Intervention Effects Of Bone Marrow-deirved Mesenchymal Stem Cells Transplantation To The Bleomycin-induced Pulmonary Fibrosis Formation Period Mice

Objective: To research the intervention effect of transplant exogenous bonemarrow-derived mesenchymal stem cells (BMSCs) on bleomycin-induced advancedpulmonary fibrosis, also defined as pulmonary fibrosis formation in mice.Methods: BMSCs were harvested with the whole bone marrow adherent cellsisolation and culture method from male6-8week-old ICR mice.160female ICR miceaged6-8week-old were randomized into4arms with40in each arm: negativecontrol arm (Group NC), positive control arm (Group PC), mesenchymal stem cellsintervention arm A (group A), mesenchymal stem cells intervention arm B (group B).Group NC was instilled with50ul PBS via nasal tube. Group PC and Group B wereinjectedbleomycin (6ug/g,50ul) via nasal tube to prepare mice pulmonary fibrosismodel. On the14th day after molding, Group A was given200ul of MSCs at theconcentration of1×107/ml via the tail intravenous infusion and from the14thday on,Group B was injected MSCs (1×107/ml,200ul) via the tail vein every other day. Allmice in each group were killed together on the28thday after molding. The wholelung tissues were weighted and partial lung tissues were taken for HE, MASSONstaining and HYP content measure. Fluorescent quanfitation PCR andImmunohistochemistry were applied to detect the expression of SP-C and AQP5inlung tissues. The DNA extracted from lung tissues was used to test the Sry gene-thegender determinative gene of male mice.Results:1.Lung coefficient: the lung coefficient obtained from group PC andgroup BMSCs A, B are higher than that from group NC. There is no significantdifference among group PC, group A and group B.2.The pathological changes:pulmonary alveolitis and fibrosis extent of group PC and group BMSCs A, B aremore severe than those of group NC. In comparison with group PC and group B, thepulmonary alveolitis and fibrosis extent of group A is less severe, but the differencehas no statistical significance.3. The content of HYP: HYP content of group PC andgroup BMSCs A, B are obviously above that of group NC. HYP content of Group Ais slightly lower than that of group PC and group B, and the difference from Group B is more obvious, which has statistical significance, and the difference from Group PCand Group B are quite small.4. Sry gene detect: Scy gene of male mice are detectedin both Group A and Group B.5. SP-C protein Expression: SP-C protein expressionof group PC and group A, B are inferior to that of group NC. That of group A ishigher than group PC, group B. No significant difference is detected between groupPC and group B.6. AQP-5protein Expression: AQP-5protein expression of groupPC and group A, B were lower than group NC. There is no significant differenceamong PC group, group A, group B.7. SP-C mRNA Expression: SP-C mRNAexpression of group PC and group BMSCs A, B were lower than group NC. That ofgroup A is higher than group PC, group B, and the difference has statisticalsignificance. But no obvious difference exists in comparison between group PC andgroup B.8. AQP5mRNA Expression: The AQP5mRNA expression of group PC,group A and group B are inferior to that of group NC. It has no significant differenceamong PC group, group A, group B.Conclusion:1. Exogenous BMSCs via the tail vein injection to Dubbobleomycin-induced pulmonary fibrosis in mouse lung tissue and can be successfullyimplemented in the local colonization.2.The fibrosis extent of the doner-derivedinjected via tail vein with single dose is lower than the simple model, but it is nostatistically significant. Currently，it cannot explain exogenous BMSCs in pulmonaryfibrosis formation period of intervention in the treatment of pulmonary fibrosis.3.There is no significant difference between the single and multiple doses ofexogenous BMSCs in pulmonary fibrosis formation period intervention.