Department of Anesthesiology and Intensive Care, Shanghai Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai 200438, P.R. China.
Department of Anesthesiology, Zhejiang Zhoushan Hospital, Zhoushan, Zhejiang 316021, P.R. China.
Mol Med Rep. 2018 Dec;18(6):5545-5551. doi: 10.3892/mmr.2018.9560. Epub 2018 Oct 16.
An increasing body of evidence has indicated that spinal microglial Toll‑like receptor 4 (TLR4) may serve a significant role in the development and maintenance of neuropathic pain (NP). In the present study, experiments were conducted to evaluate the contribution of a tetracycline inducible lentiviral‑mediated delivery system for the expression of TLR4 small interfering (si)RNA to NP in rats with chronic constriction injury (CCI). Behavioral tests, including paw withdrawal latency and paw withdrawal threshold, and biochemical analysis of the spinal cord, including western blotting, reverse transcription‑quantitative polymerase chain reaction and ELISA, were conducted following CCI to the sciatic nerve. Intrathecal administration of LvOn‑si‑TLR4 with doxycycline (Dox) attenuated allodynia and hyperalgesia. Biochemical analysis revealed that the mRNA and proteins levels of TLR4 were unregulated following CCI to the sciatic nerve, which was then blocked by intrathecal administration of LvOn‑siTLR4 with Dox. The LvOn‑siTLR4 was also demonstrated to have no effect on TLR4 or the pain response without Dox, which indicated that the expression of siRNA was Dox‑inducible in the lentivirus delivery system. In conclusion, TLR4 may serve a significant role in neuropathy and the results of the present study provide an inducible lentivirus‑mediated siRNA against TLR4 that may serve as a potential novel strategy to be applied in gene therapy for NP in the future.
越来越多的证据表明，脊髓小胶质细胞 Toll 样受体 4（TLR4）可能在神经病理性疼痛（NP）的发生和维持中发挥重要作用。本研究通过实验评估了四环素诱导的慢病毒介导的 TLR4 小干扰（si）RNA 表达载体在慢性缩窄性损伤（CCI）大鼠 NP 中的作用。CCI 坐骨神经后，进行行为学测试（包括足底缩足潜伏期和足底缩足阈值）和脊髓生化分析（包括 Western blot、逆转录-定量聚合酶链反应和 ELISA）。鞘内给予多西环素（Dox）的 LvOn-si-TLR4 减轻了痛觉过敏和痛觉超敏。生化分析显示，CCI 后脊髓 TLR4 的 mRNA 和蛋白水平上调，而鞘内给予 LvOn-siTLR4 联合 Dox 则阻断了这一过程。LvOn-siTLR4 无 Dox 时对 TLR4 或疼痛反应也没有影响，这表明该 siRNA 在慢病毒递送系统中是 Dox 诱导表达的。综上所述，TLR4 可能在神经病变中发挥重要作用，本研究结果提供了一种针对 TLR4 的诱导型慢病毒介导的 siRNA，可能为未来 NP 的基因治疗提供一种新的策略。
