Shen Hong-Bo, Huo Ze-Jun, Bai Yun-Jing, He Xiao-Juan, Li Chang-Hong, Zhao Yu-Kun, Guo Qing-Qing
Department of Traditional Chinese Medicine, Peking University Third Hospital, Beijing, 100191, China.
Division of Rheumatology, PLA Army General Hospital, Beijing, 100700, China.
Chin J Integr Med. 2018 Apr;24(4):278-283. doi: 10.1007/s11655-017-2792-2. Epub 2017 Dec 21.
To observe the effect of norcantharidin (NCTD) on collagen-induced arthritis (CIA) rats.
Sixty Sprague-Dawley(SD) rats were randomly divided into 6 groups (n=10): normal group, CIA model group(model group), NCTD low-dose group [1.35 mg/(kg•d)], NCTD middle-dose group [2.7 mg/(kg•d)], NCTD high-dose group [5.4 mg/(kg•d)] and methotrexate (MTX) group [1.8 mg/(kg/w)]. Anesthetized rats were sacrificed by luxation of cervical vertebra after 4 weeks of administration. The arthritis scores were evaluated twice a week. The pathological changes in the ankle joints of rats were observed by hematoxylin-eosin (H&E) staining. The serum levels of interleukin (IL) 1β, IL-6, tumor necrosis factor (TNF)-α, vascular endothelial growth factor (VEGF), IL-17 and transform growth factor (TGF) β were detected by enzyme linked immunosorbent assay (ELISA). The mRNA expression of retinoid-related orphan nuclear receptorγt (RORγt) and forkhead box P3 (Foxp3) in peripheral blood lymphocytes were confirmed by real-time polymerase chain reaction.
MTX and high-dose NCTD not only decreased the arthritis scores but also alleviated the pathological changes in CIA rats' ankle joints compared with the model group (P<0.05 or P<0.01). All doses of NCTD significantly inhibited the serum levels of IL-6, IL-17 and TNF-α in CIA rats (P<0.05). Only middle- and high-dose of NCTD prominently decreased serum IL-1β and TGF-β levels of CIA rats (P<0.05). However, NCTD has no effect on vascular endothelial growth factor (VEGF) level in CIA rats. The Foxp3 mRNA expression in all NCTD groups were increased significantly than in the model group (P<0.05). The mRNA expression of RORγt in NCTD high-dose group was decreased apparently in comparison with the model group (P<0.05).
NCTD showed therapeutic effect on CIA rats by inhibition of cytokines and regulation of Th17/Treg cells.
观察去甲斑蝥素（NCTD）对胶原诱导性关节炎（CIA）大鼠的影响。
将60只Sprague-Dawley（SD）大鼠随机分为6组（n = 10）：正常组、CIA模型组（模型组）、NCTD低剂量组[1.35毫克/（千克•天）]、NCTD中剂量组[2.7毫克/（千克•天）]、NCTD高剂量组[5.4毫克/（千克•天）]和甲氨蝶呤（MTX）组[1.8毫克/（千克/周）]。给药4周后，将麻醉的大鼠通过颈椎脱臼处死。每周评估两次关节炎评分。通过苏木精-伊红（H&E）染色观察大鼠踝关节的病理变化。采用酶联免疫吸附测定（ELISA）检测血清白细胞介素（IL）-1β、IL-6、肿瘤坏死因子（TNF）-α、血管内皮生长因子（VEGF）、IL-17和转化生长因子（TGF）β水平。通过实时聚合酶链反应确认外周血淋巴细胞中类视黄醇相关孤儿核受体γt（RORγt）和叉头框蛋白P3（Foxp3）的mRNA表达。
与模型组相比，MTX和高剂量NCTD不仅降低了关节炎评分，还减轻了CIA大鼠踝关节的病理变化（P < 0.05或P < 0.01）。所有剂量的NCTD均显著抑制CIA大鼠血清中IL-6、IL-17和TNF-α水平（P < 0.05）。只有中、高剂量的NCTD显著降低CIA大鼠血清IL-1β和TGF-β水平（P < 0.05）。然而，NCTD对CIA大鼠血管内皮生长因子（VEGF）水平无影响。所有NCTD组的Foxp3 mRNA表达均比模型组显著增加（P < 0.05）。与模型组相比，NCTD高剂量组的RORγt mRNA表达明显降低（P < 0.05）。
NCTD通过抑制细胞因子和调节Th17/Treg细胞对CIA大鼠显示出治疗作用。
