基于基因表达谱研究淫羊藿总黄酮干预老年大鼠海马炎性衰老的效果与机制  

Intervention outcome and mechanism of epimedium total flavonoids on aged rat hippocampus during inflamm-aging progress based on gene expression profile

作　　者：夏世金[1] 沈自尹[2] 俞卓伟[1] 邰先桃[3] 刘小雨[4] 李亚明[1] 汪海东[1] 黄建华[2] 

机构地区：[1]复旦大学附属华东医院上海市老年医学研究所,上海市200040 [2]复旦大学附属华山医院中西医结合研究所,上海市200040 [3]云南中医学院第一附属医院,云南省昆明市650011 [4]同济大学附属第十人民医院中医科,上海市200072

出　　处：《实用老年医学》2010年第1期24-27,共4页Practical Geriatrics

基　　金：国家重点基础研究发展计划(973项目)(2007CB507400);中国博士后科学基金(20060400578)

摘　　要：目的探讨炎性衰老的炎性细胞因子网络机制,及淫羊藿总黄酮(epimedium total flavonoids,EF)干预的效果与机制。方法清洁级健康雄性SD大鼠分为3组:4月龄(4 m)组、24月龄(24 m)组和24月龄加EF干预(24 m+EF)组,每组10只。自满21月龄当天开始,24 m+EF组予EF灌胃,24 m组给予等量蒸馏水灌胃,均连续90 d。取各组大鼠海马组织,应用炎症细胞因子与受体基因芯片对海马组织分别进行基因表达检测,绘制基因表达谱,筛选有表达差异的基因。海马组织匀浆,离心取上清液,运用酶联免疫吸附法检测上清液中炎性细胞因子肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)和IL-10蛋白表达。结果在海马组织中,与4 m组相比,24 m组表达上调的促炎性反应细胞因子与受体基因有9个,表达下调的抗炎性细胞因子与受体基因有6个。与24 m组比较,24 m+EF组中表达上调的抗炎性细胞因子与受体基因有8个,表达下调的促炎性细胞因子与受体基因有11个。24 m组海马组织中TNF-α和IL-6水平都显著高于4 m组(P<0.01);与24 m组比较,24 m+EF组中TNF-α和IL-6水平都明显降低,IL-10水平显著升高(P<0.01)。与4 m组相比,24 m组海马组织上清液中TNF-α/IL-10和IL-6/IL-10比值均显著升高(P<0.01);24 m+EF组TNF-α/IL-10和IL-6/IL-10比值均比24 m组显著降低(P<0.01)。结论老年大鼠海马存在促炎性细胞因子表达上调,导致促-抗炎性细胞因子网络和促-抗炎反应体系平衡失调,这可能是炎性衰老中出现高促炎性反应状态的原因和炎性衰老发生的新机制;EF可能通过下调促炎性细胞因子表达和上调抗炎性细胞因子表达,重塑促-抗炎性细胞因子网络和促-抗炎性反应体系新的平衡,达到干预炎性衰老的目的。Objective To investigate the mechanisms of inflammatory cytokines network in the development of inflamm-aging and EF intervention outcome and its mechanism.Methods Healthy male Sprague Dawley rats were divided into 3 groups： 4 m young group with 4 months rats,24 m aged group with 24 months rats treated with distilled water intragastric administration and 24 m＋EF group with 24 months rats treated with EF by intragastric administration for 3 months from 21 months,10 each group.The gene expression profiles of rat hippocampal tissues from rats in every group were analyzed by using inflammatory cytokines and receptors cDNA microarray,respectively.The differential expressed genes were screened.At the same time,the levels of inflamm-aging-related inflammatory cytokines proteins expressions of TNF-α,IL-6 and IL-10 in rat hippocampal tissues were evaluated by enzyme linked immunosorbent assay. Results In rat hippocampal tissues,compared with those in 4 m group,there were 9 differentially up-regulated expressed pro-inflammarory genes and 6 down-regulated expressed anti-inflammatory genes in 24 m group.Compared with those in 24 m group,there were 8 up-regulated expressed anti-inflammatory genes and 11 down-regulated expressed pro-inflammatory genes in 24 m＋EF group.Compared with those in 4 m group,the levels of TNF-α and IL-6 in the supernatant from rat hippocampal tissues in 24 m group increased significantly（P〈0.01）.As compared with those in 24 m group,the levels of TNF-α and IL-6 in the supernatant from rat hippocampal tissues in 24 m＋EF group decreased significantly（P〈0.01）.But the level of IL-10 in EF＋24 m group was significantly higher than that in 24 m group（P〈0.01）.The ratio of TNF-α to IL-10 and IL-6 to IL-10 in 24 m group increased significantly as compared with those in 4 m group（P〈0.01）,and decreased significantly in 24 m＋EF group as compared with those in 24 m group（P〈0.01）. Conclusions There is upregulation of mRNA and protein expressions of pro-inflammatory cytoki

关 键 词：炎性衰老 基因表达谱 炎症细胞因子与受体 酶联免疫吸附 淫羊藿总黄酮 

分 类 号：R339.38[医药卫生—人体生理学]