Preliminary Investigation Of How HMGB1 Regulates Mesenchymal Stem Cell Migration And Endothelial Differentiation

Background Mesenchymal stem cells(MSC)have multipotential differentiation and play an important role in the repair of tissue damage.In previous studies,we modified MSC(MSC-H cells)with high mobility group protein 1(HMGB1)for the treatment of radiation-induced vascular injury(RVI)in rats.We found that MSC-H cells showed strong migration and differentiation to endothelial cells(EC)in the treatment of RVI,which promoted the repair of vascular injury.Objective To investigate the mechanism of HMGB1 regulating MSC-H cell migration and differentiation to EC.Methods After EC-MSC coculture,differentiation of MSC-H cells to EC was verified by detection of CD31 expression and LDL uptake of MSC-H cells.The migration ability of MSC-H cells was analyzed by Transwell assay.Western blot was used to detect the content of HMGB1 in MSC-H cell culture supernatant.After using HMGB1 neutralizing antibody,the above method was used to detect the differentiation and migration ability of MSC-H cells.Microarray analysis was used to analyze the differentially expressed genes of MSC-H cells relative to control(MSC-C cells),and the differentially expressed genes of MSC-H cells and MSC-C cells before and after co-culture.GO analysis was used to further screen the molecules that affected the migration ability of MSC-H cells,and selected the molecules with the largest fold change.The inhibitors were used to detect the effect of the selected molecules on the migration of MSC-H cells.Results EC-MSC coculture showed that MSC-H cells had good differentiation potential to EC.The content of HMGB1 in the culture supernatant of MSC-H cells increased.After using HMGB1 neutralizing antibody,the differentiation and migration ability of MSC-H cells was inhibited.GO analysis suggested that the expression level of the Calcium voltage-gated channel subunit alpha1 H(CACNA1H)was most significantly increased,among the molecules that were differentially expressed in MSC-H cells and involved in migration.After HMGB1 knock down,the expression level of CACNA1 H decreased,and selective T-type calcium channel blockers could inhibit the migration ability of MSC-H cells.Conclusion The migration and differentiation ability of MSC-H cells modified by HMGB1 was enhanced.HMGB1 promoted the migration and differentiation of MSC-H cells through autocrine.HMGB1 promoted the migration of MSC-H cells by increasing the expression of CACNA1 H.