Department of Physiology, Institute of Acupuncture and Moxibustion, China Academy of Chinese Medical Sciences, No. 16, Nanxiaojie, Dongzhimennei, Beijing, 100700, China.
Department of Biochemistry and Molecular Biology, Institute of Acupuncture and Moxibustion, China Academy of Chinese Medical Sciences, Beijing, 100700, China.
BMC Complement Altern Med. 2018 Feb 21;18(1):74. doi: 10.1186/s12906-018-2134-8.
Cumulated evidence reveals that glial cells in the spinal cord play an important role in the development of chronic neuropathic pain and are also complicated in the analgesic effect of EA intervention. But the roles of microgliacytes and astrocytes of spinal cord in the process of EA analgesia remain unknown.
A total of 120 male Wistar rats were used in the present study. The neuropathic pain model was established by chronic constrictive injury (CCI) of the sciatic nerve. The rats were randomly divided into sham group, CCI group, and sham CCI + EA group, and CCI + EA group. EA was applied to bilateral Zusanli (ST36)-Yanlingquan (GB34). The mechanical (both time and force responses) and thermal pain thresholds (PTs) of the bilateral hind-paws were measured. The number of microgliacytes and activity of astrocytes in the dorsal horns (DHs) of lumbar spinal cord (L4-5) were examined by immunofluorescence staining, and the expression of glial fibrillary acidic protein (GFAP) protein was detected by western blot.
Following CCI, both mechanical and thermal PTs of the ipsilateral hind-paw were significantly decreased beginning from the 3rd day after surgery (P < 0.05), and the mechanical PT of the contralateral hind-paw was considerably decreased from the 6th day on after surgery (P < 0.05). CCI also significantly upregulated the number of Iba-1 labeled microgliacytes and the fluorescence intensity of glial fibrillary acidic protein (GFAP) -labeled astrocyte in the superficial laminae of DHs on bilateral sides (P < 0.05). After repeated EA, the mechanical and thermal PTs at bilateral hind-paws were significantly relieved (P < 0.05). The increased of number of microgliacytes was markedly suppressed by 2 days' EA intervention, and the average fluorescence intensity was suppressed by 2 weeks' EA. The expression of GFAP protein were down-regulated by 1 and 2 weeks' EA treatment, respectively (P < 0.05).
Repeated EA can relieve neuropathic pain and mirror-image pain in chronic neuropathic pain rats, which is probably associated with its effect in downregulating glial cell activation of the lumbar spinal cord, the microgliacyte first and astrocyte later.
累积的证据表明,脊髓中的神经胶质细胞在慢性神经病理性疼痛的发展中起着重要作用,并且在 EA 干预的镇痛作用中也很复杂。但是,脊髓中的小胶质细胞和星形胶质细胞在 EA 镇痛过程中的作用尚不清楚。
本研究共使用 120 只雄性 Wistar 大鼠。通过坐骨神经慢性缩窄性损伤(CCI)建立神经病理性疼痛模型。将大鼠随机分为假手术组、CCI 组、假手术 CCI+EA 组和 CCI+EA 组。EA 应用于双侧足三里(ST36)-悬钟(GB34)。测量双侧后爪的机械(时间和力反应)和热痛阈值(PTs)。通过免疫荧光染色检查腰椎脊髓(L4-5)背角(DHs)中小胶质细胞的数量和星形胶质细胞的活性,并通过 Western blot 检测神经胶质纤维酸性蛋白(GFAP)蛋白的表达。
CCI 后,从手术后第 3 天开始,同侧后爪的机械和热 PTs 均明显降低(P<0.05),并且从手术后第 6 天开始,对侧后爪的机械 PTs 明显降低(P<0.05)。CCI 还显著上调了双侧浅层 DHs 中 Iba-1 标记的小胶质细胞数量和神经胶质纤维酸性蛋白(GFAP)标记的星形胶质细胞的荧光强度(P<0.05)。重复 EA 后,双侧后爪的机械和热 PTs 明显缓解(P<0.05)。EA 干预 2 天后,小胶质细胞数量明显减少,2 周后平均荧光强度降低。1 周和 2 周的 EA 处理分别下调了 GFAP 蛋白的表达(P<0.05)。
重复 EA 可缓解慢性神经病理性疼痛大鼠的神经病理性疼痛和镜像疼痛,这可能与其下调脊髓胶质细胞激活有关,小胶质细胞先,星形胶质细胞后。