丝裂素活化的蛋白激酶反义寡核苷酸对p42/p44,p38和JNK激酶的靶向选择性及其对血管平滑肌细胞DNA合成的抑制作用<英文>  

Target selectivity of MAPK phosphorothioate antisense ODN on p42/p44, p38 MAPK, and JNK protein expression and its inhibitory effect on VSMC DNA synthesis

作　　者：黄韶玲[1] 丁波[2] 李江[2] 易富贤[2] 刘文兰[2] 余清声[1] 郭兆贵[2] 

机构地区：[1]广州医学院蛇毒研究所,广州中国510182 [2]湖南医科大学分子药理研究室,长沙410078

出　　处：《中国药理学报》1999年第7期613-616,共4页Acta Pharmacologica Sinica

摘　　要：目的：探讨丝裂素活化的蛋白激酶反义寡核苷酸对血清诱导的培养大鼠血和平滑肌细胞增殖的选择性及序列依赖性抑制作用。方法：用脂质体将ｐ４２－和ｐ４４－ＭＡＰＫ反义寡核苷酸转染入大鼠血管平滑肌细胞，设正义及随机寡核苷酸对照，２０％血清刺激后，设正义及随机寡核苷酸对照，２０％血清刺激后，用Ｗｅｓｔｅｒｎ Ｂｌｏｔ法测定总ｐ４４／ｐ４２－ＭＡＰＫ，ｐ３８ ＭＡＰＫ及ＪＮＫｓ蛋白水平及磷酸化ＭＡＰＫ表达。AIM: To analyze the target selective and sequence-specific inhibitory effect of mitogen-activated protein kinase (MAPK) phosphorothioate antisense oligodeoxy-nucleotides (ODN) on p42/p44, p38 MAPK, c-jun NH2-terminal protein kinases ( JNK ) protein expression, and DNA synthesis in vascular smooth muscle cell (VSMC). METHODS: Using a phos-phorothioate-protected 17-mer antisense MAPK ODN directed against the initiation of translation sites of the p42/p44 MAPK isoforms by liposomal transfection to deplete cultured rat, rabbit, and fetal calf VSMC MAP kinases. The 17-mer sense and random sequence MAPK ODN were used as controls. After liposomal transfection, cells were exposed to 20 % serum for 24 h, and then harvested in lysis buffer. P42/p44, p38 MAPK, and p46/p58 JNK protein expression were measured by Western blot. DNA synthesis was measured by [3H]thymidine incorporation. RESULTS: Treatment with MAPK antisense ODN (0.1-0.8 μmol· L-1) for 48 h reduced phosphored p42/p44 MAPK protein expression but without effect on p38 MAPK and JNK expression, and inhibited cultured rat, rabbit, and fetal calf VSMC [3H]thymidine incorporation stimulated by 20 % serum in a concentration-dependent manner. CONCLUSION: The MAPK antisense ODN target-selectively and sequence-specifically reduces the p42/p44 MAPK protein expression and concentration-dependently inhibits proliferation of rat, rabbit and fetal calf VSMC.

关 键 词：蛋白激酶 反义寡核苷酸 血管平滑肌增殖 再狭窄 

分 类 号：R972.6[医药卫生—药品] R654[医药卫生—药学]