Title:MiR-199a-5p Deficiency Promotes Artery Restenosis in Peripheral Artery Disease by Regulating ASMCs Function via Targeting HIF-1α and E2F3
Volume: 22
Issue: 5
Author(s): Duan Liu, Yexiang Jing, Guiyan Peng, Litai Wei, Liang Zheng, Guangqi Chang*Mian Wang*
Affiliation:
- Division of Vascular Surgery, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510080, China
- National-Guangdong Joint Engineering Laboratory for Diagnosis and Treatment of Vascular Diseases, The First Affiliated
Hospital of Sun Yat-sen University, Guangzhou, 510080, China
- Division of Vascular Surgery, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510080, China
- National-Guangdong Joint Engineering Laboratory for Diagnosis and Treatment of Vascular Diseases, The First Affiliated
Hospital of Sun Yat-sen University, Guangzhou, 510080, China
Keywords:
Peripheral Artery Disease, Restenosis, Arterial Smooth Muscle Cells, miR-199a-5p, HIF-1α, E2F3.
Abstract:
Background: Restenosis (RS) poses a significant concern, leading to recurrent ischemia
and the potential for amputation following intraluminal angioplasty in the treatment of Peripheral
Artery Disease (PAD). Through microRNA microarray analysis, the study detected a significant
downregulation of miR-199a-5p within arterial smooth muscle cells (ASMCs) associated
with RS.
Objective: This research aims to explore the possible function and the underlying mechanisms of
miR-199a-5p in the context of RS.
Methods: Primary ASMCs were extracted from the femoral arteries of both healthy individuals
and patients with PAD or RS. The expression levels of miR-199a-5p were assessed using both
qRT-PCR and in situ hybridization techniques. To examine the impacts of miR-199a-5p, a series
of experiments were performed, including flow cytometry, TUNEL assay, EdU assay, CCK8 assay,
Transwell assay, and wound closure assay. A rat carotid balloon injury model was employed
to elucidate the mechanism through which miR-199a-5p mitigated neointimal hyperplasia.
Results: MiR-199a-5p exhibited downregulation in RS patients and was predominantly expressed
within ASMCs. Elevated the expression of miR-199a-5p resulted in an inhibitory effect of proliferation
and migration in ASMCs. Immunohistochemistry and a dual-luciferase reporter assay uncovered
that RS exhibited elevated expression levels of both HIF-1α and E2F3, and they were identified
as target genes regulated by miR-199a-5p. The co-transfection of lentiviruses carrying
HIF-1α and E2F3 alongside miR-199a-5p further elucidated their role in the cellular responses mediated
by miR-199a-5p.
In vivo, the delivery of miR-199a-5p
via lentivirus led to the mitigation of
neointimal formation following angioplasty, achieved by targeting HIF-1α and E2F3.
Conclusion: MiR-199a-5p exhibits promise as a prospective therapeutic target for RS since it alleviates
the condition by inhibiting the proliferation and migration of ASMCs via its regulation of
HIF-1α and E2F3.
