黄连解毒汤调控炎性免疫抗类风湿性关节炎机制

Huanglian Jiedutang Regulates Inflammatory Immunity to Relieve Rheumatoid Arthritis

目的:观察黄连解毒汤对胶原诱导性关节炎(CIA)模型小鼠病理改变及免疫损伤的影响,探讨黄连解毒汤抗类风湿性关节炎的可能机制。方法:24只DBA/1小鼠,随机分为正常组、模型组、甲氨喋呤组和黄连解毒汤组,每组6只,采用Ⅱ型胶原诱导建立CIA小鼠模型,给药组给予黄连解毒汤水煎液(5 g·kg^(-1))和甲氨喋呤(0.5 mg·kg^(-1))干预,观察小鼠关节肿胀症状,每3 d进行关节炎指数评分,评分达到峰值后取材;流式细胞仪检测外周血粒细胞、单核细胞、T淋巴细胞;实时荧光定量聚合酶链式反应(Real-time PCR)检测关节炎症因子表达;微计算机断层扫描技术(Micro-CT)扫描踝关节;苏木素-伊红(HE)染色检测关节组织病理改变。结果:与正常组比较,模型组小鼠关节肿胀,关节炎指数评分明显升高(P<0.05);外周血粒细胞比例明显升高(P<0.05),单核细胞、T淋巴细胞比例明显降低(P<0.05),中性粒细胞淋巴细胞比例(NLR)显著升高(P<0.01);关节组织炎症因子肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β、IL-6表达升高(P<0.01);Micro-CT可见踝关节明显骨质破坏,病理可见到关节周围组织存在大量的炎性细胞浸润、滑膜增生。与模型组比较,黄连解毒汤组小鼠关节肿胀症状明显减轻,关节炎指数评分明显降低(P<0.05);外周血T淋巴细胞比例升高,NLR显著降低(P<0.01);关节中炎症因子TNF-α、IL-1β、IL-6的表达显著下降(P<0.01);骨破坏及关节组织病理损伤情况明显改善。结论:Ⅱ型胶原导致CIA小鼠全身和关节局部炎性免疫损伤,黄连解毒汤干预减轻炎性免疫损伤,并且对关节局部的作用更为显著,从而抑制CIA小鼠关节病变和骨破坏。

Objective:To observe the effects of Huanglian Jiedutang on pathological and immune damage in collagen-induced arthritis(CIA)model mice,and to explore the possible mechanism of Huanglian Jiedutang in relieving rheumatoid arthritis.Method:Twenty-four DBA/1 mice were randomly divided into normal group,model group,methotrexate group and Huanglian Jiedutang group,with six mice in each group.The CIA mice model were established using typeⅡcollagen induction.The administration groups were respectively treated with Huanglian Jiedutang(5 g·kg^(-1))and methotrexate(0.5 mg·kg^(-1)).The joint swelling symptoms of the mice were observed,and the arthritis index was scored every 3 days.Flow cytometry was employed to detect granulocytes,monocytes,and T lymphocytes in peripheral blood.The expression of inflammatory cytokines in joint was determined by real-time polymerase chain reaction(Real-time PCR).The ankle joint was scanned by micro-computed tomography(Micro-CT),and the histopathological changes were observed through hematoxylin-eosin(HE)staining.Result:Compared with the normal group,the modeling led to joint swelling,elevated the joint index score(P<0.05),increased the proportion of granulocytes(P<0.05)and decreased that of monocytes and T lymphocytes(P<0.01)in peripheral blood,and raised the neutrophil-tolymphocyte ratio(NLR)(P<0.01).Further,it up-regulated the expression of inflammatory cytokines such as tumor necrosis factor-α(TNF-α),interleukin(IL)-1β,and IL-6 in joint(P<0.01).Micro-CT showed obvious bone destruction in the ankle joint,and pathological examination revealed the infiltration of a large number of inflammatory cells and the synovial hyperplasia of joint tissue.Compared with the model group,Huanglian Jiedutang alleviated the symptoms of joint swelling,lowered the score of arthritis index(P<0.05),increased the proportion of T lymphocytes and lowered NLR(P<0.01).Moreover,it down-regulated the expression levels of TNF-α,IL-1β,and IL-6 in joint(P<0.01)and alleviated the bone destruction and pathological injury of joint tissue.Conclusion:TypeⅡcollagen caused systemic and local inflammatory immune damage in CIA mice.Huanglian Jiedutang alleviates such injury,especially for that in local joint,thereby inhibiting joint injury and bone destruction in CIA mice.