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

作　　者：李婷[1] 夏勇[2] 

机构地区：[1]徐州医学院心血管病研究所,江苏徐州221002 [2]徐州医学院附属医院心内科,江苏徐州221002

出　　处：《中国中药杂志》2014年第20期4040-4044,共5页China Journal of Chinese Materia Medica

摘　　要：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3~5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，Phospho-ERK（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, Phospho-ERK, 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：血管平滑肌细胞 胰岛素 大蒜素 增殖 迁移 

分 类 号：R285[医药卫生—中药学]