黄芪多糖对鸡巨噬细胞HD11的免疫调节作用研究  

Immunomodulatory effect of Astragalus polysaccharide on chicken macrophage HD11

作　　者：刘琴 王磊[1] 张康[1] 徐国伟 郭志廷[1] 王学智[2] 张景艳[1] Huub F.J.Savelkoul LIU Qin;WANG Lei;ZHANG Kang;XU Guowei;GUO Zhiting;WANG Xuezhi;ZHANG Jingyan;Huub F.J.Savelkoul(Engineering and Technology Research Center of Traditional Chinese Veterinary Medicine of Gansu Province,Lanzhou Institute of Husbandry and Pharmaceutical Sciences of CAAS,Lanzhou Gansu 730050,China;Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou Gansu 730046,China;Cell Biology and Immunology Group,Wageningen University,Wageningen 6700AH,Netherlands)

机构地区：[1]中国农业科学院兰州畜牧与兽药研究所甘肃省中兽药工程技术研究中心,甘肃兰州730050 [2]中国农业科学院兰州兽医研究所,甘肃兰州730046 [3]荷兰瓦赫宁根大学细胞生物与免疫学教研组,瓦赫宁根6700AH

出　　处：《中兽医医药杂志》2023年第4期1-7,共7页Journal of Traditional Chinese Veterinary Medicine

基　　金：国家重点研发计划(2022YFD1801104);甘肃省国际合作项目(22YF7WA032);甘肃省技术创新引导计划(22CX8NA013)。

摘　　要：调节巨噬细胞的活化是增强畜禽免疫力的有效方法。黄芪多糖(APS)作为免疫增强剂,在临床被广泛应用,但有关其对家禽巨噬细胞的调节作用机制鲜有报道。本研究用APS诱导鸡巨噬细胞HD1112 h,评价其对HD11细胞的免疫调节作用。通过CCK-8法检测细胞增殖活力;用台盼蓝检测细胞的活率;采用Griess试剂法检测细胞中NO含量;采用RT-PCR法检测细胞中细胞因子、TLRs和NF-κB p65的mRNA表达水平。结果表明,25~400μg/mL APS对鸡巨噬细胞HD11无毒性作用,能显著促进细胞增殖(P<0.05),且APS组(除400μg/mL)与LPS组无显著性差异(P>0.05)。CCK-8检测显示,50~200μg/mL APS能显著提高细胞中NO含量(P<0.05)及炎性因子IL-8和IL-10转录水平(P<0.05),且呈一定的剂量依赖性,200μg/mL APS还能显著提高炎性因子IL-1β和IL-6转录水平(P<0.05),但均对TNF-α无显著性影响(P>0.05)。此外,50~200μg/mL APS对TLR2基因表达有显著的抑制作用(P<0.05),100~200μg/mL APS对TLR4基因表达有显著的抑制作用(P<0.05),表明TLR2对APS更敏感,但200μg/mL APS能提高NF-κB p65的转录水平(P<0.05)。这些结果表明黄芪多糖作为一种免疫调节剂对鸡巨噬细胞有免疫增强作用,并表现出不同的剂量依赖效应。Regulating the activation and function of macrophages is considered an effective method for the prevention and treatment of human and animal diseases.Astragalus polysaccharide(APS),as an immune enhancer,is widely used in clinic.However,the immonoregulatory effects of APS on chicken macrophages remain largely unknown.The objective of this study is to evaluate the immunomodulatory activity of APS on HD11 macrophages by incubating HD11 macrophages with APS for 12 h.Cell proliferation and cell viability were detected by CCK-8 and trypan blue methods.Griess method was used to detect NO content,and RT-PCR method was used to determine the mRNA levels of cytokines,TLRs and NF-κB p65.The results showed that APS had no toxic effect on HD11 macrophages at 25-400μg/mL,and there was no significant difference between APS group(except for 400μg/mL)and LPS group(P>0.05).APS at 50-200μg/mL could significantly promote cell proliferation(P<0.05),NO content(P<0.05)and the expression of inflammatory factors IL-8(P<0.05)and IL-10(P<0.05)in a dose-dependent manner,without affecting TNF-α(P>0.05).Then,200μg/mL APS could significantly increase the expression of inflammatory factors IL-1β(P<0.05)and IL-6(P<0.05).In addition,50-200μg/mL APS showed a significant inhibitory effect on TLR2(P<0.05)and 100-200μg/mL APS showed a significant inhibitory effect on TLR4 gene expression(P<0.05),which is indicated that TLR2 is more sensitive than TLR4,but 200μg/mL APS can increase the gene expression of NF-κB p65(P<0.05).These results suggested that APS acts as an immunomodulator which shows differential dose-dependent effects on chicken macrophages.

关 键 词：黄芪多糖 HD11细胞 炎性因子 免疫增强 

分 类 号：S853.74[农业科学—临床兽医学]