Na~+/H~+交换器-1在窒息后血清诱导HK-2细胞损伤中作用机制的研究
|
摘要
目的:研究Na~+/H~+交换器-1(Na~+/H~+ exchanger 1,NHE1)在新生儿窒息后血清诱导人近曲肾小管上皮细胞(HK-2)损伤中的作用及机制。
方法:(1)以HK-2为研究对象;(2)以200ml/l窒息后第一天的血清作为攻击因素;(3)第一阶段实验分为2组:对照组和窒息组,探讨窒息后血清诱导人近曲肾小管上皮细胞损伤时NHE1是否参与并发挥损伤作用;(4)第二阶段实验分为:对照组,窒息组,EIPA组,探讨利用EIPA阻断NHE1活性后在窒息后血清诱导人近曲肾小管上皮细胞损伤时,对HK-2细胞的是否具有保护作用及其机制;(5)检测指标:第一阶段实验采用免疫组织化学的方法检测HK-2细胞在损伤前后的NHE1表达含量的变化,;第二阶段实验通过倒置相差显微镜观察对照组,窒息组,EIPA组的各组细胞形态,四唑盐(MTT)比色法检测HK-2细胞活力变化,生物化学法检测培养液中LDH漏出率的变化。(6)统计学处理:所有数值以均数±标准差(x±S)表示,实验组组间进行LSD检验。P<0.05为差异有显著性。采用SPSS10.0统计软件完成。
结果:(1)与对照组相比,窒息组细胞NHE1表达含量明显增加,细胞形态受损明显。(2)倒置相差显微镜下观察:对照组细胞贴壁良好,透明度大,折光性强,呈扁平的多角形,分裂相多,细胞排列紧密,连接成片,数量多;窒息组细胞生长缓慢,细胞数量比空白对照组明显减少,细胞形态发生改变,由典型的扁平多角形细胞变为不规则圆形或椭圆形,折光性减弱,轮廓增强,胞质中出现空泡,脂滴,颗粒样物质,细胞间空隙加大,连接松散,细胞间可见较多细胞碎片;(3)与对照组相比,窒息组LDH漏出率较明显升高,细胞活力(OD值)明显降低(P=0.000)。而与窒息组相比,EIPA(2μmol/l浓度)组, LDH漏出率明显降低,细胞活力(OD值)明显增高,差异具有统计学意义(P <0.05)。
结论:新生儿窒息后血清诱导HK-2细胞NHE1的表达是导致HK-2细胞损伤的重要机制。
Objective : To investigate the role of NHE1 in injury of human renal tubular cells(HK-2) induced by postasphyxial-serum in neonates.
Methods (1)Human renal proximal tubular cell(HK-2)was used as the target cel1.(2) The serum of neonates in one day after asphyxia, whose concentration were 200ml/l (volume fraction),were applied as attacking factor .(3)The experiment was designed as control group,asphyxia group,which were used to research whether NHE1 play an important role in the injury of human renal tubular cell induced by postasphyxial- serum of neonate .(4)In the second experiment was designed as control group,asphyxia group,EIPA group,which were researched whether EIPA had the protective effect . (5) The following indicators were detectied : In the first experiment,the expression of NHE1 in the HK-2 were detected by immunohistochemical method.In the second experiment,the cell viability was measured by 3-(4,5-dimethy lthiazcl-2-y1)-2,5-diphenyl–tetazoliumbromide (MTT) methods.In the first experiment,the changes of morphology were observed under inverted microscope,and the cell viability was measureed by 3-(4,5-dimethy lthiazcl-2-y1)-2,5-diphenyl -tetazolium bromide(MTT) methods,and the leakage rate of lactate dehydrogenase(LDH) was determined by biochemical methods.(6) All parameters were expressed as the mean value±standard difference( x±S), statistical analysis were carried out by the use of the least significant difference.Difference was considered significance when the p.value was less than 0.05.The statistics work were finished by spss10.0 software.
Results (1)Compared with control group,the expression of NHE1 in the HK-2 were increased significantly in asphyxia group.(2) Under inverted microscopy,HK-2 cells in control group were significantly increased,sticked to each other tightly and grew very quickly.Their adhesion were better,multyang applan,and refraction raised.The form and quantity of HK-2 cell was normal .Compared with control group,the changes in morphology of HK-2 were most serious and obvious in asphyxia group,the cells grew slowly,the mount decreased,form of the cells changed from typical multy-ang applan to off-normal round or ellipse.Refraction rate was decreased,and contour enhanced.The vacuolus,lipid droplet and granulation appeared in the kytoplasm.There was much cell debris in accrescent intercellular space.the cell viability (optical density,OD) were obviously decreased in the asphyxia group(P=0.000) ,the leakage rate of LDH were significantly increased in asphyxia group(P=0.000) ,statistical significance existed between control group and asphyxia group(P<0.05).But compared with asphyxia group, the changes in morphology of HK-2 were obviously improved in the EIPA groups ( 2μmol/l), the leakage rate of LDH were significantly decreased,statistical significance existed between EIPA group and asphyxia group(P<0.05); the cell viability were obviously increased,statistical significance existed between EIPA group and asphyxia group(P<0.05).
Conclusion :The postasphyxial-serum could induced the increased the expression of NHE1,which could play an important role in injury of renal tubular cells induced by postasphyxial-serum in neonates.
方法:(1)以HK-2为研究对象;(2)以200ml/l窒息后第一天的血清作为攻击因素;(3)第一阶段实验分为2组:对照组和窒息组,探讨窒息后血清诱导人近曲肾小管上皮细胞损伤时NHE1是否参与并发挥损伤作用;(4)第二阶段实验分为:对照组,窒息组,EIPA组,探讨利用EIPA阻断NHE1活性后在窒息后血清诱导人近曲肾小管上皮细胞损伤时,对HK-2细胞的是否具有保护作用及其机制;(5)检测指标:第一阶段实验采用免疫组织化学的方法检测HK-2细胞在损伤前后的NHE1表达含量的变化,;第二阶段实验通过倒置相差显微镜观察对照组,窒息组,EIPA组的各组细胞形态,四唑盐(MTT)比色法检测HK-2细胞活力变化,生物化学法检测培养液中LDH漏出率的变化。(6)统计学处理:所有数值以均数±标准差(x±S)表示,实验组组间进行LSD检验。P<0.05为差异有显著性。采用SPSS10.0统计软件完成。
结果:(1)与对照组相比,窒息组细胞NHE1表达含量明显增加,细胞形态受损明显。(2)倒置相差显微镜下观察:对照组细胞贴壁良好,透明度大,折光性强,呈扁平的多角形,分裂相多,细胞排列紧密,连接成片,数量多;窒息组细胞生长缓慢,细胞数量比空白对照组明显减少,细胞形态发生改变,由典型的扁平多角形细胞变为不规则圆形或椭圆形,折光性减弱,轮廓增强,胞质中出现空泡,脂滴,颗粒样物质,细胞间空隙加大,连接松散,细胞间可见较多细胞碎片;(3)与对照组相比,窒息组LDH漏出率较明显升高,细胞活力(OD值)明显降低(P=0.000)。而与窒息组相比,EIPA(2μmol/l浓度)组, LDH漏出率明显降低,细胞活力(OD值)明显增高,差异具有统计学意义(P <0.05)。
结论:新生儿窒息后血清诱导HK-2细胞NHE1的表达是导致HK-2细胞损伤的重要机制。
Objective : To investigate the role of NHE1 in injury of human renal tubular cells(HK-2) induced by postasphyxial-serum in neonates.
Methods (1)Human renal proximal tubular cell(HK-2)was used as the target cel1.(2) The serum of neonates in one day after asphyxia, whose concentration were 200ml/l (volume fraction),were applied as attacking factor .(3)The experiment was designed as control group,asphyxia group,which were used to research whether NHE1 play an important role in the injury of human renal tubular cell induced by postasphyxial- serum of neonate .(4)In the second experiment was designed as control group,asphyxia group,EIPA group,which were researched whether EIPA had the protective effect . (5) The following indicators were detectied : In the first experiment,the expression of NHE1 in the HK-2 were detected by immunohistochemical method.In the second experiment,the cell viability was measured by 3-(4,5-dimethy lthiazcl-2-y1)-2,5-diphenyl–tetazoliumbromide (MTT) methods.In the first experiment,the changes of morphology were observed under inverted microscope,and the cell viability was measureed by 3-(4,5-dimethy lthiazcl-2-y1)-2,5-diphenyl -tetazolium bromide(MTT) methods,and the leakage rate of lactate dehydrogenase(LDH) was determined by biochemical methods.(6) All parameters were expressed as the mean value±standard difference( x±S), statistical analysis were carried out by the use of the least significant difference.Difference was considered significance when the p.value was less than 0.05.The statistics work were finished by spss10.0 software.
Results (1)Compared with control group,the expression of NHE1 in the HK-2 were increased significantly in asphyxia group.(2) Under inverted microscopy,HK-2 cells in control group were significantly increased,sticked to each other tightly and grew very quickly.Their adhesion were better,multyang applan,and refraction raised.The form and quantity of HK-2 cell was normal .Compared with control group,the changes in morphology of HK-2 were most serious and obvious in asphyxia group,the cells grew slowly,the mount decreased,form of the cells changed from typical multy-ang applan to off-normal round or ellipse.Refraction rate was decreased,and contour enhanced.The vacuolus,lipid droplet and granulation appeared in the kytoplasm.There was much cell debris in accrescent intercellular space.the cell viability (optical density,OD) were obviously decreased in the asphyxia group(P=0.000) ,the leakage rate of LDH were significantly increased in asphyxia group(P=0.000) ,statistical significance existed between control group and asphyxia group(P<0.05).But compared with asphyxia group, the changes in morphology of HK-2 were obviously improved in the EIPA groups ( 2μmol/l), the leakage rate of LDH were significantly decreased,statistical significance existed between EIPA group and asphyxia group(P<0.05); the cell viability were obviously increased,statistical significance existed between EIPA group and asphyxia group(P<0.05).
Conclusion :The postasphyxial-serum could induced the increased the expression of NHE1,which could play an important role in injury of renal tubular cells induced by postasphyxial-serum in neonates.
引文
1. Aggarwal A, Kumar P, Chowdhary G, et al. Evaluation of renal functions in asphyxiated newborns[J].J Trop Pediatr. 2005 ,51(5):295-299
2.董文斌,杜逸亭,陈跃,等.TNF-α含量的改变与肾小管细胞缺氧-复氧性损伤关系的研究[J]细胞与分子免疫学杂志,2005,21(6):690-692.
3.董文斌,唐章华,陈红英,等.窒息新生儿尿中炎症细胞因子改变与肾小管损伤的关系[J].中国危重病急救医学,2000,15(2):94-96.
4. Landau M, Herz K, Padan E, et al. Model structure of the Na+/H+ exchanger 1 (NHE1): functional and clinical implications[J]. J Biol Chem. 2007, 28;282(52):37854-63.
5. Meima ME, Mackley JR, Barber DL. Beyond ion translocation: structural functions of the sodium-hydrogen exchanger isoform-1[J]. Curr Opin Nephrol Hypertens. 2007,16(4):365-72.
6.曹敏,董文斌,王明勇等.新生儿窒息后血清诱导人肾小管细胞损伤中的作用[J].实用儿科临床杂志,2006,21(17);1135-1138.
7.董文斌,曹敏,王明勇,等.新生儿窒息后血清诱导人肾小管细胞凋亡的作用[J]实用儿科临床杂志,2005,20(12):1207-1209
8.熊涛,董文斌,王明勇,等,促红细胞生成素减轻窒息新生儿血清诱导人肾小管细胞损伤[J],实用儿科临床杂志,2007,22(3):336-337
9. Kemp G, Young H, Fliegel L. Structure and function of the human Na(+)/H(+) exchanger isoform 1[J]. Channels (Austin). 2008, 23;2(5).
10. Fuster D, Moe OW, Hilgemann DW. Steady-state function of the ubiquitous mammalian Na/H exchanger (NHE1) in relation to dimer coupling models with 2Na/2H stoichiometry[J]. J Gen Physiol. 2008 ,132(4):465-80.
11. Landau M, Herz K, Padan E, et al Model structure of the Na+/H+ exchanger 1 (NHE1): functional and clinical implications[J]. J Biol Chem. 2007, 282(52):37854-63
12. Fliegel L. Regulation of the Na(+)/H(+) exchanger in the healthy and diseased myocardium[J]. Expert Opin Ther Targets. 2009 , 13(1):55-68. Review.
13. Slepkov ER, Rainey JK, Sykes BD, et al. Structural and functional analysis of the Na+/H+ exchanger[J]. Biochem J. 2007, 401(3):623-33.
14. Lee BL, Li X, Liu Y, Sykes BD, et al. Structural and functional analysis of TM XI of the nhe1 isoform of the NA+/H+ exchanger[J]. J Biol Chem. 2009 ,28.
15. Reddy T, Ding J, Li X, et al. Structural and functional characterization of transmembrane segment IX of the NHE1 isoformof the Na+/H+ exchanger[J]. J Biol Chem. 2008 , 283(32):22018-30.
16. Fliegel L. Molecular biology of the myocardial Na+/H+ exchanger[J]. J Mol Cell Cardiol. 2008 , 44(2):228-37.
17. Slepkov ER, Rainey JK, Li X. Structural and functional characterization of transmembrane segment IV of the NHE1 isoform of the Na+/H+ exchanger[J]. J Biol Chem. 2005, 280(18):17863-72.
18. Slepkov ER, Chow S, Lemieux MJ, Proline residues in transmembrane segment IV are critical for activity, expression and targeting of the Na+/H+ exchanger isoform 1[J]. Biochem J. 2004, 379(Pt 1):31-8.
19. Grenier AL, Abu-ihweij K, Zhang G, et al. Apoptosis-induced alkalinization by the Na+/H+ exchanger isoform 1 is mediated through phosphorylation of amino acids Ser726 and Ser729[J]. Am J Physiol Cell Physiol. 2008 ,295(4):C883-96.
20. Darmellah A, Rücker-Martin C, Feuvray D. et al. ERM proteins mediate the effects of Na+/H+ exchanger (NHE1) activation in cardiac myocytes. Cardiovasc Res. 2009 , 81(2):294-300.
21. Snabaitis AK, Cuello F. Avkiran M. Protein kinase B/Akt phosphorylates and inhibits the cardiac Na+/H+ exchanger NHE1[J]. Circ Res. 2008, 103(8):881-90.
22. Watts BA 3rd, George T, Good DW. The basolateral NHE1Na+/H+ exchanger regulates transepithelial HCO3- absorption through actin cytoskeleton remodeling in renal thick ascending limb[J], J Biol Chem. 2005, 280(12):11439-47.
23. Song D, Du T, Li B, et al. Astrocytic alkalinization by therapeutically relevant lithium concentrations: implications for myo-inositol depletion[J]. Psychopharmacology (Berl). 2008, 200(2):187-95.
24. Yu L, Quinn DA, Garg HG, et al. Deficiency of the NHE1 gene prevents hypoxia-induced pulmonary hypertension and vascular remodeling[J]. Am J Respir Crit Care Med. 2008 , 177(11):1276-84.
25.胡若愚,邵宏涛,董国华,等,阿米洛利同系物对老龄大鼠心肌缺血再灌注损伤的保护作用[J].实用医学杂志,2007,23(1):40-43
26. Simm A, Friedrich I, Scheubel RJ, et al. Age dependency of the cariporide-mediated cardio-protection after simulated ischemia in isolated human atrial heart muscles[J]. Exp Gerontol. 2008 ,43(7):691-9.
27. Hatch M, Freel RW. Increased colonic sodium absorption in rats with chronic renal failure is partially mediated by AT1 receptor agonism[J]. Am J Physiol Gastrointest Liver Physiol. 2008 ,295(2):G348-56.
28.张敏敏,顾勇,赖凌云,等,醛固酮可通过钠氢交换子1诱导肾小球系膜细胞外基质增生[J].中华肾脏病杂志,2006,22(8):477-482
29. Pinto V, Pinho MJ, Hopfer U, et al. Oxidative stress and the genomic regulation of aldosterone-stimulated NHE1 activity in SHR renal proximal tubular cells[J]. Mol Cell Biochem. 2008 , 310(1-2):191-201.
30.刘敬,曹海英,何纯义,等.新生儿窒息多脏器血流动力学研究[J].中华儿科学杂志,1998,36(2):69-70.
31. Fellman V, Raivio KO. Reperfusion injury as the mechanism of brain damage perinatal asphyxia [J]. Pediatr Res ,1997,38(4):599-606.
32.董文斌,陈跃,王胜会,等.血清TNF-α与窒息新生儿多器官功能不全的关系[J].中国当代儿科杂志,2003,5(5):455-456.
33.董文斌,陈红英,陈书琴,等.白细胞介素-1受体拮抗剂对新生鼠窒息后肾损伤保护作用的动态研究[J].中国危重病急救医学,2005,17(10);623-625
34.董文斌,唐章华,翟雪松,等.炎症细胞因子参与新生鼠窒息后肾损伤的作用时间[J].中华急诊医学杂志,2002,11(1):25-26
35. Friendewald JJ, Rabb H . Hamid.Rabb.Inflammatory cells in ischemic acute renal failure[J] .Kidney Int, 2004 , 66(2):486-491.
36. Maekawa N, Abe J, Shishido T,et al. Inhibiting p90 ribosomal S6kinase prevents (Na+)-H+ exchanger-mediated cardiac ischemia–reperfusion injury[J]. Circulation. 2006, 113(21):2516-23.
37. Pedersen SF, Darborg BV, Rasmussen M, et al. The Na+/H+ exchanger, NHE1, differentially regulates mitogen-activated protein kinase subfamilies after osmotic shrinkage in Ehrlich Lettre Ascites cells[J]. Cell Physiol Biochem. 2007, 20(6):735-50.
38. Gaitanaki C, Mastri M, Aggeli IK, et al. Differential roles of p38-MAPK and JNKs in mediating early protection or apoptosis in the hyperthermic perfused amphibian heart[J]. J Exp Biol. 2008 , 211(Pt 15):2524-32.
39. Wu S, Song T, Zhou S, et al. Involvement of Na+/H+ exchanger 1 in advanced glycation end products-induced proliferation of vascular smooth muscle cell[J]. Biochem Biophys Res Commun. 2008 , 375(3):384-9.
40. Cardone RA, Busco G, Greco MR, et al. HPV16 E7-dependent transformation activates NHE1 through a PKA-RhoA-induced inhibition of p38alpha[J]. PLoS ONE. 2008, 3(10):e3529.
41. Malo ME, Li L, Fliegel L, et al. Mitogen-activated protein kinase-dependent activation of the Na+/H+ exchanger is mediated through phosphorylation of amino acids Ser770 and Ser771[J]. J Biol Chem. 2007, 282(9):6292-9.
42. Luo J, Kintner DB, Shull GE. ERK1/2-p90RSK-mediatedphosphorylation of Na+/H+ exchanger isoform 1. A role in ischemic neuronal death[J]. J Biol Chem. 2007, 282(38): 28274-84.
43. Tattersall AL, Wilkins RJ. Modulation of Na+-H+ exchange isoforms NHE1 and NHE3 by insulin-like growth factor-1 in isolated bovine articular chondrocytes [J]. J Orthop Res. 2008 ,26(11):1428-33.
44. Zoltán H,Németh,,Edwin A.,Deitch, et al. Na+/H+ exchanger blockade inhibits enterocyte inflammatory response and protects against colitis[J]. Am J Physiol Gastrointest Liver Physiol , 2002 ,283: G122-G132,
45. Koliakos G, Befani C, Paletas K, Kaloyianni M, et al. Effect of endothelin on sodium/hydrogen exchanger activity of human monocytes and atherosclerosis-related functions[J]. Ann N Y Acad Sci. 2007 , 1095:274-91.
46. Magro F, Fraga S, Soares-da-Silva P,et al. Interferon-gamma-induced STAT1-mediated membrane retention of NHE1 and associated proteins ezrin, radixin and moesin in HT-29 cells[J]. Biochem Pharmacol. 2005, 70(9):1312-9.
47. Fujisawa G, Okada K, Muto S,et al. Na/H exchange isoform 1 is involved in mineralocorticoid/salt-induced cardiac injury[J]. Hypertension. 2003 ,41 (3):493-8.,
48. Leite-Dellova DC, Oliveira-Souza M, Malnic G, et al. Genomic andnongenomic dose-dependent biphasic effect of aldosterone on Na+/H+ exchanger in proximal S3 segment: role of cytosolic calcium[J]. Am J Physiol Renal Physiol. 2008 ,295(5):F1342-52.
49. Yamamuro M, Yoshimura M, Nakayama M ,et al. Direct effects of aldosterone on cardiomyocytes in the presence of normal and elevated extracellular sodium[J]. Endocrinology. 2006 ,147(3):1314-21. Epub 2005 Dec 22
50. Slepkov E, Fliegel L,et al. Regulation of expression of the Na+/H+ exchanger by thyroid hormone. Vitam Horm[J]. 2004, 69:249-69.
51. Park K, Poburko D, Wollheim C, et al. Amiloride derivatives induce apoptosis by depleting ER Ca(2+) stores in vascular endothelial cells[J]. Br J Pharmacol. 2009, 19.
52. Yamashita J, Ohkita M, Takaoka M,et al. Role of Na+/H+ exchanger in the pathogenesis of ischemic acute renal failure in mice[J]. J Cardiovasc Pharmacol. 2007 ,49(3):154-60
53. Luo J, Chen H, Kintner DB, Shull GE,,et al. Decreased neuronal death in Na+/H+ exchanger isoform 1-null mice after in vitro and in vivo ischemia[J]. J Neurosci. 2005 , 25(49):11256-68.
54. Kintner DB, Su G, Lenart B, et al. Increased tolerance to oxygen and glucose deprivation in astrocytes from Na(+)/H(+) exchanger isoform 1 null mice[J]. Am J Physiol Cell Physiol. 2004 ,287(1):C12-21
55. Hwang IK, Yoo KY, An SJ, et al. Late expression of Na+/H+ exchanger 1 (NHE1) and neuroprotective effects of NHE inhibitor in the gerbil hippocampal CA1 region induced by transient ischemia[J]. Exp Neurol. 2008, 212(2):314-23.
56. Wang Y, Luo J, Chen X, Chen H, et al. Gene inactivation of Na+/H+ exchanger isoform 1 attenuates apoptosis and mitochondrial damage following transient focal cerebral ischemia[J]. Eur J Neurosci. 2008 , 28(1):51-61.
57. Schelling JR, Abu Jawdeh BG. et al. Regulation of cell survival by Na+/H+ exchanger-1. Am J Physiol Renal Physiol[J]. 2008, 295(3):F625-32.
58. Cun L, Ronghua Z, Bin L, et al. Preconditioning with Na+/H+ exchange inhibitor HOE642 reduces calcium overload and exhibits marked protection on immature rabbit hearts[J]. ASAIO J. 2007 , 53(6):762-5
59.张本金,董文斌,唐章华,等,窒息新生儿尿中内皮素(ET一1)的变化与肾损伤的关系[J].中国优生与遗传杂志,1006—9534(2003)03—0073—02
60. Liu F, Gesek FA, et al. alpha(1)-Adrenergic receptors activate NHE1 and NHE3 through distinct signaling pathways in epithelial cells[J]. Am J Physiol Renal Physiol. 2001, 280(3):F415-25.
61. Luo J, Chen H, Kintner DB, et al. Inhibition of Na+/H+ exchangerisoform 1 attenuates mitochondrial cytochrome C release in cortical neurons following in vitro ischemia[J], Acta Neurochir Suppl. 2006, 96:244-8.
62. Sch?fer C, Ladilov YV, Sch?fer M, et,al. Inhibition of NHE protects reoxygenated cardiomyocytes independently of anoxic Ca(2+) overload and acidosis[J]. Am J Physiol Heart Circ Physiol. 2000 ,279(5):H2143-50.
63. Huc L, Sparfel L, Rissel M, Dimanche-Boitrel MT, et al. Identification of Na+/H+ exchange as a new target for toxic polycyclic aromatic hydrocarbons[J]. FASEB J. 2004 , 18(2):344-6. ,
[1] Meima ME, Mackley JR, Barber DL. Beyond ion translocation: structural functions of the sodium-hydrogen exchanger isoform-1[J]. Curr Opin Nephrol Hypertens. 2007 , 16(4):365-72.
[2] Slepkov ER, Rainey JK, Sykes BD, et al. Structural and functional analysis of the Na+/H+ exchanger[J]. Biochem J. 2007, 401(3):623-33.
[3] Slepkov ER, Rainey JK, Li X, Structural and functional characterization of transmembrane segment IV of the NHE1 isoform of the Na+/H+ exchanger[J]. J Biol Chem. 2005 , 280(18):17863-72.
[4] Slepkov ER, Chow S, Lemieux MJ, Proline residues in transmembrane segment IV are critical for activity, expression and targeting of the Na+/H+ exchanger isoform 1[J]. Biochem J. 2004, 379(Pt 1):31-8.
[5] Manucha W, Carrizo L, Ruete C, et al. Apoptosis induction is associated with decreased NHE1 expression in neonatal unilateral ureteric obstruction[J]. BJU Int. 2007 , 100(1):191-8
[6] Malo ME, Li L, Fliegel L. Mitogen-activated protein kinase-dependent activation of the Na+/H+ exchanger is mediated through phosphorylation of amino acids Ser770 and Ser771[J]. J Biol Chem. 2007, 282(9):6292-9.
[7] Luo J, Kintner DB, Shull GE,et al. ERK1/2-p90RSK-mediated phosphorylation of Na+/H+ exchanger isoform 1 A role in ischemic neuronal death[J]. J Biol Chem. 2007 ,282(38): 28274-84.
[8] Koliakos G, Befani C, Paletas K, et al. Effect of endothelin on sodium/hydrogenexchanger activity of human monocytes and atherosclerosis-related functions[J]. Ann N Y Acad Sci. 2007 , 1095:274-91.
[9] He B, Deng C, Zhang M, et al. Reduction of intracellular pH inhibits the expression of VEGF in K562 cells after targeted inhibition of the Na+/H+ exchanger. Leuk Res. 2007, 31(4):507-14.
[10] Magro F, Fraga S, Soares-da-Silva P. Interferon-gamma-induced STAT1-mediated membrane retention of NHE1 and associated proteins ezrin, radixin and moesin in HT-29 cells[J]. Biochem Pharmacol. 2005, 70(9):1312-9.
[11] Fujisawa G, Okada K, Muto S, et al. Na/H exchange isoform1 is involved in mineralocorticoid/salt-induced cardiac injury[J]. Hypertension. 2003 , 41(3):493-8.
[12] Yamamuro M, Yoshimura M, Nakayama M. Direct effects of aldosterone on cardiomyocytes in the presence of normal and elevated extracellular sodium.Endocrinology[J]. 2006, 147(3):1314-21.
[13] Slepkov E, Fliegel L. Regulation of expression of the Na+/H+ exchanger by thyroid hormone[J]. Vitam Horm. 2004, 69:249-69.
[14] Eigel BN, Hadley RW. Contribution of the Na(+) channel and Na(+)/H(+) exchanger to the anoxic rise of [Na(+)] in ventricular myocytes [J]. Am J Physiol. 1999 , 277(5 Pt 2):H1817-22.
[15] Maekawa N, Abe J, Shishido T, et al. Inhibiting p90 ribosomal S6 kinase prevents (Na+)-H+ exchanger-mediated cardiac ischemia-reperfusion injury[J]. Circulation. 2006, 113(21):2516-23.
[16] Robert SH, Semjidmaa Dashnyam, et al. Ras triggers acidosis-induced activation ofthe extracellular signal regu-lated kinase pathway in cardiac myocytes[J]. Biochem J. 2006 , 399(Pt 3): 493–501
[17] Yamashita J, Ohkita M, Takaoka M,et al. Role of Na+/H+ exchanger in the pathogenesis of ischemic acute renal failure in mice[J]. J Cardiovasc Pharmacol. 2007, 49(3):154-60
[18] Wu KL, Khan S, Lakhe-Reddy S, The NHE1 Na+/H+ exchanger recruits ezrin/radixin/moesin proteins to regulate Akt-dependent cell surviva[J]l. J Biol Chem. 2004 , 279(25):26280-6.
[19] Wu KL, Khan S, Lakhe-Reddy S,et al , Renal tubular epithelial cell apoptosis is associated with caspase cleavage of the NHE1 Na+/H+ exchanger[J]. Am J Physiol Renal Physiol. 2003 , 284(4):F829-39.
[20] Luo J, Chen H, Kintner DB,et al. Decreased neuronal death in Na+/H+ exchanger isoform 1-null mice after in vitro and in vivo ischemia[J]. J Neurosci. 2005 , 25(49):11256-68.
[21] Jung YW, Choi IJ, Kwon TH, et al. Altered expression of sodium transporters in ischemic penumbra after focal cerebral ischemia in rats[J]. Neurosci Res. 2007 , 59(2):152-9.
[22] Kintner DB, Look A, Shull GE, et al. Stimulation of astrocyte Na+/H+ exchange activity in response to in vitro ischemia depends in part on activation of ERK1/2[J]. Am J Physiol Cell Physiol. 2005 , 289(4):C934-45.
[23] Rios EJ, Fallon M, Wang J, et al. Chronic hypoxia elevates intracellularpH and activates Na+/H+ exchange in pulmonary arterial smooth muscle cells[J]. Am JPhysiol Lung Cell Mol Physiol. 2005 , 289(5):L867-74.
[24] Watts BA 3rd, George T, Good DW. The basolateral NHE1 Na+/H+ exchanger regulates transepithelial HCO3- absorption through actin cytoskeleton remodeling in renal thick ascending limb .J Biol Chem., 2005 Mar 25;280(12):11439-47.
2.董文斌,杜逸亭,陈跃,等.TNF-α含量的改变与肾小管细胞缺氧-复氧性损伤关系的研究[J]细胞与分子免疫学杂志,2005,21(6):690-692.
3.董文斌,唐章华,陈红英,等.窒息新生儿尿中炎症细胞因子改变与肾小管损伤的关系[J].中国危重病急救医学,2000,15(2):94-96.
4. Landau M, Herz K, Padan E, et al. Model structure of the Na+/H+ exchanger 1 (NHE1): functional and clinical implications[J]. J Biol Chem. 2007, 28;282(52):37854-63.
5. Meima ME, Mackley JR, Barber DL. Beyond ion translocation: structural functions of the sodium-hydrogen exchanger isoform-1[J]. Curr Opin Nephrol Hypertens. 2007,16(4):365-72.
6.曹敏,董文斌,王明勇等.新生儿窒息后血清诱导人肾小管细胞损伤中的作用[J].实用儿科临床杂志,2006,21(17);1135-1138.
7.董文斌,曹敏,王明勇,等.新生儿窒息后血清诱导人肾小管细胞凋亡的作用[J]实用儿科临床杂志,2005,20(12):1207-1209
8.熊涛,董文斌,王明勇,等,促红细胞生成素减轻窒息新生儿血清诱导人肾小管细胞损伤[J],实用儿科临床杂志,2007,22(3):336-337
9. Kemp G, Young H, Fliegel L. Structure and function of the human Na(+)/H(+) exchanger isoform 1[J]. Channels (Austin). 2008, 23;2(5).
10. Fuster D, Moe OW, Hilgemann DW. Steady-state function of the ubiquitous mammalian Na/H exchanger (NHE1) in relation to dimer coupling models with 2Na/2H stoichiometry[J]. J Gen Physiol. 2008 ,132(4):465-80.
11. Landau M, Herz K, Padan E, et al Model structure of the Na+/H+ exchanger 1 (NHE1): functional and clinical implications[J]. J Biol Chem. 2007, 282(52):37854-63
12. Fliegel L. Regulation of the Na(+)/H(+) exchanger in the healthy and diseased myocardium[J]. Expert Opin Ther Targets. 2009 , 13(1):55-68. Review.
13. Slepkov ER, Rainey JK, Sykes BD, et al. Structural and functional analysis of the Na+/H+ exchanger[J]. Biochem J. 2007, 401(3):623-33.
14. Lee BL, Li X, Liu Y, Sykes BD, et al. Structural and functional analysis of TM XI of the nhe1 isoform of the NA+/H+ exchanger[J]. J Biol Chem. 2009 ,28.
15. Reddy T, Ding J, Li X, et al. Structural and functional characterization of transmembrane segment IX of the NHE1 isoformof the Na+/H+ exchanger[J]. J Biol Chem. 2008 , 283(32):22018-30.
16. Fliegel L. Molecular biology of the myocardial Na+/H+ exchanger[J]. J Mol Cell Cardiol. 2008 , 44(2):228-37.
17. Slepkov ER, Rainey JK, Li X. Structural and functional characterization of transmembrane segment IV of the NHE1 isoform of the Na+/H+ exchanger[J]. J Biol Chem. 2005, 280(18):17863-72.
18. Slepkov ER, Chow S, Lemieux MJ, Proline residues in transmembrane segment IV are critical for activity, expression and targeting of the Na+/H+ exchanger isoform 1[J]. Biochem J. 2004, 379(Pt 1):31-8.
19. Grenier AL, Abu-ihweij K, Zhang G, et al. Apoptosis-induced alkalinization by the Na+/H+ exchanger isoform 1 is mediated through phosphorylation of amino acids Ser726 and Ser729[J]. Am J Physiol Cell Physiol. 2008 ,295(4):C883-96.
20. Darmellah A, Rücker-Martin C, Feuvray D. et al. ERM proteins mediate the effects of Na+/H+ exchanger (NHE1) activation in cardiac myocytes. Cardiovasc Res. 2009 , 81(2):294-300.
21. Snabaitis AK, Cuello F. Avkiran M. Protein kinase B/Akt phosphorylates and inhibits the cardiac Na+/H+ exchanger NHE1[J]. Circ Res. 2008, 103(8):881-90.
22. Watts BA 3rd, George T, Good DW. The basolateral NHE1Na+/H+ exchanger regulates transepithelial HCO3- absorption through actin cytoskeleton remodeling in renal thick ascending limb[J], J Biol Chem. 2005, 280(12):11439-47.
23. Song D, Du T, Li B, et al. Astrocytic alkalinization by therapeutically relevant lithium concentrations: implications for myo-inositol depletion[J]. Psychopharmacology (Berl). 2008, 200(2):187-95.
24. Yu L, Quinn DA, Garg HG, et al. Deficiency of the NHE1 gene prevents hypoxia-induced pulmonary hypertension and vascular remodeling[J]. Am J Respir Crit Care Med. 2008 , 177(11):1276-84.
25.胡若愚,邵宏涛,董国华,等,阿米洛利同系物对老龄大鼠心肌缺血再灌注损伤的保护作用[J].实用医学杂志,2007,23(1):40-43
26. Simm A, Friedrich I, Scheubel RJ, et al. Age dependency of the cariporide-mediated cardio-protection after simulated ischemia in isolated human atrial heart muscles[J]. Exp Gerontol. 2008 ,43(7):691-9.
27. Hatch M, Freel RW. Increased colonic sodium absorption in rats with chronic renal failure is partially mediated by AT1 receptor agonism[J]. Am J Physiol Gastrointest Liver Physiol. 2008 ,295(2):G348-56.
28.张敏敏,顾勇,赖凌云,等,醛固酮可通过钠氢交换子1诱导肾小球系膜细胞外基质增生[J].中华肾脏病杂志,2006,22(8):477-482
29. Pinto V, Pinho MJ, Hopfer U, et al. Oxidative stress and the genomic regulation of aldosterone-stimulated NHE1 activity in SHR renal proximal tubular cells[J]. Mol Cell Biochem. 2008 , 310(1-2):191-201.
30.刘敬,曹海英,何纯义,等.新生儿窒息多脏器血流动力学研究[J].中华儿科学杂志,1998,36(2):69-70.
31. Fellman V, Raivio KO. Reperfusion injury as the mechanism of brain damage perinatal asphyxia [J]. Pediatr Res ,1997,38(4):599-606.
32.董文斌,陈跃,王胜会,等.血清TNF-α与窒息新生儿多器官功能不全的关系[J].中国当代儿科杂志,2003,5(5):455-456.
33.董文斌,陈红英,陈书琴,等.白细胞介素-1受体拮抗剂对新生鼠窒息后肾损伤保护作用的动态研究[J].中国危重病急救医学,2005,17(10);623-625
34.董文斌,唐章华,翟雪松,等.炎症细胞因子参与新生鼠窒息后肾损伤的作用时间[J].中华急诊医学杂志,2002,11(1):25-26
35. Friendewald JJ, Rabb H . Hamid.Rabb.Inflammatory cells in ischemic acute renal failure[J] .Kidney Int, 2004 , 66(2):486-491.
36. Maekawa N, Abe J, Shishido T,et al. Inhibiting p90 ribosomal S6kinase prevents (Na+)-H+ exchanger-mediated cardiac ischemia–reperfusion injury[J]. Circulation. 2006, 113(21):2516-23.
37. Pedersen SF, Darborg BV, Rasmussen M, et al. The Na+/H+ exchanger, NHE1, differentially regulates mitogen-activated protein kinase subfamilies after osmotic shrinkage in Ehrlich Lettre Ascites cells[J]. Cell Physiol Biochem. 2007, 20(6):735-50.
38. Gaitanaki C, Mastri M, Aggeli IK, et al. Differential roles of p38-MAPK and JNKs in mediating early protection or apoptosis in the hyperthermic perfused amphibian heart[J]. J Exp Biol. 2008 , 211(Pt 15):2524-32.
39. Wu S, Song T, Zhou S, et al. Involvement of Na+/H+ exchanger 1 in advanced glycation end products-induced proliferation of vascular smooth muscle cell[J]. Biochem Biophys Res Commun. 2008 , 375(3):384-9.
40. Cardone RA, Busco G, Greco MR, et al. HPV16 E7-dependent transformation activates NHE1 through a PKA-RhoA-induced inhibition of p38alpha[J]. PLoS ONE. 2008, 3(10):e3529.
41. Malo ME, Li L, Fliegel L, et al. Mitogen-activated protein kinase-dependent activation of the Na+/H+ exchanger is mediated through phosphorylation of amino acids Ser770 and Ser771[J]. J Biol Chem. 2007, 282(9):6292-9.
42. Luo J, Kintner DB, Shull GE. ERK1/2-p90RSK-mediatedphosphorylation of Na+/H+ exchanger isoform 1. A role in ischemic neuronal death[J]. J Biol Chem. 2007, 282(38): 28274-84.
43. Tattersall AL, Wilkins RJ. Modulation of Na+-H+ exchange isoforms NHE1 and NHE3 by insulin-like growth factor-1 in isolated bovine articular chondrocytes [J]. J Orthop Res. 2008 ,26(11):1428-33.
44. Zoltán H,Németh,,Edwin A.,Deitch, et al. Na+/H+ exchanger blockade inhibits enterocyte inflammatory response and protects against colitis[J]. Am J Physiol Gastrointest Liver Physiol , 2002 ,283: G122-G132,
45. Koliakos G, Befani C, Paletas K, Kaloyianni M, et al. Effect of endothelin on sodium/hydrogen exchanger activity of human monocytes and atherosclerosis-related functions[J]. Ann N Y Acad Sci. 2007 , 1095:274-91.
46. Magro F, Fraga S, Soares-da-Silva P,et al. Interferon-gamma-induced STAT1-mediated membrane retention of NHE1 and associated proteins ezrin, radixin and moesin in HT-29 cells[J]. Biochem Pharmacol. 2005, 70(9):1312-9.
47. Fujisawa G, Okada K, Muto S,et al. Na/H exchange isoform 1 is involved in mineralocorticoid/salt-induced cardiac injury[J]. Hypertension. 2003 ,41 (3):493-8.,
48. Leite-Dellova DC, Oliveira-Souza M, Malnic G, et al. Genomic andnongenomic dose-dependent biphasic effect of aldosterone on Na+/H+ exchanger in proximal S3 segment: role of cytosolic calcium[J]. Am J Physiol Renal Physiol. 2008 ,295(5):F1342-52.
49. Yamamuro M, Yoshimura M, Nakayama M ,et al. Direct effects of aldosterone on cardiomyocytes in the presence of normal and elevated extracellular sodium[J]. Endocrinology. 2006 ,147(3):1314-21. Epub 2005 Dec 22
50. Slepkov E, Fliegel L,et al. Regulation of expression of the Na+/H+ exchanger by thyroid hormone. Vitam Horm[J]. 2004, 69:249-69.
51. Park K, Poburko D, Wollheim C, et al. Amiloride derivatives induce apoptosis by depleting ER Ca(2+) stores in vascular endothelial cells[J]. Br J Pharmacol. 2009, 19.
52. Yamashita J, Ohkita M, Takaoka M,et al. Role of Na+/H+ exchanger in the pathogenesis of ischemic acute renal failure in mice[J]. J Cardiovasc Pharmacol. 2007 ,49(3):154-60
53. Luo J, Chen H, Kintner DB, Shull GE,,et al. Decreased neuronal death in Na+/H+ exchanger isoform 1-null mice after in vitro and in vivo ischemia[J]. J Neurosci. 2005 , 25(49):11256-68.
54. Kintner DB, Su G, Lenart B, et al. Increased tolerance to oxygen and glucose deprivation in astrocytes from Na(+)/H(+) exchanger isoform 1 null mice[J]. Am J Physiol Cell Physiol. 2004 ,287(1):C12-21
55. Hwang IK, Yoo KY, An SJ, et al. Late expression of Na+/H+ exchanger 1 (NHE1) and neuroprotective effects of NHE inhibitor in the gerbil hippocampal CA1 region induced by transient ischemia[J]. Exp Neurol. 2008, 212(2):314-23.
56. Wang Y, Luo J, Chen X, Chen H, et al. Gene inactivation of Na+/H+ exchanger isoform 1 attenuates apoptosis and mitochondrial damage following transient focal cerebral ischemia[J]. Eur J Neurosci. 2008 , 28(1):51-61.
57. Schelling JR, Abu Jawdeh BG. et al. Regulation of cell survival by Na+/H+ exchanger-1. Am J Physiol Renal Physiol[J]. 2008, 295(3):F625-32.
58. Cun L, Ronghua Z, Bin L, et al. Preconditioning with Na+/H+ exchange inhibitor HOE642 reduces calcium overload and exhibits marked protection on immature rabbit hearts[J]. ASAIO J. 2007 , 53(6):762-5
59.张本金,董文斌,唐章华,等,窒息新生儿尿中内皮素(ET一1)的变化与肾损伤的关系[J].中国优生与遗传杂志,1006—9534(2003)03—0073—02
60. Liu F, Gesek FA, et al. alpha(1)-Adrenergic receptors activate NHE1 and NHE3 through distinct signaling pathways in epithelial cells[J]. Am J Physiol Renal Physiol. 2001, 280(3):F415-25.
61. Luo J, Chen H, Kintner DB, et al. Inhibition of Na+/H+ exchangerisoform 1 attenuates mitochondrial cytochrome C release in cortical neurons following in vitro ischemia[J], Acta Neurochir Suppl. 2006, 96:244-8.
62. Sch?fer C, Ladilov YV, Sch?fer M, et,al. Inhibition of NHE protects reoxygenated cardiomyocytes independently of anoxic Ca(2+) overload and acidosis[J]. Am J Physiol Heart Circ Physiol. 2000 ,279(5):H2143-50.
63. Huc L, Sparfel L, Rissel M, Dimanche-Boitrel MT, et al. Identification of Na+/H+ exchange as a new target for toxic polycyclic aromatic hydrocarbons[J]. FASEB J. 2004 , 18(2):344-6. ,
[1] Meima ME, Mackley JR, Barber DL. Beyond ion translocation: structural functions of the sodium-hydrogen exchanger isoform-1[J]. Curr Opin Nephrol Hypertens. 2007 , 16(4):365-72.
[2] Slepkov ER, Rainey JK, Sykes BD, et al. Structural and functional analysis of the Na+/H+ exchanger[J]. Biochem J. 2007, 401(3):623-33.
[3] Slepkov ER, Rainey JK, Li X, Structural and functional characterization of transmembrane segment IV of the NHE1 isoform of the Na+/H+ exchanger[J]. J Biol Chem. 2005 , 280(18):17863-72.
[4] Slepkov ER, Chow S, Lemieux MJ, Proline residues in transmembrane segment IV are critical for activity, expression and targeting of the Na+/H+ exchanger isoform 1[J]. Biochem J. 2004, 379(Pt 1):31-8.
[5] Manucha W, Carrizo L, Ruete C, et al. Apoptosis induction is associated with decreased NHE1 expression in neonatal unilateral ureteric obstruction[J]. BJU Int. 2007 , 100(1):191-8
[6] Malo ME, Li L, Fliegel L. Mitogen-activated protein kinase-dependent activation of the Na+/H+ exchanger is mediated through phosphorylation of amino acids Ser770 and Ser771[J]. J Biol Chem. 2007, 282(9):6292-9.
[7] Luo J, Kintner DB, Shull GE,et al. ERK1/2-p90RSK-mediated phosphorylation of Na+/H+ exchanger isoform 1 A role in ischemic neuronal death[J]. J Biol Chem. 2007 ,282(38): 28274-84.
[8] Koliakos G, Befani C, Paletas K, et al. Effect of endothelin on sodium/hydrogenexchanger activity of human monocytes and atherosclerosis-related functions[J]. Ann N Y Acad Sci. 2007 , 1095:274-91.
[9] He B, Deng C, Zhang M, et al. Reduction of intracellular pH inhibits the expression of VEGF in K562 cells after targeted inhibition of the Na+/H+ exchanger. Leuk Res. 2007, 31(4):507-14.
[10] Magro F, Fraga S, Soares-da-Silva P. Interferon-gamma-induced STAT1-mediated membrane retention of NHE1 and associated proteins ezrin, radixin and moesin in HT-29 cells[J]. Biochem Pharmacol. 2005, 70(9):1312-9.
[11] Fujisawa G, Okada K, Muto S, et al. Na/H exchange isoform1 is involved in mineralocorticoid/salt-induced cardiac injury[J]. Hypertension. 2003 , 41(3):493-8.
[12] Yamamuro M, Yoshimura M, Nakayama M. Direct effects of aldosterone on cardiomyocytes in the presence of normal and elevated extracellular sodium.Endocrinology[J]. 2006, 147(3):1314-21.
[13] Slepkov E, Fliegel L. Regulation of expression of the Na+/H+ exchanger by thyroid hormone[J]. Vitam Horm. 2004, 69:249-69.
[14] Eigel BN, Hadley RW. Contribution of the Na(+) channel and Na(+)/H(+) exchanger to the anoxic rise of [Na(+)] in ventricular myocytes [J]. Am J Physiol. 1999 , 277(5 Pt 2):H1817-22.
[15] Maekawa N, Abe J, Shishido T, et al. Inhibiting p90 ribosomal S6 kinase prevents (Na+)-H+ exchanger-mediated cardiac ischemia-reperfusion injury[J]. Circulation. 2006, 113(21):2516-23.
[16] Robert SH, Semjidmaa Dashnyam, et al. Ras triggers acidosis-induced activation ofthe extracellular signal regu-lated kinase pathway in cardiac myocytes[J]. Biochem J. 2006 , 399(Pt 3): 493–501
[17] Yamashita J, Ohkita M, Takaoka M,et al. Role of Na+/H+ exchanger in the pathogenesis of ischemic acute renal failure in mice[J]. J Cardiovasc Pharmacol. 2007, 49(3):154-60
[18] Wu KL, Khan S, Lakhe-Reddy S, The NHE1 Na+/H+ exchanger recruits ezrin/radixin/moesin proteins to regulate Akt-dependent cell surviva[J]l. J Biol Chem. 2004 , 279(25):26280-6.
[19] Wu KL, Khan S, Lakhe-Reddy S,et al , Renal tubular epithelial cell apoptosis is associated with caspase cleavage of the NHE1 Na+/H+ exchanger[J]. Am J Physiol Renal Physiol. 2003 , 284(4):F829-39.
[20] Luo J, Chen H, Kintner DB,et al. Decreased neuronal death in Na+/H+ exchanger isoform 1-null mice after in vitro and in vivo ischemia[J]. J Neurosci. 2005 , 25(49):11256-68.
[21] Jung YW, Choi IJ, Kwon TH, et al. Altered expression of sodium transporters in ischemic penumbra after focal cerebral ischemia in rats[J]. Neurosci Res. 2007 , 59(2):152-9.
[22] Kintner DB, Look A, Shull GE, et al. Stimulation of astrocyte Na+/H+ exchange activity in response to in vitro ischemia depends in part on activation of ERK1/2[J]. Am J Physiol Cell Physiol. 2005 , 289(4):C934-45.
[23] Rios EJ, Fallon M, Wang J, et al. Chronic hypoxia elevates intracellularpH and activates Na+/H+ exchange in pulmonary arterial smooth muscle cells[J]. Am JPhysiol Lung Cell Mol Physiol. 2005 , 289(5):L867-74.
[24] Watts BA 3rd, George T, Good DW. The basolateral NHE1 Na+/H+ exchanger regulates transepithelial HCO3- absorption through actin cytoskeleton remodeling in renal thick ascending limb .J Biol Chem., 2005 Mar 25;280(12):11439-47.