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目的:观察大鼠实验性自身免疫性脑脊髓炎(Experimental Autoimmune Encephalomyelitis,EAE)模型脑、脊髓组织转化生长因子-β(transforming growth factor-β,TGF-β)、干扰素-γ(interferon-γ,IFN-γ)、基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)免疫组化和T淋巴细胞亚群的表达,比较二黄方两种剂型(汤剂、胶囊剂)作用差异。方法:用髓鞘碱性蛋白(Myelin Basic Protein,MBP)免疫Lewis大鼠诱导EAE模型,随机分为正常组、模型组、泼尼松组、二黄汤组及二黄胶囊组,分别于免疫后15d(急性期发病)和免疫后27d(缓解期)随机选取各组大鼠,取脑和脊髓组织切片进行IFN-γ,TGF-β,MMP-9,CD4+,CD8+,CD3+,CD19+免疫组化染色。结果:与模型组比,二黄汤(免疫后27d)和二黄胶囊(免疫后15d和27d)显著提高脑组织TGF-β表达;二黄汤和胶囊均能显著降低免疫后15d,27d脑和脊髓组织IFN-γ,MMP-9表达。二黄汤、二黄胶囊组免疫后15d,27d脑和脊髓组织IFN-γ、免疫后15d脑和脊髓组织MMP-9表达降低作用强于泼尼松组。与模型组比,二黄汤组免疫后15d、27d脑组织CD4+表达显著降低、CD8+表达显著升高,CD4+/CD8+显著降低,免疫后15dCD19+表达降低;二黄胶囊组除免疫后15d脑组织CD8+与模型组无显著差异外,其他均同二黄汤组。与模型组比,二黄汤组免疫后15d、27d脊髓组织CD4+表达显著降低、CD8+表达显著升高,CD4+/CD8+显著降低,二黄胶囊组免疫后15d,脊髓组织CD4+表达显著降低、CD8+表达显著升高,CD4+/CD8+显著降低,27dCD4+表达显著降低、CD4+/CD8+显著降低。二黄汤和二黄胶囊组间比较无显著差异。结论:二黄汤、二黄胶囊可上调EAE大鼠中枢神经系统TGF-β表达,作用同泼尼松。两者均可显著降低脑和脊髓组织IFN-γ、MMP-9表达,作用优于泼尼松。二黄汤、胶囊均可降低CD4+表达、升高CD8+表达,改善CD4+/CD8+比值。二黄方对上述指标的影响,可能是其减轻EAE神经损伤,改善患者临床症状的部分机理所在。
Objective: To observe the experimental autoimmune encephalomyelitis (EAE) model in rat brain and spinal cord transforming growth factor-β (TGF-β) and interferon-γ (interferon-γ) , IFN-γ), matrix metalloproteinase-9 (MMP-9) immunohistochemistry and expression of T-lymphocyte subsets, compared the effect of two kinds of formulations (decoction, capsule). METHODS: The Lewis-induced EAE model was immunized with Myelin Basic Protein (MBP) and randomly divided into normal group, model group, prednisone group, Erhuang decoction group and Erhuang capsule group, respectively. After 15 days (onset of acute phase) and 27 days post-immunization (remission phase), rats in each group were randomly selected and brain and spinal cord sections were taken for immunohistochemistry for IFN-γ, TGF-β, MMP-9, CD4+, CD8+, CD3+, and CD19+. dyeing. RESULTS: Compared with the model group, Erhuang Decoction (27d after immunization) and Erhuang Capsule (15d and 27d after immunization) significantly increased the expression of TGF-β in brain tissue. Both Erhuang Decoction and capsules significantly reduced the brain and spinal cord at 15d and 27d after immunization. Tissue IFN-γ, MMP-9 expression. The expression of MMP-9 in brain and spinal cord tissue at the 27th day and 27th day after immunization in Erhuang Decoction and Erhuang Capsule group was lower than that in prednisone group. Compared with the model group, the expression of CD4+ was significantly decreased, CD8+ expression was significantly increased, CD4+/CD8+ was significantly decreased, and CD19+ expression was decreased at the 15th day after immunization in the Erhuang decoction group on the 15th and 27th days after immunization. The CD8+ level in the brain of the Erhuang capsule group was 15 days after immunization. There was no significant difference in the model group, the others were same with the Erhuang Decoction group. Compared with the model group, the expression of CD4+ in the spinal cord at the 15th and 27th day after immunization was significantly lower and the expression of CD8+ was significantly increased and CD4+/CD8+ was significantly decreased in the Erhuang decoction group. The expression of CD4+ in the spinal cord was significantly decreased and CD8+ expression was significantly decreased in the Erhuang capsule group at the 15th day after immunization. As a result, CD4+/CD8+ was significantly decreased, 27dCD4+ expression was significantly decreased, and CD4+/CD8+ was significantly decreased. There was no significant difference between Erhuang Tang and Erhuang Capsule group. Conclusion: Erhuang Decoction and Erhuang Capsule can up-regulate the expression of TGF-β in the central nervous system of EAE rats. The effect is similar to that of prednisone. Both of them can significantly reduce the expression of IFN-γ and MMP-9 in brain and spinal cord tissues, which is superior to prednisone. Both Erhuang Decoction and capsules can reduce the expression of CD4+, increase the expression of CD8+, and improve the ratio of CD4+/CD8+. The effect of Erhuang Fang on the above indicators may be part of the mechanism for reducing the EAE nerve damage and improving the clinical symptoms of patients.