Huang Yu-Ting, Liao Jun, Kang Xiao-Ling, Wang Di-Yi, Lin Yu-Ye, Zhang Jing-Yu, Zhang Guo-Jun, Zeng Chu-Fan, Zhang Mei-Qing, Fang Yan-Ping, Kan Yu, Wang Zhi-Fu
College of Acupuncture-moxibustion and Tuina of Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China.
Affiliated People's Hospital of Fujian University of Traditional Chinese Medicine, Fuzhou 350004.
Zhen Ci Yan Jiu. 2025 Aug 25;50(8):919-927. doi: 10.13702/j.1000-0607.20250043.
To observe the effect of electroacupuncture (EA) on mast cell (MC) activity, substance P (SP) and glial fibrillary acidic protein (GFAP) expression in the lumbar spinal dorsal horns (SDHs) and related inflammatory factors in the anterior tibial muscle, lumbar spinal cord and serum of fibromyalgia syndrome (FMS) model rats, so as to explore its mechanisms underlying improvement of FMS.
Thirty-six male SD rats were randomly divided into blank control, model and EA groups, with 12 rats in each group. The FMS model was established by injecting 2-morphine phenol ethanol sulfate into the left anterior tibial muscle. After successful modeling, EA was applied to "Zusanli"(ST36) and "Yanglingquan"(GB34) on the left hind limb for 20 min, once every other day for a total of 7 times. von Frey test was used to detect the mechanical pain threshold (MPT) of both hind limbs. H.E. staining was used to observe morphological changes of the anterior tibial muscle. Immunofluorescence staining was used to determine the expressions of SP, MC, GFAP and 5-hydroxtryptamine (5-HT) in the lumbar SDHs. Suspension chip method was used to detect the contents of inflammatory factors in the lumbar spinal cord, left anterior tibial muscle and serum.
Compared with the blank control group, the difference value of MPT between the bilateral hind-paws, immunofluorescence-positive number of SP and GFAP in the SDHs, contents of TNF-α, macrophage inflammatory protein-1α (MIP-1α), monocyte chemoattractant protein-1 (MCP-1) in the lumbar spinal cord, and the contents of MIP-α, MCP-1, interleukin (IL)-1α, granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte-colony stimulating factor (G-CSF) in the left anterior tibial muscle, as well as TNF-α, IL-1β, GM-CSF and 5-HT contents in the serum were significantly increased (<0.05), while the immunofluorescence-positive number of 5-HT and contents of 5-HT and IL-10 in the lumbar spinal cord, and IL-10 content in the left anterior tibial muscle and serum were strikingly decreased (<0.05) in the model group. In comparison with the model group, the increase of the difference value of MPT between the bilateral hind-paws, immunofluorescence-positive number of SP and GFAP, contents of TNF-α, MIP-α and MCP-1 in the lumbar spinal cord, and the contents of MIP-α, MCP-1, IL-1α, GM-CSF and G-CSF in the left anterior tibial muscle, as well as the contents of TNF-α, IL-1β, GM-CSF and 5-HT in the serum, and the decrease of the immunofluorescence-positive number of 5-HT and contents of 5-HT and IL-10 in the lumbar spinal cord, and IL-10 content in the left anterior tibial muscle and serum were reversed in the EA group (<0.05). Immunofluorescence staining displayed that no apparent changes in the number of MC in the lumbar SDHs after modeling and EA, and MCs presented degranulation in both the model group and EA group. H.E. staining showed disordered arrangement of muscle fibers, enlargement of the gaps between the muscle fibers, and obvious inflammatory infiltration of the anterior tibial muscle in the model group. Compared with the model group, the morphology of muscle fibers in the EA group was relatively complete, with orderly arrangement and reduction of the inflammatory infiltration.
EA of ST36 and GB34 can relieve pain and reduce the central and peripheral inflammatory response in FMS rats, which may be related to its functions in suppressing the expressions of SP and GFAP, and astrocytes activation in the lumbar SDHs and up-regulating the expression of 5-HT in the lumbar spinal cord.
观察电针(EA)对纤维肌痛综合征(FMS)模型大鼠腰段脊髓背角(SDHs)肥大细胞(MC)活性、P物质(SP)和胶质纤维酸性蛋白(GFAP)表达以及胫前肌、腰段脊髓和血清中相关炎症因子的影响,以探讨其改善FMS的作用机制。
将36只雄性SD大鼠随机分为空白对照组、模型组和电针组,每组12只。通过向左侧胫前肌注射2-吗啡苯酚乙醇硫酸盐建立FMS模型。造模成功后,对左侧后肢的“足三里”(ST36)和“阳陵泉”(GB34)进行电针治疗20分钟,隔日1次,共7次。采用von Frey试验检测双侧后肢的机械痛阈(MPT)。采用苏木精-伊红(H.E.)染色观察胫前肌的形态变化。采用免疫荧光染色法检测腰段SDHs中SP、MC、GFAP和5-羟色胺(5-HT)的表达。采用悬浮芯片法检测腰段脊髓、左侧胫前肌和血清中炎症因子的含量。
与空白对照组相比,模型组双侧后爪MPT差值、SDHs中SP和GFAP免疫荧光阳性数、腰段脊髓中肿瘤坏死因子-α(TNF-α)、巨噬细胞炎性蛋白-1α(MIP-1α)、单核细胞趋化蛋白-1(MCP-1)含量,左侧胫前肌中MIP-α、MCP-1、白细胞介素(IL)-1α、粒细胞-巨噬细胞集落刺激因子(GM-CSF)和粒细胞集落刺激因子(G-CSF)含量,以及血清中TNF-α、IL-1β、GM-CSF和5-HT含量均显著升高(<0.05),而腰段脊髓中5-HT免疫荧光阳性数、5-HT和IL-10含量,以及左侧胫前肌和血清中IL-10含量均显著降低(<0.05)。与模型组相比,电针组双侧后爪MPT差值、SP和GFAP免疫荧光阳性数、腰段脊髓中TNF-α、MIP-α和MCP-1含量,左侧胫前肌中MIP-α、MCP-1、IL-1α、GM-CSF和G-CSF含量,以及血清中TNF-α、IL-1β、GM-CSF和5-HT含量升高,腰段脊髓中5-HT免疫荧光阳性数、5-HT和IL-10含量,以及左侧胫前肌和血清中IL-10含量降低的情况均得到逆转(<0.05)。免疫荧光染色显示,造模和电针后腰段SDHs中MC数量无明显变化,模型组和电针组MC均出现脱颗粒现象。H.E.染色显示,模型组胫前肌肌纤维排列紊乱,肌纤维间隙增宽,有明显的炎症浸润。与模型组相比,电针组肌纤维形态相对完整,排列有序,炎症浸润减轻。
ST36和GB34电针可缓解FMS大鼠疼痛,减轻中枢和外周炎症反应,其机制可能与抑制腰段SDHs中SP和GFAP表达、星形胶质细胞激活以及上调腰段脊髓中5-HT表达有关。