黄曲霉毒素B1激活核转录因子-κB信号通路引起牛乳腺上皮细胞发生炎性反应  

Aflatoxin B1 Induces Inflammatory Response of Bovine Mammary Epithelial Cells by Activating Nuclear Transcription Factor⁃κB Signaling Pathway

作　　者：任晓丽 皇甫和平[1] 王军[1] 靳双星[1] 王俊东 石冬梅[1] REN Xiaoli;HUANGFU Heping;WANG Jun;JIN Shuangxing;WANG Jundong;SHI Dongmei(Zhengzhou Key Laboratory of Animal Nutritional Metabolic and Poisoning Diseases,College of Veterinary Medicine,Henan University of Animal Husbandry of Economy,Zhengzhou 450046,China)

机构地区：[1]河南牧业经济学院动物医药学院,郑州市动物营养代谢病与中毒病重点实验室,郑州450046

出　　处：《动物营养学报》2023年第7期4587-4595,共9页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：河南省科技攻关项目(222102110110,212102110377);河南省教育厅重点科研项目(22B230004);河南牧业经济学院校级重点学科建设项目(XJXK202202)。

摘　　要：本试验旨在研究不同浓度黄曲霉毒素B1(AFB1)对牛乳腺上皮细胞(MAC-T细胞)中核转录因子-κB(NF-κB)信号通路介导的炎症反应的影响。向MAC-T细胞中分别加入不同浓度的AFB1处理培养液培养24 h,分析AFB1对MAC-T细胞活力、乳酸脱氢酶(LDH)活性、促炎细胞因子含量和基因表达以及NF-κB信号通路相关蛋白表达的影响。结果显示:1)与对照组相比,2.5、5.0、10.0和20.0μmol/L AFB1处理MAC-T细胞均能显著或极显著降低细胞活力(P<0.05或P<0.01),显著或极显著升高培养上清液中乳酸脱氢酶(LDH)活性(P<0.05或P<0.01),且呈剂量依赖性。2)与对照组(0μmol/L AFB1)相比,5.0、10.0和20.0μmol/L AFB1处理MAC-T细胞使上清液中白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)的含量显著或极显著升高(P<0.05或P<0.01);5.0μmol/L AFB1处理使MAC-T细胞中IL-1β、IL-8和TNF-αmRNA的相对表达量显著或极显著升高(P<0.05),10.0μmol/L AFB1处理使MAC-T细胞中IL-1β、IL-6、IL-8和TNF-αmRNA的相对表达量显著或极显著升高(P<0.01),20.0μmol/L AFB1处理使MAC-T细胞中IL-1β、IL-6、IL-8和TNF-αmRNA的相对表达量极显著升高(P<0.01)。3)与对照组相比,10.0和20.0μmol/L AFB1处理使MAC-T细胞中磷酸化P65(p-P65)、磷酸化核转录因子-κB抑制因子α(p-IκBα)蛋白的相对表达量极显著升高(P<0.01)。上述结果提示,AFB1通过促进炎症相关因子表达和激活NF-κB信号通路,引起MAC-T细胞发生炎性反应。This study aimed to investigate the effects of different concentrations of aflatoxin B1(AFB1)on in⁃flammation response mediated by nuclear transcription factor⁃κB(NF⁃κB)signaling pathway in bovine mam⁃mary epithelial cells(MAC⁃T cells).The MAC⁃T cells were treated with AFB1 at different concentrations for 24 h.The effects of AFB1 on the cell viability,the lactic dehydrogenase(LDH)activity,the contents and gene expression of inflammatory cytokines,and the expression of NF⁃κB signaling pathway related proteins in MAC⁃T cells were analyzed.The results showed as follows:1)compared with the control group(0μmol/L AFB1),the MAC⁃T cells treated with 2.5,5.0,10.0 and 20.0μmol/L AFB1 could significantly or extremely significantly reduce the cell viability and increase the LDH activity in culture supernatant in a dose⁃dependent manner(P<0.05 or P<0.01).2)Compared with the control group,the contents of interleukin⁃1β(IL⁃1β),interleukin⁃6(IL⁃6),interleukin⁃8(IL⁃8)and tumor necrosis factor⁃α(TNF⁃α)in the culture supernatant of MAC⁃T cells treated with 5.0,10.0 and 20.0μmol/L AFB1 were significantly or extremely significantly in⁃creased(P<0.05 or P<0.01);moreover,the mRNA relative expression levels of IL⁃1β,IL⁃8 and TNF⁃αin MAC⁃T cells treated with 5.0μmol/L AFB1 were significantly or extremely significantly increased(P<0.05 or P<0.01),the mRNA relative expression levels of IL⁃1β,IL⁃6,IL⁃8 and TNF⁃αin MAC⁃T cells treated with 10.0μmol/L AFB1 were significantly or extremely significantly increased(P<0.05 or P<0.01),and the mRNA relative expression levels of IL⁃1β,IL⁃6,IL⁃8 and TNF⁃αin MAC⁃T cells treated with 20.0μmol/L AFB1 were extremely significantly increased(P<0.01).3)Compared with the control group,the protein rela⁃tive expression levels of phosphorylated⁃P65(p⁃P65)and phosphorylated inhibitorαof NF⁃κB(p⁃IκBα)in MAC⁃T cells treated with 10.0 and 20.0μmol/L AFB1 were extremely significantly increased(P<0.01).In conclusion

关 键 词：黄曲霉毒素B1 牛乳腺上皮细胞 NF-ΚB信号通路 炎性反应 

分 类 号：S823[农业科学—畜牧学]