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目的:观察实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)小鼠不同发病时期脑和脊髓组织中CD11b mRNA的表达变化,明确益肾达络饮的干预机制。方法:采用MOG35-55抗原免疫C57BL/6小鼠诱导EAE模型,随机选取分为正常组,佐剂组、模型组、醋酸泼尼松3.9 mg·kg-1组、益肾达络饮20 g·kg-1组,在造模后第7天开始灌胃给药,直到相应时间点进行取材。采用实时荧光定量PCR测定免疫后7 d(发病前期)、14 d(发病初期)、24 d(发病急性期)、40 d(慢性持续期)4个不同时间点EAE小鼠脑和脊髓组织中CD11b mRNA表达的变化。结果:与正常组比较,在免疫后14,24,40d,模型组EAE小鼠大脑中CD11b mRNA表达分别升高至2.39±1.09,3.20±1.14,1.58±0.36(P<0.05)。与模型组比较,在免疫后40 d激素组和益肾达络饮组EAE小鼠大脑中CD11b mRNA的表达水平明显降低(P<0.05);与正常组比较,激素组、益肾达络饮组在免疫后40 d CD11b mRNA在EAE小鼠大脑中表达水平无显著改变;在免疫后14 d激素组EAE小鼠脊髓中CD11b mRNA的表达水平明显降低。结论:单核细胞是实验性自身免疫性脑脊髓炎的潜在调节细胞;中药复方益肾达络饮干预EAE的作用机制与中枢神经系统单核细胞黏附分子CD11b的调节密切相关。
OBJECTIVE: To observe the change of CD11b mRNA expression in brain and spinal cord in different stages of experimental autoimmune encephalomyelitis (EAE) mice and to clarify the mechanism of Yishen Daluo drinking intervention. Methods: C57BL / 6 mice were immunized with MOG35-55 antigen to induce EAE model. The mice were randomly divided into normal group, adjuvant group, model group, prednisone acetate 3.9 mg · kg-1 group, Yishen Daluo drinking 20 g · Kg-1 group, and started gavage administration on the 7th day after model establishment until the corresponding time point. Real-time fluorescence quantitative PCR was used to detect the expression levels of EAE in brain and spinal cord tissue of the EAE mice at 7 d (early onset), 14 d (early onset), 24 d (acute onset) and 40 d (chronic duration) CD11b mRNA expression changes. Results: Compared with the normal group, the expression of CD11b mRNA in the brain of model group EAE mice increased to 2.39 ± 1.09, 3.20 ± 1.14 and 1.58 ± 0.36 (P <0.05) at 14, 24 and 40 days after immunization. Compared with the model group, the expression of CD11b mRNA in the brain of EAE mice on the 40th day after immunization was significantly decreased (P <0.05), while the hormone group and Yishen Daluo Drink The expression of CD11b mRNA in the EAE mice brain did not change significantly at 40 d after immunization. The expression of CD11b mRNA in the spinal cord of EAE mice at 14 d after immunization was significantly decreased. CONCLUSION: Monocyte is a potential regulator of experimental autoimmune encephalomyelitis. The mechanism of Yishen Daluowan Decoction in intervention of EAE is closely related to the regulation of monocyte adhesion molecule CD11b in central nervous system.