RESUMO
There was no effective measures can be obtained at present to reverse or prevent airway remodeling. We investigated the therapeutic effect of Erythropoietin (EPO) gene modified mesenchymal stem cells (MSCs) on asthmatic airway remodeling and the possible underlied molecular mechanisms. EPO gene was transfected into MSCs via lentivirus vector. The transfected cells (EPO-MSCs) were identified by flow cytometry and the EPO secreting function was detected by PCR and Western blot. MSCs or EPO-MSCs were administrated to albumin (OVA)-induced chronic asthmatic mouse model via tail veins. The asthmatic phenotype was analyzed. Number of cells in bronchoalveolar lavage fluid (BALF) was counted using a hemocytometer. Histological findings of airways were evaluated by microscopic examination. The concentrations of interleukin 4(IL-4), interleukin 5(IL-5), and interleukin 13(IL-13) in lung homogenate were determined by ELISA. The activation state of transforming growth factor-ß 1 (TGF-ß1), Transforming growth factor beta-activated kinase 1 (TAK1), and p38 Mitogen Activated Protein Kinase (p38MAPK) signaling was detected by Real-Time PCR and Western blotting. EPO-MSCs were successfully constructed. EPO-MSCs showed a more potently suppressive effect on local asthmatic airway inflammation and the level of IL-4, IL-5, and IL-13 in lung tissue than MSCs. Moreover, the numbers of goblet cells, the thicknesses of smooth muscle layer, collagen density, percentage of proliferating cell nuclear antigen positive (PCNA+ ) mesenchymal cells, and von Willebrand factor positive(vWF+ ) vessels were also significantly inhibited by EPO-MSCs. Furthermore, EPO-MSCs could downregulate the expression of TGF-ß1, TAK1, and p38MAPK in lung tissue both in mRNA level and in protein level. EPO gene modified MSCs may more efficiently attenuate asthmatic airway remodeling, which maybe related with the downregulation of TGF-ß1-TAK1-p38MAPK pathway activity.
Assuntos
Remodelação das Vias Aéreas/efeitos dos fármacos , Asma/terapia , Modelos Animais de Doenças , Eritropoetina/farmacologia , Células-Tronco Mesenquimais/citologia , Animais , Asma/genética , Asma/metabolismo , Asma/patologia , Líquido da Lavagem Broncoalveolar/citologia , Eritropoetina/genética , Regulação da Expressão Gênica , Terapia Genética , Interleucinas/metabolismo , Lentivirus/genética , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos BALB CRESUMO
OBJECTIVE: To investigate the molecular mechanism of carbapenem resistance in the clinical isolates of Acinetobacter baumannii from Xi'an and their profile of carbapenemase production. METHODS: A total of 146 Acinetobacter baumannii strains were isolated from 6 general hospitals in Xi'an. Antimicrobial susceptibility test was performed for all the strains, followed by detection of imipenem resistance using E-test for metallo-beta-lactamase (MBL) and NaCl inhibition test for OXA type carbapenemase. Bla(OXA-23)and bla(OXA-58) were amplified by PCR, and the positive products were sequenced. RESULTS: From the collected strains, 15 non-repetitive imipenem-resistant Acinetobacter baumannii strains were identified, among which 14 yielded negative results in E-test for MBL production. All the resistant strains showed increased sensitivity to imipenem after NaCl inhibition, suggesting the presence of carbapenemase production. Eleven of the strains harbored OXA -23 type gene and 1 harbored OXA -58 type gene. The concordance rate of the results by NaCl inhibition test and PCR was 85.7%. CONCLUSIONS: Production of OXA-type carbapenemase is the most important reason for carbapenem resistance in Acinetobacter baumannii in Xi'an. The OXA-58 type gene is a novel carbapenemase genotype in China. NaCl inhibition test is a convenient and cost-effective method for detecting carbapenemase in Acinetobacter baumannii.
