Menstrual Blood-derived Stem Cells Mitigate Pulmonary Fibrosis And The Underlving Mechanism

Idiopathic Pulmonary fibrosis is an interstitial lung disease with unknown etiology and featured as interstitial fibrosis,the pathogenesis of which is unclear,and there is currently a lack of diagnostic criteria.The prognosis of patients is poor.Recently,mesenchymal stem cells have shown great potential in regenerative medicine due to their immunomodulatory function and multidirectional differentiation potential,and they are increasingly being used in the treatment of diseases.Menstrual blood-derived stem cells(MenSCs)are a new source of mesenchymal stem cells,derived from menstrual blood,with a wide range of sources,easy access and no ethical concerns.Previous studies have found that MenSCs have a good therapeutic effect in acute lung injury and liver fibrosis,we further explored the effects of MenSCs on pulmonary fibrosis.In this study,we explored the effects of MenSCs on Bleomycin-induced pulmonary fibrosis.We found that transplantation of MenSCs in BLM-induced pulmonary fibrosis mice increased the lung’s dry-to-wet ratio,decreased lung collagen deposition and alveolar lavage fluid total protein.As the same time MenSCs transplantation decreased inflammatory cells and inflammatory factors.In addition,we found that MenSCs also significantly inhibited the apoptosis of lung epithelial cells induced by BLM,and the expression of BCL2 in lung tissue was increased while cleaved caspase3 and Bax was decreased.Furthermore,we found that MenSCs can migrate more to injured lung,but rarely differentiate into lung epithelial cells.Moreover,transwell co-culture experiments showed that MenSCs inhibited the proliferation and differentiation of lung fibroblasts.Meanwhile,we also found that MenSCs inhibited BLM-induced apoptosis of lung epithelial cells through CCK8 experiments,plate cloning experiments,apoptosis and cell cycle experiments.In addition,immunofluorescence experiments,migration and invasion experiments showed that MenSCs inhibited the epithelia-mesenchymal transition process of lung epithelia cells.In order to further clarify the underlying mechanism of MenSCs in pulmonary fibrosis,we collected the co-cultured supernatant of MenSCs and lung epithelial cells(MLE-12),and detected the secretion of cytokines through antibody chips.The results showed that MenSCs significantly reduced the expression of RANTES,GM-CSF,MIP-1γ,MCP-5,Eotaxin1,CCL1 and GITR.At the same time,ELISA experiments further verified the results of the antibody chip.In conclusions,our results showed that MenSCs significantly inhibited lung epithelial cell damage caused by BLM in vivo and in vitro.Meanwhile,MenSCs inhibited the proliferation and differentiation of lung fibroblasts in vitro.Moreover,MenSCs inhibited the inflammation in mice with pulmonary fibrosis.Furthermore,MenSCs reduced the expression of RANTES,GM-CSF,MIP-1γ,MCP-5,Eotaxin1,CCL1 and GITR factors to alleviate apoptosis and EMT of MLE-12 induced by BLM.In summary,our results show that MenSCs alleviated the process of pulmonary fibrosis through anti-inflammatory and anti-apoptosis,which lays a theoretical basis for future clinical applications.