| 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-α mRNA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=0 μg/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1 μg/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。 |
| 关键词: 奶牛 棕榈酸 淋巴细胞 自噬 炎症 |
| DOI：10.11841/j.issn.1007-4333.2021.01.05 |
| 投稿时间：2020-02-14 |
| 基金项目: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) |
|
| Palmitic acid induces autophagy to regulate lymphocytic inflammatory pathways in dairy cows |
|
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) |
| Abstract: |
| 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 LC3B, Beclin1, mTOR, UKL1, SQSTM1, IL-1β, IL-6 and TNF-α in lymphocytes were detected by qRT-PCR. The cell activity and release of proinflammatory cytokines TNF-α, IL-6 and IL-1β were 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=0 μg/mL), the expression levels of ULK1 and LC3B 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-1β and IL-6 mRNA expression were significantly increased(P<0. 01), while TNF-α was not expressed. After the addition of 3-MA, SQSTM1 and Beclin1 were significantly increased, while LC3B mRNA expression was significantly decreased(P<0. 01); IL-1β and IL-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. |
| Key words: cow palmitic acid lymphocyte autophagy inflammation |
