刺五加多糖对鸡淋巴细胞体外增殖及细胞因子mRNA表达水平的影响  

Effect of Acanthopanax plus polysaccharide on chicken lymphocyte proliferation and mRNA expression level of cytokines in vitro

作　　者：张英楠 徐晶 刘洪章 张桂山 杨树宝 ZHANG Yingnan;XU Jing;LIU Hongzhang;ZHANG Guishan;YANG Shubao(College of Life Science,Changchun Sci-Tech University,Changchun Jilin 130600,China;College of Public Health,Jilin Medical University,Jilin Jilin 132013,China;Basic Medical College,Jilin Medical University,Jilin Jilin 132013,China;College of Animal Science and Technology,Jilin Agricultural University,Changchun Jilin 130118,China)

机构地区：[1]长春科技学院生命科学学院,吉林长春130600 [2]吉林医药学院公共卫生学院,吉林吉林132013 [3]吉林医药学院基础医学院,吉林吉林132013 [4]吉林农业大学动物科技学院,吉林长春130118

出　　处：《中兽医医药杂志》2022年第2期1-6,共6页Journal of Traditional Chinese Veterinary Medicine

基　　金：吉林省教育厅“十三五”科学技术项目(JJKH20200355KJ,JJKH20201293JY);国家自然科学基金资助项目(31372391)。

摘　　要：研究刺五加多糖(ASPS)对鸡淋巴细胞体外增殖及细胞因子mRNA表达水平的影响。将不同浓度的ASPS添加到体外分离培养的血液淋巴细胞中,采用噻唑蓝(MTT)法检测淋巴细胞增殖变化,以实时荧光定量PCR方法检测淋巴细胞细胞因子IFN-γ、IL-2、IL-4、IL-10、IL-1β、IL-6、iNOS和TNF-αmRNA表达水平。结果显示,ASPS可以单独或协同植物血凝素(PHA)或脂多糖(LPS)促进淋巴细胞体外增殖,并且ASPS在质量浓度为200μg/mL时效果最佳。另外,ASPS能够促进淋巴细胞细胞因子mRNA的表达,当200μg/mL的ASPS与淋巴细胞体外共培养至24 h时,IFN-γ、IL-2、IL-1β、IL-6、iNOS和TNF-α的mRNA表达量均最高,而IL-4和IL-10 mRNA的相对表达量则以ASPS质量浓度为400μg/mL和培养时间48 h条件下最高,说明IL-4和IL-10 mRNA的表达滞后,而且呈剂量依赖性。结果表明,ASPS能够提高鸡淋巴细胞的体外增殖功能,促进Th1和Th2细胞因子分泌,这为进一步揭示刺五加多糖的免疫调节机制奠定了基础。The objective was to study the effects of Acanthopanax plus polysaccharide(ASPS)on the proliferation of chicken blood lymphocytes and the mRNA expression of cytokines in vitro.Different concentrations of ASPS were added into blood lymphocytes isolated and cultured in vitro.The proliferation of lymphocytes was detected by thiazolium blue(MTT)assay.The mRNA expression levels of cytokines IFN-γ,IL-2,IL-4,IL-10,IL-1β,IL-6,iNOS and TNF-α in lymphocytes were detected by real-time fluorescence quantitative assay.The results showed that ASPS could promote lymphocyte proliferation alone or in combination with PHA or LPS,and the best effect of ASPS was found at 200μg·mL^(-1).In addition,ASPS could promote the expression of cytokine mRNA in lymphocytes.Except for IL-4 and IL-10,the mRNA expression levels of IFN-γ,IL-2,IL-1β,IL-6,iNOS and TNF-α were the highest when the ASPS were co-cultured with lymphocytes at 200μg·mL^(-1) for 24 h in vitro.However,the relative expression levels of IL-4 and IL-10 mRNA were the highest when ASPS concentration was 400μg·mL^(-1) and incubation time was 48 h,indicating that the expression of IL-4 and IL-10 mRNA was delayed and dependent on high dose.The results indicated that ASPS could improve the proliferation function of lymphocytes and the secretion of Th1 and Th2 cytokines in vitro,which laid a foundation for further revealing the immunomodulatory mechanism of ASPS.

关 键 词：刺五加多糖 淋巴细胞增殖 细胞因子 MRNA 鸡 

分 类 号：S853.6[农业科学—临床兽医学]