阿里红多糖对小鼠腹腔巨噬细胞体外抗弓形虫感染细胞因子分泌的影响  

Effects of Fomes officinalis polysaccharides on cytokine secretion of mouse peritoneal macrophages in response to a Toxoplasma infection in vitro

作　　者：高剑[1] 李莹花 唐云剑 王宇靖 齐瑞瑞 帕丽达·阿不力孜[3] 马秀敏[1] GAO Jian;LI Ying-hua;TANG Yun-jian;WANG Yu-jing;QI Rui-rui;PALIDA Ablizi;MA Xiu-min(College of Basic Medicine,Xinjiang Medical University,Urumqi 830011,China;First College of Clinical Medicine,Xinjiang Medical University;College of Pharmacy,Xinjiang Medical University)

机构地区：[1]新疆医科大学基础医学院,新疆乌鲁木齐830011 [2]新疆医科大学第一临床医学院,乌鲁木齐 [3]新疆医科大学药学院

出　　处：《中国病原生物学杂志》2020年第10期1137-1143,共7页Journal of Pathogen Biology

基　　金：新疆维吾尔自治区科技厅科技计划联合基金项目(No.2016D01C186)。

摘　　要：目的探索新疆植物药阿里红粗多糖(fomes officinalis polysaccharides,FOPS)提取物对小鼠腹腔巨噬细胞体外抗弓形虫TgRH株速殖子感染细胞因子分泌的影响。方法建立刚地弓形虫RH速殖子与小鼠腹腔巨噬细胞接种比例为1∶5的体外感染模型,实验设空白对照组、感染对照组和高(400μg/ml)、中(200μg/ml)、低(100μg/ml)3个药物剂量的药物对照组及感染模型治疗组。以流式微球捕获芯片技术(cytometric bead array,CBA)检测各组细胞感染虫体和药物作用6、12、24、36、48 h培养上清中IL-6、IL-10、TNF、INF、IL-12p70及MCP-1细胞因子水平。结果各实验组培养上清中IL-10和INF在实验过程均无表达。空白对照组感染48 h时TNF-α、IL-12p70及MCP-1略升高。感染组IL-6随感染时间延长水平显著升高,至感染48 h达(881.05±6.99)pg/ml,与空白对照组比较差异有统计学意义(F=235.3,P<0.05);TNF-α至感染48h时达(292.30±2.63)pg/ml,高于空白对照组(F=52.45,P<0.05);MCP-1和IL-12p70至感染48 h时与空白对照组差异无统计学意义(F值分别为2.624和3.518,P均>0.05)。各剂量FOPS作用于空白小鼠腹腔巨噬细胞,培养上清中MCP-1和TNF-α随作用时间延长而显著升高,至24 h时达到高峰,之后下降。各剂量药物组与空白对照组在药物作用24 h时组间MCP-1和TNF-α水平差异有统计学意义(F分别为41.95和231.4,P均<0.05);TNF-α表达水平与药物浓度升高成正比,MCP-1在培养上清中表达水平为M-FOP>L-FOP>H-FOP>空白对照组。高剂量H-FOP作用于感染巨噬细胞36 h内IL-6表达均较低,48 h略有升高,M-FOP组和L-FOP组IL-6表达趋势与感染对照组相似,但各感染时间IL-6表达水平均低于感染对照组(F=461.9,P<0.05)。各药物组TNF-α表达水平于药物作用24 h时升高,36 h时显著降低,48 h又迅速升高,各药物组与感染对照组比较差异有统计学意义(F=163600,P<0.05)。高剂量H-FOP组作用24 h细胞培养上清中TNF-α为(1624.426±6.54)pObjective To investigate the effects of a crude extract of Fomes officinalis polysaccharides(FOPS)on cytokine secretion of mouse peritoneal macrophages in response to a Toxoplasma infection in vitro.Methods An in vitro model of infection with T.gondii RH tachyzoites was established with an inoculation ratio of 1:5 for the parasite and mouse peritoneal macrophages.Animals were divided into the following groups:a blank control group,an infection control group,3 control groups receiving either a high(400μg/ml),medium(200μg/ml),or low(100μg/ml)dose of FOPS,and 3 infected groups receiving a high,medium,or low dose of FOPS.Cell culture supernatants were collected from each group after 6 h,12 h,24 h,36 h,and 48 h,and the levels of expression of the cytokines IL-6,IL-10,TNF,INF,IL-12 p70,and MCP-1 in the supernatant from each group were determined using a cytometric bead array(CBA).Results IL-10 and INF-were not expressed in the supernatants from each group during the entire experiment.Forty-eight h after infection,the levels of TNF-α,MCP-1,and IL-12 p70 rose slightly in the blank control group.The level of IL-6 in the infection control group increased significantly as time passed after infection time,reaching 881.05±6.99 pg/ml 48 h after infection.This differed significantly from the level in the blank control group(F=235.3,P<0.05).The level of TNF-reached 292.30±2.63 pg/ml 48 h after infection and was slightly higher than the level in the control group(F=52.45,P<0.05),while the levels of MCP-1 and IL-12 p70 did not differ significantly from those in the blank control group 48 h after infection(F:2.624 and 3.518,respectively,P>0.05).When each dose of FOPS acted on mouse peritoneal macrophages,the levels of MCP-1 and TNF-increased significantly,peaked at 24 h,and then decreased.Levels in each treatment group and the blank control group differed significantly 24 h after treatment(F:41.95 and 231.4,respectively,P<0.05).The level of TNF-increased as the drug concentration increased,but the level of MCP-1 in the cultur

关 键 词：阿里红多糖 刚地弓形虫 小鼠腹腔巨噬细胞 细胞因子 流式微球捕获芯片技术 

分 类 号：R382.5[医药卫生—医学寄生虫学]