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背景:通痹灵是由《金匮要略》中桂枝芍药知母汤加减而成,运用于临床治疗类风湿关节炎已十多年,拟从白细胞介素2及白细胞介素2受体α链(CD25)的角度,从体内和体外两个方面,阐明其调控细胞免疫以及抗类风湿关节炎的作用机制。目的:从体内体外两个方面,研究通痹灵对于胶原诱导性关节炎大鼠关节滑膜原代细胞分泌白细胞介素2,以及通痹灵总碱对于白细胞介素2受体的α链CD25表达的影响。设计:以实验动物为研究对象的完全随机设计,对照实验研究。单位:一所大学的金匮教研室,及另一所大学的生命科学院组织移植与免疫中心。材料:实验于2001-04/07在广州中医药大学第一附属医院中心实验室完成。清洁级Wistar大鼠36只,雄性,购自中山大学医学院实验动物中心,动物合格证号2000A050。体质量(110g±20)g。分为正常组,造模组,甲氨喋呤高剂量组,甲氨喋呤低剂量组,复方通痹灵高剂量组,复方通痹灵低剂量组,每组6只。方法:体内实验,采用胶原诱导性关节炎大鼠动物模型,容积法测量足肿胀度,原代滑膜细胞培养,放免法检测培养液上清,观察通痹灵对滑膜内白细胞介素2的影响;体外实验,分离大鼠淋巴细胞,佛波醇酯或刀豆蛋白体外刺激,双色免疫荧光标记,流式细胞仪检测,观察通痹灵总碱对CD3+,CD25+表达的影响。主要观察指标:复方通痹灵?
BACKGROUND: Tongqiaoling is composed of Guizhi Shaoyao Zhimu Decoction in “The Golden Chamber”. It has been used clinically for treatment of rheumatoid arthritis for more than ten years and is intended to be derived from interleukin 2 and interleukin 2 receptor alpha. The perspective of the chain (CD25), both in vivo and in vitro, elucidates the mechanisms that regulate cellular immunity and resistance to rheumatoid arthritis. OBJECTIVE: To study the effect of Tongbiling on the secretion of interleukin 2 from primary synoviocytes of collagen-induced arthritis rats in vivo and in vitro, and Tongzhuling total base for interleukin 2 receptor alpha chain CD25 The effect of expression. Design: Completely randomized design of experimental animals and controlled experimental studies. Units: A university’s Jin-Jin teaching and research section, and another university’s life sciences organization transplantation and immunization center. MATERIALS: The experiment was performed at the Central Laboratory of the First Affiliated Hospital of Guangzhou University of Chinese Medicine from April 2004 to July 2007. Thirty-six clean Wistar rats, male, were purchased from the Laboratory Animal Center of Sun Yat-Sen University School of Medicine, Animal Certificate No. 2000A050. Body weight (110g ± 20) g. They were divided into normal group, model group, high-dose methotrexate group, low-dose methotrexate group, high-dose Tongfuling group, and low-dose Tongfuling group, with 6 rats in each group. METHODS: In vivo experiments using rat model of collagen-induced arthritis, volumetric method for measuring foot swelling, primary synovial cell culture, radioimmunoassay for supernatant detection, and observation of Tongluling for intra-synovial interleukin-2 In vitro experiments, isolated rat lymphocytes, phorbol esters or concanavalin in vitro stimulation, two-color immunofluorescence labeling, flow cytometry, to observe the effect of Tongqiaoling total base on CD3 +, CD25 + expression. MAIN OUTCOME MEASURES: Compound Tongweiling?