LU Jinye,GU Beibei,LU Wei,et al.Rapamycin attenuates Escherichia coli induced inflammatory response in rat mammary epithelial cells[J].Journal of Nanjing Agricultural University,2019,42(4):740-745.[doi:10.7685/jnau.201806033]





Rapamycin attenuates Escherichia coli induced inflammatory response in rat mammary epithelial cells
卢劲晔 顾蓓蓓 卢炜 刘静
江苏农牧科技职业学院, 江苏 泰州 225300
LU Jinye GU Beibei LU Wei LIU Jing
Jiangsu Agri-animal Husbandry Vocational College, Taizhou 225300, China
Escherichia coliinflammation responsesrapamycinmammary epithelial cells
[目的]阐明雷帕霉素(Rapa)对大肠杆菌诱发的乳腺感染的保护作用机制,为临床上寻找乳腺炎防控新方法提供理论依据。[方法]原代培养大鼠乳腺上皮细胞,待细胞贴壁后,试验组采用100 nmol·L-1的Rapa处理4 h后,细胞经感染比(MOI)为10的大肠杆菌处理2 h后用细胞刮刀刮取细胞,收集细胞上清液,用于Toll样受体4(TLR-4)及髓样分化因子(MyD88)蛋白、促炎性细胞因子表达及活性氧(ROS)释放检测。[结果]大肠杆菌刺激原代培养的乳腺上皮细胞后,TLR-4和MyD88蛋白表达显著升高;而Rapa预处理可降低MyD88蛋白表达;同时,大肠杆菌刺激后,乳腺上皮细胞中促炎性细胞因子,如肿瘤坏死因子α(TNF-α)、白介素1β(IL-1β)的表达及ROS的释放均显著升高。雷帕霉素能下调促炎性细胞因子的释放,显著降低ROS的释放量。[结论]Rapa抑制MyD88信号通路后可减轻大肠杆菌诱导的大鼠乳腺炎症反应。
[Objectives]The present study aimed to investigate the effect of rapamycin(Rapa)on inflammatory response in Escherichia coli(E.coli)induced rat mastitis model.[Methods]Primary mammary epithelial cells were separated and when cells fused to 80%,the monolayers in treated group were treated with 100 nmol·L-1 Rapa for 4 h. Then cells were infected with E.coli in logarithmic phase at a multiplicity of infection(MOI)of 10 for 2 h at 37℃. Cells and the supernatant were collected.[Results]The protein level of Toll like receptor 4(TLR-4)and Myeloid differentiation factor 88(MyD88)significantly increased in E.coli stimulated primary rat mammary epithelial cells while the elevated expression of MyD88 was significantly suppressed by Rapa pretreatment compared with the control. The pro-inflammatory cytokines level,such as TNF-α,IL-1β expression was all significantly up-regulated by E.coli stimulation. Besides that,ROS releasing and the level of MDA also increased and the anti-oxidant indices of SOD and T-AOC increased by E.coli stimulation. However,Rapa treatment effectively attenuated E.coli-induced inflammatory responses by suppressing the over-expression of pro-inflammatory cytokines,decreasing the release of ROS and strengthening the anti-oxidant system.[Conclusions]The treatment of Rapa abolished E.coli-induced inflammatory response by inactivating the MyD88 signaling pathway.


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更新日期/Last Update: 1900-01-01