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目的探究马索罗酚(NDGA)对实验性自身免疫性脑脊髓炎(EAE)小鼠血脑屏障的保护作用及其机制。方法将27只C57BL/6小鼠随机分为正常组、EAE组与NDGA组,应用MOG35-55诱导EAE模型。小鼠发病后神经功能评分达1分时开始给予NDGA组NDGA 10 mg(/kg·d)腹腔注射,EAE组及正常对照组给予5%DMSO 10 ml/(kg·d)腹腔注射。给药20 d后比较各组小鼠神经功能评分,应用荧光显微镜观察伊文斯兰染色后各组小鼠血脑屏障通透性改变情况,激光共聚焦显微镜观察各组小鼠claudin-5、occludin、ZO-1表达情况,Real-time PCR检测各组小鼠NF-κB及MMP-9 mRNA水平。结果与正常对照组相比,EAE组伊文斯兰染色后荧光漏出范围较大,紧密连接蛋白连续性遭到破坏,NF-κB、MMP-9 mRNA水平均上调;与EAE组相比,NDGA组神经功能评分显著降低(P<0.05),伊文斯兰染色后荧光范围较小且强度较弱,紧密连接蛋白连续性破坏减轻,NF-κB、MMP-9 mRNA水平下调(P<0.05)。结论马索罗酚可以保护EAE小鼠血脑屏障免遭破坏,延缓疾病进展,其机制可能是保护紧密连接蛋白完整性以及抑制MMP-9、NF-κB表达。
Objective To investigate the protective effect and mechanism of daunorubicin (NDGA) on the blood-brain barrier in experimental autoimmune encephalomyelitis (EAE) mice. Methods Twenty-seven C57BL / 6 mice were randomly divided into normal group, EAE group and NDGA group. MOG35-55 induced EAE model. The mice were injected with NDGA NDGA 10 mg (/ kg · d) intraperitoneally once the score of neurological function reached 1 point. The rats in EAE group and normal control group were given intraperitoneal injection of 5% DMSO 10 ml / (kg · d). After 20 days of administration, neurological scores of mice in each group were compared. The changes of permeability of blood-brain barrier in each group were observed by fluorescence microscopy. The expressions of claudin-5, occludin , ZO-1 expression, Real-time PCR detection of NF-κB and MMP-9 mRNA levels in each group. Results Compared with the normal control group, the fluorescence intensity of Evans blue in EAE group was larger than that in normal control group, the continuity of tight junction protein was destroyed, and the levels of NF-κB and MMP-9 mRNA were up-regulated. Compared with EAE group, The score of neurological function was significantly lower (P <0.05). The fluorescence range of Ivodinsilan staining was weaker and the intensity was weaker. The destruction of tight junction protein and the decrease of NF-κB and MMP-9 mRNA were also observed (P <0.05). Conclusion Marosalol can protect the blood-brain barrier of EAE mice from damage and delay the progression of the disease. The mechanism may be that it protects the integrity of tight junction protein and inhibits the expression of MMP-9 and NF-κB.