Cilostazol inhibits proliferation and induces apoptosis in rat vascular smooth muscle cells through Rb-p53-p21 pathways
10.3760/cma.j.issn.0253-3758.2013.01.011
- VernacularTitle:西洛他唑通过Rb-p53-p21通路抑制大鼠血管平滑肌细胞体外增殖
- Author:
Shou-Li WANG
1
;
Ming-Fang HUANG
;
Fei SUN
;
Zhao YIN
;
Li-Liang CHEN
;
Guo-Bin FENG
Author Information
1. 解放军第三0六医院心血管内科
- Keywords:
Muscle,smooth,vascular;
Cell proliferation;
Cilostazol
- From:
Chinese Journal of Cardiology
2013;41(1):48-53
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effects and related mechanisms of cilostazol on rat vascular smooth muscle cells (VSMCs)proliferation.Methods VSMCs were treated with DMEM (control) and various doses of cilostazol (1.0 × 10-7,2.5 × 10-7,5.0 × 10-7,7.5 × 10-7 and 1.0 × 10-6 mol/L) for 13 d (cell counting) or 72 h.Proliferation of VSMCs was investigated by cell-counting,MTT and flow cytometry analysis.Cell apoptosis was determined by TUNEL staining.mRNA and protein expressions of cell cycle regulatory proteins,such as Rb,p53 and p21 were detected by RT-PCR and Western blot,respectively.Results Cilostazol inhibited VSMCs proliferation and induced VSMCs arrest at G1 phase in a dose-dependent manner.High dose of cilostazol (7.5 × 10-7 and 1.0 × 10-6 mol/L) induced VSMCs apoptosis.p53 mRNA expression in 2.5 × 10-7 mol/L to 7.5 × 10-7 mol/L groups as well as 1.0 × 10-6 mol/L group (3.22 ±0.45 vs.1.75 ±0.32) and p53 protein expression in 7.5 × 10-7 mol/L group and 1.0 × 10-6 mol/L group (0.53 ± 0.11 vs.0.18 ± 0.06) were significantly upregulated after 72 h culture (all P＜0.05 vs.control).Low dose of cilostazol (1.0 × 10-7,2.5 × 10-7 and 5.0 × 10-7 mol/L)significantly upregulated p21 mRNA expression compared to control group (1.86 ± 0.19,2.20 ± 0.24 and 2.10 ± 0.18 vs.1.210 ± 0.18,all P ＜ 0.05).Similarly,Rb mRNA expression was significantly upregulated in 1.0 × 10-7,2.5 × 10-7 and 5.0 × 10 7 mol/L groups (0.89 ±0.07 vs.0.38 ±0.04)compared with control group (all P ＜ 0.05).However,high dose cilostazol (7.5 × 10-7 and 1.0 × 10-6mol/L) significantly downregulated p21 mRNA expression (0.81 ±0.09 vs.1.21 ±0.18,0.36 ±0.10 vs.1.2t ±0.18,all P ＜0.05 vs.control) and Rb mRNA expression (0.12 ±0.02 and 0.11 ±0.02 vs.0.38± ± 0.04,all P ＜ 0.05 vs.control).p21 and Rb protein expressions also upregulated at low concentrations of cilostazol and downregulated at high concentrations of cilostazol.Conclusion Cilostazol could inhibit the proliferation of rat VSMCs through modulating Rb-p53-p21 pathway and induce VSMCs apoptosis through upregulating p53.
