Department of Anesthesiology, Tangshan Workers' Hospital, Tangshan, China.
Department of Anesthesiology, Tangshan Hongci Hospital, Tangshan, China.
Ann Palliat Med. 2021 Jun;10(6):6873-6882. doi: 10.21037/apm-21-669.
microRNA-138 (miRNA-138) might have a promising therapeutic effect in the Neuropathic pain (NP). We aim to investigate the effects of miRNA-138 on NP and explore its underlying mechanism.
we performed a partial sciatic nerve ligation (pSNL) surgery in rats to induce pain and inflammation. Rats were administrated by intrathecal injection of lentiviral (LV)-mediated miRNA-138. Mechanical withdrawal threshold (MWT) and paw withdrawal thermal latency (PWTL) were measured to evaluate the pain degree. The expression levels of miRNA-138, toll-like receptor 4 (TLR4), tumor necrosis factor-alpha (TNF-α), interleukin-β (IL-β), and IL-6 in the spinal cord were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Western blotting was performed to measure the expressions of macrophage inflammatory protein-1 alpha (MIP-1α) and C-C chemokine receptor type 1 (CCR1). Next, the mechanism of miRNA-138 on NP was investigated by intrathecal injection of CCR1 inhibitor or MIP-1α neutralizing antibody. Inflammatory factors, MWT, and PWTL were also measured on day 7.
Intrathecal injection of miRNA138 significantly reduced MWT and PWTL. qRT-PCR showed that miRNA138 mimic group significantly reduced the level of TLR4, TNF-α, Il-β, and IL-6 on day 7. Western blotting showed that the protein expressions of MIP-1α and CCR1 in pSNL + miRNA138 mimic group were significantly decreased on day 7. In addition, the miRNA138 inhibitor inversely increased MWT, PWTL and inflammatory cytokines. Further, the effect of miRNA138 inhibitor all were significantly reversed by CCR1 inhibitor or MIP-1α neutralizing antibody.
Intrathecal injection of miRNA-138 can remarkably alleviate NP in rats with a pSNL, which may be achieved by suppressing the TLR4 and MIP-1α/CCR1 signaling pathways.
微小 RNA-138（miRNA-138）在神经病理性疼痛（NP）中可能具有有前景的治疗作用。我们旨在研究 miRNA-138 对 NP 的影响，并探索其潜在机制。
我们在大鼠中进行部分坐骨神经结扎（pSNL）手术以诱导疼痛和炎症。大鼠通过鞘内注射慢病毒（LV）介导的 miRNA-138 进行给药。通过机械撤足阈值（MWT）和足底撤足热潜伏期（PWTL）测量来评估疼痛程度。通过实时定量聚合酶链反应（qRT-PCR）检测脊髓中 miRNA-138、Toll 样受体 4（TLR4）、肿瘤坏死因子-α（TNF-α）、白细胞介素-β（IL-β）和白细胞介素-6（IL-6）的表达水平。通过蛋白质印迹法测量巨噬细胞炎性蛋白-1α（MIP-1α）和 C-C 趋化因子受体 1（CCR1）的表达。然后，通过鞘内注射 CCR1 抑制剂或 MIP-1α 中和抗体来研究 miRNA-138 对 NP 的作用机制。第 7 天还测量了炎性因子、MWT 和 PWTL。
鞘内注射 miRNA138 可显著降低 MWT 和 PWTL。qRT-PCR 显示 miRNA138 模拟组在第 7 天显著降低了 TLR4、TNF-α、IL-β 和 IL-6 的水平。蛋白质印迹法显示 pSNL+miRNA138 模拟组在第 7 天 MIP-1α 和 CCR1 的蛋白表达明显降低。此外，miRNA138 抑制剂可逆转增加 MWT、PWTL 和炎性细胞因子。进一步，miRNA138 抑制剂的作用均被 CCR1 抑制剂或 MIP-1α 中和抗体显著逆转。
鞘内注射 miRNA-138 可显著减轻 pSNL 大鼠的 NP，这可能是通过抑制 TLR4 和 MIP-1α/CCR1 信号通路实现的。
