CXCR4对PDGF-BB诱导的血管平滑肌细胞增殖、迁移的影响及白藜芦醇的抑制作用
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摘要
目的研究趋化因子受体4(Chemokine Receptor Type 4,CXCR4)对血管平滑肌细胞(Vascular Smooth Muscle Cells,VSMCs)增殖、迁移的影响及白藜芦醇(Resveratrol,RSV)的抑制作用。方法采用体外培养VSMCs,分为空白对照组、模型组[血小板源性生长因子-BB (PDGF-BB),10 ng·mL~(-1)]、AMD3100(CXCR4特异性活性抑制剂)不同浓度给药组(1、5、10μg·mL~(-1))、RSV不同浓度给药组(50、100、150μmol·L~(-1))。采用MTT法检测细胞增殖情况;Transwell细胞穿膜实验检测VSMCs迁移能力;Western Blot法检测VSMCs的CXCR4、增殖细胞核抗原(Proliferating Cell Nuclear Antigen, PCNA)及基质金属蛋白酶2(MatrixMetalloproteinase 2,MMP2)蛋白表达情况。结果与空白对照组比较,模型组的VSMCs增殖、迁移显著增强(P<0. 01),CXCR4、PCNA及MMP2蛋白表达显著增加(P<0.05,P<0.01)。与模型组比较,AMD3100不同浓度给药组均可显著抑制PDGF-BB诱导的VSMCs增殖、迁移(P<0.05,P<0.01);RSV不同浓度给药组均可显著抑制PDGF-BB诱导的VSMCs增殖、迁移和CXCR4蛋白表达(P<0.05,P<0.01)。结论 CXCR4参与了PDGF-BB诱导的VSMCs增殖、迁移;RSV可抑制VSMCs的增殖、迁移,可能与其抑制CXCR4蛋白表达有关。
Objective To evaluate the effect of CXCR4(Chemokine Receptor Type 4)on proliferation and migration of vascular smooth muscle cells(VSMC) and the inhibitory effect of RSV(resveratrol). Methods VSMCs were cultured and divided into control group,model group(10 ng·mL~(-1) PDGF-BB),CXCR4 specific inhibitor AMD3100 group(1,5,10 μg·mL~(-1))and RSV group(50,100,150 μmol·L~(-1)). Western Blot was used to detect the effect of PDGF-BB on the expression of CXCR4, PCNA(Proliferating Cell Nuclear Antigen) and MMP2(Matrix Metalloproteinase 2). MTT assay was used to detect the proliferation of VSMCs induced by PDGF-BB,and transwell test was used to detect the migration ability of VSMCs. Results Compared with the control group,PDGF-BB could significantly promote the proliferation and migration of VSMCs(P < 0.01)and the expression of CXCR4,PCNA and MMP2(P < 0.05,P < 0.01). Compared with the model group,different concentrations of AMD3100 significantly inhibited PDGF-BB-induced cell proliferation and migration(P < 0.05, P < 0.01). Compared with the model group,different concentrations of RSV significantly inhibited PDGF-BB-induced cell proliferation,migration and CXCR4 protein expression(P < 0.05,P < 0.01). Conclusion CXCR4 is involved in the proliferation and migration of VSMCs induced by PDGF-BB. RSV can inhibit the proliferation and migration of VSMCs by inhibiting the expression of CXCR4.
Objective To evaluate the effect of CXCR4(Chemokine Receptor Type 4)on proliferation and migration of vascular smooth muscle cells(VSMC) and the inhibitory effect of RSV(resveratrol). Methods VSMCs were cultured and divided into control group,model group(10 ng·mL~(-1) PDGF-BB),CXCR4 specific inhibitor AMD3100 group(1,5,10 μg·mL~(-1))and RSV group(50,100,150 μmol·L~(-1)). Western Blot was used to detect the effect of PDGF-BB on the expression of CXCR4, PCNA(Proliferating Cell Nuclear Antigen) and MMP2(Matrix Metalloproteinase 2). MTT assay was used to detect the proliferation of VSMCs induced by PDGF-BB,and transwell test was used to detect the migration ability of VSMCs. Results Compared with the control group,PDGF-BB could significantly promote the proliferation and migration of VSMCs(P < 0.01)and the expression of CXCR4,PCNA and MMP2(P < 0.05,P < 0.01). Compared with the model group,different concentrations of AMD3100 significantly inhibited PDGF-BB-induced cell proliferation and migration(P < 0.05, P < 0.01). Compared with the model group,different concentrations of RSV significantly inhibited PDGF-BB-induced cell proliferation,migration and CXCR4 protein expression(P < 0.05,P < 0.01). Conclusion CXCR4 is involved in the proliferation and migration of VSMCs induced by PDGF-BB. RSV can inhibit the proliferation and migration of VSMCs by inhibiting the expression of CXCR4.
引文
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[2]王晓华,付超,张鑫,等.SDF-1/CXCR4-R7轴与动脉粥样硬化关系研究进展[J].中国实验诊断学,2015,19(5):1601-1605.
[3]ABI-YOUNES S,SAUTY A,MACH F,et al.The stromal cellderived factor-1 chemokine is a potent platelet agonist highly expressed in atherosclerotic plaques[J].Circulation Research,2000,86(2):131-138.
[4]LI L X,ZHANG X F,BAI X,et al.SDF-1 promotes ox-LDLinduced vascular smooth muscle cell proliferation[J].Cell Biology International,2013,37(19):988-994.
[5]CHO S,NAMKOONG K,SHIN M,et al.Cardiovascular protective effects and clinical applications of resveratrol[J].Journal of Medicinal Food,2017,20(4):323-334.
[6]安梅,周瑾,陈晓宇.白藜芦醇药理学作用的研究进展[J].肿瘤药学,2014,4(4):242-246.
[7]周志明,冯义柏,周玉杰.白藜芦醇对血管平滑肌细胞增殖的影响[J].中华老年心脑血管病杂志,2004,6(5):335-337.
[8]CHAO H H,JUAN S,LIU J C,et al.Resveratrol inhibits angiotensinII induced endothelin-1 gene expression and subsequent proliferation rat aortic smooth muscle cells[J].Eur J Pharmacol,2005,515(1-3):1-9.
[9]李建坤,王顺祥,徐卓,等.白藜芦醇抑制缺氧诱导的肝癌细胞株HepG2侵袭并降低CXCR4表达[J].海峡药学,2018,30(8):13-15.
[10]陈鹏,赵丽丽,聂茜,等.贴块法培养大鼠主动脉血管平滑肌细胞条件的优化[J].河北医科大学学报,2014,35(10):1220-1222.
[11]HUANG M,DUHADAWAY J B,PRENDERGAST G C,et al.RhoB regulates PDGFR-beta trafficking and signaling in vascular smooth muscle cells[J].Arterioscler Thromb Vasc Biol,2007,27(12):2597-2605.
[12]SUN X Q,CHENG G C,HAO M G,et al.CXCL12/CXCR4/CXCR7 chemokine axis and cancer progression[J].Cancer Metastasis Reviews,2010,29(4):709-722.
[13]XIA N,DAIBER A,FORSTERMANN U,LI H.Antioxidant effects of resveratrol in the cardiovascular system[J].Br JPharmacol,2017,174(12):1633-1646.