黄芪多糖在LPS诱导的DF-1细胞炎症反应中的抗炎作用及其调节机制  

Anti-inflammatory effect of Astragalus polysaccharides on LPS-induced DF-1 cell inflammation and its regulatory mechanism

作　　者：刘丹华 张瑞莉[1] 田旭 吕晓萍[1] 高雪丽[1] 郑世民[1] 刘超男[1] LIU Danhua;ZHANG Ruili;TIAN Xu;LYU Xiaoping;GAO Xueli;ZHENG Shimin;LIU Chaonan(Heilongjiang Key Laboratory for Laboratory Animals and Comparative Medicine,College of Veterinary Medicine,Northeast Agricultural University,Harbin 150030,China)

机构地区：[1]东北农业大学,动物医学学院,黑龙江省实验动物与比较医学重点实验室,黑龙江哈尔滨150030

出　　处：《中国兽医学报》2021年第1期143-149,共7页Chinese Journal of Veterinary Science

基　　金：高等学校博士学科点科研基金资助项目(20122325120005)。

摘　　要：为探讨黄芪多糖(Astragalus polysaccharide,APS)在LPS(lipopolysaccharide,LPS)诱导的鸡胚成纤维细胞系DF-1(chicken embryonic fibroblast cell line,DF-1)炎症模型中对IL-1β和TNF-α表达的影响,以及细胞因子信号转导抑制因子3(suppressers of cytokine signaling 3,SOCS3)对炎症信号通路NF-κBp65和p38MAPK的调节机制。将DF-1细胞分为对照组(C)、LPS组(L)、APS组(A)以及APS+LPS组(A+L),分别在LPS刺激后6,12,24,48,72 h检测各组细胞IL-1β、TNF-α、NF-κBp65、p38MAPK和SOCS3 mRNA和蛋白水平的变化。研究发现,当LPS终质量浓度为2 mg/L时可诱导DF-1细胞出现明显的炎症反应,而APS终质量浓度为100 mg/L时为本试验最佳抗炎浓度。与对照组相比,APS组炎性因子IL-1β和TNF-αmRNA表达及蛋白含量在6~72 h均显著升高(P<0.05);与LPS组相比,APS+LPS组SOCS3 mRNA表达在6~72 h均显著升高(P<0.05),NF-κBp65、IL-1β和TNF-αmRNA表达在6~72 h均显著降低(P<0.05),而p38MAPK变化不显著(P>0.05)。在DF-1细胞中,APS单独处理可以促进IL-1β和TNF-α等细胞因子释放而增强免疫;APS和LPS共处理时,APS通过抑制炎性因子IL-1β和TNF-α的释放发挥抗炎作用,这种抑制作用与高表达的SOCS3对NF-κBp65过度激活密切相关。To investigate the role of Astragalus polysaccharide in LPS-induced DF-1 cell inflammation model,and the regulatory mechanism of suppressers of cytokine signaling 3(SOCS3)on inflammatory factor signaling pathways NF-κBp65 and p38 MAPK.In this experiment,DF-1 cells were randomly divided into 4 groups,which were control group(C),LPS group(L),APS group(A)and APS+LPS group(A+L).The changes of IL-1β,TNF-α,NF-κBp65,p38 MAPK and SOCS3 mRNA and protein levels in each group were detected at 6,12,24,48 and 72 h after LPS stimulation.The study found that when the final LPS concentration was 2 mg/L,DF-1 cells could induce a significant inflammatory response;when the final APS concentration was 100 mg/L,the optimal anti-inflammatory concentration was achieved;compared with the control group,the expression of inflammatory factors IL-1βand TNF-αmRNA and protein content in the APS group were significantly increased between 6 and 72 hours(P<0.05);compared with the LPS group,in the APS+LPS group,SOCS3 mRNA expression was significantly increased from 6 to 72 hours(P<0.05),NF-κBp65,IL-1βand TNF-αmRNA expressions were significantly decreased from 6 to 72 hours(P<0.05),and the change of p38 MAPK was not significantly(P>0.05).These results showed that APS treatment alone can promote immune enhancement by promoting cytokine release in DF-1 cells;When APS and LPS are processed together,APS can significantly inhibit the release of inflammatory factors IL-1βand TNF-α,and this inhibition is closely related to the regulation of NF-κBp65 overactivation by overexpressing SOCS3.

关 键 词：IL-Β TNF-α NF-ΚBP65 P38MAPK SOCS3 黄芪多糖 DF-1细胞 

分 类 号：S831.7[农业科学—畜牧学]