In vivo modulation of the rat cytochrome P450 1A1 by double-stranded phosphorothioate oligodeoxynucleotides.
In vivo modulation of the rat cytochrome P450 1A1 by double-stranded phosphorothioate oligodeoxynucleotides.
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双链硫代磷酸寡脱氧核苷酸对大鼠细胞色素 P450 1A1 的体内调节。
DOI：
10.1006/taap.1995.1221
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Iversen,P
中科院分区：
文献类型：
--
作者：
Tracewell,W;Desjardins,J;Iversen,P
CYP 1A 1基因表达受已知的控制CYP 1A 1转录抑制和诱导的顺式和反式作用元件调节。在Sprague-Dawley大鼠中检查了与CYP 1A 1负调控元件（NRE）序列相同的双链硫代磷酸寡核苷酸（dsODN）的影响。采用了两种策略：（i）形成双链ODN（ODN 1）的两个单链互补的25-mer ODN和（ii）形成dsODN发夹的54-碱基自身互补的ODN（ODN 2）。评估具有乱序NRE序列的dsODN发夹作为对照（ODN 3）。单次静脉注射1.7 mg ODN 1后，Zoxazolamine麻痹时间（CYP 1A 1活性的体内标志物）从生理盐水处理大鼠的184 ± 18 min缩短至103 ± 12.5 min。肝微粒体EROD（CYP 1A 1/2活性的体外标志物）从生理盐水处理动物的210 ± 10 pmol分别增加至静脉内ODN 1和静脉内ODN 2后的703 ± 73和623 ± 89 pmol试卤灵/mg蛋白/min。ODN 1的活性不改变CYP 2 E1和2B 1/2的标志物PNP羟基化和PROD。ODN 2没有显著改变PNP，但显著改变PROD。ODN 3在任何测定中均未引起任何显著变化。在ZX麻痹和EROD中，在静脉注射后观察到ODN 1诱导的反应，但在ip注射ODN 1后没有观察到。ODN 1和HPO处理的大鼠肝微粒体的蛋白质印迹分析也显示CYP 1A 1蛋白质增加。这些数据表明，双链ODN模拟顺式作用的NRE在体内诱导CYP 1A 1在没有其他外源性物质。
CYP1A1 gene expression is regulated by known cis- and transacting elements controlling inhibition and induction of CYP1A1 transcription. The influence of a double-stranded phosphorothioate oligonucleotide (dsODN) with sequence identical to the CYP1A1 negative regulatory element (NRE) was examined in Sprague-Dawley rats. Two strategies were employed: (i) two single-stranded complementary 25-mer ODNs that form a double-stranded ODN (ODN1) and (ii) a 54-base, self-complementary ODN which forms a dsODN hairpin (ODN2). A dsODN hairpin with scrambled NRE sequence was evaluated as a control (ODN3). Zoxazolamine paralysis times, an in vivo marker of CYP1A1 activity, were reduced from 184 ± 18 min in saline-treated rats to 103 ± 12.5 min 24 hr after a single 1.7-mg ODN1 iv injection. Liver microsomal EROD, an in vitro marker of CYP1A1/2 activity, was increased from 210 ± 10 pmol in saline-treated animals to 703 ± 73 and 623 ± 89 pmol resorufin/mg protein/min after iv ODN1 and iv ODN2, respectively. ODN1′s activity did not change PNP hydroxylation and PROD, markers of CYPs 2E1 and 2B1/2. ODN2 did not significantly change PNP but did significantly alter PROD. The ODN3 did not cause any significant changes in any assay measured. The ODN1-induced responses in ZX paralysis and EROD were observed post-iv injection, but not following ip injection of ODN1. Western blot analysis of ODN1- and HPO-treated rat liver microsomes also revealed increased in CYP1A1 protein. These data indicate double-stranded ODNs mimic the cis-acting NRE in vivo inducing CYP1A1 in the absence of other xenobiotics.