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目的探讨雷公藤内酯醇(Tri)对实验性自身免疫性脑脊髓炎(EAE)小鼠脊髓中基质金属蛋白酶9(MMP-9)表达的影响。方法雌性C57BL/6小鼠随机分为EAE组(n=28)、Tri治疗组(n=20)和佐剂组(n=20)。EAE组和Tri治疗组用髓鞘少突胶质细胞糖蛋白多肽(MOG35～55)免疫建立EAE模型,佐剂组以等量生理盐水代替。Tri治疗组于免疫后d5开始按100μg·kg-1·d-1腹腔注射Tri。观察3组小鼠发病情况,采用5分制评分法进行症状评分。发病后18～20d取脑和脊髓行HE染色观察病理改变。RT-PCR和Westernblot检测3组脊髓中MMP-9的mRNA及蛋白表达。结果EAE平均最高症状评分为(2.5±s0.8)分,较Tri治疗组(1.2±0.3)有显著差异(P<0.01)。HE染色观察EAE小鼠脊髓的灰、白质均有明显的炎性细胞浸润。与佐剂组相比,EAE组小鼠脊髓中MMP-9的mRNA及蛋白表达与激活明显增多(P<0.01),且MMP-9的激活水平与症状评分呈显著正相关(r=0.827,P=0.001);Tri治疗组脊髓中MMP-9的mRNA表达和蛋白激活较EAE组减少,差异显著(P<0.01)。结论Tri可治疗EAE,其作用机制之一可能是下调MMP-9的表达、减少MMP-9的激活。
Objective To investigate the effect of triptolide on the expression of matrix metalloproteinase 9 (MMP-9) in the spinal cord of experimental autoimmune encephalomyelitis (EAE) mice. Methods Female C57BL / 6 mice were randomly divided into EAE group (n = 28), Tri treatment group (n = 20) and adjuvant group (n = 20). EAE group and Tri treatment group with myelin oligodendrocyte glycoprotein (MOG35 ~ 55) immune model established EAE, adjuvant group with an equal volume of saline instead. Tri group at the d5 after immunization by 100μg · kg-1 · d-1 intraperitoneal injection of Tri. The incidence of the mice in the three groups was observed, and the symptom scores were scored by 5-point scale. The brain and spinal cord were taken from 18 to 20 days after onset to observe the pathological changes by HE staining. The mRNA and protein expression of MMP-9 in three groups of spinal cord were detected by RT-PCR and Western blot. Results The mean symptom score of EAE was (2.5 ± s 0.8) points, which was significantly lower than that of Tri treatment group (1.2 ± 0.3) (P <0.01). Hematoxylin and eosin (HE) staining showed obvious inflammatory cell infiltration in the gray and white matter of spinal cord of EAE mice. Compared with the adjuvant group, the mRNA and protein expression and activation of MMP-9 in EAE group were significantly increased (P <0.01), and the activation level of MMP-9 was positively correlated with symptom score (r = 0.827, P = 0.001). The mRNA and protein expressions of MMP-9 in the spinal cord of Tri-treated group decreased significantly compared with EAE group (P <0.01). Conclusion Tri can treat EAE. One of its mechanisms may be down-regulating the expression of MMP-9 and decreasing the activation of MMP-9.