Effect of antisense mitogen-activated protein kinase oligonucleotides on rat vascular smoo

• 作 者: 黄韶玲 丁波 余清声 郭兆贵
• 机 构: 湖南医科大学分子药理研究室
• 出 处: Acta Pharmacologica Sinica
• 发表日期: 2004年期
• 关键词: 血管平滑肌细胞 antisense 起始部位 mitogen 髓磷脂碱性蛋白 表皮生长因子 核苷酸类 抑制作用 胸腺嘧啶 转染

目的：探讨丝裂素活化的蛋白激酶（ＭＡＰＫ）反义寡核苷酸（ＯＤＮ）对表皮生长因子（ＥＧＦ）诱导的培养大鼠血管平滑肌细胞增生的抑制作用．方法：用脂质体将ｐ４２和ｐ４４ＭＡＰＫＯＤＮ０．２μｍｏｌ·Ｌ－１转染入大鼠血管平滑肌细胞，设正义及随机ＯＤＮ为对照，用ＷｅｓｔｅｒｎＢｌｏｔ法结合Ｐ８１滤纸法以髓磷脂碱性蛋白为底物测定ＭＡＰＫ活性．［３Ｈ］胸腺嘧啶核苷酸掺入测定平滑肌细胞ＤＮＡ合成．结果：ＭＡＰＫ０ＤＮ能明显抑制ＥＧＦ诱导的ＭＡＰＫ蛋白表达及ＭＡＰＫ活性，并明显抑制血管平滑肌细胞的［３Ｈ］胸腺嘧啶核苷酸掺入．结论：针对ｐ４２和ｐ４４ＭＡＰＫ起始部位设计的１７ｍｅｒＯＤＮ能有效抑制ＥＧＦ诱导的血管平滑肌细胞的增生．

AIM: To investigate the inhibitory effect of mitogen-activated protein kinase (MAPK) antisense oligonucleotide (ODN) on epidermal growth factor (EGF) -induced proliferation of cultured rat vascular smooth muscle cells. Methods: p42 and p44MAPKODN0.2μmol·L1 were transfected into rat vascular smooth muscle cells by lipofectamine. The sense and random ODNs were used as control. Western Blot method and P  81 filter paper were used to detect myelin basic Protein as a substrate for measuring MAPK activity. [3H] thymidine incorporation assay for smooth muscle cell DNA synthesis. Results: MAPK0DN could significantly inhibit the expression of MAPK and MAPK induced by EGF, and significantly inhibit the incorporation of [3H] thymidine into vascular smooth muscle cells. Conclusion: 17merODN designed for p42 and p44MAPK initiating sites can effectively inhibit EGF-induced proliferation of vascular smooth muscle cells.