论文部分内容阅读
目的:探讨Ca~(2+)-钙调蛋白依赖性货白激酶(丝裂素活化的蛋白激酶)(CCDPK)在生长因子诱导体外培养大鼠血管平滑肌细胞增殖中的作用及反义CCDPK寡脱氧核苷酸(ODN)对球囊损伤后大白鼠血管内膜增生的抑制作用。方法:利用脂质体转染17-mer CCDPK反义ODN进入培养的血管甲滑肌细胞以抑制CCDPK活性,设正义及随机ODN作对照。用蛋白质印迹法测定CCDPK表达。[~3H]胸腺嘧啶核苷酸掺入测定平滑肌细胞DNA合成。用2F球囊导管造成大白鼠颈动脉再狭窄模型,利用多聚胶F127-ODN系统由血管外膜部位给药。于损伤后2周取样,固定及HE染色观察内膜增生情况。FITC标记的ODN观察体内外给药方法的分布及吸收情况。结果:CCDPK反义ODN能明显抑制PDGFF及ET诱导的CCDPK蛋白表达及[~3H]胸腺嘧啶核苷酸掺入。在大鼠颈动脉再狭窄模型,能明显抑制血管内膜增生。结论:CCDPK介导了PDGF及ET诱导的血管平滑肌细胞增殖。针对p42-和p44-CCDPK起始部位设计的17-mer反义ODN能有效抑制生长因子诱导的血管平滑肌细胞的增殖及球囊损伤大鼠血管内膜增生。
AIM: To investigate the role of CCDPK (Ca2 +) - calmodulin - dependent PKCK in the growth of rat vascular smooth muscle cells induced by growth factors and the antisense CCDPK oligo Inhibitory effect of deoxynucleotide (ODN) on intimal hyperplasia of balloon injured balloon in rats. Methods: The 17-mer CCDPK antisense ODN was transfected into cultured vascular smooth muscle cells to inhibit CCDPK activity. Lipofectamine 2000 was used as a control. CCDPK expression was measured by Western blotting. [~ 3H] thymidine incorporation assay for DNA synthesis in smooth muscle cells. The model of carotid restenosis in rats was induced by 2F balloon catheter and the vascular adventitial site was administered by using the polymer F127-ODN system. At 2 weeks after injury, samples were taken and fixed and HE staining to observe the intimal hyperplasia. FITC labeled ODN to observe the distribution and absorption of in vitro and in vivo administration methods. Results: CCDPK antisense ODN significantly inhibited CCDPK protein expression and [~ 3H] thymidine incorporation induced by PDGFF and ET. In rat carotid artery restenosis model, can significantly inhibit intimal hyperplasia. Conclusion: CCDPK mediates the proliferation of vascular smooth muscle cells induced by PDGF and ET. 17-mer antisense ODN designed for p42- and p44-CCDPK initiation sites effectively inhibits growth factor-induced proliferation of vascular smooth muscle cells and intimal hyperplasia in balloon-injured rats.