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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
- 陆军军医大学学报 2016年38卷第3期 页码：307-313
- 作者机构：
- 作者简介：
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DOI：
中图分类号： R322.21;R289.5;R684.305-
纸质出版：2016
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乾灿,王勇,匡梅,吴红,王恒.羌活二黄汤对大鼠佐剂性关节炎的疗效及其机制[J].陆军军医大学学报(原第三军医大学学报),2016,38(03):307-313.
Qian Can,Wang Yong,Kuang Mei,Wu Hong,Wang Heng.Efficacy and mechanism of Qianghuo Erhuang Decoction in treatment of adjuvant-induced arthritis in rats[J].J Amry Med Univ (J Third Mil Med Univ),2016,38(03):307-313.
乾灿,王勇,匡梅,吴红,王恒.羌活二黄汤对大鼠佐剂性关节炎的疗效及其机制[J].陆军军医大学学报(原第三军医大学学报),2016,38(03):307-313. DOI：
Qian Can,Wang Yong,Kuang Mei,Wu Hong,Wang Heng.Efficacy and mechanism of Qianghuo Erhuang Decoction in treatment of adjuvant-induced arthritis in rats[J].J Amry Med Univ (J Third Mil Med Univ),2016,38(03):307-313. DOI：
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），P0.01；24 d（3.00±1.33)，28 d（0.67±0.67），P0.01
]
，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)，P0.01；24 h（0.26±0.03)，48 h（0.25±0.03），72 h（0.27±0.01），P0.01
]
，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）%，P0.01
]
，而Th17百分比明显增高[（1.84±0.45）%，P0.01
]
，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）%，P0.01
]
，Th17百分比下降[（0.98±0.35）%，P0.05
]
。与模型组相比，中药中、低剂量组无明显治疗效应（P0.05）。 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。
Abstract
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 ra
ts 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
P0.01) and high-dose QED group (24 d: 3.00±1.33
28 d: 0.67±0.67
P0.01) 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
P0.01; the high-dose QED group 24 h: 0.26±0.03
48 h: 0.25±0.03
72 h: 0.27±0.01
P0.01) 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)%
P0.01
]
was decreased
while that of Th17 cells [(1.84±0.45)%
P0.01
]
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)%
P0.01
]
and an decrease of percentage of Th17 cells [(0.98±0.35)%
P0.05
]
. The moderate- and low-dose QED had no therapeutic effect when compared with the AA group (P0.05). Conclusion High dose of QED (18 g/kg) exerts certain therapeutic effect for AA
which might be due to its correcting the imbalance of CD4
+
T lymphocytes subsets
Treg/Th17.
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