山茱萸新苷对EAE中枢神经系统MCP-1表达及CD68阳性细胞浸润的影响  

The Impact of Cornuside on the Expression of MCP-1 and the Infiltration of CD68 Positive Cells in the Central Nervous System in EAE

作　　者：张荣博[1] 徐彬[1] 朱敏姿[1] 周未莹[1] 吴忧[1] 梁顺利[1] 章水晶[1] 李铮[2] 袁强[2] 

机构地区：[1]浙江中医药大学附属第二医院,杭州310005 [2]浙江中医药大学

出　　处：《浙江中医药大学学报》2016年第6期482-487,共6页Journal of Zhejiang Chinese Medical University

基　　金：浙江省中医药科技计划项目(2013ZB063)~~

摘　　要：[目的]研究山茱萸新苷对实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)大鼠中枢神经系统单核细胞趋化蛋白-1(monocyte chemoattractant protein-1,MCP-1)表达及CD68阳性细胞浸润的影响。[方法]制备豚鼠全脊髓匀浆免疫抗原,皮下注射至Lewis大鼠,建立EAE模型。设正常对照组、EAE组、山茱萸新苷组、波尼松组,每天神经功能评分,待症状达到高峰处死实验动物,用RT-PCR、Western Blot法比较各组实验动物中枢神经系统MCP-1 m RNA及蛋白的表达,免疫组织化学染色法比较各组实验动物中枢神经系统CD68阳性细胞浸润情况。[结果]正常对照组、EAE组、山茱萸新苷组、波尼松组大鼠MCP-1 m RNA的相对表达量分别为(11.265±2.928)、(401.373±55.398)、(124.987±20.244)、(75.465±7.766),MCP-1蛋白相对表达量分别为(7.458±2.570)、(24.155±1.420)、(19.568±0.863)、(17.458±1.630),CD68阳性指数分别为0%、(41.93±12.25)%、(16.08±8.70)%、(5.38±2.88)%。使用单因素方差分析法,MCP-1 m RNA的相对表达量、MCP-1蛋白相对表达量、CD68阳性指数组间差异显著,均有统计学意义(F=199.734、66.081、35.565,均P=0.000)。山茱萸新苷组与EAE组在神经功能评分、MCP-1 m RNA的相对表达量、MCP-1蛋白相对表达量、CD68阳性指数表达差异均有统计学意义(P=0.002、0.000、0.003、0.013)。[结论]山茱萸新苷可改善EAE大鼠神经功能,抑制EAE大鼠中枢神经系统MCP-1表达及CD68阳性细胞浸润。[Objective] To explore the impact of cornuside on the expression of MCP-1 and the infiltration of CD68 positive ceils in the central nervous system in EAE. [Methods] An EAE model was established by injecting the guinea pig spinal cord homogenate antigen in subcutaneous tissue of Lewis rats. The rats were randomly divided into normal control group, EAE group, cornuside group, and prednisone group. The neurological function scores were assessed every day. At last, these animals were killed when the symptoms reached the peak. The expressions of MCP-1 mRNA and MCP-1 protein in the central nervous system of each group were compared by using real-time quantitative PCR（RT-PCR） and Western Blot. staining was used to compare the infiltration of CD68 positive cells in the central nervous system of experimental animals in each group. [Results] The relative expression of MCP-I mRNA by RT-PCR of normal control group, EAE group, cornuside group, prednisone group rats was respectively（11.265＋2.928） （401.373±55.398）,（124.987±20.244）,（75.465±7.766）, the relative expression of MCP-1 protein by Western Blotting was respectively（7.458±2.570）, （24.155±1.420）,（19.568±0.863）,（17.458±1.630）, and the CD68 positive indexes were respectively 0%,（41.93±12.25）%,（16.08±8.70）%,（5.38±2.88）%. Using one-way analysis of variance, there were significant statistical differences of the relative expression of MCP-1 mRNA, the relative expression of MCP-I protein and CD68 positive index between the groups（F=199.734, 66.081, 35.565, all P=0.000）. The difference in neurological function score, the relative expression of MCP-1 mRNA, the relative expression of MCP-1 protein and CD68 positive index between cornuside group and EAE group had statistical significance（P=0.002, 0.000, 0.003, 0.013）. [Conclusion] Cornuside can improve neural function of EAE rats, also can inhibit the expression of MCP-1 and the infiltration of CD68 positive cells in the central nervous system of EAE rats.

关 键 词：EAE 多发性硬化 山茱萸新苷 MCP-1 CD68 

分 类 号：R331[医药卫生—人体生理学]