雌二醇对原代小胶质细胞铁相关蛋白表达的影响
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摘要
目的探讨雌二醇(E2)对原代小胶质细胞铁相关蛋白表达的影响。方法原代培养Wistar大鼠腹侧中脑的小胶质细胞,分离纯化后将细胞分为对照组和E2组,其中E2组培养液内加入E2(终浓度为10nmol/L),对照组培养液内加入等体积的二甲基亚砜,两组作用时间均为24h。采用蛋白质免疫印迹实验检测两组细胞二价金属离子转运蛋白1(DMT1)、膜铁转运蛋白1(FPN1)和铁调节蛋白1(IRP1)的表达。结果与对照组相比,E2组细胞DMT1的表达水平显著增高(t=2.937,P<0.05),FPN1的表达水平显著降低(t=2.294,P<0.05);两组IRP1的表达水平比较差异无显著性(P>0.05)。结论 10nmol/L的E2可引起原代小胶质细胞DMT1的表达增加、FPN1的表达降低,且这种变化与IRP1调节无关。
Objective To investigate the effect of estradiol(E2)on the expression of iron-related proteins in primary cultured microglial cells. Methods Primary cultured microglial cells from the ventral midbrain of Wistar rats were divided into control group and E2 group after isolation and purification.The cells in the E2 group were treated with E2 at a final concentration of10 nmol/L for 24 hand those in the control group were treated with an equal volume of dimethyl sulfoxide for 24 h.Western blot was used to measure the expression of divalent metal transporter 1(DMT1),ferroportin 1(FPN1),and iron regulatory protein 1(IRP1)in cells. Results Compared with the control group,the E2 group had significantly higher expression of DMT1(t=2.937,P<0.05)and significantly lower expression of FPN1(t=2.294,P<0.05).There was no significant difference in the expression of IRP1 between the two groups(P>0.05). Conclusion E2 at a concentration of 10 nmol/L can increase the expression of DMT1 and reduce the expression of FPN1 in primary cultured microglial cells,which is not associated with IRP1 regulation.
Objective To investigate the effect of estradiol(E2)on the expression of iron-related proteins in primary cultured microglial cells. Methods Primary cultured microglial cells from the ventral midbrain of Wistar rats were divided into control group and E2 group after isolation and purification.The cells in the E2 group were treated with E2 at a final concentration of10 nmol/L for 24 hand those in the control group were treated with an equal volume of dimethyl sulfoxide for 24 h.Western blot was used to measure the expression of divalent metal transporter 1(DMT1),ferroportin 1(FPN1),and iron regulatory protein 1(IRP1)in cells. Results Compared with the control group,the E2 group had significantly higher expression of DMT1(t=2.937,P<0.05)and significantly lower expression of FPN1(t=2.294,P<0.05).There was no significant difference in the expression of IRP1 between the two groups(P>0.05). Conclusion E2 at a concentration of 10 nmol/L can increase the expression of DMT1 and reduce the expression of FPN1 in primary cultured microglial cells,which is not associated with IRP1 regulation.
引文
[1]SHEFTEL A,STEHLING O,LILL R.Iron-sulfur proteins in health and disease[J].Trends in Endocrinology and Metabolism,2010,21(5):302-314.
[2]GOZZELINO R,JENEY V,SOARES M P.Mechanisms of cell protection by heme oxygenase-1[J].Annual Review of Pharmacology and Toxicology,2010,50:323-354.
[3]EVSTATIEV R,GASCHE C.Iron sensing and signalling[J].Gut,2012,61(6):933-952.
[4]KELL D B.Towards a unifying,systems biology understanding of large-scale cellular death and destruction caused by poorly liganded Iron:Parkinson’s,Huntington’s,Alzheimer’s,prions,bactericides,chemical toxicology and others as examples[J].Archives of Toxicology,2010,84(11):825-889.
[5]KUMAR H,LIM H W,MORE S V,et al.The role of free radicals in the aging brain and Parkinson’s disease:convergence and parallelism[J].International Journal of Molecular Sciences,2012,13(8):10478-10504.
[6]DIANA R H,IRIS C Y,GHEORGHE D P,et al.Alterations in brain transition metals in huntington disease:an evolving and intricate story[J].Archives of Neurology,2013,69(7):887-893.
[7]陈瑛,竺飞燕,王炼,等.DMT1在多巴胺能神经元变性中的作用研究[J].中国病理生理杂志,2011,27(2):350-356.
[8]FARINA M,AVILA D S,DA ROCHA J B,et al.Metals,oxidative stress and neurodegeneration:a focus on iron,manganese and mercury[J].Neurochemistry International,2013,62(5):575-594.
[9]SCHIPPER H M.Neurodegeneration with brain iron accumulation-clinical syndromes and neuroimaging[J].Biochimica et Biophysica Acta,2012,1822(3):350-360.
[10]BRUCE-KELLER A J,KEELING J L,KELLER J N,et al.Antiinflammatory effects of estrogen on microglial activation[J].Endocrinology,2000,141(10):3646-3656.
[11]DREW P D,CHAVIS J A.Female sex steroids:effects upon microglial cell activation[J].Journal of Neuroimmunology,2000,111(1/2):77-85.
[12]VEGETO E,BELCREDITO S,GHISLETTI S,et al.The endogenous estrogen status regulates microglia reactivity in animal models of neuroinflammation[J].Endocrinology,2006,147(5):2263-2272.
[13]VEGETO E,BONINCONTRO C,POLLIO G,et al.Estrogen prevents the lipopolysaccharide-induced inflammatory response in microglia[J].The Journal of Neuroscience,2001,21(6):1809-1818.
[14]XU Manman,TAN Xu,LI Na,et al.Differential regulation of estrogen in iron metabolism in astrocytes and neurons[J].Journal of Cellular Physiology,2018.doi:10.1002/jcp.27188.
[15]GUNSHIN H,MACKENZIE B,BERGER U V,et al.Cloning and characterization of a mammalian proton-coupled metal-ion transporter[J].Nature,1997,388(6641):482-488.
[16]HUANG E,ONG W Y,CONNOR J R.Distribution of divalent metal transporter-1in the monkey basal ganglia[J].Neuroscience,2004,128(3):487-496.
[17]SALAZAR J,MENA N,HUNOT S,et al.Divalent metal transporter 1(DMT1)contributes to neurodegeneration in animal models of Parkinson’s disease[J].PNAS,2008,105(47):18578-18583.
[18]MCKIE A T,MARCIANI P,ROLFS A,et al.A novel duodenal iron-regulated transporter,IREG1,implicated in the basolateral transfer of iron to the circulation[J].Molecular Cell,2000,5(2):299-309.
[19]ABBOUD S,HAILE D J.A novel mammalian iron-regulated protein involved in intracellular Iron metabolism[J].Journal of Biological Chemistry,2000,275(26):19906-19912.
[20]DONOVAN A,BROWNLIE A,ZHOU Y,et al.Positional cloning of zebrafish ferroportin1identifies a conserved vertebrate Iron exporter[J].Nature,2000,403(6771):776-781.
[21]CAMASCHELLA C.Understanding iron homeostasis through genetic analysis of hemochromatosis and related disorders[J].Blood,2005,106(12):3710-3717.
[22]CHRISTOVA T,TEMPLETON D M.Effect of hypoxia on the binding and subcellular distribution of Iron regulatory proteins[J].Molecular and Cellular Biochemistry,2007,301(1/2):21-32.
[23]WANG Jun,XU Huamin,YANG Haidong,et al.Rg1reduces nigral iron levels of MPTP-treated C57BL6mice by regulating certain iron transport proteins[J].Neurochemistry International,2009,54(1):43-48.
[24]JIANG Hong,SONG Ning,XU Huamin,et al.Up-regulation of divalent metal transporter 1in 6-hydroxydopamine intoxication is IRE/IRP dependent[J].Cell Research,2010,20(3):345-356.
[25]QIAN Yi,YIN Chunyang,CHEN Yue,et al.Estrogen contributes to regulating iron metabolism through governing ferroportin signaling via an estrogen response element[J].Cellular Signalling,2015,27(5):934-942.
[26]RASO G M,IRACE C,ESPOSITO E A,et al.Ovariectomy and estrogen treatment modulate iron metabolism in rat adipose tissue[J].Biochemical Pharmacology,2009,78(8):1001-1007.
[2]GOZZELINO R,JENEY V,SOARES M P.Mechanisms of cell protection by heme oxygenase-1[J].Annual Review of Pharmacology and Toxicology,2010,50:323-354.
[3]EVSTATIEV R,GASCHE C.Iron sensing and signalling[J].Gut,2012,61(6):933-952.
[4]KELL D B.Towards a unifying,systems biology understanding of large-scale cellular death and destruction caused by poorly liganded Iron:Parkinson’s,Huntington’s,Alzheimer’s,prions,bactericides,chemical toxicology and others as examples[J].Archives of Toxicology,2010,84(11):825-889.
[5]KUMAR H,LIM H W,MORE S V,et al.The role of free radicals in the aging brain and Parkinson’s disease:convergence and parallelism[J].International Journal of Molecular Sciences,2012,13(8):10478-10504.
[6]DIANA R H,IRIS C Y,GHEORGHE D P,et al.Alterations in brain transition metals in huntington disease:an evolving and intricate story[J].Archives of Neurology,2013,69(7):887-893.
[7]陈瑛,竺飞燕,王炼,等.DMT1在多巴胺能神经元变性中的作用研究[J].中国病理生理杂志,2011,27(2):350-356.
[8]FARINA M,AVILA D S,DA ROCHA J B,et al.Metals,oxidative stress and neurodegeneration:a focus on iron,manganese and mercury[J].Neurochemistry International,2013,62(5):575-594.
[9]SCHIPPER H M.Neurodegeneration with brain iron accumulation-clinical syndromes and neuroimaging[J].Biochimica et Biophysica Acta,2012,1822(3):350-360.
[10]BRUCE-KELLER A J,KEELING J L,KELLER J N,et al.Antiinflammatory effects of estrogen on microglial activation[J].Endocrinology,2000,141(10):3646-3656.
[11]DREW P D,CHAVIS J A.Female sex steroids:effects upon microglial cell activation[J].Journal of Neuroimmunology,2000,111(1/2):77-85.
[12]VEGETO E,BELCREDITO S,GHISLETTI S,et al.The endogenous estrogen status regulates microglia reactivity in animal models of neuroinflammation[J].Endocrinology,2006,147(5):2263-2272.
[13]VEGETO E,BONINCONTRO C,POLLIO G,et al.Estrogen prevents the lipopolysaccharide-induced inflammatory response in microglia[J].The Journal of Neuroscience,2001,21(6):1809-1818.
[14]XU Manman,TAN Xu,LI Na,et al.Differential regulation of estrogen in iron metabolism in astrocytes and neurons[J].Journal of Cellular Physiology,2018.doi:10.1002/jcp.27188.
[15]GUNSHIN H,MACKENZIE B,BERGER U V,et al.Cloning and characterization of a mammalian proton-coupled metal-ion transporter[J].Nature,1997,388(6641):482-488.
[16]HUANG E,ONG W Y,CONNOR J R.Distribution of divalent metal transporter-1in the monkey basal ganglia[J].Neuroscience,2004,128(3):487-496.
[17]SALAZAR J,MENA N,HUNOT S,et al.Divalent metal transporter 1(DMT1)contributes to neurodegeneration in animal models of Parkinson’s disease[J].PNAS,2008,105(47):18578-18583.
[18]MCKIE A T,MARCIANI P,ROLFS A,et al.A novel duodenal iron-regulated transporter,IREG1,implicated in the basolateral transfer of iron to the circulation[J].Molecular Cell,2000,5(2):299-309.
[19]ABBOUD S,HAILE D J.A novel mammalian iron-regulated protein involved in intracellular Iron metabolism[J].Journal of Biological Chemistry,2000,275(26):19906-19912.
[20]DONOVAN A,BROWNLIE A,ZHOU Y,et al.Positional cloning of zebrafish ferroportin1identifies a conserved vertebrate Iron exporter[J].Nature,2000,403(6771):776-781.
[21]CAMASCHELLA C.Understanding iron homeostasis through genetic analysis of hemochromatosis and related disorders[J].Blood,2005,106(12):3710-3717.
[22]CHRISTOVA T,TEMPLETON D M.Effect of hypoxia on the binding and subcellular distribution of Iron regulatory proteins[J].Molecular and Cellular Biochemistry,2007,301(1/2):21-32.
[23]WANG Jun,XU Huamin,YANG Haidong,et al.Rg1reduces nigral iron levels of MPTP-treated C57BL6mice by regulating certain iron transport proteins[J].Neurochemistry International,2009,54(1):43-48.
[24]JIANG Hong,SONG Ning,XU Huamin,et al.Up-regulation of divalent metal transporter 1in 6-hydroxydopamine intoxication is IRE/IRP dependent[J].Cell Research,2010,20(3):345-356.
[25]QIAN Yi,YIN Chunyang,CHEN Yue,et al.Estrogen contributes to regulating iron metabolism through governing ferroportin signaling via an estrogen response element[J].Cellular Signalling,2015,27(5):934-942.
[26]RASO G M,IRACE C,ESPOSITO E A,et al.Ovariectomy and estrogen treatment modulate iron metabolism in rat adipose tissue[J].Biochemical Pharmacology,2009,78(8):1001-1007.