反义c-myc寡核苷酸对AVP诱导鼠血管平滑肌细胞增殖的抑制作用

The Despression of Antisense c myc Oligodeoxynucleotides on AVP Inducing Proliferation of Rat Vascular Smooth Muscle Cells

目的：探讨ＡＶＰ对鼠血管平滑肌细胞（ＶＳＭＣ）增殖的影响和反义ｃ－ｍｙｃ寡核苷酸对ＡＶＰ作用的干预效应。方法：以培养的ＳＤ大鼠ＶＳＭＣ为模型，采用培养细胞计数、蛋白质定量和细胞周期分析方法及ｍ ＲＮＡ打点杂交技术，动态观察ＡＶＰ对ＶＳＭＣ增殖的影响和反义ｃ－ｍｙｃ寡核苷酸的干预效果。结果：（１）ＡＶＰ促ＶＳＭＣ数目增加，在４８ｈ 时与对照组比较，有统计学意义（Ｐ＜ ０．０５）；（２）ＡＶＰ促ＶＳＭＣ的Ｓ期百分率增加，与对照组比较有非常显著性差异（Ｐ＜ ０．０１）；（３） ＡＶＰ促单个ＶＳＭＣ的蛋白质含量增加，在４８ｈ 时与对照组比较，差异非常显著（Ｐ＜ ０．０１）；（４）ＡＶＰ使ＶＳＭＣ 的ｃ－ｍｙｃ 的ｍ ＲＮＡ杂交信号比对照组明显地增强；反义ｃ－ｍｙｃ 寡核苷酸与对照组比较，ＶＳＭＣ的ｃ－ｍｙｃ的ｍ ＲＮＡ杂交信号明显地减弱；（５）反义ｃ－ｍｙｃ寡核苷酸与ＡＶＰ共同作用组的ＶＳＭＣ数目和细胞Ｓ期百分率均小于ＡＶＰ组和对照组，并均有非常显著性差异（Ｐ＜ ０．０１）；（６）反义ｃ－ｍｙｃ寡核苷酸与ＡＶＰ共同作用组ＶＳＭＣ的ｃ－ｍｙｃ癌基因ｍ ＲＮＡ杂交信号强度明显地高于对照组，而低于ＡＶＰ组；（７）反义ｃ－ｍｙｃ寡核苷酸?

Aim: To investgate the effect of AVP on rat smooth muscle cells (VSMCs) proliferation and observe the interference effect of antisence c myc oligodeoxynucleotides on AVP action. Methods:By means of the cell counting method,protein content measurement, cell cycle analysis and mRNA dot blot hybridzation techniques, cultured VSMCs derived from the aorta of Sprague Dawley (SD) rat were used to dynamically observe the effect of AVP on VSMCs proliferation and interference effect of antisense c myc oligodeoxynucleotides. Results:(1) the number of VSMCs induced by AVP was significantly increased in comparision with that of the control at 48h ( P< 0.05); (2) the percentage of S stage of VSMCs induced by AVP was significantly increased in comparision with that of the control ( P< 0.01); (3) protein content of single VSMC induced by AVP was significantly increased in comparision with that of the control at 48h ( P< 0.01); (4) compared with the control, AVP could enhance the level of c myc mRNA expression, but antisence c myc could decrease the level of c myc mRNA expression; (5) the cell number and the percentage of S stage VSMCs incubated with antisense c myc and AVP together decreased more remarkably than those of the control and AVP group ( P< 0.01); (6) the level of c myc mRNA expression of VSMCs incubated with antisense c myc and AVP was higher than that of the control, but lower than that of AVP group;(7) protein content of single VSMC incubated with antisense c myc and AVP together was remarkly higher than that of the control ( P< 0.01), but was not different from that of AVP group ( P> 0.05). Conclusion:The effect of AVP induced VSMCs proliferation and hypertrophy was related to c myc overexpression, antisense c myc oligonucleotides specially inhibited c myc expression and AVP induced VSMCs proliferation. But antisense c myc could not inhibit the effect of AVP induced VSMCs protein synthesis. It suggests that VSMCs protein synthesis was independent of the c myc oncogene expression and that antisense c myc could not completely reverse the vascular remodeling of EH. The clinical value of antisense c myc in preventing and treating EH is limited.