Objective To explore the effects of allicin on proliferation of vascular smooth muscle cells (VSMCs) in rabbits induced by homocysteine (Hcy) and the possible underling mechanisms. Methods The primary VSMCs cultured by attachment-block in vitro were induced by Hcy to establish cell proliferating model and divided into six groups: control,Hcy stimulation (Hcy 1 mmol/L ) , high (Allicin 10 μg/ml +Hcy 1 mmol/L ) ,middle ( Allicin 5 μg/ml +Hcy 1 mmol/L ) , low dose allicin ( Allicin 2. 5 μg/ml +Hcy 1 mmol/L) ,folic acid ( folic acid 5 mmol/L +Hcy 1 mmol/L) groups. Activity of cell proliferation was measured by MTT colorimetric method. The PCNA expression was detected by immunocytochemical technique. The method of Real-time PCR was used to observe the mRNA expression of c-myc.Results After Hcy stimulation, the activity of cell proliferation, the expressions of c-myc mRNA and PCNA were obviously higher than those of control group( P ＜0. 01 ). The activity of cell proliferation, the expressions of comyc mRNA and PCNA were lower in high, middle and low dose allicin groups than those of Hcy stimulation group, presenting dose dependent ( P ＜ 0.01 or P ＜ 0.05 ), the better effect in high and middle dose allicin groups than folic acid group ( P ＜ 0. 01 or P ＜0. 05 ). Conclusions Allicin could obviously inhibit the VSMCs proliferation induced by Hcy, and the mechanism may be related to down-regulating the expression of c-myc gene.%目的 探讨大蒜素(Allicin)对同型半胱氨酸(Hcy)诱导的血管平滑肌细胞(VSMCs)增殖的影响及其可能机制.方法 组织贴块法体外原代培养兔主动脉VSMCs,由Hcy诱导建立细胞增殖模型,将细胞随机分为6组,对照组、Hcy刺激组( Hcy 1 mmol/L)、Allicin大剂量组(Allicin 10 μg/ml+ Hcy 1 mmol/L)、Allicin中剂量组(Allicin 5 μg/ml+ Hcy 1 mmol/L)、Allicin小剂量组(Allicin 2.5 μg/ml+ Hcy1 mmol/L)、叶酸组(叶酸5 mmol/L+ Hcy 1 mmol/L).通过MTT比色法检测细胞的增殖活度,免疫细胞化学技术检测增殖细胞核抗原(PCNA)的表达及RT PCR技术观察对c-myc mRNA表达的影响.结果 与对照组相比,Hcy 刺激组的增殖活度、PCNA及c-myc mRNA的表达均显著升高(P<0.01);与Hcy刺激组比较,Allicin大、中、小剂量组细胞增殖活度减低,PCNA及c-myc mRNA的表达均下降(P<0.01),并呈现剂量依赖性,且Allicin大、中剂量组的抑制作用优于叶酸组(P<0.01或P<0.05).结论 Allicin可以明显抑制Hcy诱导的VSMCs的增殖,部分机制可能与下调c-myc基因的表达有关.
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