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不同波形电针对急性痛风性关节炎大鼠抗炎及镇痛机制的研究

The Research on Mechanisms of Anti-Inflammation and Analgesia for Rats of Acute Cerebral Infarction by Scalp-Ponint Penetrative Acupancture

【作者】 金弘

【导师】 孙忠人;

【作者基本信息】 黑龙江中医药大学 , 针灸推拿学, 2009, 博士

【摘要】 目的:明确不同波形电针治疗急性痛风性关节炎的疗效,揭示其抗炎及镇痛机制,优化急性痛风性关节炎的电针治疗方案,指导临床治疗。方法:采用Coderre等经典造模方法建立急性痛风性关节炎大鼠模型。随机分成5组:空白组、模型组、电针疏波组、电针密波组、电针疏密波组。各组随机分成1d、3d、5d三个时间点,选取“足三里”、“三阴交”穴进行电针治疗。并于治疗1、3、5天后,分别测定各组大鼠踝关节肿胀度和痛阈变化。分别采用比色法、荧光分光光度法、ELISA酶联免疫法,检测关节滑膜组织中疼痛介质、炎症介质、细胞因子含量变化。通过病理学检测、免疫组织化学和原位杂交等技术手段探讨其病理形态学变化及相关蛋白和基因表达的变化,评价其抗炎及镇痛效果。并进行统计学分析。结果:(1)电针疏密波组能显著减轻踝关节肿胀度(P<0.05,P<0.01),表明电针有明显的抗炎消肿作用;(2)HE染色显示:电针疏密波组可使大鼠关节炎症细胞浸润明显减轻,滑膜细胞增生明显被抑制,肉芽组织增生明显减少;(3)电针密波组能提高实验性大鼠痛阈(P<0.05,P<0.01),降低外周疼痛介质K~+、DA、5-HT的含量(P<0.05,P<0.01),表明电针有明显的镇痛作用;(4)在病变关节局部,电针能显著降低急性痛风性关节炎大鼠滑膜组织IL-1β、IL-8、TNF-α含量(P<0.05,P<0.01),升高IL-4的含量(P<0.05,P<0.01),并降低关节滑膜内COX-2mRNA的表达(P<0.05,P<0.01),可促使急性痛风性关节炎大鼠滑膜内HSP-70蛋白表达显著增加(P<0.05,P<0.01),从而抑止滑膜组织炎症发生。结论:电针“足三里”和“三阴交”穴具有明显的抗炎及镇痛作用。(1)电针不同波形均能显著抑制急性痛风性关节炎大鼠踝关节炎性细胞浸润及滑膜细胞的增生,减轻踝关节肿胀度,其抗炎消肿作用以电针疏密波为最佳。(2)电针不同波形均能提高急性痛风性关节炎大鼠的痛阈,降低外周疼痛介质K~+、DA、5-HT的含量,其镇痛作用以电针密波为最佳。(3)电针不同波形均能明显下调大鼠滑膜组织中IL-1β、IL-8、TNF-α的含量,增加IL-4的含量,减轻踝关节局部炎症反应,且以电针疏密波为最佳。(4)电针不同波形均能有效抑制急性痛风性关节炎大鼠足踝关节滑膜内COX-2mRNA的表达,上调HSP-70蛋白表达,从而减轻踝关节局部炎症反应,减轻关节损伤,且以电针疏密波为最佳。

【Abstract】 Objective To define the therapeutic effects of electroacupuncture in different wave formsfor acute gouty arthritis,to reveal its mechanisms of anti-inflammation andanalgesia,to optimize treatment program of electroacupuncture for acutegouty arthritis, and to guide the clinical treatment.Methods Coderre’s Classical methods have been used to establish the model ofacute gouty arthritis in rats . Wistar rats were divided randomly into fivegroups: blank group, model group, group of electroacupuncture in Shu-wave,group of electroacupuncture in dense-wave, group of electroacupuncture inShu and dense wave. Each group was separated into 1d, 3d, 5d as subgroup forinvestigation. Select "Zusanli", "Sanyinjiao" to be electroacupuncture points.The rats were measured by changes of ankle swelling degree and painthreshold in 1 , 3 , 5 days’ treatment.Colorimetry,, fluorescencespectrophotometry, and ELISA enzyme-linked immunosorbent assay havebeen used respectively to detect the changes of pain medium,inflammatorymediators,and cytokine levels in synovial tissue. Through the detection ofpathology, immunohistochemistry, in situ hybridization and other techniquesto explore the pathological changes and the changes of associated protein andgene expression to evaluate their anti-inflammatory and analgesic effects.Statistical analysis has been done based on the these results.Results(1) Group of electroacupuncture in Shu and dense wave can significantlyreduce the degree of ankle swelling(P<0.05, P<0.01), which shows thatelectroacupuncture has obvious anti-inflammatory and repercussive roles;(2) Theresults of HE staining showed that group of electroacupuncture in Shuand dense wave can significantly reduce inflammatory cell infiltration,inhibit synovial cell proliferation, and decrease proliferation of granulationtissue in rats joints; (3) group of electroacupuncture in dense-wave can increase the pain thresholdof experimental rats (P<0.05, P<0.01), reduce the content of peripheral painmediums, such as K~+, DA and 5-HT(P<0.05, P<0.01), which shows thatelectroacupuncture has significant analgesic effect;(4) In the local lesion of joints, electroacupuncture can significantly reducethe content of IL-1β, IL-8, TNF-a, increase IL-4 content, decrease theexpression of COX-2mRNA, and significantly promote the expression ofHSP-70 protein in synovial tissues of acute gouty arthritis rats (P<0.05,P<0.01),thus restrain the occurrence of synovial tissue inflammation.Conclusion Electroacupuncture on "Zusanli" and "Sanyinjiao" point hasobvious anti-inflammatory and analgesic effects.(1) Electroacupuncture of different waveforms can significantly inhibitinflammatory cell infiltration and synovial cell proliferation, and reduceankle swelling in rats of acute gouty arthritis, and its anti-inflammatory andrepercussive roles are best in group of electroacupuncture in Shu and densewave.(2) Electroacupuncture of different waveforms can improve pain thresholdand reduce the content of peripheral pain medium,such as K +, DA and 5-HT,and its analgesic effect is best in group of electroacupuncture in dense-wave.(3) Electroacupuncture of different waveforms can significantly reduce thecontent of IL-1β, IL-8 and TNF-a, increase IL-4 content, reduce the localinflammatory response in synovial tissues of ankle in rats, and group ofelectroacupuncture in Shu and dense wave is the best.(4) Electroacupuncture of different waveforms can effectively inhibitexpression of COX-2mRNA, increase HSP-70 protein expression, so as toreduce the local inflammatory response of ankle joint and reduce jointdamage in synovial tissues of acute gouty arthritis rats, and group ofelectroacupuncture in Shu and dense wave is the best.

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