2,4,6-三硝基苯磺酸诱导的结肠炎模型小鼠内脏疼痛和穴位致敏的外周神经机制。

Xin-Yan Gao, Nan Zhang, Kun Liu, Han-Qing Xi, Yun Liu, Xun He, Shu Han, Bing Zhu
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引用次数: 0

摘要

目的通过对背根神经节(DRG)神经元进行活体钙成像,探讨内脏痛及相关躯体(穴位)致敏的神经机制:将 8 只 BALB/c 小鼠随机分为对照组和模型组,每组 4 只。结肠炎模型由 2,4,6-三硝基苯磺酸(TNBS)结肠灌注诱发,每天一次,连续 7 天。对照组小鼠接受每天一次的生理盐水结肠灌注,连续 7 天。静脉注射EB后,观察两组小鼠躯体神经源性炎症(皮肤渗出埃文斯蓝 [EB])的部位和面积。疼痛行为学试验:将16只C57BL/6J小鼠随机分为对照组和模型组,每组8只,用Von Frey灯丝刺激结肠炎小鼠的转归躯体反应区,记录10次以内的回避和爪抽反应次数。将 24 只 Pirt-GCaMP6s 转基因小鼠随机平均分为对照组和结肠炎模型组,进行体内 DRG 钙成像测试。使用共聚焦荧光显微镜观察L6或L4 DRG神经元对结肠直肠扩张(CRD)、下背部刷毛或后爪机械刺激的反应:与对照组相比,结肠炎模型组(PPPPPPirt-GCaMP6s转基因小鼠)后爪和背部下方区域的EB渗出斑面积增大,在刷洗L6 DRG神经元感受野(背部下方)皮肤后,结肠炎模型组刷洗激活的DRG神经元以及小、中、大尺寸神经元的荧光强度显著高于对照组(PPPPPP结论:结肠炎可能会导致体表敏感性降低:结肠炎可能会导致同一神经节段和相邻神经节段的体表敏化以及有反应的腰部DRG神经元数量和活性的增加,其中与直肠结肠处于同一神经节段的L6 DRG神经元的反应器数量和钙荧光信号强度增加,而与直肠结肠相邻水平的L4 DRG神经元的反应器数量增加,这表明外周神经敏化可能存在不同的机制。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Peripheral neural mechanism of visceral pain and acupoint sensitization in 2, 4, 6-trinitrobenzene sulfonic acid-induced colitis model mice.

Objectives: To explore the neural mechanism of visceral pain and related somatic (acupoints) sensitization by using in vivo calcium imaging of dorsal root ganglia (DRG) neurons.

Methods: Eight BALB/c mice were randomly divided into control and model groups, with 4 mice in each group. The colitis model was induced by colorectal perfusion of 2, 4, 6-trinitrobenzene sulfonic acid (TNBS) once daily for 7 days. Mice of the control group received colorectal perfusion of normal saline once daily for 7 days. The location and area of the somatic neurogenic inflammation (cutaneous exudation of Evans blue [EB]) of the 2 groups of mice were observed after intravenous injection of EB. For pain behavioral tests, sixteen C57BL/6J mice were randomly divided into control and model groups, with 8 mice in each group, and a Von Frey filament was used to stimulate the referred somatic reactive regions in colitis mice, and the number of avoidance and paw withdraw reaction within 10 tests was recorded. For in vivo DRG calcium imaging tests, 24 Pirt-GCaMP6s transgenic mice were randomly and equally divided into control group and colitis model group. The responses of the neurons in L6 or L4 DRG to colorectal distension (CRD), lower back brushing, or mechanical stimulation at the hindpaw were observed using confocal fluorescence microscope.

Results: Compared with the control group, the area of EB exudation spot in the hindpaw and lower back regions was increased in the colitis model group (P<0.05), and the avoidance or paw withdraw numbers induced by Von Frey stimulation at the lower back and hindpaw were increased (P<0.01, P<0.05), indicating that colitis induced regional skin (acupoints) sensitization in the lower back and hindpaw regions. Compared with the control group, the percentage of L6 DRG neurons activated by 60 mm Hg CRD in the colitis model mice were apparently increased (P<0.01), the activated neurons mainly involved the medium-sized DRG neurons (P<0.01). In Pirt-GCaMP6s transgenic mice, following brushing the skin of the receptive field (lower back) of L6 DRG neurons, the fluorescence intensity of the brushing-activated DRG neurons and small, medium and large-sized neurons were significantly higher in the colitis model group than those in the control group (P<0.001, P<0.01, P<0.05). After brushing and clamping the skin of the right hindpaw (receptive field of L4 DRG neurons), the percentages of the activated L4 DRG neurons were obviously higher in the colitis model group than those in the control group (P<0.01, P<0.05), while there were no significant changes in the proportion of small, medium and large-sized neurons between the control and colitis model groups.

Conclusions: Colitis may lead to body surface sensitization at the same and adjacent neuro-segments as well as to an increase of the number and activity of the responsive lumbar DRG neurons, among which the L6 DRG neurons at the same neuro-segment as the rectum colon showed an increase in the number of responders and intensity of calcium fluorescence signal while L4 DRG neurons at the level adjacent to the rectum colon showed an increase in the number of responders, suggesting that there may be different mechanisms of peripheral neural sensitization.

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