【摘要】 背景:近年来,类风湿性关节炎(rheumatoidarthritis,RA)发病机制及病理变化研究等方面取得了明显进展,但在RA治疗学方面中医药具有独特的优势。目的:研究通痹灵的有效部位总生物碱对细胞增殖及T淋巴细胞转铁蛋白受体(CD71)的影响,探讨通痹灵调节细胞免疫的作用机制。设计:以细胞为研究对象,完全随机分组设计,探索性研究。单位:一所中医药大学医院六内科,一所大学生命科学院组织移植与免疫中心。材料:本实验于2002-07/2003-08在广州中医药大学第一附属医院中心实验室(国家中医药管理局三级实验室)完成。实验选用清洁级雄性SD大鼠10只。方法:分离大鼠腹股沟淋巴结淋巴细胞进行培养,刀豆蛋白刺激72h,MTT法检测通痹灵总碱对淋巴细胞增殖的影响。佛波醇酯或刀豆蛋白刺激48h,流式细胞术检测T淋巴细胞CD71的表达率。主要观察指标:通痹灵总碱对淋巴细胞增殖及T细胞活化的影响。结果:不同浓度的通痹灵总碱可明显抑制刀豆蛋白刺激下的淋巴细胞增殖;佛波醇酯和刀豆蛋白激活的T淋巴细胞CD71+表达率明显高于空白对照组(P<0.01),刀豆蛋白刺激下不同浓度的通痹灵总碱(50,100,200mg/L)CD3+CD71+表达率犤(62.03±1.51)%,25.28(±1.57)%,20.29±1.72)%犦明显低于BPS阳性对照组犤(62.03(±1.28)%犦(P<0.05);佛波酯醇刺激下10更多还原

【Abstract】 BACKGROUND:Researches on the pathogenesis and pathological changes of rheumatoid arthritis(RA) have achieved significant progress in recent years. But traditional Chinese medicine(TCM) has unique advantage in RA therapy.OBJECTIVE:To study effects of overall alkali in tongbiling(TBL) on the proliferation of lymphocytes and the transferrin receptor of T lymphocytes(CD71) to explore the mechanism of TBL on the modulation of cell immunity. DESIGN:A completely randomized grouping design and an explorative study by employing cells as subjects.SETTING:Sixth internal medicine department of a TCM university,center of tissue transplantation and immunology college of life science of a university PARTICIPANTS:The study was conducted in the central laboratory (tertiary laboratory of National TCM Administrator) of the first affiliated hospital of Guangzhou TCM medical university between July 2002 and August 2003.Ten clean male SD rats were selected. METHODS:Lymphocyte was separated from rat inguinal lymph node for culture.Concanavalin(ConA) was used for 72 hour stimulation. The impacts of overall alkali TBL on lymphocyte proliferation were tested by MTT. The expression of T lymphocyte CD71 was tested by flow cytometer after 48 hour stimulation of phorbol 12,13- dibutyrate(PDB) or ConA. MAIN OUTCOME MEASURES:The impacts of overall alkali TBL on lymphocyte proliferation and T cell activation.RESULTS:Different concentration of overall alkali TBL could significantly inhibit the proliferation of lymphocytes under ConA stimulation.PDB and ConA activated T lymphocyte CD71+ expressions were significantly higher than that of blank control group(P< 0.01). CD3+ CD71+ expressions [(62.03± 1.51)% ,(25.28± 1.57)% ,(20.29± 1.72)% ]activated by ConA under different concentration of overall alkali TBL(50,100,200 mg/L) were significantly lower than(72.03± 1.28)% of BPS positive control group(P< 0.05). CD3+ CD71+ expressions activated by PDB under 100 mg/L and 200 mg/L of overall alkali TBL were significantly lower than that of phosphate buffer solution (PBS )positive control group(P< 0.05). Different concentration of overall alkali TBL had significant down regulated effects on CD71 expression in T lymphocyte activated by PDB or ConA and there was also a significant dose effect relationship(P< 0.05).The inhibition on ConA activated CD71 expression was stronger than that of PDB. CONCLUSION:Overall alkali TBL can inhibit the abnormal proliferation of T lymphocyte and its mechanism might be realized through its inhibition on transferrin receptor.更多还原