淫羊藿、巴戟天中药提取物对恒河猴M0、M1型单核衍生巨噬细胞基因表达的影响  

Influence of Extracts from Herba Epimedii and Radix Morindae Officinalis on Gene Expression of M0 and M1 Type Monocyte-derived Macrophages in Rhesus Monkeys

作　　者：成业[1] 陈颂[1] 王晓玉[1] 熊思艺 符林春[1] 

机构地区：[1]广州中医药大学热带医学研究所,广东广州510405

出　　处：《广州中医药大学学报》2016年第4期520-525,共6页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：国家科技重大专项资助项目(编号:2014ZX10005002)

摘　　要：【目的】探讨中药淫羊藿、巴戟天提取物(中药注射液喘可治的组成)对恒河猴单核衍生的巨噬细胞在未极化(M0)、M1极化条件下基因表达的影响,分析其可能的免疫调节的作用机制。【方法】采用密度梯度离心法分离中国恒河猴外周血单个核细胞(PBMCs),采用贴壁法纯化出单核细胞,并以粒细胞巨噬细胞集落刺激因子(GM-CSF)刺激5 d以使单核细胞分化成巨噬细胞;然后不加入极化因子(M0)或以干扰素γ(IFN-γ,继续培养2 d)诱导巨噬细胞极化为M1表型,同时分别加入淫羊藿、巴戟天提取物进行干预;然后提取m RNA并逆转录后,采用实时荧光定量PCR检测CCR5、CD4、CTLA-4、Fox P3、IDO、IL-10、TGF-β等基因表达量。【结果】与空白对照组比较,加入巴戟天提取物培养的M0巨噬细胞基因表达上调不明显;经淫羊藿提取物培养的M0巨噬细胞,CCR5基因表达量上调4.21倍,TGF-β上调7.66倍,Fox P3、IL-10轻度上调,而CD4、CTLA-4、IDO等基因表达改变倍数无明显变化。经巴戟天提取物刺激的M1型巨噬细胞,CTLA-4基因表达量上调3.22倍,Fox P3上调3.69倍,CCR5、CD4、IL-10、IDO基因改变倍数均无明显变化,TGF-β未测出;而由淫羊藿提取物刺激的M1型巨噬细胞,IL-10的表达量上调11.83倍,Fox P3上调4.55倍,IDO、CCR5、CD4、CTLA-4基因改变倍数无明显变化,TGF-β未测出。【结论】巴戟天和淫羊藿提取物培养能够上调M1型巨噬细胞IL-10、Fox P3、CTLA-4等抑炎基因表达,淫羊藿提取物能够上调M0巨噬细胞CCR5和TGF-β等基因表达,促进病原体清除和组织修复,从而发挥多效免疫调节作用。Objective To study the influence of the extracts from Herba Epimedii （Yinyanghuo）and Radix Morindae Officinalis （Bajitian）, the main ingredients of Chinese herbal injection Chuankezhi, on the gene expression of unpolarized （M0）and polarized （M1）monocyte-derived macrophages in rhesus monkeys, and to analyze the immunoregulatory mechanisms. Methods Peripheral blood mononuclear cells （PBMCs） in rhesus monkeys were isolated by density-gradient centrifugation method, and were purified by adherent culture method. The purified mononuclear cells were induced to differentiate into M0 macrophages following stimulation with granulocyte-macrophage colony-stimulating factor （GM-CSF） for 5 days, and then the M0 macrophages were induced into M1 macrophages without polarization factor added or with IFN-T added for 2 days. After M1 macrophages were treated with the extracts from Yinyanghuo or Bajitian, the total mRNA was extracted and then was used for reverse transcription to cDNA. The gene expression of CCR5, CD4, CTLA -4, FoxP3, IDO , interleukine （IL）-10 and transforming growth factor（ TGF ）-β was measured by real-time polymerase chain reaction （ PCR ）. Results Compared with the blank control group, the gene expression of MO macrophages cultured with Bajitian extract showed no obvious change, while the gene expression of CCR5 in M0 macrophages cultured with Yinyanghuo extract was up-regulated by 4.21 folds, TGF-β was up-regulated by 7.66 folds, FoxP3 and IL-10 were up-regulated mildly, and no obvious changes were shown in the up-regulation folds of CD4, CTLA-4 or IDO. The gene expression of CTLA-4 in M1 macrophages cultured with Bafitian extract was up-regulated by 3.22 folds, FoxP3 was up-regulated by 3.69 folds, no obvious changes were shown in the up- regulation folds of CCR5, CD4, IL-10 or IDO, and TGF-13 was undetectable. In M1 macrophage treated with Yinyanghuo extract, IL-10 gene expression was up-regulated by 11.83 folds, FoxP3 was up-regulated by 4.55 folds, no obvious changes w

关 键 词：巨噬细胞 淫羊藿 巴戟天 免疫调节 基因表达调控 恒河猴 

分 类 号：R285.5[医药卫生—中药学]