therapy , Myocardium/metabolism , Transcriptome , Animals , Female , Gene Expression Profiling , Gene Expression Regulation , Inflammation Mediators/blood , Male , Mice , Mice, Inbred C57BL , Myocardial Infarction/immunology , Myocardial Infarction/metabolism , Myocardium/pathology , Oligonucleotide Array Sequence Analysis , Oxygen Consumption , Physical Exertion , Ventricular RemodelingABSTRACT
Recently, the P2X(7) receptor has been reported to be associated with chronic inflammatory and neuropathic pain. Because Rheedia longifolia extract has analgesic and anti-inflammatory activity, we evaluated the in vitro inhibitory potential of methanol extract and fractions from its leaves on the P2X(7) purinergic receptor. The activity of P2X(7) was studied with a dye uptake assay and with the whole-cell patch clamp technique in mouse peritoneal macrophages treated with methanol extract of R. longifolia leaves and fractions. The dye uptake was evaluated by flow cytometry and fluorescence microscopy. The R. longifolia extract and some fractions showed an inhibitory effect on the P2X(7) purinergic receptor in a dose-dependent manner. The ethyl acetate fraction exhibited the most potent inhibitory effects. The methanol extract and the butanol fraction showed the same inhibitory effects, despite their lower potency compared with the other fractions. The R. longifolia extract and some of its fractions may be anti-inflammatory because of their inhibitory effect on the P2X(7) receptor. Further investigation is needed to determine the pattern of inhibition and selectivity. Chromatographic analysis indicated the presence of bisflavonoids in the methanol extract fractions. A member of this chemical family is the most probable active compound responsible for the P2X(7) inhibitory effects present in the R. Longifolia extract and fractions.
Subject(s)
Clusiaceae/chemistry , Drug Discovery , Macrophages, Peritoneal/drug effects , Plant Extracts/pharmacology , Plant Leaves/chemistry , Purinergic P2X Receptor Antagonists/pharmacology , Receptors, Purinergic P2X7/chemistry , Animals , Anti-Inflammatory Agents, Non-Steroidal/analysis , Anti-Inflammatory Agents, Non-Steroidal/chemistry , Anti-Inflammatory Agents, Non-Steroidal/isolation & purification , Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Cell Membrane Permeability/drug effects , Cell Survival/drug effects , Cells, Cultured , Chemical Fractionation , Flavonoids/adverse effects , Flavonoids/analysis , Flavonoids/isolation & purification , Flavonoids/pharmacology , Fluorescent Dyes/metabolism , Macrophages, Peritoneal/cytology , Macrophages, Peritoneal/enzymology , Macrophages, Peritoneal/immunology , Male , Mice , Patch-Clamp Techniques , Plant Extracts/adverse effects , Plant Extracts/chemistry , Plant Extracts/isolation & purification , Purinergic P2X Receptor Antagonists/analysis , Purinergic P2X Receptor Antagonists/chemistry , Purinergic P2X Receptor Antagonists/isolation & purification , Solvents/chemistryABSTRACT
RATIONALE: Aerobic conditioning improves exercise capacity and decreases symptoms in patients with asthma. However, its benefits in the context of allergic airway inflammation are poorly understood. OBJECTIVES: To evaluate the effects of two intensities of aerobic exercise on airway inflammation and remodeling in a model of chronic allergic lung inflammation. METHODS: Mice were subjected to chronic ovalbumin (OVA) sensitization and to 4 weeks of low (OVA+Low) or moderate (OVA+Mod) exercise training in a treadmill. Airway inflammation and remodeling and expression of helper T-cell type 1 and 2 cytokines were evaluated. MEASUREMENTS AND MAIN RESULTS: OVA-induced allergic airway inflammation and remodeling were characterized by an increase in collagen (288%), elastic fiber (56%), smooth muscle (380%), and epithelial (402%) contents (P < 0.001) when compared with the control group. OVA+Low and OVA+Mod groups presented a decrease in bronchoalveolar lavage fluid eosinophils (respectively, 84 and 75%; P < 0.01) and airway walls (respectively, 94 and 58%; P < 0.001) when compared with the OVA group. OVA+Low and OVA+Mod groups also presented a reduction in the number of peribronchial inflammatory cells expressing IL-4 (respectively, 85 and 75%; P < 0.01) and IL-5 (respectively, 88 and 89%; P < 0.01) when compared with the OVA group. Aerobic conditioning did not change the expression of either IFN-gamma or IL-2 by inflammatory cells or plasma levels of IgE or IgG1. OVA+Low and OVA+Mod groups presented an increase in the expression of IL-10 (P < 0.001). Low and moderate aerobic conditioning also reduced airway remodeling in OVA-sensitized mice when compared with the OVA group. CONCLUSIONS: We concluded that low and moderate aerobic exercise decreases airway inflammation and remodeling in a murine model of asthma.
