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目的:探讨Ca~(2+)-钙调蛋白依赖性蛋白激酶(CCDPK)在碱性成纤维细胞生长因子(bFGF)诱导体外培养大鼠血管平滑肌细胞(VSMC)增殖中的作用及反义CCDPK寡核苷酸的抑制作用。方法:利用CCDPK特异性抑制剂PD98059或17-mer CCDPK反义寡核苷酸抑制CCDPK活性,用Western blot法测定p44-/p42-CCDPK蛋白表达及磷酸化。[~3H]胸腺嘧啶核苷酸掺入测定SMC DNA合成。结果:PD98059及CCDPK反义寡核苷酸明显抑制bFGF诱导的CCDPK蛋白表达和CCDPK活性以及[~3H]胸腺嘧啶核苷酸掺入。结论:CCDPK介导了bFGF诱导的VSMC增殖。针对p42-和p44-CCDPK起始部位设计的17-mer反义寡核苷酸使p44-和p42-CCDPK蛋白表达下调,能有效抑制bFGF诱导的VSMC的增殖。
AIM: To investigate the role of CCDPK in the proliferation of rat vascular smooth muscle cells (VSMCs) induced by basic fibroblast growth factor (bFGF) and the effect of CCDPK Inhibition of oligonucleotides. METHODS: CCDPK activity was inhibited by CCDPK inhibitor PD98059 or 17-mer CCDPK antisense oligonucleotide. The protein expression and phosphorylation of p44- / p42-CCDPK were determined by Western blot. [~ 3H] thymidine incorporation assay for SMC DNA synthesis. Results: PD98059 and CCDPK antisense oligodeoxynucleotides significantly inhibited the expression of CCDPK and CCDPK induced by bFGF and the incorporation of [~ 3H] thymidine. Conclusion: CCDPK mediates bFGF-induced VSMC proliferation. 17-mer antisense oligonucleotides designed for the p42- and p44-CCDPK initiating sites down-regulated the expression of p44- and p42-CCDPK proteins and effectively inhibited bFGF-induced VSMC proliferation.