Bradykinin-pretreated human cardiac-specific c-kit + cells enhance exosomal miR-3059-5p and promote angiogenesis against hindlimb ischemia in mice

Abstract

Aims: Protection of cardiac function following myocardial infarction was largely enhanced by bradykinin-pretreated cardiac-specific c-kit+ (BK-c-kit+) cells, even without significant engraftment, indicating that paracrine actions of BK-c-kit+ cells play a pivotal role in angiogenesis. Nevertheless, the active components of the paracrine actions of BK-c-kit+ cells and the underlying mechanisms remain unknown. This study aimed to define the active components of exosomes from BK-c-kit+ cells and elucidate their underlying protective mechanisms. Methods and results: Matrigel tube formation assay, cell cycle, and mobility in human umbilical vein endothelial cells (HUVECs) and hindlimb ischemia (HLI) in mice were applied to determine the angiogenic effect of condition medium (CM) and exosomes. Proteome profiler, microRNA sponge, Due-luciferase assay, microRNA-sequencing, qRT-PCR, and Western blot were used to determine the underlying mechanism of the angiogenic effect of exosomes from BK-c-kit+. As a result, BK-c-kit+ CM and exosomes promoted tube formation in HUVECs and the repair of HLI in mice. Angiogenesis-related proteomic profiling and microRNA sequencing revealed highly enriched miR-3059-5p as a key angiogenic component of BK-c-kit+ exosomes. Meanwhile, loss- and gain-of-function experiments revealed that the promotion of angiogenesis by miR-3059-5p was mainly through suppression of TNFSF15 (VEGI)-inhibited effects on vascular tube formation, cell proliferation and cell migration. Moreover, enhanced angiogenesis of miR-3059-5p-inhibited TNFSF15 has been associated with Akt/Erk1/2/Smad2/3-modulated signaling pathway. Conclusion: Our results demonstrated a novel finding that BK-c-kit+ cells enrich exosomal miR-3059-5p to suppress TNFSF15 and promote angiogenesis against hindlimb ischemia in mice.

Deletion of endothelial IGFBP5 protects against ischaemic hindlimb injury by promoting angiogenesis

“…Age‐matched male control mice (IGFBP5 f/f ) and endothelial cell (EC)‐specific IGFBP5‐KO (IGFBP5 EKO ) mice aged from 8 to 12 weeks were used in this study. Unilateral HLI was induced, as previously reported 28 . Briefly, mice were anesthetised by isoflurane vaporisation.…”

NEDD4L is a promoter for angiogenesis and cell proliferation in human umbilical vein endothelial cells

“…After treatment, the migration ability of the HUVECs was tested by using the Transwell assay as previously reported. [22][23][24][25] Briefly, a total of 2 × 10 4 HUVECs (passage 3-8) were suspended in 200 μL ECM medium (0% FBS) and seeded in a modified Boyden chamber with an 8 μm-pore polycarbonate membrane (Corning, USA) for 30 min. The lower chambers were placed with 600 μL ECM medium (2.5% FBS).…”