红景天苷对小鼠腹腔巨噬细胞体外增殖、凋亡、吞噬、ROS和NO产生的影响  

Effects of salidroside on proliferation,apoptosis,phagocytosis,ROS and NO production of murine peritoneal macrophages in vitro

作　　者：叶莎莎[1] 曾耀英[1] 尹乐乐[1] 

机构地区：[1]暨南大学组织移植与免疫实验中心,广东广州510632

出　　处：《细胞与分子免疫学杂志》2011年第3期237-241,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家重点基础研究发展计划(973)资助项目(2006CB504201;2004CB720100);国家自然科学基金资助项目(30500466);广东省自然科学基金资助项目(5300419)

摘　　要：目的:研究红景天苷(Sal)对小鼠腹腔巨噬细胞体外增殖、凋亡、吞噬、胞内活性氧簇(ROS)及分泌一氧化氮(NO)的影响,初步探讨其对小鼠腹腔巨噬细胞的免疫调节作用。方法:无菌分离小鼠腹腔巨噬细胞,并制备单细胞悬液,以不同终浓度(80μmol/L、160μmol/L及320μmol/L)的Sal和巨噬细胞共培养4 h,再以脂多糖(LPS)和γ-干扰素(IFN-γ)进行共刺激。利用MTT比色法检测Sal对巨噬细胞体外增殖的影响。用放线菌酮(CHX)诱导巨噬细胞凋亡,用Sytox G reen染色结合荧光酶标仪检测Sal对CHX诱导巨噬细胞凋亡的影响。用流式细胞术(FCM)检测Sal对巨噬细胞吞噬功能的影响。用2-7-二氯氢化荧光素乙二脂(H2DCFDA)染色法结合荧光酶标仪检测Sal对胞内ROS产生的影响;用G riess反应检测Sal对巨噬细胞分泌NO的影响。结果:MTT比色法检测显示,终浓度为80、160、320μmol/L的Sal均可显著促进LPS+IFN-γ刺激巨噬细胞增殖(P<0.05)。荧光酶标仪检测Syto xG reen染色法的结果显示,160μmol/L的Sal可抑制CHX诱导的巨噬细胞凋亡(P<0.01)。FCM结果显示,各浓度的Sal均能促进单纯药物组和实验药物组LPS+IFN-γ刺激巨噬细胞的吞噬功能(P<0.05)。用荧光酶标仪检测DH2DCFDA染色结果表明,各浓度的Sal对LPS+IFN-γ刺激的巨噬细胞胞内ROS的产生均具有显著的抑制作用(P<0.01)。Griess反应检测NO含量的结果显示,各浓度的Sal对LPS+IFN-γ刺激巨噬细胞产生NO均具有促进作用(P<0.05)。结论:Sal对LPS和IFN-γ刺激的巨噬细胞增殖具有显著的促进作用,对CHX诱导的巨噬细胞凋亡具有显著的抑制作用,对静息态和活化态的巨噬细胞的吞噬功能均有增强作用,并能减少LPS和IFN-γ活化的巨噬细胞胞内ROS的产生;但能促进LPS和IFN-γ活化的巨噬细胞NO的分泌。AIM： To investigate the effects of salidroside（Sal） on proliferation,apoptosis,phagocytosis,the production of ROS and NO of murine peritoneal macrophages in vitro as well as its immunoregulation.METHODS： The single cell suspension of murine peritoneal macrophages was prepared under sterile condition,then co-cultured with different concentrations of Sal（80,160 and 320 μmol/L）for 4 hours prior to stimulation with LPS and IFN-γ,the proliferation of macrophages was measured by MTT colorimetry.The effect of Sal on the apoptosis of Sytox Green-labelled peritoneal macrophages induced by CHX was detected by Fluorescence enzyme-labelled meter.FCM was used to detect the effect of Sal on phagocytosis of peritoneal macrophages.Fluorescence enzyme-labelled meter was used to measure the effects of Sal on ROS of H2DCFDA-labelled macrophages induced by LPS and IFN-γ.Griess Gragent was used to detect the role of Sal in production of NO in peritoneal macrophages activated by LPS and IFN-γ.RESULTS： MTT result demonstrated that Sal could promote the proliferation of peritoneal macrophages activated by LPS and IFN-γ at the final concentrations of 80,160,320 μmol/L,respectively（P〈0.05）.The result of Fluorescence enzyme-labelled meter detected showed that Sal at the final concentration of 160 μmol/L could inhibit apoptosis of peritoneal macrophages induced by CHX（P〈0.01）.FCM analysis showed that different concentrations of Sal significantly promoted the phagocytosis of peritoneal macrophages which include un-activated and activated by LPS and IFN-γ（P〈0.05）.Fluorescence enzyme-labelled meter showed that Sal could reduce the production of ROS in activated peritoneal macrophages induced by LPS and IFN-γ（P〈0.05）.Sal also increased the production of NO in activated peritoneal macrophages induced by LPS and IFN-γ（P〈0.05）.CONCLUSION： Sal can promote proliferation of peritoneal macrophages stimulated by LPS and IFN-γ,and it can inhibit apoptosis of peritoneal macrophages induced by CH

关 键 词：红景天苷 腹腔巨噬细胞 增殖 凋亡 吞噬 活性氧 一氧化氮 

分 类 号：R282.17[医药卫生—中药学]