Basic Research of Gene Therapy for Intractable Pain with siRNA

为了研究BDNF的哪个外显子参与炎症性疼痛，我们量化了大鼠DRG中炎症诱导的BDNF外显子的表达。大鼠左后爪足底表面皮下注射完全弗氏佐剂（CFA）。在fa处理下，大鼠同侧drg外显子I的表达显著增加。我们的结果支持炎症增加总BDNF mRNA。此外，我们的研究结果表明，外显子I在炎症后BDNF mRNA的增加中起重要作用。因此，我们针对BDNF的外显子I进行siRNA敲除。我们为BDNF的exson I创造了两种不同的sirna。在注射CFA前10天插入鞘内导管。在注射CFA之前，对BDNF外显子I的两种不同sirna进行了两次检测。给药后疼痛行为减少，但未完全抑制。两种sirna均未完全抑制DRG中BDNF的表达。在未来，我们必须研究应该施用多少剂量的siRNA。我们还必须检查siRNA的使用时机。

To investigate which exon of BDNF is involved in inflammatory pain, we quantitated infIammation-induced expression of BDNF exons in the rat DRG. Rats received subcutaneous injections of complete Freund's adjuvant (CFA) into plantar surface of the left hind paw. InCFA-treated rats, the expressions of exon I in ipsilateral DRGincreased significantly. Our results support that inflammation increases total BDNF mRNA. Furthermore, our results indicate that exon I plays important role in increase in BDNF mRNA after inflammation.Therefore, we targeted the exon I of BDNF for knock-down with siRNA. We created the two different siRNAs for exson I of BDNF. The intrathecal catheter was inserted 10 days before CFA injection. The two different siRNAs for the exon I of BDNF twice before CFA injection.The pain behaviors were decreased by the administration of siRNAs, but not completed suppressed. The expression of BDNF was not completed suppressed in DRG after the administration of the two siRNAs.In the future, We have to investigate how much doses of siRNA should be administered. We also have to check the timing when siRNA should be administerd.