Osthole Alleviates Neointimal Hyperplasia in Balloon-Induced Arterial Wall Injury by Suppressing Vascular Smooth Muscle Cell Proliferation and Downregulating Cyclin D1/CDK4 and Cyclin E1/CDK2 Expression

Abstract

Percutaneous coronary intervention (PCI) is the most widely used therapy for treating ischemic heart disease. However, intimal hyperplasia and restenosis usually occur within months after angioplasty. Modern pharmacological researchers have proven that osthole, the major active coumarin of Cnidium monnieri (L.) Cusson, exerts potent antiproliferative effects in lung cancer cells, the human laryngeal cancer cell line RK33 and TE671 medulloblastoma cells, and its mechanism of action is related to cell cycle arrest. The goal of the present study was to observe the effect of osthole on vascular smooth muscle cell (VSMC) proliferation using platelet-derived growth factor-BB (PDGF-BB)-stimulated VSMCs isolated from rats and vascular balloon injury as models to further elucidate the molecular mechanisms underlying this activity. We detected the relative number of VSMCs by the MTT assay and EdU staining and examined cell cycle progression by flow cytometry. To more deeply probe the mechanisms, the protein expression levels of PCNA, the cyclin D1/CDK4 complex and the cyclin E1/CDK2 complex in balloon-treated rat carotid arteries and the mRNA and protein expression levels of the cyclin D1/CDK4 and cyclin E1/CDK2 complexes in VSMCs were detected by real-time RT-PCR and western blotting. The data showed that osthole significantly inhibited the proliferation of VSMCs induced by PDGF-BB. Furthermore, osthole caused apparent VSMC cycle arrest early in G0/G1 phase and decreased the expression of cyclin D1/CDK4 and cyclin E1/CDK2. Our results demonstrate that osthole can significantly inhibit PDGF-BB-induced VSMC proliferation and that its regulatory effects on cell cycle progression and proliferation may be related to the downregulation of cyclin D1/CDK4 and cyclin E1/CDK2 expression as well as the prevention of cell cycle progression from G0/G1 phase to S phase. The abovementioned mechanism may be responsible for the alleviation of neointimal hyperplasia in balloon-induced arterial wall injury by osthole.

Yohimbine, an α2-Adrenoceptor Antagonist, Suppresses PDGF-BB-Stimulated Vascular Smooth Muscle Cell Proliferation by Downregulating the PLCγ1 Signaling Pathway

“…We noted that YOH administration significantly attenuated PDGF-BB-induced CDK2 and PCNA upregulation in the VSMCs. A previous study revealed that CRISPR/Cas9-mediated knockout of CDK2 induced G0/G1 arrest and apoptosis in A375 melanocytes [ 40 ], and several other studies have shown a substantial downregulation of CDK2 in G0/G1-arrested VSMCs [ 39 , 41 , 42 ]. This, thus, explains why the expression of PCNA, a classic biomarker of proliferative cells, was suppressed in the YOH-treated MOVAS-1 cells.…”

Yohimbine Inhibits PDGF-Induced Vascular Smooth Muscle Cell Proliferation and Migration via FOXO3a Factor

“…Specifically, YHB inhibits the phosphorylation of p38 MAPK and mTOR, among the various signaling molecules activated by PDGF-BB, resulting in the suppression of PCNA, cyclin D1, CDK4, and cyclin E expression. These cell cycle regulatory proteins are overexpressed in PDGF-BB-induced VSMCs and promote cell proliferation [30,31]. Additionally, PDGF-BB activates many downstream signaling molecules related to cell growth, survival, and proliferation, such as the PI3K, Akt, PLC-γ1, and ERK1/2 pathways [32].…”

Icariside II Attenuates Vascular Remodeling Via Wnt7b/CCND1 Axis

“…16 Additionally, numerous studies have demonstrated that targeting CCND1 is useful for treating neointimal formation. 9,17,18 The Wnt genes were first characterized in 1978 in mutant wingless Drosophila melanogaster. 19 Since then, the Wnt family has been implicated in a myriad of physiological processes, including cell proliferation, differentiation, and migration.…”

“…The cell culture methods used for the primary VSMCs have been described previously. 9 The cells were grown in Dulbecco's modified eagle medium (DMEM) supplemented with 10% fatal bovine serum, 100 U/mL penicillin, and 100 μg/mL streptomycin at 37°C in a humidified incubator containing 5% CO 2 . The VSMCs purity was confirmed by immunofluorescence with α-SMA antibody.…”