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目的:探讨丝裂素活化蛋白激酶(MAPK)反义寡核苷酸(ODN)对血管紧张素Ⅱ(Ang Ⅱ)诱导的心肌成纤维细胞c-myc基因及其细胞增殖的抑制效应。方法:MAPK反义ODN转染培养新生大鼠心肌成纤维细胞,Western Blot法结合p-81滤纸法测定MAPK活性;Northern Blot法检测c-myc mRNA的表达;[~3H]TdR掺入和[~3H]脯氨酸接入测定细胞DNA和胶原蛋白的合成。结果:MAPK反义ODN显著抑制Ang Ⅱ诱导的MAPK蛋白表达及其活性;显著抑制c-myc基因的表达以及细胞DNA和胶原蛋白的合成。结论:MAPK反义ODN特异性下调MAPK的活性,有效抑制了Ang Ⅱ诱导的c-myc基因的表达以及心肌成纤维细胞的增殖和胶原蛋白的合成。
Objective: To investigate the inhibitory effect of mitogen-activated protein kinase (MAPK) antisense oligonucleotide (ODN) on c-myc gene and cell proliferation induced by angiotensin Ⅱ (Ang Ⅱ) in cardiac fibroblasts. Methods: MAPK activity was detected by Western Blot and p-81 filter paper. The expression of c-myc mRNA was detected by Northern Blot. [~ 3H] TdR incorporation and [ ~ 3H] Proline Accession for the Determination of Cell DNA and Collagen Synthesis. Results: MAPK antisense ODN significantly inhibited Ang Ⅱ-induced MAPK protein expression and activity; significantly inhibited c-myc gene expression and cell DNA and collagen synthesis. CONCLUSION: MAPK antisense ODN specifically down-regulates the activity of MAPK and effectively inhibits the expression of c-myc gene induced by Ang Ⅱ and the proliferation and collagen synthesis of cardiac fibroblasts.