三叶青提取物对人树突状细胞功能的影响  

Effects of Tetrastigma hemsleyanum extract on human dendritic cells function

作　　者：廖雨琴 许青 祝宇翀 单宇 杨阳 刘军权 黄丹婷 孙香香 Liao Yuqin;Xu Qing;Zhu Yuchong;Shan Yu;Yang Yang;Liu Junquan;Huang Danting;Sun Xiangxiang(Hangzhou Kingmed Medical Laboratory Co.,Ltd.,Hangzhou 310053,China;Department of Clinical Laboratory,the 71st Group Military Hospital of the Chinese People's Liberation Army,Xuzhou 221004,China)

机构地区：[1]杭州金域医学检验所有限公司,310053 [2]中国人民解放军陆军第七十一集团军医院检验科,徐州221004

出　　处：《中华临床实验室管理电子杂志》2019年第3期150-155,共6页Chinese Journal of Clinical Laboratory Management(Electronic Edition）

基　　金：南京军区医学科技创新课题(MS039)

摘　　要：目的探讨三叶青提取物TH-b对树突状细胞(dendritic cells,DC)功能的影响。方法分离健康人外周血中单个核细胞,用粒细胞-巨噬细胞集落刺激因子(granulocyte-macrophage colony stimulating factor,GM-CSF)、白介素-4(interleukin-4,IL-4)、γ-干扰素(interferon-γ,IFN-γ)和脂多糖(lipopolysaccharide,LPS)诱导培养DC。用念珠菌孢子法检测经TH-b作用后的DC吞噬能力;将经TH-b作用后的DC与同种外周血淋巴细胞进行混合培养,以检测淋巴细胞增殖率;采用流式细胞术检测DC表面标记CD195和TH-b作用后DC的表面标记CD80、CD83、CD86、HLA-DR;用酶免疫法检测TH-b作用后DC的培养上清液中的IL-12含量。结果培养7 d后DC CD195表达水平(85.3%)明显高于培养前(2.4%)。三叶青提取物TH-b部分对DC吞噬能力有作用明显,TH-b浓度为0.62μg/ml时吞噬能力最强。各种TH-b浓度作用后DC的细胞表面CD80、CD83、CD86和HLA-DR表达均明显升高,以0.62μg/ml最明显,分别为88.56%±8.16%、86.75%±7.59%、99.34%±8.24%、96.15%±8.27%,显著高于对照组的71.38%±7.31%、43.61%±4.29%、93.41%±8.34%、79.26%±5.12%,差异有统计学意义(P<0.05)。TH-b作用后的DC与淋巴细胞在1:1、1:10、1:100的比例下,对同种外周血淋巴细胞具有强烈的激发和促增殖作用,以1:1最明显。TH-b作用DC 48 h后,DC分泌的IL-12均明显升高,在0.62μg/ml时IL-12分泌达最高值(526.3pg/ml),显著高于对照组(260.1 pg/ml)(P<0.05),并有浓度窗现象。结论低浓度三叶青提取物TH-b部分能够显著促进人DC吞噬功能,促进DC表面标志物CD80和CD86的表达,提高DC成熟标记和IL-12分泌能力。Objective To investigate the effect of TH-b fraction of Tetrastigma hemsleyanum on the function of dendritic cells(DC).Methods Peripheral blood mononuclear cells were isolated from healthy donors.DC were induced by granulocyte-macrophage colony stimulating factor(GM-CSF),interleukin-4(IL-4),interferon-gamma(IFN-γ)and lipopolysaccharide(LPS).The phagocytosis ability of DC treated with TH-b was detected by candida spore method,the proliferation rate of lymphocyte was detected by mixed culture of DC treated by TH-b and peripheral blood lymphocyte.Surface marker CD195 and markers(CD80,CD83,CD86 and HLA-DR)of DC treated by TH-b were detected by flow cytometry.The concentration of IL-12 in DC culture supernatant was measured after TH-b treatment by enzyme immunoassay.Results After 7 days culture,the expression of CD195 on DC was significantly increased compare to the base line(85.3%vs.2.4%).The Tetrastigma hemsleyanum extract TH-b increased DC phagocytosis function,and 0.62μg/ml was the optimal TH-b concertration to DC phagocytic ability.The expression of CD80,CD83,CD86,and HLA-DR on the surface of DC increased significantly after concluture with TH-b,The ratios of these cell markers expressed on DC were most obvious when treated with 0.62 ug/ml TH-b,and were 88.56%±8.16%,86.75%±7.59%,99.34%±8.24%,96.15%±8.27%respectively,which were 71.38%±7.31%,43.61%±4.29%,93.41%±8.34%and 79.26%±5.12%in the control group.The treated DC(with TH-p)could strongly stimulate and promoted the proliferation of human peripheral blood lymphocyte,when coculture with lymphocyte in the ratio of 1:1,1:10 and 1:100.The 1:1 ratio was the optimal one.After 48 hours culture with TH-b,IL-12 secreted by DC increased significantly.And when treated with 0.62 ug/ml TH-b,DC secreted the highest level of IL-12(526.3 pg/ml),which was significantly higher than that of the control group(260.1 pg/ml)(P<0.05),and there was a concentration window phenomenon.Conclusion Low concentration of TH-b could significantly promote the phagocytosis of human DC,pro

关 键 词：三叶青提取物 树突状细胞 吞噬 混合淋巴细胞反应 

分 类 号：R73[医药卫生—肿瘤]