芍甘附子汤加味对CIA寒证大鼠Egr2/Egr3及其信号通路表达的影响

Effects of Shaogan Fuzi Decoction on expression of Egr2/Egr3 and the signal pathway in collagen-induced arthritis rats with syndrome of cold

目的:研究芍甘附子汤加味对胶原诱导性关节炎(CIA)寒证大鼠Egr2/Egr3及其信号通路表达的影响,并探讨Egr2/Egr3在类风湿关节炎(RA)发病中的作用。方法:110只雌性Wistar大鼠被随机分为7组:正常组、模型组、对照组[甲氨蝶呤(MTX)组、雷公藤组]和芍甘附子汤加味组(大、中、小剂量组)。正常组常规饲养,其余各组在复制CIA寒证大鼠模型后分别MTX、雷公藤多苷片及芍甘附子汤加味(大、中、小剂量)灌胃,干预4周,每周评价各组大鼠炎症指数。4周后测定ELISA法检测大鼠外周血IL-6、TGF-β表达;RT-q PCR检测大鼠膝关节滑膜Egr2/Egr3 m RNA、SOCS1/SOCS3 m RNA、STAT1/STAT3 m RNA表达。实验结束后测定大鼠脏器指数。结果:各组大鼠AI评分均呈下降趋势,但各组间差异无统计学意义。HE染色提示各给药组及模型组存在不同程度的滑膜增生、炎性细胞浸润以及骨侵蚀等,各给药组较模型组均有所减轻,模型组和各给药组脾脏指数显著高于正常组(P<0.05),大、中剂量组胸腺指数较模型组显著下降(P<0.05)。大鼠滑膜组织模型组和各治疗组Egr2/Egr3 m RNA、SOCS1/SOCS3 m RNA相对表达量较正常组升高,其中芍甘附子汤中剂量组Egr2/Egr3 m RNA表现尤为突出(P<0.05)。CIA造模后大鼠外周血TGF-β、IL-6水平均显著高于正常组水平,各药物干预组治疗4周后较模型组均有下降尤以TGF-β更为明显。结论:Egr2/Egr3参与了RA发病过程的调控环节,对于疾病的早期诊断及早期干预有提示作用。

Objective： To study the effect of Shaogan Fuzi Decoction on the expression of Egr2/Egr3 and its signal pathway in collagen-induced arthritis（CIA） rats and to explore the role of Egr2/Egr3 in the pathogenesis of rheumatoid arthritis. Methods： 110 female Wistar rats were randomly divided into 7 groups： normal group, model group, control group（methotrexate（MTX） group, Tripterygium wilfordii group） and Shaogan Fuzi Decoction group（ high, medium, low dose）. The rats in the control group were treated with MTX, Tripterygium wilfordii tablets. Shaogan Fuzi Decoction group were treated with high, medium and small dose Shaogan Fuzi Decoction, and the rats in each group were treated rats for 4 weeks. Inflammation index of rats in each group was evaluated weekly. After 4 weeks, the expression of IL-6 and TGF-beta in peripheral blood of rats. were detected by ELISA, the expression of Egr2/Egr3 mRNA, SOCS1/SOCS3 mRNA and STAT1/STAT3 mRNA in rat synovium were detected by RT-qPCR. At the end of the experiment, the rat organ index was measured. Results： The scores of AI were decreased in each group, but there was no significant difference between the two groups. HE staining indicated that there were different levels of synovial hyperplasia, inflammatory cell infiltration and bone erosion both in the treated group and the model group, but compared to model group, the inflammation of each treated group were relieved. The spleen index of model group and each treated group was significantly higher than that of normal group, while there was no significant difference among each treated group and model group. The expression of Egr2/Egr3 mRNA and SOCS1/SOCS3 mRNA in the synovial tissue of model group and treated group were higher than those in the normal group. The expression of Egr2/Egr3 mRNA in the middle dose group was significantly higher than that in the control group（P〈0.05）. The levels of TGF-β and IL-6 in the peripheral blood of rats in model group were significantly higher than those of the normal group, and the levels of TGF-β and IL-6 in the treated group were significantly decreased after 4 weeks of treatment. Conclusion： Egr2/Egr3 is involved in the regulation of RA and plays an important role in the early diagnosis and early intervention of disease.