Immunoneutralization of c-Fos using intrathecal antibody electroporation attenuates chronic constrictive injury-induced hyperalgesia and regulates preprodynorphin expression in rats
Immunoneutralization of c-Fos using intrathecal antibody electroporation attenuates chronic constrictive injury-induced hyperalgesia and regulates preprodynorphin expression in rats
复制标题
DOI：
10.1097/00000542-200310000-00029
复制
发表时间：
2003-10-01
期刊：
影响因子：
8.8
通讯作者：
Lin, CR
中科院分区：
文献类型：
--
作者：
Lan, CY;Tan, PH;Lin, CR
背景：体内电穿孔已成功用于将 DNA、RNA、寡核苷酸和蛋白质引入细胞中，用于实验和治疗目的。作者在体外和体内评估了电穿孔介导的 c-Fos 抗体疗法对神经性疼痛的疗效。方法。首先，作者研究了鞘内 c-Fos 抗体电穿孔对转染 p-AP-Luc 质粒并用 100 muM 谷氨酸激活的培养脊髓神经元细胞中激活蛋白 (AP-1) 启动子活性的抑制作用。 c-Fos的抑制作用；根据相对荧光素酶活性评估抗体电穿孔对AP-1启动子活性的调节。其次，使用 c-Fos 抗体对患有慢性缩窄性损伤的大鼠进行神经性疼痛的电穿孔治疗。在慢性缩窄性损伤之前以及随后的偶数天（直至第14天（包括第14天））测量热伤害感受阈值，以评估和比较鞘内电穿孔的治疗效果。通过蛋白质印迹分析和免疫组织化学分析评估时间进程。通过评估鞘内 c-Fos 电穿孔后第 2 天和第 10 天的强啡肽原 mRNA 和强啡肽来测量促痛感受基因表达。结果：c-Fos 抗体的共转染显着降低了谷氨酸诱导的 AP-1 活性。 c-Fos 鞘内电转移；抗体减弱脊髓强啡肽水平，表现为慢性缩窄性损伤受影响肢体的疼痛阈值显着升高。结论：本研究表明，通过鞘内电穿孔将抗体转移到大鼠脊髓中是研究脊柱相关疾病内源性因素功能的有用方法。
Background: In vivo electroporation has been successfully used for the introduction of DNA, RNA, oligonucleotides, and proteins into cells for experimental and therapeutic purposes. The authors evaluated the efficacy of electroporation-mediated c-Fos antibody therapy for neuropathic pain in vitro and in vivo.Methods. First, the authors studied the inhibitory effects of intrathecal c-Fos antibody electroporation on the activating protein (AP-1) promoter activity in cultured spinal neuronal cells transfected with p-AP-Luc plasmid and activated with 100 muM glutamate. The inhibitory effect of c-Fos; antibody electroporation in the regulation of AP-1 promoter activity was assessed according to the relative luciferase activity. Second, rats with chronic constrictive injury underwent electroporation treatment for neuropathic pain using c-Fos antibody. Thermal nociceptivc thresholds were measured before chronic constrictive injury and then on even-numbered days, up to and including day 14, to assess and compare the therapeutic effects of intrathecal electroporation. The time course was assessed by Western blot analysis and by inummohistochemical analysis. Pronociceptivc gene expression was measured by assessing prodynorphin mRNA and dynorphin peptides on days 2 and 10 after intrathecal c-Fos electroporation.Results: Cotransfection of c-Fos antibody significantly decreased glutamate-induced AP-1 activity. Intrathecal electrotransfer of c-Fos; antibody attenuated spinal dynorphin levels, as manifested by significantly elevated pain thresholds in the chronic constrictive injury-affected limbs.Conclusion: This study shows that transfer of antibody into rat spinal cords by intrathecal electroporation is a useful method to study the function of endogenous factors of spinal-related disorders.