韧粘素反义寡核苷酸对血管平滑肌细胞增殖的抑制作用

Inhibitory Effect of Antisense Tenascin Oligonucleotides on Proliferation of Cultured Vascular Smooth Muscle Cells

目的探讨韧粘素 (TN)反义寡核苷酸 (ODN)对培养大鼠血管平滑肌细胞 (VSMC)增殖的抑制作用。方法采用细胞计数及快速竞争性逆转录聚合酶链反应 (RT -PCR)等方法 ,观察不同剂量 ( 5、10、2 0 μg/ml)TN反义ODN和正义ODN对体外培养大鼠VSMC的增殖以及TNmRNA表达水平的影响。 结果TN反义ODN可有效抑制大鼠血管平滑肌细胞的增殖 ,作用呈剂量依赖性 (反义组 5、10、2 0 μg/ml剂量细胞计数分别为 10 5/ml× ( 4 .99± 0 .2 1)、( 4 .0 8± 0 .14 )、( 2 .85± 0 .39) ,对照组为 10 5/ml× ( 5 .95± 0 .41) ,(P <0 .0 5 ) ;TN反义ODN亦可下调TNmRNA表达水平 (反义组 0 .32± 0 .0 5 ,对照组为 0 .5 2± 0 .0 8;P <0 .0 1) ;而TN正义ODN则无上述作用。 结论TN反义ODN可能通过下调TNmRNA表达水平而抑制血管平滑肌细胞的增殖。

Objective To study the inhibitory effect of antisense tenascin(TN) oligonucleotides(ODN) transfection on the growth of cultured rat (vascular smooth muscle cells,VSMC). Methods Different doses(5?10?20μg/ml)of TN antisense/sense ODN were transfected into cultured VSMC of rats.Cell count was used to evaluate cell hyperplasia,and rapid competitive RT-PCR was used to detect TN mRNA expression of VSMC. Results Antisense TN ODN inhibited proliferation of VSMC in a dose-dependant manner [cell numbers of AS group were respectively 10 5/ml×(4.99±0.21)?(4.08± 0.14)?( 2.85±0.39) vs CON group 5.95±0.41(P<0.05)].TNmRNA expression was obviously suppressed by antisense TN ODN(AS group 0.32±0.05 vs CON group 0.52±0.08(P<0.01).In contrast,sense TN ODN had no significant effect on cell proliferation and expression of TN mRNA. Conclusion Antisense TN ODN might inhibit proliferation of the cultured rat VSMC by down-regulating TNmRNA expression.