AG490对凝血酶诱导VSMCs增殖及Bcl-2表达的影响  

Effect of AG490 on the VSMCs Proliferation and the Expression of Bcl-2 Protein Induced by Thrombin

作　　者：方正旭[1] 朱亮[1] 李鸣媛[1] 王伶[2] 刘东[2] 程晓曙[2] 

机构地区：[1]南昌大学第四附属医院心内科,南昌330003 [2]南昌大学第二附属医院心内科,南昌330006

出　　处：《南昌大学学报（医学版）》2010年第3期9-11,15,共4页Journal of Nanchang University：Medical Sciences

基　　金：江西省卫生厅科研课题(20063138)

摘　　要：目的探讨Jak2的特异性抑制剂AG490对凝血酶诱导大鼠血管平滑肌细胞(Vascular smooth musclecells,VSMCs)增殖作用及与Bcl-2蛋白表达的影响。方法采用培养大鼠胸主动脉4-10代的VSMCs的单细胞悬液用于实验。实验分3组:对照组(DMSO)、凝血酶组(0.5 kU.L-1)和凝血酶(0.5 kU.L-1)+AG490(50 mmol.L-1)组。采用MTS法测定3组0、1、12、24、72 h VSMCs细胞增殖率;Western blot测定Bcl-2含量。结果①与对照组比较,凝血酶组在各时间段均有明显促VSMCs增殖作用(P<0.05),增殖曲线呈双峰样改变(1 h和24 h);与凝血酶组比较,AG 490+凝血酶组有效地降低了凝血酶诱导VSMCs 1 h及24 h峰值(均P<0.05)。②凝血酶组Bcl-2蛋白表达在72 h明显增加,且1、12、24、48 h表达量也有增加;与凝血酶组比较,AG490+凝血酶组的Bcl-2蛋白表达量在72 h高峰明显被抑制(P<0.01),并且6、12、244、8 h表达量也有所减少。结论①凝血酶促VSMCs的增殖在时间上呈双峰样改变;AG490能抑制凝血酶早期快速及后期促VSMCs增殖作用。②AG490使得VSMC中Bcl-2蛋白表达下调,增加VSMCs的凋亡。Objective To investigate the effect of AG490 on the VSMCs proliferation and the expression of Bcl-2 protein induced by thrombin.Methods Growth-arrested VSMCs（4-10 generation） were used in the experiments.Three groups were included：control（DMSO）,thrombin（0.5 kU·L-1）,thrombin（0.5 kU·L-1） plus AG490（50 mmol·L-1） respectively.Cell proliferation was quantified by MTS method at 0,1,6,12,24,48,72 h respectively.The expression of Bcl-2 protein in cells was investigated by Western blot.Results ①Compared with control group,cell proliferation was significantly enhanced by thrombin at selective time（P0.05）.The proliferation curve had two peaks,one at 1 hour,and another at 24 h.Compared with thrombin group,AG490 attenuated the thrombin-induced cell proliferation at 1 h（P0.05） and at 24 h（P0.05）.②The expression of Bcl-2 protein in thrombin group increased significantly at 72 h（P0.01） and at 1,12,24,48 h respectively.Compared with thrombin group,the Bcl-2 protein expression of AG490 group was decreased at 72 h（P0.01） and at 6,12,24,48 h respectively.Conclusion ①The cell proliferation induced by thrombin has two peaks at 1 hr and 24 hr.The inhibition effect of AG490 in thrombin-induced VSMCs proliferation was not only quickly and transient but also torpid and late.②AG490 can induce the apoptosis of VSMCs by down-regulating the expression of Bcl-2 protein.

关 键 词：凝血酶 血管平滑肌细胞 Bcl-2 增殖 凋亡 

分 类 号：R-33[医药卫生]