摘要
为探讨体外棕榈酸(Palmitic acid,PA)是否可以通过自噬调节奶牛淋巴细胞炎症信号通路的激活,分离健康奶牛淋巴细胞,采用3-MA(3-Methyladenin,细胞自噬抑制剂)和不同质量浓度PA作用于淋巴细胞,收集细胞及上清,利用qRT-PCR检测淋巴细胞LC3B、Beclin1、mTOR、UKL1、SQSTM1、IL-1β、IL-6和TNF-αmRNA表达情况;利用CCK-8和酶联免疫吸附法分别测定细胞活性和促炎细胞因子TNF-α、IL-6和IL-1β的释放量。CCK-8结果表明:PA显著抑制淋巴细胞活性(P<0.01),添加自噬抑制剂3-MA后细胞活性显著增强(P<0.01);qRT-PCR结果表明:与对照组(PA=0μg/mL)相比,PA处理组(不含3-MA)ULK1和LC3B mRNA表达极显著增加(P<0.01),mTOR、Beclin1和SQSTM1 mRNA表达极显著降低(P<0.01);IL-1β和IL-6 mRNA表达显著增强(P<0.01),而TNF-α未表达;添加3-MA后,与1μg/mL PA处理组相比较,SQSTM1和Beclin1极显著增加,而LC3B mRNA表达极显著降低(P<0.01);IL-1β和IL-6 mRNA表达显著下调(P<0.01);PA增加促炎因子IL-6和IL-1β的释放(P<0.01),显著抑制TNF-α释放(P<0.01),而含有3-MA的PA混合处理组则显著降低(P<0.01)。综上,PA可通过自噬调节奶牛淋巴细胞炎症信号通路的激活。
To investigate whether palmitic acid(PA)in dairy cows in vitro can regulate the activation of inflammatory signaling pathway through autophagy,lymphocytes isolated from healthy dairy cows were treated with 3-MA and different concentrations of PA and cells and supernatants were collected.The expressions of LC3 B,Beclin1,mTOR,UKL1,SQSTM1,IL-1β,IL-6 and TNF-αin lymphocytes were detected by qRT-PCR.The cell activity and release of proinflammatory cytokines TNF-α,IL-6 and IL-1βwere measured by CCK-8 and ELISA,respectively.The CCK-8 results showed that PA significantly inhibited lymphocyte activity(P<0.01),and its cell activity was significantly enhanced after 3-MA treatment(P<0.01);Compared with the control group(PA=0μg/mL),the expression levels of ULK1 and LC3 B mRNA in the PA treatment group(excluding 3-MA)were significantly increased(P<0.01),mTOR,Beclin1,and SQSTM1 mRNA were significantly decreased(P<0.01),IL-1βand IL-6 mRNA expression were significantly increased(P<0.01),while TNF-αwas not expressed.After the addition of 3-MA,SQSTM1 and Beclin1 were significantly increased,while LC3 B mRNA expression was significantly decreased(P<0.01);IL-1βandIL-6 mRNA expression were significantly down-regulated(P<0.01);PA increased the release of proinflammatory cytokines IL-6 and IL-1β(P<0.01),significantly inhibited the release of TNF-α(P<0.01),while it was significantly decreased in 3-MA+PA mixed treatment(P<0.01).In conclusion,PA can regulate the activation of lymphocyte inflammatory signaling pathway in cows through autophagy.
作者
李树森
杨晓燕
李莹
邓清华
薛江东
王新波
张玉明
杜立银
LI Shusen;YANG Xiaoyan;LI Ying;DENG Qinghua;XUE Jiangdong;WANG Xinbo;ZHANG Yuming;DU Liyin(College of Animal Science and Technology,Inner Mongolia University for Nationalities,Tongliao 028000,China)
出处
《中国农业大学学报》
CAS
CSCD
北大核心
2021年第1期43-50,共8页
Journal of China Agricultural University
基金
国家自然科学基金项目(31260626、31760752)
内蒙古自治区肉牛疾病防控工程技术研究中心开放课题(MDK2017021)
内蒙古自治区自然科学基金项目(2018LH03009)。
关键词
奶牛
棕榈酸
淋巴细胞
自噬
炎症
cow
palmitic acid
lymphocyte
autophagy
inflammation
作者简介
第一作者:李树森,硕士研究生,E-mail:shusenlil995@163.com;通讯作者:张玉明,讲师,主要从事动物代谢免疫学研究,E-mail:zhangyuminglll7@163.com;通讯作者:杜立银,教授,主要从事动物临床免疫学研究,E-mail:dly2000@aliyun.com。
