Gene knockdown with intrathecal siRNA of NMDA receptor NR2B subunit reduces formalin-induced nociception in the rat
Gene knockdown with intrathecal siRNA of NMDA receptor NR2B subunit reduces formalin-induced nociception in the rat
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DOI：
10.1038/sj.gt.3302376
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发表时间：
2005-01-01
期刊：
影响因子：
5.1
通讯作者：
Cheng, JT
中科院分区：
文献类型：
--
作者：
Tan, PH;Yang, LC;Cheng, JT
n -甲基- d -天冬氨酸（NMDA）受体的激活，在脊髓水平上，已经在一些动物模型中被证明在促进伤害感受中起重要作用。然而，由于NMDA受体亚型的非选择性作用，NMDA拮抗剂作为镇痛药的使用受到严重副作用的限制。最近的发现表明，将小干扰rna （sirna）转染到动物细胞中会导致特定基因的有效、持久的转录后沉默。因此，我们研究了鞘内注射靶向NMDA- r2b受体亚基蛋白（NR2B）受体（NMDA受体的一个亚基）的sirna对疼痛的调节作用。结果表明，real-time PCR和Western blotting结果显示，靶向NR2B亚基的siRNA不仅可以降低NR2B mRNA及其相关蛋白的表达，还可以消除福尔马林诱导的大鼠疼痛行为。注射5mg siRNA-NR2B后，mRNA和蛋白的作用分别在第3天和第7天达到峰值。这些数据证明了itt sirna在全动物行为背景下研究功能性基因表达以治疗慢性疼痛的可行性。
N-methyl-D-aspartate (NMDA) receptor activation, at the level of the spinal cord, has been shown to play an important role in the facilitation of nociception in several animal models. However, the use of NMDA antagonists as analgesics is limited by serious side effects due to nonselective effects among the NMDA receptor subtypes. Recent discoveries revealed that the transfection of small interfering RNAs (siRNAs) into animal cells resulted in the potent, long-lasting, post-transcriptional silencing of specific genes. Thus, we investigated the effect of intrathecal (i.t.) injection of siRNAs targeting NMDA-R2B receptor subunit protein (NR2B) receptors, a subunit of NMDA receptor, for the modulation of pain. The results indicate that the use of siRNA targeting the NR2B subunit not only decreased the expression of NR2B mRNA and its associated protein, as demonstrated by real-time PCR and Western blotting, but also abolished formalin-induced pain behaviors in rat model. The peak effect occurred on day 3 for mRNA and day 7 for its protein, following i.t. injection of 5 mg of siRNA-NR2B. These data prove the feasibility of i.t. siRNAs in the investigation of functional gene expression in the context of whole animal behavior for the management of chronic pain.