大车前苷抑制猪肺炎支原体诱导的炎性反应的作用初探  

Preliminary Study on the Inhibitory Effect of Plantamajoside on Inflammatory Response Induced by Mesomycoplasma hyopneumoniae

作　　者：王吉英 向书涵 李志强 李炻漾 张磊[3] 谢青云 熊祺琰[3,5] 邵国青 冯志新[3,5] 于岩飞[3,5] WANG Jiying;XIANG Shuhan;LI Zhiqiang;LI Shiyang;ZHANG Lei;XIE Qingyun;XIONG Qiyan;SHAO Guoqing;FENG Zhixin;YU Yanfei(School of Basic Medical Sciences,Hunan University of Medicine,Huaihua 418000,China;College of Animal Science and Technology,Huaihua Vocational and Technical College,Huaihua 418099,China;Institute of Veterinary Medicine,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China;College of Veterinary Medicine,Hunan Agricultural University,Changsha 410125,China;College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China;College of Animal Science and Technology,Hunan Agricultural University,Changsha 410128,China)

机构地区：[1]湖南医药学院基础医学院,怀化418000 [2]怀化职业技术学院动物科技学院,怀化418099 [3]江苏省农业科学院兽医研究所,南京210014 [4]湖南农业大学动物医学院,长沙410125 [5]南京农业大学动物医学院,南京210095 [6]湖南农业大学动物科学技术学院,长沙410128

出　　处：《畜牧兽医学报》2025年第4期1958-1968,共11页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：湖南省自然科学基金项目(2022JJ50321);湖南省教育厅科学研究项目(22B1055);江苏省科协青年科技人才托举工程(JSTJ-2023-XH031);江苏省农业自主创新资金[CX(22)1011];怀化市科学技术局项目(2020R2206)。

摘　　要：猪肺炎支原体(Mesomycoplasma hyopneumoniae, Mhp)在呼吸道上皮细胞定植,引起猪气管、支气管及肺组织的炎性损伤,使猪饲料转化率降低,日增重下降,给养猪业造成严重的经济损失。大车前苷(Plantamajoside, PMS)为一种常用的抗炎中药,其在抑制Mhp感染引起的炎性反应中的作用未知。本研究拟通过Mhp感染PK-15细胞和小鼠为模型,探讨大车前苷在体外及体内条件下对Mhp诱导的炎性反应的影响及其相关机制。通过不同浓度的PMS作用PK-15细胞,确定PMS对细胞形态和活力无影响的最高浓度。随后采用该最高浓度的PMS的作用于Mhp感染的PK-15细胞,通过RT-qPCR检测PK-15细胞中肿瘤坏死因子α(tumor necrosis factor-α, TNF-α)、白细胞介素-6(interleukin-6, IL-6)等炎性相关因子的mRNA表达变化。结果显示,与细胞对照组相比,Mhp感染组细胞中的IL-6、TNF-α、NF-κB和IκBα的mRNA水平显著升高,而PMS的作用可降低Mhp感染时这种mRNA水平的升高。进一步通过Western blot和免疫荧光分析发现PMS可抑制Mhp诱导的NF-κB P65磷酸化,抑制NF-κB P65的核易位。小鼠感染模型的相关研究发现PMS可改善Mhp诱导的小鼠肺脏病变,使炎性细胞减少。同时,大车前苷可显著降低血清中TNF-α的含量。以上结果表明,PMS可能通过NF-κB通路改善Mhp诱导的炎性反应,为防治猪肺炎支原体感染新方法的提出提供重要的理论依据。Mycoplasma hyopneumoniae(Mhp)colonizes to respiratory epithelial cells,causing the inflammatory injury of trachea,bronchus and lungs in pigs,which reduced the feed conversion rate and daily weight gain of pigs,resulting in serious economic losses to the swine industry.plantamajoside(PMS),as a commonly used anti-inflammatory traditional Chinese medicine,its role in inhibiting the inflammatory response caused by Mhp infection is unknown.The aim of this study was to investigate the effect and related mechanisms of PMS on inflammatory response induced by Mhp in vitro and in vivo.Mhp infected PK-15 cells and mice were used as infection models,respectively.First,PK-15 cells were treated with different concentrations of PMS to determine the maximum concentration of PMS that had no effect on cell morphology and viability.This highest concentration of PMS was then applied to PK-15 cells which infected with Mhp.The mRNA expression of tumor necrosisfactor-alpha(TNF-α),interleukin-6(IL-6)and other inflammatory factors in PK-15 cells was detected by RT-qPCR.The results showed that the mRNA levels of IL-6,TNF-α,NF-κB and IκBαwere significantly increased in PK-15 cells which infected with Mhp compared with the control group,while PMS could reduce the effect.Western blot and immunofluorescence were used to further analysis and the results showed that PMS could inhibited the phosphorylation of NF-κB P65 and IκBα,and inhibited the nuclear translocation of NF-κB P65,which induced by Mhp.Related studies on mouse models have found that PMS can improve the lung lesions,reduce the inflammatory cells of lung which induced by Mhp.At the same time,PMS can significantly reduce the content of TNF-αin serum.These results suggest that PMS may ameliorate Mhp-induced inflammatory response through NF-κB pathway,which provides important theoretical basis for novel prevention and treatment of Mhp infection.

关 键 词：大车前苷(PMS) 猪肺炎支原体(Mhp) 肺炎 核因子-ΚB 

分 类 号：S853.74[农业科学—临床兽医学] S852.62[农业科学—兽医学]