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[电针对关节炎大鼠局部皮肤CB2受体阳性细胞免疫反应性的影响]

[Effects of electroacupuncture on the immunoreactivity of focal cutaneous CB2 receptor positive cells in arthritis rats].

作者信息

Li Jun-Jun, Chen Ru-Man, Liu Lei, Wang Shu-Ying, Yu Ping, Xie Yong, Li Man, Shi Jing

机构信息

Department of Neurobiology, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430030, China.

出版信息

Zhen Ci Yan Jiu. 2007 Feb;32(1):9-15.

PMID:17580433
Abstract

OBJECTIVE

To study the effects of electroacupuncture (EA) on the immunoreactivity of focal cutaneous cannabinoid receptor 2 (CB2) positive cells in adjuvant arthritis (AA) rats.

METHODS

A total of 48 adult female SD rats were randomly divided into control group (n = 12), model group (n = 12), acupoint group (n = 12), and non-acupoint group (n = 12). Arthritis model was established by hypodermic injection of complete Freund's adjuvant (CFA, 50 microL) into the left ankle joint. EA (2/ 15 Hz, 1 mA) was applied to "Huantiao" (GB 30) and "Yanglingquan" (GB 34) or two control points 5 mm left to GB30 and GB34 on the diseased side for 30 min, once every other day from the second day on after injection of CFA. Behavioral performance (pain test score) was assesed by using dorsiflexion and plantarflexion pain tests. On the 6th and 16th day after injection of CAF, the animals were anesthetized with 20% urethane (1 g/kg) for collecting the focal skin and subcutaneous tissue samples which were cut into sections (5 microm) to be stained with HE (haematoxylin & cosin) method and immunohistochemical technique respectively for observing changes of the focal cells of the inflamatory tissue and the immunoactivity of the focal cutaneous CB receptor positive cells.

RESULTS

  1. In comparison with control group, the scores of both dorsiflexion and plantarflexion pain tests in model, acupoint and non-acupoint groups increased evidently after modeling (P < 0.05). Compared with model group, the scores of dorsiflexion test on the 3rd day and 5th day and plantarflexion test on the 5th day in EA-acupoint group were considerably lower (P < 0.05), and the scores of the two tests on the 3rd day and 5th day of acupoint group were also markedly lower than those of non-acupoint group (P < 0.05), suggesting a marked pain-relief after EA; while no significant differences were found between non-acupoint and model groups. 2) HE staining showed that the inflammatory cells in the dermal layer of the focus of acupoint group were evidently fewer than those of model and non-acupoint groups on the 6th day and 16th day after modeling. 3) The immunohistochemical results revealed that compared with control group, on the 6th day, the percentages of CB2 receptor positive cell area in the focus of model, non-acupoint and acupoint groups were significantly higher (P < 0.05), and that of acupoint group was markedly higher than those of model and non-acupoint groups (P < 0.05), suggesting further upregulation of the expression of CB2 receptor positive cells; no significant differences of percentages of CB2 receptor immunoreaction positive cell area among model, non-acupoint and acupoint groups were found on the 16th day (P > 0.05).

CONCLUSION

EA of GB30 and GB34 can raise the immunoactivity of cutaneous CB2 receptor positive cells in the inflammatory tissue which maybe contribute to its effects in relieving inflammatory pain and suppressing focal inflammation and adjusting the balance of nociception and antinociception in AA rats.

摘要

目的

研究电针对佐剂性关节炎(AA)大鼠局部皮肤大麻素受体2(CB2)阳性细胞免疫反应性的影响。

方法

将48只成年雌性SD大鼠随机分为对照组(n = 12)、模型组(n = 12)、穴位组(n = 12)和非穴位组(n = 12)。通过将完全弗氏佐剂(CFA,50 μL)皮下注射到左踝关节建立关节炎模型。将电针(2/15 Hz,1 mA)施加于患侧的“环跳”(GB 30)和“阳陵泉”(GB 34)或GB30和GB34左侧5 mm处的两个对照点,每次30分钟,自注射CFA后的第二天起每隔一天进行一次。采用背屈和跖屈疼痛试验评估行为表现(疼痛测试评分)。在注射CAF后的第6天和第16天,用20%乌拉坦(1 g/kg)麻醉动物,收集局部皮肤和皮下组织样本,切成切片(5 μm),分别用苏木精-伊红(HE)染色法和免疫组织化学技术染色,观察炎症组织局部细胞的变化以及局部皮肤CB受体阳性细胞的免疫活性。

结果

1)与对照组相比,模型组、穴位组和非穴位组建模后背屈和跖屈疼痛试验评分均明显升高(P < 0.05)。与模型组相比,电针穴位组第3天和第5天的背屈试验评分以及第5天的跖屈试验评分明显较低(P < 0.05),穴位组第3天和第5天的两项试验评分也明显低于非穴位组(P < 0.05),表明电针后疼痛明显减轻;而非穴位组与模型组之间无显著差异。2)HE染色显示,建模后第6天和第16天,穴位组病灶真皮层的炎症细胞明显少于模型组和非穴位组。3)免疫组织化学结果显示,与对照组相比,第

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