[Effect of electroacupuncture on the expression of Iba-1 in spinal dorsal horn and hippocampus of chronic pain rats with knee osteoarthritis].

Jie Zheng, Feng Yang, Pu-Wei Yuan, Jing-Jie Fan, Xiao-Hui Zheng, Rui Zhu, Yang-le Li
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Abstract

OBJECTIVE To observe the effect of electroacupuncture(EA) on pain-ralated behaviors, morphology of hippocampus, concentrations of inflammatory cytokines and expression of ionized calcium binding adapter molecule 1(Iba-1) in dorsal horn of the spinal cord and the hippocampus, and brain-derived neurotrophic factor (BDNF) in hippocampus of rats with knee osteoarthritis (KOA), so as to explore the mechanism of EA in improving chronic pain of KOA. METHODS Forty SD rats were randomly divided into blank group, saline group, model group and EA group, with 10 rats in each group. Monosodium iodoacetate(MIA, 80 mg/mL, 50 μL) was injected into the left knee joint cavity of rats in the model group and EA group to establish the chronic pain model of KOA, while the same volume of normal saline was injected into the left knee joint cavity of rats in the saline group. Rats in the EA group received EA stimulation(2 Hz/100 Hz, 1-2 mA) at left "Yanglingquan"(GB34) and "Neixiyan"(EX-LE4) for 15 min, 14 d after MIA injection. The treatment was given once daily, 5 d as 1 session and 2 sessions of treatment were required. Methanical withdrawl threshold(MWT) and weight-bearing capacity tests on left hind limbs were carried out 1 d before, 7 d,14 d, 20 d and 26 d after MIA injection. At the 27th day, rats were sacrificed and HE staining was used to observe the morphology of hippocampal CA1 area. Concentrations of interleukin(IL)-1β and tumor necrosis factor(TNF)-α in the left L3-L5 spinal dorsal horn and hippocampal CA1 area were detected by ELISA, the expressions of Iba-1 in the spinal dorsal horn and hippo-campal CA1 area were detected by immunofluorescence, and the expression of BDNF in left hippocampal CA1 area was detected by Western blot. RESULTS The HE staining results of the hippocampal CA1 area showed reduced number of neurons, unclear cell contour and boundary between nucleus and cytoplasm, and nuclear pyknosis in the model group, which was relatively milder in the EA group. Compared with the blank group, MWT and weight-bearing capacity of rats' left hind limbs, and expression of BDNF protein in hippocampal CA1 area were significantly decreased (P<0.01), while the contents of IL-1β and TNF-α, the expression of Iba-1 in spinal dorsal horn and hippocampal CA1 area were significantly increased (P<0.01) in the model group. In comparison with the model group, MWT and weight-bearing capacity of rats' left hind limbs, and protein expression of BDNF in hippocampal CA1 area were significantly increased (P<0.01), while the contents of IL-1β and TNF-α, and the expression of Iba-1 protein in spinal dorsal horn and hippocampal CA1 area were significantly decreased after EA intervention(P<0.01). CONCLUSION EA at GB34 and EX-LE4 can alleviate the pain-related behaviors of KOA rats. The mechanism might be related to the inhibition of inflammatory reaction mediated by microglia in spinal dorsal horn and hippocampus, and the up-regulation of BDNF expression in hippocampus.
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[电针对慢性疼痛大鼠膝骨关节炎脊髓背角和海马Iba-1表达的影响]。
目的:观察电针(EA)对膝骨关节炎(KOA)大鼠疼痛相关行为、海马形态、脊髓背角及海马炎症因子浓度、离子钙结合适配分子1(Iba-1)表达及海马脑源性神经营养因子(BDNF)的影响,探讨电针(EA)改善KOA慢性疼痛的机制。方法:将40只SD大鼠随机分为空白组、生理盐水组、模型组和EA组,每组10只。模型组和EA组大鼠左膝关节腔内注射碘乙酸钠(MIA, 80 mg/mL, 50 μL),建立KOA慢性疼痛模型,生理盐水组大鼠左膝关节腔内注射等量生理盐水。EA组大鼠在MIA注射后14 d,分别在“阳陵泉”(GB34)和“内痔炎”(EX-LE4)左侧进行EA刺激(2 Hz/100 Hz, 1-2 mA) 15 min。治疗方法为每日1次,5 d为1次,需2次治疗。分别于注射MIA前1 d、7 d、14 d、20 d、26 d进行机械脱断阈值(MWT)和左后肢负重测试。第27天处死大鼠,HE染色观察海马CA1区形态。ELISA法检测左L3-L5脊髓背角和海马CA1区白细胞介素(IL)-1β和肿瘤坏死因子(TNF)-α浓度,免疫荧光法检测脊髓背角和海马CA1区Iba-1的表达,Western blot法检测左海马CA1区BDNF的表达。结果:海马CA1区HE染色结果显示,模型组神经元数量减少,细胞轮廓及核质边界不清,核固缩,EA组相对较轻。与空白组比较,大鼠左后肢MWT、负重能力及海马CA1区BDNF蛋白表达均显著降低(ppppp)。结论:GB34、EX-LE4部位EA可减轻KOA大鼠的疼痛相关行为。其机制可能与脊髓背角和海马小胶质细胞抑制炎症反应,上调海马BDNF表达有关。
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