标题：Gene therapy by lentivirus-mediated RNA interference targeting extracellular-regulated kinase alleviates neuropathic pain in vivo
作者：Jia, Lei; Zhang, Yang; Qu, Yu-Juan; Huai, Juan; Wei, Hui; Yue, Shou-Wei
作者机构：[Jia, Lei; Zhang, Yang; Qu, Yu-Juan; Huai, Juan; Wei, Hui; Yue, Shou-Wei] Shandong Univ, Dept Phys Med & Rehabil, Qilu Hosp, 107 Wenhua Xi Rd, Jinan 2 更多
通讯作者地址：[Yue, SW]Shandong Univ, Dept Phys Med & Rehabil, Qilu Hosp, 107 Wenhua Xi Rd, Jinan 250012, Shandong, Peoples R China.
来源：JOURNAL OF CELLULAR BIOCHEMISTRY
关键词：extracellular-regulated kinase; gene therapy; neuropathic pain; RNA; interference; transient receptor potential vanilloid 4
摘要：Backgrounds Neuropathic pain is an abnormal pain, which is related to the activation of extracellular-regulated kinase (ERK) signaling. This study was to investigate the effects of ERK knockdown via lentivirus-mediated RNA interference on allodynia in rats with chronic compression of the dorsal root ganglia (DRG) and to uncover the potential mechanisms. Methods The model of chronic compression of the dorsal root ganglia (CCD) was established in rats by surgery. Gene silence was induced by injecting rats with lentivirus expressing ERK short hairpin RNA (shRNA). Behavioral test was performed by calculating paw withdrawal mechanical threshold (PWMT) and thermal paw withdrawal latency (TPWL). Results We firstly generated lentivirus expressing ERK shRNA to downregulate ERK gene expression both in vitro and in vivo by using Western blot analysis and quantitative reverse transcription polymerase chain reaction. In CCD, ERK mRNA, and protein levels in DRG neurons were dramatically increased, accompanied with decreased PWMT and TPWL. Lentivirus-mediated RNA interference decreased ERK gene expression in DRG neurons and normalized the PWMT and TPWL in CCD rats, but not in rats infected with lentivirus expressing negative control shRNA. Further, calcium responses of DRG neurons to the hypotonic solution and 4 alpha-phorbol 12,13-didecanoate were enhanced in CCD rats, which were suppressed by lentivirus-mediated ERK gene silence. Finally, the levels of transient receptor potential vanilloid 4 gene expressions in DRG neurons and L4 to L5 spinal cord isolated from CCD rats were dramatically upregulated, which were reversed by lentivirus-mediated ERK gene knockdown. Conclusion Lentivirus-mediated RNA interference (RNAi) silencing targeting ERK might reverse CCD-induced neuropathic pain in rats through transient receptor potential vanilloid 4.