补肾益髓方促进实验性自身免疫性脑脊髓炎小鼠小胶质细胞向M2极化的作用

Effect of Bu Shen Yi Sui Formula on promoting polarization of microglia to M2 in mice with experimental autoimmune encephalomyelitis

目的观察补肾益髓方(Bu Shen Yi Sui Formula,BSYS)促进实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)小鼠小胶质细胞向M2极化的作用。方法 80只C57BL/6小鼠采用数字表法随机分为正常对照组、模型、醋酸泼尼松及BSYS组。各造模组小鼠以髓鞘少突胶质细胞糖蛋白(myelin oligodendroglia glycoprotein,MOG)35-55诱导EAE模型。给予BSYS方干预后,观察小鼠的神经功能评分。于造模第23天和第40天分别取材,HE染色观察小鼠脑和脊髓病理变化,免疫荧光法测定小胶质细胞M1、M2型标志物CD86、Arg-1表达,Western blotting法检测过氧化物酶体增生物激活受体-γ(peroxisome proliferator activated receptor-γ,PPAR-γ)的表达,酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)法测定抗炎因子干扰素-β(interferon-β,INF-β)和转化生长因子-β1(transforming growth factor-β1,TGF-β1)的含量。结果与模型组相比,BSYS组小鼠神经功能评分和炎性细胞浸润评分显著降低(P <0. 05);脑及脊髓组织中CD86表达下降(P <0. 05),Arg-1表达升高(P <0. 05),抗炎因子INF-β、TGF-β1的表达显著升高(P <0. 05),PPAR-γ蛋白表达显著升高(P <0. 001)。结论 BSYS对EAE小鼠有神经保护作用,其作用可能与上调PPAR-γ表达,促进小胶质细胞向M2极化有关。

Objective To observe the effect of Bu Shen Yi Sui Formula(BSYS)on promoting the polarization of microglia to M2in experimental autoimmune encephalomyelitis(EAE)mice.Methods Eighty C57BL/6mice were randomly divided into normal control,model,prednisone acetate and BSYS group.The EAE model in mice were induced by myelin oligodendroglia glycoprotein(MOG)35-55.Each mice was intragastrically administered by BSYS,and then the neurological function scores of the mice were recorded,the pathological changes in the brain and spinal cord of mice were observed by HE staining,the expressions of CD86and Arg-1,M1and M2markers,in microglia were determined by immunofluorescence,and the peroxisome proliferator activated receptor-γ(PPAR-γ)was detected with Western blotting.The expression of anti-inflammatory factor interferon-β(INF-β)and transforming growth factor-β1(TGF-β1)was determined with enzyme-linked immunosorbent assay.Results Compared with the model group,the neurological function scores and the inflammatory cell infiltration scores of the mice in the BSYS group were significantly lower(P<0.05).The expression of CD86was decreased in microglia(P<0.05),and Arg-1was increased(P<0.05).The anti-inflammatory factors INF-βand TGF-β1in brain tissue were significantly increased(P<0.05).PPAR-γprotein in brain tissue was also found to be significantly elevated(P<0.001).Conclusion BSYS showed neuroprotective effects on EAE mice,which may be related to up-regulation of PPAR-γexpression and promotion of microglia to M2polarization.