盐酸青藤碱治疗性给药对胶原诱导性关节炎小鼠的保护作用

Study on the Protective Effect of Therapeutic Administration of Sinomenine Hydrochloride on Experimental CIA Mice

目的探讨盐酸青藤碱治疗性给药对胶原诱导性关节炎(CIA)小鼠的保护作用。方法采用尾根部注射鸡II型胶原建立C57BL/6小鼠关节炎模型。于第2次免疫注射后第7天,将造模成功小鼠随机分为模型组、盐酸青藤碱组(100 mg·kg^(-1))、地塞米松组(2 mg·kg^(-1)),同时设置正常组。给药组小鼠分别腹腔注射相应治疗药物,每天1次,连续治疗33 d,正常组和模型组小鼠腹腔注射等体积无菌生理盐水。给药周期内,每3 d进行1次小鼠足趾关节炎指数评分;采用ELISA法检测外周血中TNF-α、IL-6炎症因子水平;采用HE染色法观察关节滑膜炎性细胞浸润情况;采用Micro-CT扫描分析骨质破坏情况。结果与正常组相比,模型组小鼠足趾关节炎指数评分显著升高,血清中TNF-α和IL-6炎症因子水平显著升高(P<0.001);关节滑膜部位炎性细胞浸润明显增多、骨质破坏明显,膝、踝关节病理总评分均显著升高(P<0.001)。与模型组相比,盐酸青藤碱干预治疗后,能显著降低足趾关节炎指数评分及血清中TNF-α、IL-6炎症因子水平(P<0.05,P<0.01),并能阻止关节滑膜部位炎性细胞浸润,减轻骨质破坏,降低膝、踝关节病理总评分(P<0.01)。结论盐酸青藤碱治疗性给药可阻止CIA小鼠的关节骨质破坏,其作用机制与减轻关节滑膜炎症有关。

Objective To investigate the protective effect of sinomenine hydrochloride on collagen induced arthritis in mice. Methods C57 BL/6 mouse model of arthritis was established by injecting chicken type II collagen into the base of tail. On the seventh day after the second immune injection,the successful modeled RA mice were randomly divided into model group,sinomenine hydrochloride group(100 mg·kg(-1)),and dexamethasone group(2 mg·kg(-1)). At the same time, the mice without any treatment were taken as normal control group. Mice in the sinomenine hydrochloride group and dexamethasone group were intraperitoneally injected with sinomenine hydrochloride and dexamethasone, respectively. Mice in normal group and model group were intraperitoneally injected with equal volume of sterile normal saline. The treatment was carried out once a day consecutively for 33 days. During the treatment period, the arthritis indexes of mice were scored every 3 days, the levels of TNF-α and IL-6 in peripheral blood were detected by ELISA, HE staining was used to observe the synovial inflammatory cell infiltration,and Mcro-CT scanning was used to analyze the bone destruction. Results Compared with the normal group, toe arthritis index score of the model group mice significantly increased, serum levels of inflammatory factors, TNF-α and IL-6, were significantly increased(P<0.001);the inflammatory cell infiltration at joint synovial site was obvious increased, with obvious bone destruction, and the total score of knee and ankle joint pathology was significantly increased(P<0.001). Compared with the model group, sinomenine hydrochloride intervention treatment significantly reduced toe arthritis index score(P<0.05) and serum TNF-α and IL-6 inflammatory factor expression levels(P<0.05,P<0.01),prevented inflammatory cells infiltrating the synovium of the joints,alleviated bone destruction,and reduced the total pathological score of the knee and ankle joint(P<0.01). Conclusion Sinomenine hydrochloride can prevent the destruction of joint bones of CIA mice, and its mechanism may be related to the reduction of synovial inflammation.