杜仲提取物对类风湿关节炎大鼠免疫功能及TLR2/NF-κB/TNF信号通路的影响

The Mechanism of Eucommia Extract on Immune Function Indexes and TLR2/NF-kB/TNF Signaling Pathway in Rheumatoid Arthritis Rats

目的观察杜仲提取物对类风湿关节炎大鼠免疫功能及TLR2/NF-κB/TNF信号通路的影响。方法将35只SD大鼠分为正常对照组、模型组、杜仲低剂量组(杜仲提取物200 mg/kg)、氨甲蝶呤组(氨甲蝶呤0.75 mg/kg)、杜仲高剂量组(杜仲提取物800 mg/kg),每组7只,连续给药3周。采用冷水游泳+足跖部皮内注射CFAO制备类风湿关节炎模型。记录各组大鼠关节肿胀程度及关节指数评分,ELISA检测血清免疫球蛋白水平;流式细胞仪检测T淋巴细胞水平;HE染色观察关节病变;Western blotting检测大鼠踝关节滑膜组织中Toll样受体2(TLR2)、核转录因子-κB(NF-κB)、肿瘤坏死因子-α(TNF-α)蛋白表达水平。结果与正常对照组相比,模型组大鼠左后足肿胀度、关节指数评分及免疫球蛋白水平、TLR2、NF-κB、TNF-α升高,CD4+、CD4+CD25+Treg水平降低(P<0.05),与模型组相比,杜仲低剂量组、氨甲蝶呤组、杜仲高剂量组大鼠左后足肿胀度、关节指数评分及免疫球蛋白水平、TLR2、NF-κB、TNF-α降低,CD4+、CD4+CD25+Treg水平升高(P<0.05),杜仲低剂量组与氨甲蝶呤组差异无统计学意义(P>0.05),与氨甲蝶呤组相比,左后足肿胀度、关节指数评分及免疫球蛋白水平、TLR2、NF-κB、TNF-α降低,CD4+、CD4+CD25+Treg水平升高(P<0.05)。结论杜仲提取物可通过调节大鼠血清免疫球蛋白及T淋巴细胞水平,抑制类风湿TLR2、NF-κB信号通路,降低炎性水平。

Objective:To investigate the effects of eucommia extract on immune function and TLR2/NF-kB/TNF signaling pathway in rats with rheumatoid arthritis.Methods:A total of 35 SD rats were divided into the control group(CG),rheumatoid arthritis rat model group(MG),model rats treated with eucommia extract low-dose(EELD)group,model rats treated with methotrexate intervention(EEMD)group,and model rats were treated with eucommia extract high-dose(EEHD)group,with 7 rats in each group.The degree of joint swelling and joint index score were recorded.Serum immunoglobulin levels were detected by ELISA.The level of T lymphocytes was detected by flow cytometry.HE staining was used to observe joint lesions.The protein expressions of TLR2,NF-kB and TNF in the synovial tissue of ankle joint were detected by Western blotting.Results:Compared with the CG group,the left hind foot swelling degree,joint index score,immunoglobulin water,TLR2,NF-kB,TNF-a and CD4+and CD4+CD25+Treg levels of the MG group were increased,and the levels of CD4+and CD4+CD25+Treg were decreased(P<0.05).The left hind foot swelling degree,joint index score,immunoglobulin water,TLR2,NF-kB and TNF-a of rats in the EELD,EEMD and EEHD groups were decreased,while CD4+and CD4+CD25+Treg levels were increased(P<0.05).There was no significant difference between the EELD and EEMD groups(P>0.05).Compared with the EEMD group,the left hind foot swelling degree,joint index score,immunoglobulin water,TLR2,NF-kB,TNF-a were decreased,and CD4+and CD4+CD25+Treg levels were increased(P<0.05).Conclusion:Eucommia extract may play a protective role by regulating the levels of serum immunoglobulin and T lymphocytes,inhibiting the TLR2 and NF-kB signaling pathways in rheumatoid arthritis,reducing the inflammatory level and improving rheumatoid arthritis.