中国中药杂志

2014, v.39(20) 4040-4044

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大蒜素抑制胰岛素诱发血管平滑肌细胞增殖和迁移实验研究
Experimental study on effect of allicin in inhibiting insulin-induced vascular smooth muscle cell proliferation and migration

李婷;夏勇;
LI Ting;XIA Yong;Institute of Cardiovascular Disease,Xuzhou Medical College;Department of Cardiology,the Affiliated Hospital of Xuzhou Medical College;

摘要(Abstract):

目的:探讨大蒜素(allicin)抑制胰岛素(insulin)诱导的血管平滑肌细胞(VSMCs)增殖和迁移及其作用机制。方法:采用组织贴块法培养大鼠的VSMCs,并进行平滑肌α-肌动蛋白(α-SMA)免疫荧光鉴定,取35代细胞进行实验。建立胰岛素刺激的VSMCs模型。实验分5组:对照组、胰岛素组、大蒜素组、ERK抑制剂PD98059组、大蒜素+PD98059组。噻唑蓝(CCK8)比色法检测VSMCs的增殖;细胞计数法检测VSMCs的迁移;免疫印迹实验(Western blotting)测定total ERK,PhosphoERK(p-ERK),PCNA蛋白表达的情况。结果:原代培养的VSMCs生长良好,光镜下呈长梭形及"峰与谷"样生长。α-SMA免疫荧光显示培养的细胞具有典型的VSMCs特征。胰岛素能刺激VSMCs的增殖和迁移,且以100 nmol·L-1浓度时效果最明显。大蒜素预处理能显著抑制胰岛素刺激的VSMCs的增殖和迁移,且呈剂量依赖性。PD98059、大蒜素+PD98059预处理亦能显著抑制胰岛素刺激的VSMCs的增殖和迁移。胰岛素可明显促进VSMCs表达p-ERK,PCNA蛋白。大蒜素能显著抑制VSMCs表达p-ERK,PCNA蛋白,且呈剂量依赖性。结论:大蒜素能显著抑制胰岛素诱导的VSMCs的增殖与迁移,其作用机制可能是抑制ERK信号通路的激活。
Objective: To investigate the effect of allicin in inhibiting insulin-induced vascular smooth muscle cell( VSMC)proliferation and migration. Method: The tissue explant method was adopted to culture rat' s VSMCs,and the immunofluorescence method was used to identify α-SMA. Cells from the third to fifth generations were selected in the experiment. The insulin-induced VSMC model was established. The experiment was carried out in five groups: the control group,the insulin group,the allicin group,the ERK inhibitors PD98059 group( 20 μmol·L- 1) and the PD98059 + allicin group. VSMCs' proliferation was determined by CCK8 colorimetric method,while its migration was detected by cell counting; The western blotting was used to detect total ERK,PhosphoERK,PCNA protein's expression. Result: Primary cultured VSMCs grew well in the spindle shape under the lightmicroscope,with peak and valley. α-SMA immunofluorescence results showed that the cultured cells had typical VSMCs' features. Insulin could stimulate VSMCs' proliferation and migration,with the best effect at the concentration of 100 nmol · L- 1. The pretreatment with allicin could significantly inhibit VSMCs' proliferation and migration induced by insulin in a dose-dependent manner. The pretreatment with PD98059 and allicin + PD98059 could inhibit VSMCs' proliferation and migration induced by insulin remarkably as well. Insulin could significantly accelerate VSMCs' expression of such proteins as p-ERK,PCNA. Contrarily,allicin could notably inhibit VSMCs' expression of such proteins as p-ERK,PCNA in a dose-dependent manner. Conclusion: Allicin could significantly inhibit VSMCs' proliferation and migration induced by insulin,which may be related to the inhibition of the activation of ERK signal path.

关键词(KeyWords): 血管平滑肌细胞;胰岛素;大蒜素;增殖;迁移
vascular smooth muscle cell;insulin;allicin;proliferation;migration

Abstract:

Keywords:

基金项目(Foundation):

作者(Authors): 李婷;夏勇;
LI Ting;XIA Yong;Institute of Cardiovascular Disease,Xuzhou Medical College;Department of Cardiology,the Affiliated Hospital of Xuzhou Medical College;

参考文献(References):

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