摘要
本试验旨在研究表没食子儿茶素没食子酸酯(EGCG)通过核因子-κB(NF-κB)通路对大肠杆菌诱导型奶牛乳腺上皮细胞炎性损伤的影响。试验选用原代奶牛乳腺上皮细胞(pbMEC)为研究对象,分别用不同浓度的EGCG[0(对照)、20、40、60、80和100μmol/mL]培养12 h,测定细胞活力,确定以60μmol/mL的EGCG进行后续试验。将培养好的细胞分为3个组,分别为对照组、大肠杆菌处理组(1×10^(7))CFU大肠杆菌处理细胞6 h)和EGCG+大肠杆菌处理组(先用60μg/mL EGCG处理细胞24 h,再用1×10~7CFU大肠杆菌处理细胞6 h),每组设3个重复。结果表明:1)EGCG对大肠杆菌抑制的细胞增殖有一定的保护作用。2)EGCG显著降低了大肠杆菌诱导的pbMEC中丙二醛生成量的升高(P<0.05),明显降低了大肠杆菌诱导的pbMEC中活性氧含量的升高。3)EGCG显著降低了大肠杆菌诱导的pbMEC中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、诱导型一氧化氮合酶(iNOS)和Toll样受体4(TLR4)mRNA相对表达量的升高(P<0.05),显著降低了大肠杆菌诱导的pbMEC中TLR4、磷酸化p65(p-p65)和髓样分化因子88(MyD88)蛋白表达量的升高(P<0.05),并且明显降低了p-p65在大肠杆菌诱导的pbMEC中胞质和胞核中的表达。结果提示,EGCG能够抑制大肠杆菌诱导的奶牛乳腺上皮细胞炎症因子以及NF-κB通路相关蛋白的表达,对乳腺上皮细胞炎症反应以及氧化应激具有一定的缓解作用。
The aim of this study was to investigate the effects of epigallocatechin gallate(EGCG)on Escherichia coli induced inflammatory injury in bovine mammary epithelial cells via nuclear factor-κB(NF-κB)pathway.Primary bovine mammary epithelial cells(pbMEC)were selected and incubated with different concentrations of EGCG[0(control),20,40,60,80 and 100μmol/mL]for 12 h to determine the cell viability,and 60μmol/mL EGCG was determined for subsequent experiments.The cultured cells were divided into three groups as control group,Escherichia coli treatment group(treated with 1×10~7CFU Escherichia coli for 6 h)and EGCG+Escherichia coli treatment group(treated with 60μg/mL EGCG for 24 h,and then treated with 1×10^(7) CFU Escherichia coli for 6 h),with three replicates in each group.The results showed as follows:1)EGCG had a certain protective effect on cell proliferation inhibited by Escherichia coli.2)EGCG significantly decreased the increase of malondialdehyde production in Escherichia coli induced pbMEC(P<0.05),and obviously decreased the increase of reactive oxygen species content in Escherichia coli induced pbMEC.3)EGCG significantly decreased the increase of mRNA relative expression levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),inducible nitric oxide synthase(iNOS)and Toll-like receptor 4(TLR4)(P<0.05),significantly decreased the increase of the protein expressions of TLR4,phospho-p65(p-p65)and myeloid differentiation factor 88(MyD88)(P<0.05),and obviously decreased the increase of p-p65 expression in cytoplasm and nucleus in Escherichia coli induced pbMEC.The results suggest that EGCG can inhibit the expression of inflammatory factors and NF-κB pathway related proteins induced by Escherichia coli,and alleviate the inflammatory response and oxidative stress of mammary epithelial cells to a certain extent.[Chinese Journal of Animal Nutrition,2022,34(11):7370-7380]
作者
徐天乐
朱浩
刘润
梁艳
曹海南
杨章平
XU Tianle;ZHU Hao;LIU Run;LIANG Yan;CAO Hainan;YANG Zhangping(Institutes of Agricultural Science and Technology Development(Joint International Research Laboratory of Agriculture&Agri-Product Safety),Yangzhou University,Yangzhou 225000,China;College of Animal Science and Technology,Yangzhou University,Yangzhou 225000,China)
出处
《动物营养学报》
CAS
CSCD
北大核心
2022年第11期7370-7380,共11页
CHINESE JOURNAL OF ANIMAL NUTRITION
基金
国家自然科学基金项目(31872324)
国家自然科学基金青年项目(32102731)
教育部农业与农产品安全国际合作联合实验室开放课题(JILAR-KF202204)。
