薯蓣皂苷对实验性变态反应性脑脊髓炎模型小鼠损伤的保护作用及机制研究  

Study on the Protective Effect and Mechanism of Dioscin on the Injury of Experimental Allergic Encephalomyelitis in Mice

作　　者：夏凡林 沈磊莹 曹阳[3] 林子晶[3] XIA Fan-lin;SHEN Lei-ying;CAO Yang;LIN Zi-jing(Shanghai Urban Construction Vocational College,Shanghai 201415,China;The Ninth People’s Hospital Affiliated to Shanghai Jiaotong University,Shanghai 200011,China;The Second Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]上海城建职业学院,上海201415 [2]上海交通大学附属第九人民医院,上海200011 [3]重庆医科大学附属第二医院,重庆400016

出　　处：《中医药导报》2021年第3期13-18,共6页Guiding Journal of Traditional Chinese Medicine and Pharmacy

摘　　要：目的:探讨薯蓣皂苷治疗实验性变态反应性脑脊髓炎(EAE)的效果及其可能机制。方法:40只成年雌性C57BL/6小鼠随机分为对照组、模型组和薯蓣皂苷低、高剂量组,以髓鞘碱性蛋白+完全弗氏佐剂构建EAE小鼠模型。LFB和HE染色观察髄鞘和浸润细胞;免疫荧光染色检测T细胞活化标志物(CD4)、巨噬细胞活化标志物(F4/80)、星形胶质细胞标志物(GFAP)、小胶质细胞标志物(IBA-1)的表达;ELISA法检测小鼠血清IFN-γ、IL-17、IL-22、IL-4、NLRP3的表达水平;Western blotting法检测脊髓组织中总蛋白NF-κB、p-NF-κB(细胞核和细胞质)、IFN-γ、IL-17、IL-22、IL-4、NLRP3蛋白的表达水平,采用Trans AMTM NF-κB p65活性检测试剂盒检测NF-κB的活性。结果:与对照组比较,模型组小鼠脊髓组织中CD4、F4/80、GFAP和IBA-1的表达明显升高(P<0.05);与模型组比较,薯蓣皂苷低、高剂量组CD4、F4/80、GFAP和IBA-1的表达明显降低(P<0.05)。与对照组比较,模型组小鼠血清和脊髓组织中IFN-γ、IL-17、IL-22、NLRP3的蛋白表达水平均明显升高,而IL-4水平明显降低(P<0.05);薯蓣皂苷低、高剂量组IFN-γ、IL-17、IL-22、NLRP3的蛋白表达水平均明显低于模型组,而IL-4水平明显高于模型组,且呈剂量依赖性(P<0.05)。与对照组比较,模型组小鼠脊髓细胞质NF-κB明显降低、细胞核NF-κB明显升高(P<0.05);与模型组比较,薯蓣皂苷低、高剂量组小鼠脊髓细胞质NF-κB明显升高、细胞核NF-κB明显降低(P<0.05)。与对照组比较,模型组细胞核中NF-κB p65与DNA结合活性明显增加(P<0.05);薯蓣皂苷低、高剂量组细胞核NF-κB p65与DNA结合活性明显低于模型组(P<0.05)。结论:薯蓣皂苷可能通过抑制p-NF-κB的活化而改善外周血和脊髓组织中炎症因子的表达水平,进而改善EAE小鼠脊髓病理损伤。Objective:To investigate the effect of dioscin on experimental allergic encephalomyelitis(EAE)and its possible mechanism.Methods:A total of 40 adult female C57 BL/6 mice were randomly divided into control group,model group and dioscin low-dose group and dioscin high-dose group.EAE models were constructed with myelin basic protein and complete Freund’s adjuvant.LFB and HE staining were used to observe the iliac sheath and infiltrating cells.Immunofluorescence staining was used to detect the expression of T cell activation markers(CD4),macrophage activation markers(F4/80),astrocyte markers(GFAP)and microglia marker(IBA-1).The expression levels of IFN-γ,IL-17,IL-22,IL-4 and NLRP3 in serum were detected by ELISA.The expression levels of NF-κB,p-NF-κB(nucleus and cytoplasm),IFN-γ,IL-17,IL-22,IL-4 and NLRP3 in spinal cord were detected by Western blotting.The activity of NF-κB was detected by Trans AMTM NF-κB p65 assay kit.Results:Compared with the control group,the expressions of CD4,F4/80,GFAP and IBA-1 in the spinal cord of the model group were significantly increased(P<0.05).Compared with the model group,the expressions of CD4,F4/80,GFAP and IBA-1 in the dioscin low and high-dose groups were significantly decreased(P<0.05).Compared with the control group,the protein expression levels of IFN-γ,IL-17,IL-22 and NLRP3 in serum and spinal cord tissue of the model group were significantly increased,while the level of IL-4 was significantly decreased(P<0.05).The protein expression levels of IFN-γ,IL-17,IL-22 and NLRP3 in the dioscin low and high-dose groups were significantly lower than those in the model group,while the level of IL-4 was significantly higher than that in the model group in a dose-dependent manner(P<0.05).Compared with the control group,NF-κB in the cytoplasm of spinal cord was significantly decreased and NF-κB in the nucleus was significantly increased in the model group(P<0.05).Compared with the model group,NF-κB in bone marrow cytoplasm was significantly increased and NF-κB in nucleus was

关 键 词：薯蓣皂苷 实验性变态反应性脑脊髓炎 炎症 核因子ΚB 小鼠 

分 类 号：R285.5[医药卫生—中药学]