| Objective:To study the changes of Wnt5a/JNK signal pathway in the lung tissue of asthmatic rats, and human placenta mesenchymal stem cells transplantation on inflammatory reaction and airway remodeling in lung tissue of asthma rats, and the expression of Wnt5a/JNK signal pathway.Methods:A total of 36 healthy male SD rats were randomly divided into 3 groups, 12 rats in each group:normal control group, asthma group, human placenta derived mesenchymal stem cell transplantation group (hPMSCs).By ovalbumin (OVA) sensitizing and stimulating rats to make asthma model.The collection of bronchoalveolar lavage fluid and the total cell and eosinophil counts, determination of IL-4 and OVA-specific IgE in serum, lung tissue HE staining to confirm asthma model preparation success.Isolation of human placenta derived mesenchymal stem cells with collagenase enzyme digestion method.Through the observation of cell morphology, the expression of specific surface molecules, osteogenic and adipogenic induction to prove that the cell were human placenta derived mesenchymal stem cells and used for transplantation therapy.After the last challenge,airway pathological changes were observed by means of HE staining, the expression of Wnt5a mRNA in lung tissue were examined by quantitative real-time PCR (qRT-PCR), and the expression of p-c-Jun, p-JNK protein in lung tissue were measured by Western blotting.Result:1, The establishment of OVA sensitized rat asthma model:OVA atomization excitation process, rats showed the lips and tip cyanosis, dyspnea, standing instability, incontinence. With increasing excitation days, symptoms gradually worsened. Inflammatory cytokines IL-4 and OVA specific IgE antibody in plasma was increased, the total number of cells and eosinophils in bronchoalveolar lavage fluid was increase.2,Human placenta mesenchymal stem cell isolation, culture and the multipotent differentiation capacity:isolation and culture of human placenta derived mesenchymal stem cells by enzymatic digestion method. Flow cytometry was used to detect cell surface molecules The results showed that the cell surface high expression CD73, CD90,CD105 significantly and low expression molecular CD34,CD45,HLA-DR.In vitro experiments showed that human placenta derived mesenchymal stem cells could different into osteoblasts and adipocytes.3,Compared with the control group, thickended bronchial mucosa with more obvious chrysanthemum fold, narrower airway, and inflammatory cells infiltrated around trachea and pulmonary vessels were observed in asthma rats. The inner airway wall and smooth muscle layer were much thicker in rats of asthma group than those of control group. hPMSCs transplantation group, the pathological changes of asthma group improved obviously. The thickness of bronchial smooth muscle thickness was significantly decreased4, The level of Wnt5a mRNA and p-c-Jun, p-JNK protein expression in lung and blood of asthmatic rats were higher compared with the control group (P<0.05). Compared with asthma group, the expression of Wnt5a mRNA and p-JNK? p-c-Jun in lung tissue was reduced in hPMSCs transplantation group (P< 0.05).Conclusion:1, OVA sensitized can successfully reproduce the asthma model. The inflammation and airway remodeling can relieve in asthma rats by the transplantation of hPMSCs.2, successful extraction of human placenta derived mesenchymal stem cells cultured by enzyme digestion, and they could different into osteoblasts and adipocytes.3, the Wnt5a/JNK signal transduction pathway in asthma rats lung tissue was activated. Transplantation of human placenta mesenchymal stem cells inhibited Wnt5a/JNK. HPMSCs reduces inflammation and airway remodeling in rats with asthma, the Wnt5a/JNK pathway may be one of the effective mechanism of hPMSCs transplantation for the treatment of asthma... |
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