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目的探讨组蛋白去乙酰化酶(HDAC)对大鼠血管平滑肌细胞(VSMC)增殖及迁移的影响。方法采用DNA重组技术构建抑制HDAC基因RNAi载体,将其转染到分离培养的Wistar大鼠VSMC中抑制其表达,并检测其细胞生物学改变。结果测序结果表明所构建的RNAi载体包含有正确的片段插入。将该载体转染VSMC后,经荧光定量PCR、免疫印迹检测其HDAC1基因表达受到明显抑制,最大抑制效率为55%,且组蛋白H3乙酰化水平明显升高。RNAi抑制HDAC1蛋白表达后,VSMC倍增时间增加、细胞迁移率下降。结论HDAC基因RNAi抑制大鼠VSMC的增殖与迁移,提示组蛋白去乙酰化在VSMC的增殖与迁移中发挥重要的作用。
Objective To investigate the effect of histone deacetylase (HDAC) on the proliferation and migration of rat vascular smooth muscle cells (VSMCs). Methods RNAi vector containing HDAC gene was constructed by DNA recombination technique. The recombinant plasmid was transfected into VSMC of isolated Wistar rats to inhibit the expression of the RNAi vector and detect the change of cell biology. Results The sequencing results showed that the constructed RNAi vector contained correct insertions. After the vector was transfected into VSMC, the expression of HDAC1 gene was significantly inhibited by fluorescence quantitative PCR and Western blotting. The maximum inhibitory efficiency was 55% and the histone H3 acetylation level was significantly increased. RNAi inhibited the expression of HDAC1, VSMC doubling time increased, cell migration decreased. Conclusion HDAC gene RNAi can inhibit the proliferation and migration of rat VSMCs, suggesting that histone deacetylation plays an important role in VSMC proliferation and migration.