The Study Of Menstrual Blood-derived Stem Cells(MenSCs)in The Therapy Of Acute Lung Injury(ALI)

Mesenchymal stem cells(MSCs)are adult stem cells derived from mesoderm,which has the characteristics of self-renewal and multipotent differentiation ability,and can play an important role in regenerative medicine.Menstrual blood-derived mesenchymal stem cells(MenSCs),a member of mesenchymal stem cells,are extracted from female menstrual blood and have the advantages of convenient and no side effects.Compared with other sources of mesenchymal stem cells,MenSCs have no ethical and traumatic problems while the same genetic background can be achieved.At present,MenSCs have been used in the treatments of various diseases such as myocardial infarction,type I diabetes and other diseases.Many infectious diseases such as H7N9,sepsis and some bacteria,especially Gram-negative infection could cause acute lung injury.However,it has not yet been used in the therapy of acute lung injury(ALI).In this study,we will discuss the effects of MenSCs in the therapy of ALI.First,we found that the isolated cells exhibited a spindle-shaped,fibroblast-like morphology and are CD29+,CD44+,CD73+,CD90+,CD105+ and CD34-,CD45-,CDllb-,CD14-,CD19-,CD79a-,CD117-,HLA-DR-,demonstrating they are a kind of MSCs.We also confirmed that MenSCs have a good ability of proliferation in vitro and can be differentiated into osteoblasts and adipocytes.In addition,we found that MenSCs are positive for CD 140b,CD 194,CD221,and CD 184,the receptors of chemokines or cytokines with chemotaxis.It indicates that MenSCs can be stimulated by a variety of cytokines and migrated to the injury,playing an important role in targeted therapies.Then we explored the effects of MenSCs in the therapy of ALI.In vitro experiments,we have shown that MenSCs can migrate to bronchial epithelial cells BEAS-2B and protect them from LPS injury.After two weeks of induction,the shape of the cells had changed from spindle-like to polygonal-like,similar to that of lung epithelial cells and they can express pulmonary surfactant protein SPB,SPC and SPD.In LPS-mediated ALI model mice,X-ray and histopathological sections confirmed the effects of MenSCs in attenuating the inflammation of ALI and reducing the damage of injury lungs.After MenSCs administrated,the dry/wet ratio and expressions of P-catenin protein were elevated while the total proteins in BALF were reduced.These results suggested that the lung permeabilities of mice were improved after MenSCs transplantation.Further,we examined the effect of MenSCs on pulmonary inflammation by detecting the inflammatory cells and MPO activity.Meantimely,we also found that MenSCs can increase the expression of PCNA and reduce the expression of Caspase-3,promoting the repair of damaged lungs.Finally,we explored the possible mechanisms of MenSCs in the therapy of acute lung injury.We found that after MenSCs transplantation,the expressions of anti-inflammatory cytokines IL-10,TGF-? increased and the expressions of pro-inflammatory cytokine IL-? and IL-6 decreased in BALF,serum and lung tissue,indicating that MenSCs may inhibite inflammation through immune regulation.Meantimely,we found that the expressions level of KGF,VE-cadherin and ?-catenin was increased after MenSCs transplantation,indicating that MenSCs could promote the expression of adhesion protein,form adhesion junction,promote cell growth and repair.What is more,we detected the expression of PI3K,p-gsk3?,gsk3?,p-Src and Src proteins in lung tissue using western blot and found that PI3K,gsk3p/p-catenin and Src may be involved in lung repair.These results suggested that MenSCs can play a therapeutic role in LPS-mediated acute lung injury in mice.