| Objective: To study the mechnisum and changes of Th17、 Treg and effect of human placenta mesenchmal stem cells transplantation in lung tissue and blood of capillary bronchitis rats caused by respiratory syncytial virus.Methods: The rats were randomly divided into normal group, RSV-B group and hpMSCs goup. The model of RSV-B was estabolished successfully by RSV intranasal dripping, and the pathological changes of lung tissues were detected by HE staining; the level of interleukin 4(IL-4), IgE and ECP of plasma were detected by ELISA. All of that are used to congfirm RSV bronchitis modle success.Human placenta mesenchymal stem cells were isolated by collagenase enzyme digestion method. The cells were proved to be Human placenta mesenchymal stem cells by specific surface molecules, adipogenic and osteogenic induction and cell morphology, then they would be transplanted into rats for therapy. After 8 weeks,pathological changes in lung tissues were observed using hematoxylin and eosin staining. The levels of Th17 and Treg subsets were determined via flow cytometry.The related cytokines such as IL-17. IL-23、IL-10、TGF-β were measured using enzyme-linked immunosorbent assay. The expression levels of related transcription factors, such as RORyt and FOXP3, were examined using real-time quantitative polymerase chain reaction and Western blot analysis.Results1. The bronchitis model was successfully established by using nasal drip with RSV.2. The isolation, culture and the multipotent differentiation capacity of human placenta mesenchymal stem cell: human placenta mesenchymal stem cell could be isolated and cultured by enzymatic digestion method. The cell surface molecules were detected by Flow cytometry. We found that the cells could highly expressed CD73, CD95, CD 105 and lowly expressed CD34、CD45、HLA-DR. Mesenchymal stem cells derived from human placenta have the capacity to differentiate into osteoblasts and adipocytes.3. The percentage of Treg cells in plasma were lower in the RSV-B group than in the NC group (P<0.05); The percentage of Treg cells in plasma were higher in the hpMSCs goup than in the RSV-B group (P<0.05). By contrast, the percentage of Th17 cells in plasma were higher in the RSV-B group than in the NC group (P<0.05);The percentage of Th17 cells in plasma were lower in the hpMSCs goup than in the RSV-B group (P<0.05).4. Compared with the NC group, the RSV-B group exhibited pulmonary interstitial hyperemia and edema,inflammatory cell infiltration,wide alveolar septa,and bronchial collapse and deformation. While the the pathological changes of hpMSCs goup improved obviously compared with RSV-B group.5. The concentrations of Th17-related cytokines IL-17, IL-23 in plasma were higher in the RSV-B group than in the NC group (P<0.05); The cytokines IL-17, IL-23 in plasma were lower in the hpMSCs goup than in the RSV-B group (P<0.05). By contrast, the concentrations of Treg-related cytokines IL-10, TGF- β in plasma were lower in the RSV-B group than in the NC group (P<0.05); the concentrations of Treg-related cytokines IL-10,TGF- β in plasma were higher in the hpMSCs goup than in the RSV-B group (P<0.05).6.Compared with the NC group, the level of RORγt mRNA and RORyt protein was higher in the RSV-B group than in the NC group (P<0.05); the level of RORyt mRNA and ROR γ t protein was lower in the hpMSCs goup than in the RSV-B group(P<0.05). By contrast, the level of and FOXP3 mRNA and FOXP3 protein was lower in the RSV-B group than in the NC group (P<0.05); the level of and FOXP3 mRNA and FOXP3 protein was higher in the hpMSCs goup than in the RSV-B group(P<0.05).Conclusion 1.The bronchitis model was successfully established by using nasal drip with RSV, and the the pathological changes of RSV bronchitis was decreased by hpMSCs transplation.2. Mesenchymal stem cells derived from human placenta successfully extracted by enzyme digestion could different into osteoblasts and adipocytes.3. There was an imbalance between Th17 and Treg in blood of RSV bronchitis rats,which shows a higher ROR expression and a lower FOXP3 expression in the capillary bronchitis rats caused by RSV. Then we could conclud that the imbalance in Th17/Treg are involved in the pathogenesis of RSV bronchiolitis.4.Mesenchymal stem cells derived from human placentamay could suppress airway inflammation in the capillary bronchitis rats caused by RSV by regulating the expression of related factors related Th17 and Treg. |
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