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作 者:何水净 丛妍 何秀 王开露 裴秀丛 HE Shuijing;CONG Yan;HE Xiu;WANG Kailu;PEI Xiucong(Postgraduate Majoring in Toxicology,School of Public Health,Shenyang Medical College,Shenyang 110034,China;Department of Epidemiology and Health Statistics,School of Public Health,Guizhou Medical University;Department of Toxicology,School of Public Health,Shenyang Medical College)
机构地区:[1]沈阳医学院公共卫生学院,辽宁沈阳110034 [2]贵州医科大学公共卫生与健康学院流行病与卫生统计学系 [3]沈阳医学院公共卫生学院毒理学教研室
出 处:《沈阳医学院学报》2023年第6期582-587,597,共7页Journal of Shenyang Medical College
基 金:辽宁省教育厅科学研究项目(No.LJKZ1145);辽宁省科学技术计划创新能力提升联合基金(No.2022-NLTS-14-02);沈阳医学院研究生课题(No.Y20220523)。
摘 要:目的:探讨Raf/MEK/ERK和Ca^(2+)/CaN信号通路在双酚A (BPA)调控巨噬细胞分泌IL-10中的作用。方法:以小鼠单核白血病巨噬细胞(RAW264.7)为靶细胞,2μg/ml脂多糖(LPS)为激活剂,采用1、10、100μmol/L BPA染毒。以10、 30μmol/L钙离子通道阻断剂(Ver), 0.25、 0.5μmol/LCaN阻断剂(FK506)和10、 40μmol/LERK抑制剂(PD98059)作为干预剂加入100μmol/LBPA,分别作用细胞4、12、24h。采用实时定量PCR技术测定ARaf、CRaf、MEK、ERK1、ERK2、NFATp、NFATc及IL-10的基因表达,采用ELISA方法检测细胞上清液IL-10的蛋白含量。结果:与对照组(0μmol/L BPA)相比,2μg/ml LPS组IL-10基因和蛋白表达水平升高,CRaf、MEK、ERK1、ERK2、NFATp和NFATc基因表达水平升高;与LPS组比较,100μmol/LBPA组IL-10的基因与蛋白表达水平降低,ARaf、CRaf、MEK、ERK1、ERK2、NFATp和NFATc基因表达水平升高;与100μmol/LBPA组比较,Ver组和FK506组IL-10基因和蛋白表达水平升高,NFATp和NFATc基因表达水平降低,而高浓度PD98059组IL-10基因和蛋白表达水平降低,NFATc表达水平降低而NFATp表达水平升高,差异均有统计学意义。结论:本实验条件下,Raf/MEK/ERK和Ca^(2+)/CaN信号通路共同调控NFAT,进而介导了BPA对巨噬细胞分泌细胞因子IL-10的影响,具体机制还需进一步探讨。Objective:To investigate the role of Ca^(2+)/CaN and Raf/MEK/ERK signaling pathways in the regulation of IL-10 secretion by BPA in macrophages.Methods:Mouse mononuclear leukemia macrophages(RAW264.7)were used as target cells,2μg/ml lipopolysaccharide(LPS)was used as activator,1,10 and 100μmol/L BPA was used as poisoning agents.And 10,30μmol/L calcium channel blocker(Ver),0.25 and 0.5μmol/L CaN blocker(FK506),10 and 40μmol/L ERK inhibitor(PD98059)were added with 100μmol/L BPA as intervention agents,then the cells were treated for 4,12 and 24 h,respectively.The gene expressions of ARaf,CRaf,MEK,ERK1,ERK2,NFATp,NFATc and IL-10 were detected by real-time PCR.The protein content of IL-10 in cell supernatant was detected by ELISA.Results:Compared with the control group,the mRNA and protein expressions of IL-10,as well as the mRNA expressions of CRaf,MEK,ERK1,ERK2,NFATp and NFATc were significantly increased in 2μg/ml LPS group.Compared with LPS group,the mRNA and protein expressions of IL-10 were decreased,and the mRNA expressions of ARaf,CRaf,MEK,ERK1,ERK2,NFATp and NFATc were increased in 100μmol/L BPA group.Compared with 100μmol/L BPA group,the mRNA and protein expressions of IL-10 were increased,and the mRNA expressions of NFATp and NFATc were decreased in Ver group and FK506 group.The mRNA and protein expressions of IL-10 were decreased,the expression of NFATc was decreased and the expression of NFATp was increased in high concentration of PD98059 group. The differences were all statistically significant. Conclusion:Raf/MEK/ERK and Ca^(2+)/CaN signaling pathways jointly regulate NFAT,thereby mediating the effect of BPA on the secretion of IL-10 inmacrophages,but the specific mechanism needs to be further explored.
关 键 词:双酚A 白细胞介素-10 巨噬细胞 Raf/MEK/ERK Ca^(2+)/CaN
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