产前PFOA暴露对新生鼠肺组织免疫细胞功能的影响
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摘要
目的通过动物模型与实验方法探究产前全氟辛酸(PFOA)暴露对新生鼠肺组织细胞免疫功能的影响。方法建立产前PFOA暴露(1、5、10 mg/kg)孕鼠模型,并设置正常对照组,持续每天灌胃至分娩。出生d1的小鼠(PND1)断尾检测血糖值,并收集新鲜肺组织用于生化法测定肺匀浆中乳酸脱氢酶(LDH)水平,酶联免疫吸附测定(ELISA)法测定肺匀浆中白细胞介素6(IL-6)和干扰素γ(IFN-γ)含量,免疫荧光法检测分析肺组织细胞中CD29和CD73阳性细胞数量,蛋白免疫印迹(western blotting)法检测肺组织细胞中CD29和CD73蛋白水平的表达。结果与对照小鼠比较,产前PFOA干预组中PND1小鼠的血糖有升高趋势(P>0.05),肺匀浆中LDH,IL-6,IFN-γ含量增加(P<0.05),肺组织细胞中CD29和CD73阳性细胞显著减少(P<0.05),同时肺组织细胞中CD29和CD73蛋白表达相应下调(P<0.05),体现出剂量依赖性。结论产前PFOA暴露对新生鼠肺组织细胞免疫功能有抑制效应,其分子机制可能与钝化血糖依赖性的CD29/73信号通路有关。
Objective To investigate the effects of prenatal perfluorooctanoic acid(PFOA) exposure on cellular immune function in neonatal rat lung tissue via animal models and experimental methods. Methods To establish a prenatal PFOA exposure(1,5,10 mg/kg,w/w) pregnant mice model, and to set up a normal control group, all groups being for continue daily until delivery. The postnatal day 1(PND1) was tested for blood glucose from tail-cutting blood, and fresh lung tissue was collected for biochemical determination of lactate dehydrogenase(LDH) levels in lung homogenate. The levels of interleukin-6(IL-6) and interferon gamma(IFN-γ) in homogenate were detected by enzyme-linked immunosorbent assay(ELISA). The cell counts of CD29 and CD73 positive cells in lung tissue cells were assayed through immunofluorescence stains.Western blotting tests were used to detect intracellular expression of CD29 and CD73 protein levels in the lung. Results Compared with the control mice, the blood glucose of PND1 mice in the prenatal PFOA intervention group increased(P>0.05), and the levels of LDH,IL-6 and IFN-γ in the lung homogenate increased(P<0.05). The number of CD29 and CD73 positive cells in the cells was significantly decreased(P<0.05), and the expression of CD29 and CD73 protein in lung tissue cells was down-regulated(P <0.05), showing dose-dependently. Conclusion Prenatal PFOA exposure has an inhibitory effect on cellular immune function in lung tissue of neonatal mice, and its molecular mechanism may be related to the inactivation of glycemic-dependent CD29/73 signaling pathway.
Objective To investigate the effects of prenatal perfluorooctanoic acid(PFOA) exposure on cellular immune function in neonatal rat lung tissue via animal models and experimental methods. Methods To establish a prenatal PFOA exposure(1,5,10 mg/kg,w/w) pregnant mice model, and to set up a normal control group, all groups being for continue daily until delivery. The postnatal day 1(PND1) was tested for blood glucose from tail-cutting blood, and fresh lung tissue was collected for biochemical determination of lactate dehydrogenase(LDH) levels in lung homogenate. The levels of interleukin-6(IL-6) and interferon gamma(IFN-γ) in homogenate were detected by enzyme-linked immunosorbent assay(ELISA). The cell counts of CD29 and CD73 positive cells in lung tissue cells were assayed through immunofluorescence stains.Western blotting tests were used to detect intracellular expression of CD29 and CD73 protein levels in the lung. Results Compared with the control mice, the blood glucose of PND1 mice in the prenatal PFOA intervention group increased(P>0.05), and the levels of LDH,IL-6 and IFN-γ in the lung homogenate increased(P<0.05). The number of CD29 and CD73 positive cells in the cells was significantly decreased(P<0.05), and the expression of CD29 and CD73 protein in lung tissue cells was down-regulated(P <0.05), showing dose-dependently. Conclusion Prenatal PFOA exposure has an inhibitory effect on cellular immune function in lung tissue of neonatal mice, and its molecular mechanism may be related to the inactivation of glycemic-dependent CD29/73 signaling pathway.
引文
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[2]高宏平,常薇,张磊,等.纺织品中全氟辛烷磺酸及全氟辛酸的测定及其研究进展[J].产业用纺织品,2011,29(9):41-44.
[3]Yatim KM,Lakkis FG.A brief journey through the immune system[J].Clinical Journal of the American Society of Nephrology,2015,10(7):1274-1281.
[4]王玉.全氟辛烷磺酸(PFOS)和全氟辛酸(PFOA)对BALB/c小鼠免疫系统毒性效应研究[D].武汉:华中农业大学,2011.
[5]Zhang HJ,Fang WD,Wang DD,et al.The role of interleukin family in perfluorooctanoic acid(PFOA)-induced immunotoxicity[J].Journal of Hazardous Materials,2014,280:552-560.
[6]Qin XY,Xie GJ,Wu XM,et al.Prenatal exposure to perfluorooctanoic acid induces nerve growth factor expression in cerebral cortex cells of mouse offspring[J].Environmental Science and Pollution Research,2018,25(19):18914-18920.
[7]Wu XM,Liang MQ,Yang Z,et al.Effect of acute exposure to PFOAon mouse liver cells in vivo and in vitro[J].Environmental Science and Pollution Research,2017,24(31):24201-24206.
[8]Wu X,Xie G,Xu X,et al.Adverse bioeffect of perfluorooctanoic acid on liver metabolic function in mice[J].Environmental Science and Pollution Research International,2018,25(5):4787-4793.
[9]Augoff K,Hryniewicz-Jankowska A,Tabola R.Lactate dehydrogenase 5:an old friend and a new hope in the war on cancer[J].Cancer Letters,2015,358(1):1-7.
[10]Hunter CA,Jones SA.IL-6 as a keystone cytokine in health and disease[J].Nature Immunology,2015,16(5):448-457.
[11]Lazear HM,Nice TJ,Diamond MS.Interferon-λ:Immune functions at barrier surfaces and beyond[J].Immunity,2015,43(1):15-28.
[12]Ginsberg MH.Integrin activation[J].BMB Reports,2014,47(12):655-659.
[13]Allard D,Allard B,Gaudreau P,et al.CD73-adenosine:a nextgeneration target in immuno-oncology[J].Immunotherapy,2016,8(2):145-163.