菊苣酸对磷酰胺氮芥诱导巨噬细胞免疫抑制的调节作用  

Regulatory effect of chicoric acid on immunosuppression of macrophages induced by phosphoramide mustard

作　　者：冯晨婧 白莉霞 杨亚军[2] 刘希望[2] 李世宏[2] 秦哲[2] 葛闻博 许笑 李剑勇[2] 李存 FENG Chenjing;BAI Lixia;YANG Yajun;LIU Xiwang;LI Shihong;QIN Zhe;GE Wenbo;XU Xiao;LI Jianyong;LI Cun(Tianjin City Key Laboratory of Agricultural Animal Breeding and Healthy Husbandry,College of Animal Science and Veterinary Medicine,Tianjin Agricultural University,Tianjin 300392,China;Key Laboratory of New Animal Drug Project of Gansu Province/Key Laboratory of Veterinary Pharmaceutical Development of Ministry of Agriculture and Rural Affairs,Lanzhou Institute of Husbandry and Pharmaceutical Science,Chinese Academy Agriculture Sciences,Lanzhou 730050,China)

机构地区：[1]天津农学院动物科学与动物医学学院天津市农业动物繁育与健康养殖重点实验室,天津300392 [2]中国农业科学院兰州畜牧与兽药研究所农业农村部兽用药物创制重点实验室/甘肃省新兽药工程重点实验室,甘肃兰州730050

出　　处：《中国兽医学报》2024年第2期319-326,共8页Chinese Journal of Veterinary Science

基　　金：甘肃省青年科技基金资助项目(21JR7RA031);中国农业科学院基本科研业务费专项所级统筹基金资助项目(1610032021013)。

摘　　要：用3μmol/L磷酰胺氮芥(phosphoramide mustard,PM)诱导RAW264.7细胞24 h,建立体外细胞免疫抑制模型;给予不同剂量(25、50、100μmol/L)的菊苣酸(chicoric acid,CA)干预细胞24 h,考察CA对免疫抑制的调节作用。利用CCK-8法检测细胞活力,中性红吞噬法检测细胞吞噬能力,ELISA试剂盒测定细胞IL-1β和IL-6表达,NO含量测定试剂盒测定细胞NO水平,实时荧光定量PCR测定IL-1β、IL-6和iNOS mRNA的相对表达量。结果显示,3μmol/L PM诱导RAW264.7细胞24 h后,巨噬细胞活力与吞噬能力显著降低(P<0.05),NO、IL-1β与IL-6的表达以及相应的mRNA相对表达量均显著下降(P<0.05),巨噬细胞免疫抑制模型成功建立;给予不同剂量(25、50、100μmol/L)的CA干预24 h后,巨噬细胞活力与吞噬能力显著增强(P<0.05),巨噬细胞NO、IL-1β与IL-6分泌水平以及相应的mRNA相对表达量明显升高(P<0.05)。结果表明,PM可以有效建立体外巨噬细胞免疫抑制模型,CA可能通过增强巨噬细胞活力与吞噬能力,促进炎症因子表达发挥免疫增强作用,从而提高机体免疫功能。本研究建立体外巨噬细胞免疫抑制模型,探究CA对PM诱导RAW264.7细胞免疫抑制的调节作用,为将CA开发为新型兽用免疫增强剂提供了理论基础。This study aims to establish a macrophage immunosuppression model in vitro and explore the regulatory effect of chicoric acid(CA)on phosphamide mustard(PM)-induced immunosuppression of RAW264.7 cells,providing a theoretical basis for the development of CA as a new veterinary immune enhancer.The cellular immunosuppression model was established by inducing with a concentration of 3μmol/L PM for 24 h in RAW264.7 cells.Then cells were treated with different concentrations of CA(25,50,100μmol/L)for 24 h to investigate the regulatory effects of CA on immunosuppression.Cell viability was determined by CCK-8 method,cellular phagocytic capacity was determined by neutral red phagocytosis method,IL-1βand IL-6 expression were determined by ELISA kit,NO content assay kit was used to measure NO level,and real-time PCR was used to determine the relative expression of IL-lβ,IL-6 and iNOS mRNA.The results showed that macrophage viability and phagocytic capacity were significantly reduced(P<0.05)after RAW264.7 cells were induced by 3μmol/L PM for 24 h,the expressions of NO,IL-1β,IL-6 and the corresponding relative expression of mRNA were decreased significantly(P<0.05),which revealed the macrophage immunosuppression model was successfully established.After treatment for24 h with different concentrations of CA(25,50,100μmol/L),macrophage viability and phagocytic capacity were significantly enhanced(P<0.05),and the secretion levels of NO,IL-1β,IL-6 and the corresponding relative expression of mRNA were increased(P<0.05)in macrophages.To sum up,PM can effectively establish an immunosuppressive model of macrophages in vitro,CA may enhance the immune function of the body by improving the vitality and phagocytic ability of macrophages,promoting the expression of inflammatory factors,and exerting an immune enhancing effect.

关 键 词：菊苣酸 磷酰胺氮芥 巨噬细胞 免疫抑制 免疫增强 

分 类 号：S859.7[农业科学—临床兽医学]