羌活二黄汤对大鼠佐剂性关节炎的疗效及其机制  

Efficacy and mechanism of Qianghuo Erhuang Decoction in treatment of adjuvant-induced arthritis in rats

作　　者：乾灿 王勇[1] 匡梅[2] 吴红[1] 王恒[1] 

机构地区：[1]第三军医大学西南医院中西医结合科,重庆400038 [2]第三军医大学药学系药理学教研室,重庆400038

出　　处：《第三军医大学学报》2016年第3期307-313,共7页Journal of Third Military Medical University

摘　　要：目的观察羌活二黄汤对佐剂型关节炎大鼠的治疗效果,并从Treg/Th17细胞角度探讨其作用机制。方法选取雄性SD大鼠,常规建立佐剂型关节炎(adjuvant arthritis,AA)大鼠模型,造模成功后设正常对照组,模型组,甲氨蝶呤组,中药(羌活二黄汤)高剂量组(18 g/kg)、中剂量组(9 g/kg)及低剂量组(4.5 g/kg)。于关节肿胀第1天开始定时灌胃给药,在4周内连续观察各组大鼠关节炎指数评分的变化,4周后处死大鼠,留取踝关节作常规苏木精-伊红(HE)染色,CCK-8检测脾淋巴细胞增殖,流式细胞术检测大鼠脾淋巴细胞Treg/Th17的百分比。结果与模型组比较,甲氨蝶呤组、中药高剂量组能够降低大鼠关节炎指数评分[24 d(1.33±1.56),28 d(0.17±0.28),P<0.01;24 d(3.00±1.33),28 d(0.67±0.67),P<0.01],明显抑制脾淋巴细胞的增殖[24 h(0.25±0.03),48 h(0.25±0.03),72 h(0.27±0.01),P<0.01;24 h(0.26±0.03),48 h(0.25±0.03),72 h(0.27±0.01),P<0.01],减轻大鼠踝关节滑膜组织炎症。与正常对照组比较,模型组脾淋巴细胞Treg百分比降低[(5.75±0.46)%,P<0.01],而Th17百分比明显增高[(1.84±0.45)%,P<0.01],呈失衡状态。经高剂量中药干预后,大鼠脾淋巴细胞Treg百分比明显上升[(11.19±2.01)%,P<0.01],Th17百分比下降[(0.98±0.35)%,P<0.05]。与模型组相比,中药中、低剂量组无明显治疗效应(P>0.05)。结论18 g/kg羌活二黄汤对大鼠佐剂性关节炎具有一定的治疗效果。其机制可能与纠正CD4+T淋巴细胞亚群Treg/Th17的失衡有关。Objective To observe the efficacy of Qianghuo Erhuang Decoction（ QED,a set prescription of traditional Chinese herbs） in the treatment of adjuvant-induced arthritis in rats and explore the mechanism by its effect on Treg/Th17 cells. Methods Male SD rats were induced to adjuvant arthritis（ AA）model. There were 6 groups of rats in this study,including normal control group（ oral taking of normal saline）,AA group（ normal saline）,AA ＋ methotrexate group（ MTX,0. 4 mg/kg per week）,AA ＋ high-,moderate- and low-dose QED groups（ daily dose： 4. 5,9 and 18 g/kg,respectively）. From the first day of joints swelling,the rats were given with corresponding agents through oral taking or intragastric injection. The changes of arthritis index score were observed during the following 4 consecutive weeks. Hematoxylin-eosin（ HE） staining was used to evaluate joint histological changes. The proliferation of splenic lymphocytes was detected ex vivo by CCK-8 assay,and the percentages of T helper 17（ Th17） and T regulatory（ Treg） cells in the spleen were measured by flow cytometry. Results The arthritis index score was significantly decreased in the MTX group（ 24 d： 1. 33 ± 1. 56,28 d： 0. 17 ± 0. 28,P〈0. 01） and high-dose QED group（ 24 d： 3. 00 ±1. 33,28 d： 0. 67 ± 0. 67,P〈0. 01） than the AA group,and significant inhibited proliferation of splenic lymphocytes（ the MTX group 24 h： 0. 25 ± 0. 03,48 h： 0. 25 ± 0. 03; 72 h： 0. 27 ± 0. 01,P〈0. 01; the high-dose QED group 24 h： 0. 26 ± 0. 03,48 h： 0. 25 ± 0. 03,72 h： 0. 27 ± 0. 01,P〈0. 01） and amelioration of synovial tissue inflammation were observed in the 2 groups. Compared with the normal control,the percentage of Treg cells [（ 5. 75 ± 0. 46） %,P〈0. 01]was decreased,while that of Th17 cells [（ 1. 84 ±0. 45） %,P〈0. 01] was increased,and their ratio was in an imbalanced state. High-dose QED resulted in an increase of percentage of Treg cells [（ 11. 19 ± 2. 01） %,P〈0. 01] and an decr

关 键 词：羌活二黄汤 佐剂型关节炎 脾淋巴细胞 Th17 TREG 

分 类 号：R322.21[医药卫生—人体解剖和组织胚胎学] R289.5[医药卫生—基础医学]