| Introduction:Our previous study reported that bradykinin(BK)can regulate the proliferation and migration in cardiac c-Kit+cells,but whether BK can promotes the angiogenesis effects of exosomes in cardiac c-Kit+cells has not been reported.In this study,we hypothesis that BK promotes the angiogenic activity of the exosome from cardiac c-Kit+ cells by regulating miRNA expression.Methods:BK-treated cardiac c-Kit+ cells medium was collected and treated on human umbilical vein endothelial cells(HUVECs)for detecting the angiogenic activity.The exosomes in cardiac c-Kit+ cells medium were separated with ultracentrifugation.In addition,the effects of exosomes on angiogenesis are explored in HUVECs with matrigel assay in vitro and hind limbs ischemic mice in vivo.We also detected the angiogenesis-related proteins and microRNA in the exosome to explore the related regulator involved in the angiogenesis effect of BK.Results:The results showed that BK-treated cardiac c-Kit+ cells medium have more pro-angiogenesis effect on HUVECs compared to the control cardiac c-Kit+medium.Then we separated the exosome from the BK or vehicle treated cardiac c-Kit+medium.Interestingly,the exosome from BK-treated cells medium also have more pro-angiogenesis in HUVECs,but excluded exosome medium from two groups have no effect on HUVECs angiogenesis.This angiogenesis effect was confirmed in hind limbs ischemic mice by laser doppler speckle system.In order to explore the pro-angiogenic activity related protein or microRNA,we applied protein chips and microRNA-sequences.We found that the angiogenesis related protein such as bFGF,EGF,PDGF in exosome showed no differences in two groups.While in the miRNA-sequences,we found that miR-3059-5p were significant increased and indicate that it play an important role in the BK-promoted c-Kit+cells exosome angiogenic activity.Conclusions:BK promotes angiogenic activity by regulating the expression of miRNA-3059-5p in c-Kit+ cell exosome. |
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