| Research topic: Mesenchymal stem cells(BMSC)- secreting keratinocyte growth factor(KGF) in the repair of lipopolysaccharide(LPS)- induced alveolar epithelial cells(MLE12) in mice and its mechanismBackground: Acute respiratory distress syndrome is that is caused by infection, mechanical stimulation, shock, blood transfusion causes pulmonary capillary endothelium and alveolar epithelium in diffuse damage, with acute refractory hypoxemia as the clinical manifestations of acute respiratory failure, at present there is no effective treatment improves the rate of the mortality of ARDS. Animal and cellular studies showed that bone marrow mesenchymal stem cells(bone marrow mesenchymal stem cells and bone marrow mesenchymal stem cells(BMSCs) by homing, graft regeneration effect, immune regulation, regulating alveolar fluid clearance rate, change the pulmonary vascular endothelial permeability and other mechanisms to promote repair of lung injury and early ARDS treatment to provide new ideas. Previous studies found that MSC can significantly repair LPS induced ARDS in mouse lung epithelial injury, also accompanied with alveolar irrigation lotion KGF levels were significantly increased and KGF can influence lung epithelial cell growth, differentiation and molding, it can be concluded that MSC may through paracrine KGF repair ARDS lung skin injury. In recent years, many studies found that the rage for lung epithelial role in the restoration. Therefore, RAGE may be the key molecule regulating effect.Objective: This study intends to in vitro BMSC and alveolar epithelial cell co culture model based on and release of rage by MSC intervention on acute respiratory distress syndrome(ARDS) beside the BMSC secretion mechanism of keratinocyte growth factor on alveolar epithelial repair, and epithelial cells of rage beside the BMSC secretion KGF on acute respiratory distress syndrome(ARDS) in alveolar epithelial injury repair plays a role, to better understand MSc therapy for acute respiratory distress syndrome(ARDS) mechanism, for the future to improve the therapeutic effect of MSC on acute respiratory distress syndrome(ARDS) provide new research foundation.Research methods 1 Establishment of LPS induced MLE12 cell damage model, in vitro indirect co culture model and cell treatment Cell groups are as follows: the blank group: mle12 cultured alone; alveolar epithelial cells injury: mle12 with LPS incubation; indirect co culture control group: mle12 with MSC indirect co culture; indirect co culture group injury: mle12 and MSc indirectly co cultured with LPS incubation; KGF group: mle12 join LPS incubation incubation, join the KGF; rage antibody group: mle12 and MSc co culture LPS plus anti rage antibody was added. 2.Specimen collection and detection: indirect co cultured for 1D, 3D or 7d after collect mle12 cells. By Western blot method determination of rage in cells, ZO-1 content, collecting three time points of culture medium, the ELISA method was used for the determination of KGF in supernatant of culture, the rage of the content. 3. Statistical methods: using SPSS statistical software for statistical analysis, variables were number + standard difference(mean+SD said, the measurement data of multiple group comparison using the Bonferroni correction test analysis, P < 0.05 suggested that the difference was statistically significant.Result:1.MSC has repair effect on lung epithelial injury. In the three time points of 1D, 3D, 7d, mle12 surface in each group, the expression of ZO-1. Western blot showed injury group compared with the blank control group compared to the expression of ZO-1 protein content decreased(P < 0.05); indirect co culture injury group than indirect injury group compared with ZO-1 protein expression was increased(P < 0.05) and KGF group compared with the injury group compared to the expression of ZO-1 protein was significantly increased(P < 0.05); 2 MSC repair epithelial damage, may by the side of the secretion of KGF. In the three time points of 1D, 3D, 7d, conditions in each group training base KGF expression, ELISA results showed that injury group than in the control group compared to the culture medium supernatant factor KGF expression decreased(P< 0.05); indirect co culture injury group than that in the injury group compared, culture supernatant of KGF for sub expression increased(P < 0.05). 3.MSC repair of KGF adjacent to epithelial injury may be related to the regulation of RAGE expression in epithelial cells. In 1D, 3D time groups of epithelial cells and the expression of rage. Western blot showed indirect co culture injury group than that in the injury group compared, rage protein expression was increased(P < 0.05); injury group, on the basis of the added KGF and KGF group than that in the injury group compared. Rage protein expression was significantly increased(P < 0.05). In the three time points of 1D, 3D, 7d, each condition culture medium RAGE expression, ELISA results showed that injury group than the control group compared to culture medium supernatant factor RAGE expression decreased(P < 0.05); indirect co culture injury group than in the injured group compared, culture medium supernatant factor RAGE expression volume increased(P < 0.05); injury group, on the basis of the added KGF and KGF group than that in the injury group compared, culture supernatant factor RAGE expression is increased significantly.Conclusion 1.BMSC induces MLE12 cell damage in LPS cells. 2.BMSC secreted by KGF plays an important role in the repair of MLE-12 induced LPS cell damage, which is the main determinant of alveolar epithelial damage repair. 3 BMSC KGF repair of LPS induced MLE-12 cell damage may be achieved by regulating the expression of RAGE in alveolar epithelial cells, so RAGE may be a key molecule in the alveolar epithelial damage in MSC KGF repair ARDS. |
PDF Full Text Request