孕期和哺乳期壬基酚暴露对后代小鼠神经免疫系统改变的影响及相关机制研究  

Effects of pregnancy and lactation nonylphenol exposure on the changes of neuroimmune function in offspring mice and its related mechanisms

作　　者：史许锋 刘侃[1] 宋婉玉 王莉[1] 武海英[1] Shi Xufeng;Liu Kan;Song Wanyu;Wang Li;Wu Haiying(Department of Obstetric,Henan Provincial People's Hospital,Zhengzhou 450003,China)

机构地区：[1]河南省人民医院产科,郑州450003

出　　处：《中华微生物学和免疫学杂志》2021年第3期174-181,共8页Chinese Journal of Microbiology and Immunology

摘　　要：目的探讨孕期和哺乳期壬基酚(nonylphenol,NP)暴露对后代小鼠脑组织Th17/Treg细胞平衡的影响及其相关机制研究。方法30只C57BL/6孕鼠随机分为3组:对照组(饮用蒸馏水)和NP处理组(饮用0.2μg/ml或2.0μg/ml NP水溶液)。使用ELISA试剂盒测量仔鼠血清中的TNF-α、IFN-γ和IL-17水平,流式细胞术分析仔鼠脾脏Treg、Th17细胞频率,定量RT-PCR分析仔鼠脑组织中RORγt、Foxp3 mRNA表达,Western blot分析仔鼠脑组织中RORγt、Foxp3和PI3K/Akt/mTOR信号通路相关蛋白质表达,免疫荧光分析仔鼠脑组织中离子钙结合衔接分子1(Iba1)表达情况。结果与对照组相比,NP暴露增加了雄性后代小鼠的血清IL-17、TNF-α水平(P<0.05),降低了IFN-γ水平(P<0.05)。流式细胞术分析显示,NP暴露(0.2μg/ml或2.0μg/ml)的雄性后代小鼠脾脏中Th17细胞的百分比较对照组显著增加,而Tregs细胞的百分比降低。与对照组相比,NP暴露(0.2μg/ml或2.0μg/ml)的雄性后代小鼠脑组织中Foxp3蛋白质水平显著降低,RORγt蛋白质水平显著升高(P<0.05)。在qRT-PCR分析中也观察到类似的mRNA表达结果。在孕期和哺乳期暴露于剂量增加的NP期间,雄性后代小鼠脑组织中mTOR(p-mTOR)及其上游相关调节因子[PI3K、p-Akt(Ser473)、p-Akt(Thr308)]的蛋白质表达水平均升高(P<0.05)。免疫荧光分析显示,与对照组相比,NP暴露(0.2μg/ml或2.0μg/ml)的雄性后代小鼠脑组织中Iba1阳性细胞的数量显著增加(P<0.05)。结论孕期和哺乳期小鼠暴露于NP可能通过神经免疫轴影响后代神经元的发育/功能,增加后代神经发育障碍的风险。Objective To investigate the effects of pregnancy and lactation nonylphenol(NP)exposure on the balance of Treg/Th17 cells in the brain of offspring mice and the related mechanisms.Methods Thirty pregnant C57BL/6 mice were randomly divided into three groups:control group(drinking distilled water),and NP-treated groups(drinking 0.2μg/ml or 2.0μg/ml NP water solution).ELISA kit was used to analyze the levels of TNF-α,IFN-γ and IL-17,flow cytometry was used to analyze the frequency of Treg and Th17 cells in spleen,quantitative RT-PCR was used to analyze the RORγt,Foxp3 mRNA,Western blot was used to analyze the protein expression of RORγt,Foxp3 and PI3K/Akt/mTOR signal pathway,and immunofluorescence was used to analyze the expression of Iba1 in the brain tissue of offspring mice.Results Compared with the control group,NP exposure increased the serum levels of IL-17 and TNF-α in male offspring mice(P<0.05),and decreased the levels of IFN-γ(P<0.05).Flow cytometry analysis showed that the percentage of Th17 cells in the spleen of male offspring mice exposed to NP(0.2μg/ml or 2.0μg/ml)was significantly higher than that of the control group,while the percentage of Tregs cells was lower.Compared with the control group,the expression levels of Foxp3 proteins in the brain tissue of male offspring mice exposed to NP(0.2μg/ml or 2.0μg/ml)was significantly lower,accompanied by a dramatic increase in RORγt protein levels(P<0.05).Similar mRNA expression was also observed in qRT-PCR analysis.The protein expression levels of mTOR(p-mTOR)and its upstream related regulators[PI3K,p-Akt(Ser473),p-Akt(Thr308)]in the brain of male offspring mice increased gradually during the period of exposure to NP(P<0.05).Immunofluorescence analysis showed that compared with the control group,the number of Iba1 positive cells in brain tissue of male offspring mice exposed to NP(0.2μg/ml or 2.0μg/ml)increased significantly(P<0.05).Conclusions Maternal exposure to NP during pregnancy and lactation may affect the development/function of neu

关 键 词：孕期 哺乳期 小鼠 壬基酚 Th17/Treg细胞平衡 PI3K/Akt/mTOR信号通路 

分 类 号：R114[医药卫生—卫生毒理学]