萘醌衍生物抗血管平滑肌细胞增殖的活性研究
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摘要
目的探索一种合成的萘醌衍生物对血小板衍生生长因子BB(PDGF-BB)刺激的大鼠主动脉血管平滑肌细胞株A10细胞增殖的影响,并初步阐明其作用机制。方法通过细胞增殖实验、[~3H]胸腺嘧啶核苷掺入实验、细胞周期进程分析、免疫印迹分析等实验手段,研究此合成萘醌衍生物对PDGF-BB诱导的A10细胞周期进展和细胞周期主要信号转导途径的影响。结果合成萘醌衍生物浓度依赖性地降低了S期细胞比例,升高G_0/G_1期细胞比例,减少了DNA合成。合成萘醌衍生物预处理24 h,抑制了PDGF-BB诱发的A10细胞增殖,最大抑制率为44.4%。1μmol·L~(-1)的合成萘醌衍生物明显降低了由PDGF-BB诱导的ERK1/2、Akt、磷脂酶Cγ1、PDGFβ受体的磷酸化水平(P<0.01)。结论此合成的萘醌衍生物通过阻止A10细胞由G_0/G_1期进入S期,显示出抑制PDGF-BB诱导的血管平滑肌细胞增殖的活性,其作用机制可能与下调ERK1/2、Akt、磷脂酶Cγ1、PDGFβ受体的磷酸化水平有关。
Aim To explore the effect of a synthetic naphthoquinone derivative on the proliferation of rat aortic vascular smooth muscle cells(RAVSMCs) stimulated by platelet-derived growth factor BB(PDGF-BB) and to clarify its mechanism underlying the anti-proliferative effect.Methods The influence of the synthetic naphthoquinone derivative on cell cycle progression and main signal transduction pathway of cell cycle induced by PDGF-BB were investigated by cell proliferation assay,[~3H]thymidine incorporation test, cell cycle process analysis and immunoblotting assay.Results S phase cell percentage in the cell cycle was reduced, while G_0/G_1 phase cell percentage was increased by the synthetic naphthoquinone in a dose-dependent(0.1, 0.5, 1 μmol·L~(-1)) manner. Moreover, DNA synthesis also decreased. The inhibitory rate of PDGF-BB-induced RAVSMCs proliferation achieved the maximum of 44.4% after 24 h pre-treatment by the synthetic naphthoquinone derivative at the concentration of 1 μmol·L~(-1). The phosphorylation of extracellular regulated kinase 1/2(ERK1/2), Akt, phospholipase C(PLC) γ1 and PDGF receptor β(PDGFRβ) induced by PDGF-BB was significantly decreased( P<0. 01) by addtion of 1 μmol · L-1 synthetic naphthoquinone derivative. Conclusions The synthetic naphthoquinone derivative exhibits anti-proliferation activity against RAVSMCs induced by PDGF-BB via blocking the transformation of G0/G1 phase cell into S-phase cell in the cell cycle. The mechanisms might be related to its down-regulatory effect on phosporalation level of ERK1/2,Akt,PLCγ1 and PDGFRβ.
Aim To explore the effect of a synthetic naphthoquinone derivative on the proliferation of rat aortic vascular smooth muscle cells(RAVSMCs) stimulated by platelet-derived growth factor BB(PDGF-BB) and to clarify its mechanism underlying the anti-proliferative effect.Methods The influence of the synthetic naphthoquinone derivative on cell cycle progression and main signal transduction pathway of cell cycle induced by PDGF-BB were investigated by cell proliferation assay,[~3H]thymidine incorporation test, cell cycle process analysis and immunoblotting assay.Results S phase cell percentage in the cell cycle was reduced, while G_0/G_1 phase cell percentage was increased by the synthetic naphthoquinone in a dose-dependent(0.1, 0.5, 1 μmol·L~(-1)) manner. Moreover, DNA synthesis also decreased. The inhibitory rate of PDGF-BB-induced RAVSMCs proliferation achieved the maximum of 44.4% after 24 h pre-treatment by the synthetic naphthoquinone derivative at the concentration of 1 μmol·L~(-1). The phosphorylation of extracellular regulated kinase 1/2(ERK1/2), Akt, phospholipase C(PLC) γ1 and PDGF receptor β(PDGFRβ) induced by PDGF-BB was significantly decreased( P<0. 01) by addtion of 1 μmol · L-1 synthetic naphthoquinone derivative. Conclusions The synthetic naphthoquinone derivative exhibits anti-proliferation activity against RAVSMCs induced by PDGF-BB via blocking the transformation of G0/G1 phase cell into S-phase cell in the cell cycle. The mechanisms might be related to its down-regulatory effect on phosporalation level of ERK1/2,Akt,PLCγ1 and PDGFRβ.
引文
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[7] Sun Y Y,Qin S S,Cheng Y H,et al. MicroRNA expression profile and functional analysis reveal their roles in contact inhibition and its disruption switch of rat vascular smooth muscle cells[J]. Acta Pharmacol Sin, 2018, 39(5):885-92.
[8] Li Y,Anand-Srivastava M B. Implication of multiple signaling pathways in the regulation of angiotensin II induced enhanced expression of Giα proteins in vascular smooth muscle cells[J]. Can J Physiol Pharmacol, 2012, 90(8):1105-16.
[9] Seo K W,Lee S J,Kim Y H,et al. Mechanical stretch increases MMP-2 production in vascular smooth muscle cells via activation of PDGFR-β/Akt signaling pathway[J]. PLoS One, 2013, 8(8):e70437.
[10] Chistiakov D A, Orekhov A N, Bobryshev Y V. Vascular smooth muscle cell in atherosclerosis[J]. Acta Physiol, 2015, 214(1): 33-50.
[11] Dong X, Hu H, Fang Z, et al. CTRP6 inhibits PDGF-BB-induced vascular smooth muscle cell proliferation and migration[J]. Biomed Pharmacother, 2018, 103: 844-50.
[2] 黄警,黄燮南,张纾, 等.人参总皂苷对 PDGF-BB所致血管平滑肌细胞增殖周期的影响[J]. 中国药理学通报, 2010, 26(6):787-91.[2] Huang J, Huang X N, Zhang S, et al. Effect of total ginsenosides on the cell cycle of rat vascular smooth muscle cell proliferation induced by PDGF-BB[J]. Chin Pharmacol Bull, 2010, 26(6):787-91.
[3] Kim T J, Jeon J, Jin Y R, et al. Effects of KTJ740, a novel antithrombotic agent, on platelet-derived growth factor-induced rat aortic smooth muscle cell proliferation and cell cycle progression[J]. J Cardiovasc Pharmacol, 2007, 49(5): 280-6.
[4] Zhang X, Hu W, Wu F, et al. Shikonin inhibits TNF-α-induced growth and invasion of rat aortic vascular smooth muscle cells[J]. Can J Physiol Pharmacol, 2015, 93(8): 615-24.
[5] Kim J H, Jin Y R, Park B S, et al. Luteolin prevents PDGF-BB-induced proliferation of vascular smooth muscle cells by inhibition of PDGF beta-receptor phosphorylation[J]. Biochem Pharmacol, 2005, 69(12): 1715-21.
[6] Tong L,Qi G. Crocin prevents plateletderived growth factor BB-induced vascular smooth muscle cells proliferation and phenotypic switch[J]. Mol Med Rep, 2018, 17(6):7595-602.
[7] Sun Y Y,Qin S S,Cheng Y H,et al. MicroRNA expression profile and functional analysis reveal their roles in contact inhibition and its disruption switch of rat vascular smooth muscle cells[J]. Acta Pharmacol Sin, 2018, 39(5):885-92.
[8] Li Y,Anand-Srivastava M B. Implication of multiple signaling pathways in the regulation of angiotensin II induced enhanced expression of Giα proteins in vascular smooth muscle cells[J]. Can J Physiol Pharmacol, 2012, 90(8):1105-16.
[9] Seo K W,Lee S J,Kim Y H,et al. Mechanical stretch increases MMP-2 production in vascular smooth muscle cells via activation of PDGFR-β/Akt signaling pathway[J]. PLoS One, 2013, 8(8):e70437.
[10] Chistiakov D A, Orekhov A N, Bobryshev Y V. Vascular smooth muscle cell in atherosclerosis[J]. Acta Physiol, 2015, 214(1): 33-50.
[11] Dong X, Hu H, Fang Z, et al. CTRP6 inhibits PDGF-BB-induced vascular smooth muscle cell proliferation and migration[J]. Biomed Pharmacother, 2018, 103: 844-50.