罗浮山风湿膏药通过抑制TLR4/TNF-α信号通路缓解小鼠神经病理性疼痛

傅玉芳; 谭伟玲; 李小翠; 林荣钿; 刘叔文; 叶玲

doi:10.12122/j.issn.1673-4254.2025.11.01

南方医科大学学报 ›› 2025, Vol. 45 ›› Issue (11) : 2285 -2296. DOI: 10.12122/j.issn.1673-4254.2025.11.01

罗浮山风湿膏药通过抑制TLR4/TNF-α信号通路缓解小鼠神经病理性疼痛

傅玉芳 ¹ ,
谭伟玲 ¹^,² ,
李小翠 ¹ ,
林荣钿 ³ ,
刘叔文 ¹^,⁴^,⁵ ,
叶玲 ¹^,⁵

作者信息 +

LuoFuShan Rheumatism Plaster ameliorates neuropathic pain in mice by suppressing TLR4/TNF-α signaling

Yufang FU ¹ ,
Weiling TAN ¹^,² ,
Xiaocui LI ¹ ,
Rongtian LIN ³ ,
Shuwen LIU ¹^,⁴^,⁵ ,
Ling YE ¹^,⁵

Author information +

文章历史 +

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摘要

目的探究罗浮山风湿膏药（LFS）对神经病理性疼痛（NP）的治疗效果与作用机制。方法构建坐骨神经慢性压迫性损伤（CCI）小鼠模型，分别以低、中、高剂量（2.2、4.4、8.8 cm²）的LFS连续干预14 d。通过检测机械刺激缩足反射阈值（MWT）和热刺激缩足反射潜伏期（PWL），血浆炎症因子IL-6、TNF-α水平，以及坐骨神经组织病理学分析评估其治疗效果；采用网络药理学与分子对接技术筛选LFS抗NP的关键靶点及通路；通过RT-qPCR和免疫组化验证相关靶点和通路；最后测定心脏、肝脏、肾脏脏器系数及功能损伤标志物评价安全性。结果与CCI模型组相比，LFS可剂量依赖性地升高MWT与PWL，降低血浆炎症因子IL-6、TNF-α水平，减轻坐骨神经炎症的损伤程度（P<0.05）。网络药理学分析显示，LFS含378种活性成分，可作用于279个NP相关基因，主要富集于TLR和TNF信号通路。分子对接结果表明，LFS关键成分槲皮素和熊果酸可与TLR4和TNF-α靶点稳定结合。与CCI 模型组相比，LFS 可剂量依赖性地下调脊髓组织中Tlr4 和Tnf-α的mRNA 表达水平，小鼠坐骨神经中TLR4和TNF-α蛋白的表达亦同步降低（P<0.05）。安全性评估显示，各剂量LFS组脏器系数及心肝肾损伤标志物与CCI组和假手术组差异无统计学意义（P>0.05）。结论 LFS通过抑制TLR4/TNF-α通路介导的神经炎症缓解NP，且具有良好的安全性。

Abstract

Objective To explore the therapeutic effect of LuoFuShan Rheumatism Plaster (LFS) on neuropathic pain (NP) and its molecular mechanism. Methods Mouse models of sciatic nerve chronic constriction injury (CCI) were treated with low, medium, and high doses (2.2, 4.4, and 8.8 cm², respectively) of LFS by topical application for 14 consecutive days. The therapeutic effects were assessed by evaluating the mechanical withdrawal threshold (MWT), paw withdrawal latency (PWL), plasma IL-6 and TNF-α levels, and histopathology of the sciatic nerve. Network pharmacology and molecular docking were used to identify the key targets and signaling pathways. The key targets were verified by RT-qPCR and immunohistochemistry. The biosafety of LFS was evaluated by measuring the organ indices and damage indicators of the heart, liver, and kidneys. Results Compared with the CCI group, LFS dose-dependently increased MWT and PWL, reduced plasma IL-6 and TNF-α levels, and alleviated sciatic nerve inflammation in the mouse models. Network pharmacology identified 378 bioactive compounds targeting 279 NP-associated genes enriched in TLR and TNF signaling. Molecular docking showed that quercetin and ursolic acid in LFS could stably bind to TLR4 and TNF‑α. In the mouse models of sciatic nerve CCI, LFS significantly downregulated the mRNA expression levels of Tlr4 and Tnf-α in the spinal cord in a dose-dependent manner and lowered the protein expressions of TLR4 and TNF-α in the sciatic nerve. LFS treatment did not cause significant changes in the organ indices or damage indicators of the heart, liver and kidneys as compared with those in the CCI model group and sham-operated group. Conclusion LFS alleviates NP in mice by suppression of TLR4/TNF-α-mediated neuroinflammation with a good safety profile.