反义寡脱氧核苷酸对脯氨酰4-羟化酶mRNA表达的抑制作用

The inhibitory effect of antisense oligodeoxynucleotides on prolyl 4 hydroxylase mRNA expression

目的：观察反义寡脱氧核苷酸（ＯＤＮ）对脯氨酰４－羟化酶（Ｐ４Ｈ）两种亚基ｍ ＲＮＡ 表达的影响。方法：采用定量ＲＴ－ＰＣＲ技术检测培养的人皮肤成纤维细胞中Ｐ４Ｈ ｍ ＲＮＡ的相对含量。结果：不同浓度（０．１，０．２，０．５，１．０ μｍ ｏｌ／Ｌ）反义ＯＤＮ处理组，人皮肤成纤维细胞中Ｐ４Ｈ ｍ ＲＮＡ表达水平均显著低于对照组及错配ＯＤＮ 处理组；其中０．１，０．２ μｍ ｏｌ／Ｌ反义ＯＤＮ作用２０ ｈ 时，抑制效应达到最大，５０～７０ ｈ 后，逐渐恢复到对照组水平。正义ＯＤＮ与错配ＯＤＮ对Ｐ４Ｈ ｍ ＲＮＡ表达则无明显抑制作用。结论：反义ＯＤＮ 对Ｐ４Ｈ 两种亚基的ｍ ＲＮＡ表达均有显著和特异的抑制作用。

Objective: To observe the effects of antisense oligodeoxynucleotides (ODN) on the 2 subunits of prolyl 4 hydroxylase (P4H) gene expression. Methods: Quantities of P4H mRNA in the cultured human skin fibroblasts was determined by quantitative RT PCR technology. Results: The mRNA levels of antisense OND(0.1,0.2,0.5,1.0 μmol/L) group were significantly lower than that of control and missense groups. The effective period of antisense ODN varied in 20~40 h and the greatest inhibition appeared at 20 h. After 50~70 h the inhibitory effectiveness reduced as the control. No significant inhibition was seen in the sense and missense ODN groups. Conclusion: The antisense ODN could inhibit gene expression of P4H (α and β subunit) significantly and specifically.