火针对类风湿性关节炎大鼠踝关节JNK、p38丝裂原活化蛋白激酶的影响

Effect of Fire-needle Intervention on JNK and p38MAPK in Ankle-joint of Rheumatoid Arthritis Rats

目的观察火针对类风湿性关节炎（RA）模型大鼠踝关节c-junN末端激酶（c-jun N-terminal kinease,JNK）和p38丝裂原活化蛋白激酶（p38MAPK）表达的影响,探讨火针治疗RA的作用机制。方法采用雄性SD大鼠40只,随机分为正常组、模型组、火针组、药物（甲氨蝶呤,MTX）组,每组各10只。模型组、火针组、药物组大鼠分别给予建立佐剂性关节炎模型,正常组不造模。火针组点刺腰部夹脊穴和阿是穴,共治疗2次。分别于实验第1、2、4、6、8天检测大鼠右后足足跖肿胀度。实验第9天取血测大鼠血清TNF-α和IL-1β,取踝关节滑膜组织做关节组织病理学切片观察及局部滑膜组织蛋白JNK、p38的免疫组化,Western blot法测定局部滑膜组织JNK、p38的蛋白表达的影响。结果实验第2天,模型组、火针组、药物组足跖肿胀度均与正常组有明显差异（P〈0.01）,实验第8天,火针组的足跖肿胀度明显低于模型组（P〈0.05）;模型组大鼠血清TNF-α和IL-1β（P〈0.05）含量高于正常组,滑膜细胞增生及炎性细胞浸润明显增多,而火针组、药物组较模型组血清TNF-α和IL-1β（P〈0.05）含量有降低,滑膜细胞增生及炎性细胞浸润减少;与正常组比较,模型组大鼠局部滑膜组织JNK、p38的表达均有所升高（P〈0.05-0.01）,与模型组比较,火针组、药物组JNK水平均表现为明显下降（P〈0.05）,这种改变在相应的免疫组化镜下图片中均有所体现。结论火针治疗可以有效地调节丝裂原活化蛋白激酶（MAPK）信号系统中JNK、p38蛋白活性的表达,降低MAPK活性,这可能是火针治疗RA起效的作用机制之一。

OBJECTIVE To observe the influence of fire-needle therapy on JNK and p38 MAPK in ankle-joint of rheumatoid arthritis（RA）rats to explore its underlying mechanism in RA treatment.METHODS 40 male SD rats were randomly divided into normal group,model group,fire-needle group and medication（methotrexate,MTX）group,with 10 cases in each group.Fire-needle was applied to Jiaji（EX-B2）and Ashi point every other day,twice altogether.The right hindpaw swelling volume of rats were detected on the 1st,2nd,4th,6th and 8th day,respectively.The pathological changes in synovial tissue of the right ankle joint were observed by optical microscope.The expression of JNK and p38 was detected by immunohistochemical method and Western blot.RESULTS In comparison with the control group,the swelling paw volumes at the 2nd day significantly increased of the rest 3groups（P〈0.01）.Compared to the model group,the swelling paw volumes on the 8th day in fire-needle group decreased significantly（P〈0.05）.Serum TNF-αand IL-1β（P〈0.05）contents increased in model group comparing with normal group,while decreased in fire-needle group and medication group comparing with model group.The pathological states of synovial tissue of the right ankle joint in both fire-needle and medication groups were relatively lighter than those of the model group.In comparison with normal group,the expression of JNK and p38 in local synovial tissue of the right ankle joint increased significantly in model group（P〈0.05-0.01）.Compared with model group,the expression of JNK and p38 of fire-needle and medication group both decreased significantly（P〈0.05）.These changes were also found in photographs made by immunohistochemical method.CONCLUSION Fire-needle therapy can dowm-regulate the expression of JNK and p38 in local synovial tissue of the right ankle join in MAPK signal transduction pathways,which is the possible mechanism of fireneedle therapy for RA treatment.