Peng Jie, Ma Jiahui, Yang Xue, He Huan, Wu Haopeng, Ma Tongtong, Lu Jianhua
Department of Anesthesiology, Guangzhou General Hospital of Guangzhou Military Command of Chinese PLA, Guangzhou 510010, Guangdong, China.
Department of Anesthesiology, Second Affiliated Hospital, Guangzhou University of TCM, Guangzhou 510120, Guangdong, China.
Pain Res Manag. 2018 Jan 14;2018:7436060. doi: 10.1155/2018/7436060. eCollection 2018.
We constructed a water-soluble lipopolymer (WSLP) as a nonviral gene carrier to deliver siRNA targeting NR2B. The cytotoxicity and serum stability of WSLP loaded with siRNA were evaluated, and the knockdown efficiency of WSLP/NR2B-siRNA in PC12 cells was examined. The results showed that WSLP could protect the loading siRNAs from enzymatic degradation in serum and exhibit low cytotoxicity to cells. After transfection, WSLP/NR2B-siRNA complexes reduced the NR2B transcriptional level by 50% and protein level by 55% compared to control siRNA. Moreover, 3 days after intrathecal injection of WSLP/NR2B-siRNA complexes into rats, the NR2B protein expression decreased significantly to 58%, compared to control treatment ( < 0.01). Injection of WSLP with scrambled siRNA or of polyethylenimine (PEI) with NR2B-siRNA did not show this inhibitory effect. Additionally, injection of WSLP/NR2B-siRNA complexes significantly relieved inflammatory pain in rats at 3, 4, and 5 days with reduced MWT and decreased TWL scores, while injection of WSLP with scrambled siRNA or of PEI with NR2B-siRNA did not. These results demonstrated that WSLP can efficiently deliver siRNA targeting NR2B to PC12 cells and relieve pain in rats with chronic inflammatory pain.
我们构建了一种水溶性脂质聚合物（WSLP）作为非病毒基因载体，用于递送靶向NR2B的小干扰RNA（siRNA）。评估了负载siRNA的WSLP的细胞毒性和血清稳定性，并检测了WSLP/NR2B-siRNA在PC12细胞中的敲低效率。结果表明，WSLP可以保护负载的siRNA在血清中不被酶降解，并且对细胞表现出低细胞毒性。转染后，与对照siRNA相比，WSLP/NR2B-siRNA复合物使NR2B转录水平降低了50%，蛋白质水平降低了55%。此外，将WSLP/NR2B-siRNA复合物鞘内注射到大鼠体内3天后，与对照处理相比，NR2B蛋白表达显著下降至58%（P<0.01）。注射携带乱序siRNA的WSLP或携带NR2B-siRNA的聚乙烯亚胺（PEI）均未显示出这种抑制作用。此外，注射WSLP/NR2B-siRNA复合物在第3、4和5天显著减轻了大鼠的炎性疼痛，机械缩足阈值（MWT）降低，热缩足潜伏期（TWL）评分下降，而注射携带乱序siRNA的WSLP或携带NR2B-siRNA的PEI则没有这种效果。这些结果表明，WSLP可以有效地将靶向NR2B的siRNA递送至PC12细胞，并减轻慢性炎性疼痛大鼠的疼痛。
