Tissue factor pathway inhibitor promotes the migration, proliferation and colonization of MSCs by regulating CXCL12/CXCR4 signaling pathway
MSCs (mesenchymal stem cells) can promote the structural and functional recovery of lung injury, but most of MSCs do not retain in the injury sites. TFPI (tissue factor pathway inhibitors) can promote stem/progenitor cell migration and homing. The objective of this study was to explore investigate the feasibility and efficacy of TFPI combined therapy with MSCs to alleviate lung injury and the possible mechanism. rhTFPI or (and) MSCs were administered via tail vein to LPS-induced ALI mice. Lung H&E, coagulation function, TFPI, TNF-a, IL-1ß, CXCL12 and CXCR4 in BALF and plasma, 7-day survival rate, and colonization of MSC in lung were observed. Transmembrane migration assay, CCK-8 and EdU assay were used to determine the migration, viability and proliferation of MSCs. RNA sequencing was performed to find differentially expressed genes. The lentivirus transfection technique was used to infect MSCs to overexpession and knockdown TFPI of MSCs. The data demonstrated that rhTFPI could promote cell viability, proliferation and migration of MSCs to improve lung injury. RNA sequence indicated that CXCL12 was in top 40 DEGs and up-regulated after the combination treatment of rhTFPI and MSCs. LPS dampened TFPI, CXCL12/CXCR4 levels in a dose- and time- dependent manner, but can restored by rhTFPI. The level of TFPI expressed by MSCs affected the levels of CXCL12/CXCR4 in MSCs, and migration, proliferation and colonization of MSCs. The results indicated that TFPI may promote the migration, proliferation and colonization of MSCs by regulating CXCL12/CXCR4 signaling pathway. Overall design: Fresh lung tissue was prepared and added Trizol for lysis and total RNA extraction. mRNA library was prepared, and sequencing was conducted using VAHTS Universal V6 RNA-seq Library Prep Kit. Differential expression analysis was performed using the DESeq2. Q value < 0.05 and foldchange > 2 or foldchange < 0.5 was set as the threshold for significantly differential expression gene (DEGs). GO and KEGG pathway enrichment analysis of DEGs were performed to screen the significant enriched term using R (v3.2.0). Protein-protein interaction (PPI) analysis was performed for target genes.