同型半胱氨酸诱导人血管平滑肌细胞组织因子的表达

Induction of Tissue Factor by Homocysteine in Cultured Human Vascular Smooth Muscle Cells

目的研究同型半胱氨酸(Hcy)诱导人血管平滑肌细胞(VSMCs)表达组织因子(TF)的作用。方法采用组织贴块法培养人脐动脉 VSMCs,应用α-actin 免疫组化法鉴定细胞；将不同浓度的 Hcy 与 VSMCs 孵育,采用半定量 RT-PCR 检测细胞 TF mRNA 表达,流式细胞技术(FCM)检测细胞表面 TF 蛋白水平。结果终浓度分别为0、10、100、 500、1000μmol/L 的 Hcy 作用4 h,Hcy 浓度为10μmol/L 时,TF 表达开始升高,500μmol/L 达峰值,Hcy 呈剂量依赖性诱导 VSMCs 表达 TF mRNA；100μmol/L Hcy 作用0、1、2、3、4、8、24 h,2 h 时 TF 表达明显增加,8 h 达峰值,24 h 表达回到基础水平,Hcy 呈时间依赖性诱导 VSMCs 表达 TF mRNA；VSMCs 细胞膜表面有低水平的 TF 蛋白表达,终浓度分别为0、 10、100、500、1000μmol/L Hcy 作用4 h,10μmol/L 即可诱导 VSMCs 表达 TF 蛋白,1000μmol/L 达高峰,Hcy 呈剂量依赖性诱导 VSMCs 表达 TF 蛋白；终浓度为100μmol/L Hcy 作用0、1、2、4、8、16、24 h,2 h 时 TF 开始表达升高,Hcy 呈时间依赖性诱导 VSMCs 表达 TF 蛋白。结论 Hcy 呈时间依赖性和剂量依赖性诱导 VSMCs 表达 TF,可能是 Hcy 导致 AS 和冠心病等疾病发生发展的重要机制。

Objective To investigated the expression of tissue factor （TF） induced by homoeysteine （Hey） in cultured human vascular smooth muscle cells （VSMCs）. Methods Human umbilical artery VSMCs were cultured by tissue explanting method, and were incubated with different dosage of Hey. RT-PCR was used to measure the expression of TF mRNA, and flow eytometry （FCM） was used to detect the expression of TF on the surface of VSMCs. Results VSMCs incubated with 0,10,100,500,1 000 μmoL/L Hey for 4 h and incubated with 100μmol/L Hey for 0,1,2,3,4,8,24 h . The data showed that Hey could induce VSMCs expressing TF mRNA in a dose- and time-dependent manner. Hey could also induce VSMCs expressing TF protein on the cell surface in a dose- and time-dependent manner. Conclusion Hey could induce human VSMCs expressing TF in mRNA and protein level. These results suggest that Hey may play an important role in atherogenesis and thrombosis.