Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, China.
Science and Technology Innovation Center, Guangzhou University of Chinese Medicine, Guangzhou, 510405, China.
J Ethnopharmacol. 2022 May 10;289:115051. doi: 10.1016/j.jep.2022.115051. Epub 2022 Jan 29.
Until now, inflammatory pain, especially ones with central sensitization in the spinal cord, is far from effectively treated. Yu-Xue-Bi Tablets (YXB) is a patented medicine, which has been widely applied for inflammatory pain. However, its therapeutic characteristics and mechanism remain unknown.
This study is designed to evaluate the analgesic characteristics and explore the underlying mechanism of YXB in the inflammatory pain model induced by Complete Freund's Adjuvant (CFA).
The analgesic effects were measured by Von Frey test. The expression of calcitonin gene-related peptide (CGRP) was quantified by immunofluorescence. The expression of immune factors was analyzed via Luminex assay. The further quantifications of C-C Motif chemokine ligand 3 (CCL3) were verified by Enzyme-linked immunosorbent assay (ELISA). The transmigration of macrophage and activation of microglia were evaluated by immunofluorescence. Spinal injections of purified CCL3, CCR1 antagonist (J113863) and CCR5 antagonist (Maraviroc) were used to clarify roles of CCL3 assumed in the pharmacological mechanism of YXB.
In CFA mice, YXB ameliorated the mechanical allodynia in dose and time dependent way, suppressed the central sensitization in dose dependent way. In the L5 spinal cord, YXB downregulated the expression of macrophage M1 pro-inflammatory factors TNFRI and CCL3, inhibited the transmigration of circulating macrophage and the activation of microglia. Purified CCL3 led to the transmigration of macrophage, activation of microglia, central sensitization, and mechanical allodynia in the Sham mice. Inhibitors of CCR1 and CCR5 attenuated above symptoms in CFA mice. Purified CCL3 blocked YXB mediated down regulation of CCL3, inhibition of macrophage transmigration, but not activation of microglia.
YXB exerts the analgesic effects by inhibiting CCL3-mediated peripheral macrophage transmigrate into spinal cord. This study provided a novel approach for inflammatory pain treatment and new insight into the pharmacological action of YXB.
到目前为止，炎症性疼痛，特别是脊髓中枢敏化引起的疼痛，还远未得到有效治疗。愈伤灵痹片（YXB）是一种专利药物，已广泛用于治疗炎症性疼痛。然而，其治疗特点和机制尚不清楚。
本研究旨在评估 YXB 在完全弗氏佐剂（CFA）诱导的炎症性疼痛模型中的镇痛特征，并探讨其潜在机制。
采用 Von Frey 测试测量镇痛效果。免疫荧光法定量测定降钙素基因相关肽（CGRP）的表达。通过 Luminex 分析检测免疫因子的表达。酶联免疫吸附试验（ELISA）进一步验证 C-C 基序趋化因子配体 3（CCL3）的定量。免疫荧光法评估巨噬细胞迁移和小胶质细胞激活。鞘内注射纯化的 CCL3、CCR1 拮抗剂（J113863）和 CCR5 拮抗剂（Maraviroc），以阐明 CCL3 在 YXB 药理学机制中的作用。
在 CFA 小鼠中，YXB 呈剂量和时间依赖性改善机械性痛觉过敏，呈剂量依赖性抑制中枢敏化。在 L5 脊髓中，YXB 下调巨噬细胞 M1 促炎因子 TNFRI 和 CCL3 的表达，抑制循环巨噬细胞的迁移和小胶质细胞的激活。纯化的 CCL3 导致 Sham 小鼠的巨噬细胞迁移、小胶质细胞激活、中枢敏化和机械性痛觉过敏。CCR1 和 CCR5 的抑制剂可减轻 CFA 小鼠的上述症状。纯化的 CCL3 阻断了 YXB 介导的 CCL3 下调、巨噬细胞迁移抑制，但不抑制小胶质细胞激活。
YXB 通过抑制 CCL3 介导的外周巨噬细胞迁移到脊髓中发挥镇痛作用。本研究为炎症性疼痛治疗提供了一种新方法，并为 YXB 的药理学作用提供了新的见解。
