益肾达络饮对EAE小鼠p-JNK1/2,JNK1/2,COX-2的影响

Effect of Yishen Daluo Decoction on p-JNK1/2,JNK1/2 and COX-2 in EAE Mice

目的：研究益肾达络饮对实验性自身免疫性脑脊髓炎（EAE）小鼠磷酸化c-Jun氨基末端激酶1/2（p-JNK1/2）,c-Jun氨基末端激酶1/2（JNK1/2）及环氧合酶-2（COX-2）的影响。方法：将48只雌性SJL小鼠（8-12周龄）随机分为4组,即正常组、模型组、益肾达络饮组和泼尼松组,每组各12只。在模型组、益肾达络饮组和泼尼松组每只小鼠上腹部,分两点ih抗原乳剂200μL[含髓鞘蛋白脂质蛋白（PLP）139-151150μg,H37 Ra 400μg],记为第1天;分别在第1天和第3天给予每只小鼠尾静脉注射百日咳杆菌液100μL（含百日咳杆菌0.6×106个）。造模的第7天给药,正常组和模型组给予生理盐水10 m L·kg-1ig,益肾达络饮组给予益肾达络饮生药量20 g·kg-1ig,泼尼松组给予泼尼松药液3.9 mg·kg-1ig,共干预15 d。免疫后每天记录各组小鼠的神经功能评分;Western blot法测定EAE小鼠脑组织内p-JNK1/2,JNK1/2,COX-2的表达。结果：与正常组比较,模型组EAE小鼠的神经功能评分明显升高（P〈0.01）,p-JNK1,p-JNK2和COX-2的表达水平明显升高（P〈0.05）;与模型组比较,益肾达络饮组和泼尼松组明显降低EAE小鼠的神经功能评分（P〈0.05）,明显降低p-JNK1,p-JNK2和COX-2蛋白表达的水平（P〈0.05,P〈0.01）;泼尼松组COX-2的表达水平低于益肾达络饮组（P〈0.05）。结论：JNK1/2通路和COX-2与EAE的发病有关,益肾达络饮可通过降低p-JNK2和COX-2的表达降低EAE小鼠的神经功能评分,但其具体的机制需进一步探讨。

Objective： To study the effect of Yishen Daluo decoction on phosphated c-Jun N-terminal kinase 1 and 2（ p-JNK1 /2）,c-Jun N-terminal kinase 1 and 2（ JNK1 /2） and cyclooxygenase 2（ COX-2） in mice with experimental autoimmune encephalomyelitis（ EAE）. Method： Totally 48 female SJL mice（ aged 8-12weeks） were randomly divided into 4 groups,the normal group,the model group,the Chinese medicine（ CM）group and the Western medicine（ WM） group,with 12 mice in each group. Each mouse in model,CM and WM groups was injected subcutaneously with 200 μL antigen emulsion [including 150 μg proteolipid protein（ PLP）139-151 and 400 μg H37 Ra] at two sites of epigastrium on the first day,100 μL Bordetella pertussis liquid（ containing 6 × 105 B. pertussis） was injected to each mouse by caudal vein at the 1stand 3rddays. At the 7thday,10 m L·kg- 1of saline was orally given to each mouse in normal and model groups,20 g·kg- 1of crude Yishen Daluo decoction was orally given to the CM group,and 3. 9 mg·kg- 1of prednisone was orally given to the WM group for15 days. After the immunization,the neurological score for each group was recorded every day. Western blot was used to detect the expressions of p-JNK1 /2,JNK1 /2 and COX-2 in cerebral tissues of EAE mice. Result： The neurologicalscore significantly increased in EAE mice of the model group compared with the model group（ P〈0. 01）. The expressions of p-JNK1 /2 and COX-2 in the model group increased（ P〈0. 01）. Compared with the model group,CM and WM groups showed obvious decreases in the neurological score（ P〈0. 05） and expressions of p-JNK1 /2,JNK1 /2（ P〈0. 05,P〈0. 01）,and the WM group showed a lower COX-2 expression than the CM group（ P〈0. 05）. Conclusion： The JNK1 /2 pathway and COX-2 are correlated with the onset of EAE. Yishen Daluo decoction can reduce the neurological score of EAE by decreasing the expressions p-JNK2 and COX-2. However,further studies shall be conducted for its specific mechanism.