摘要
旨在探究异绿原酸C(ICAC)对乳腺炎及NF-κB炎性通路的抑制作用。本研究以脂多糖(LPS)分别刺激牛乳腺上皮细胞(MAC-T)和小鼠乳腺组织建立体外和体内炎性模型,采用MTT方法筛选ICAC处理MAC-T细胞的适宜浓度,ELISA检测炎性因子(IL-1β、IL-6和TNF-α)和炎性介导因子(COX-2和iNOS)的表达水平,Western blot方法检测NF-κB信号通路关键蛋白(IκB和p65)的表达水平和磷酸化水平。结果发现:1)ICAC在20~100 mg·L^(-1)的浓度下对MAC-T细胞没有显著的生长抑制作用(P>0.05);2)1 mg·L^(-1)的LPS处理引起MAC-T细胞中IL-1β、IL-6的蛋白质表达量极显著高于对照组(P<0.01),ICAC处理(20、50、80 mg·L^(-1))极显著下调了MAC-T细胞中IL-1β、IL-6、TNF-α、COX-2和iNOS的表达量(P<0.01);3)小鼠腹腔注射的ICAC(60、80、100 mg·kg^(-1))均降低了LPS诱导的CD3的表达,小鼠乳腺组织T淋巴细胞浸润减少;4)ICAC显著降低了LPS诱导的MAC-T细胞和小鼠乳腺组织中p65和IκB的磷酸化水平(P<0.05),抑制了p65的核转移。上述结果说明,异绿原酸C通过NF-κB炎性通路抑制MAC-T细胞和小鼠乳腺组织的炎性反应,但是异绿原酸C在奶牛乳腺炎防治中的应用有待进一步研究。
The research aims to investigate the inhibitory effect of isochlorogenic acid C(ICAC)on mastitis and NF-κB inflammatory pathway.In this study,bovine mammary epithelial cells(MAC-T)and mouse mammary tissue stimulated with lipopolysaccharide(LPS)were used as in vitro and in vivo inflammatory model,respectively.MTT method was used to screen the optimal concentrations of ICAC for MAC-T cell treatment.The expression levels of inflammatory factors(IL-1β,IL-6 and TNF-α)and inflammatory mediators(COX-2 and iNOS)were detected using ELISA,whereas the expression and phosphorylation levels of key proteins(IκB and p65)in NF-κB signaling pathway were detected by Western blot.The results showed that:1)ICAC did not significantly inhibit the growth of MAC-T cells at the concentrations from 20 to 100 mg·L^(-1)(P>0.05);2)The expression levels of IL-1βand IL-6 protein in MAC-T cells treated with 1 mg·L^(-1) LPS were significantly higher than those in untreated group(P<0.01),but ICAC(20,50,80 mg·L^(-1))significantly down-regulated the expression of IL-1β,IL-6,TNF-α,COX-2 and iNOS in MAC-T cells(P<0.01);3)Intraperitoneal inject ICAC(60、80、100 mg·kg^(-1))reduced LPS-induced CD3 expression and T lymphocyte infiltration in mammary tissues of mice;4)ICAC significantly reduced the phosphorylation of p65 and IκB in LPS-induced MAC-T cells and in mouse mammary tissue(P<0.05),and it inhibited the nuclear transfer of p65.These results indicate that ICAC inhibits the inflammatory response in MAC-T cells and mouse mammary tissue through NF-κB inflammatory pathway,but the role of ICAC in the prevention and treatment of bovine mastitis needs further investigation.
作者
陈喜宏
路桂聪
王浩磊
苟少校
玉永雄
林涛
蒋曹德
CHEN Xihong;LU Guicong;WANG Haolei;GOU Shaoxiao;YU Yongxiong;LIN Tao;JIANG Caode(Chongqing Engineering Research Center for Herbivores Resource Protection and Utilization,College of Animal Science and Technology,Southwest University,Chongqing 400715,China;Guang’an Feed Industry Management Station,Guang’an 638000,China)
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2023年第9期3931-3940,共10页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
重庆民生专项创新项目(cstc2017shms-kjfp0114,cstc2017shms-zdyfx0045)
中央高校基本科研业务费(XDJK2017A003)
国家自然科学基金(32171673)。
