钠/钙交换蛋白SLC24A6在实验性颅内出血导致脑损伤过程中的作用及机制研究
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摘要
目的探究钾依赖钠/钙交换蛋白-6(SLC24A6)参与大鼠脑出血后继发性脑损伤的作用和可能机制。方法建立SD大鼠脑出血模型,检测大鼠脑出血后尾状核SLC24A6表达及其介导的细胞内钙浓度([Ca~(2+)]_i)随时间变化的情况,观察SLC24A6在正常氧浓度和低氧条件下对[Ca~(2+)]_i的调控。结果脑出血早期,尾状核SLC24A6蛋白和SLC24A6mRNA水平均降低,在脑出血后3 d降至最低水平,5和7 d后轻微升高。脑出血后早期,[Ca~(2+)]_i增加,于脑出血后3 d达最高水平,5和7 d时逐步下降。正常氧浓度下,转染SLC24A6导致HEK293[Ca~(2+)]_i升高。结论 SLC24A6通过抑制钙超载在脑出血后脑损伤中起保护作用。
Aim Intracerebral hemorrhage(ICH) can lead to tragic disability and mortality. Accumulating evidence has been shown that sodium calcium exchanger(NCX) may contribute to secondary injury of stroke. A novel member of NCX, SLC24A6, was reported, which was involved in brain damage in rats after ICH. Methods The time course of expression of SLC24A6 and its mediated intracellular calcium concentration in caudate nucleus tissues of rats after ICH were examined. And the roles of SLC24A6 in regulating calcium concentration under normoxic or hypoxic conditions was also investigated. Results Both m RNA and protein levels of SLC24A6 decreased in hemorrhagic caudate nucleus tissues early after ICH, and reached to the lowest levels at day 3, and then slightly increased at day 5 and day 7. On the contrary, intracellular calcium concentration increased early after ICH, peaking at day 3 followed by declines at day 5 and day 7. Under normoxic conditions, transfection of SLC24A6 led to a significant increase of intracellular calcium concentration in HEK 293 cells. SLC24A6 overexpression, however, inhibited calcium overload and protected cells from death under hypoxic conditions. Conclusion These data suggested that SLC24A6 could exert a protective role in brain damage after ICH via inhibiting calcium overload.
Aim Intracerebral hemorrhage(ICH) can lead to tragic disability and mortality. Accumulating evidence has been shown that sodium calcium exchanger(NCX) may contribute to secondary injury of stroke. A novel member of NCX, SLC24A6, was reported, which was involved in brain damage in rats after ICH. Methods The time course of expression of SLC24A6 and its mediated intracellular calcium concentration in caudate nucleus tissues of rats after ICH were examined. And the roles of SLC24A6 in regulating calcium concentration under normoxic or hypoxic conditions was also investigated. Results Both m RNA and protein levels of SLC24A6 decreased in hemorrhagic caudate nucleus tissues early after ICH, and reached to the lowest levels at day 3, and then slightly increased at day 5 and day 7. On the contrary, intracellular calcium concentration increased early after ICH, peaking at day 3 followed by declines at day 5 and day 7. Under normoxic conditions, transfection of SLC24A6 led to a significant increase of intracellular calcium concentration in HEK 293 cells. SLC24A6 overexpression, however, inhibited calcium overload and protected cells from death under hypoxic conditions. Conclusion These data suggested that SLC24A6 could exert a protective role in brain damage after ICH via inhibiting calcium overload.
引文
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[14]Zheng M,Zhu H,Gong Y,et al.Involvement of GMRP1,a novel mediator of Akt pathway,in brain damage after intracerebral hemorrhage[J].Int J Clin Exp Pathol,2013,6:224-229
[15]Cai X,Lytton J.Molecular cloning of a sixth member of the K+-dependent Na+/Ca2+exchanger gene family,NCKX6[J].J Biol Chem,2004,279:5867-5876
[16]Bano D,Young KW,Guerin CJ,et al.Cleavage of the plasma membrane Na+/Ca2+exchanger in excitotoxicity[J].Cell,2005,120:275-285
[17]Pignataro G,Tortiglione A,Scorziello A,et al.Evidence for a protective role played by the Na+/Ca2+exchanger in cerebral ischemia induced by middle cerebral artery occlusion in male rats[J].Neurophar macology,2004,46:439-448
[18]Pignataro G,Gala R,Cuomo O,et al.Two sodium/calcium exchanger gene products,NCX1 and NCX3,play a major role in the development of permanent focal cerebral ischemia[J].Stroke,2004,35:2566-2570
[19]Secondo A,Staiano RI,Scorziello A,et al.BHK cells transfected with NCX3 are more resistant to hypoxia followed by reoxygenation than those transfected with NCX1 and NCX2:Possible relationship with mitochondrial membrane potential[J].Cell Calcium,2007,42:521-535
[20]Cuomo O,Gala R,Pignataro G,et al.A critical role for the potassiumdependent sodium-calcium exchanger NCKX2 in protection against focal ischemic brain damage[J].J Neurosci,2008,28:2053-2063
[2]Zazulia AR,Diringer MN,Derdeyn CP,et al.Progression of mass effect after intracerebral hemorrhage[J].Stroke,1999,30:1167-1173
[3]Gong Y,Xi G,Hu H,et al.Increase in brain thrombin activity after experimental intracerebral hemorrhage[J].Acta Neurochir Supp,2008,105:47-50
[4]Xi G,Hua Y,Bhasin RR,et al.Mechanisms of edema formation after intracerebral hemorrhage:effects of extravasated red blood cells on blood flow and blood-brain barrier integrity[J].Stroke,2001,32:2932-2938
[5]Gong Y,Xi GH,Keep RF,et al.Complement inhibition attenuates brain edema and neurological deficits induced by thrombin[J].Acta Neurochir Suppl,2005,95:389-392
[6]Zhang X,Li H,Hu S,et al.Brain edema after intracerebral hemorrhage in rats:the role of inflammation[J].Neurol India,2006,54:402-407
[7]Orrenius S,Zhivotovsky B,Nicotera P.Regulation of cell death:the calcium-apoptosis link[J].Nat Rev Mol Cell Biol,2003,4:552-565
[8]Lee JM,Zipfel GJ,Choi DW.The changing landscape of ischaemic brain injury mechanisms[J].Nature,1999,399:A7-14
[9]Li MH,Inoue K,Si HF,et al.Calcium-permeable ion channels involved in glutamate receptor-independent ischemic brain injury[J].Acta Pharmacol Sin,2011,32:734-740
[10]Blaustein MP,Lederer WJ.Sodium/calcium exchange:its physiological implications[J].Physiol Rev,1999,79:763-854
[11]Cai X,Lytton J.The cation/Ca(2+)exchanger superfamily:phylogenetic analysis and structural implications[J].Mol Biol Evol,2004,21:1692-1703
[12]Palty R,Ohana E,Hershfinkel M,et al.Lithium-calcium exchange is mediated by a distinct potassium-independent sodium-calcium exchanger[J].J Biol Chem,2004,279:25234-25240
[13]Tsoi M,Rhee KH,Bungard D,et al.Molecular cloning of a novel potassium-dependent sodium-calcium exchanger from rat brain[J].JBiol Chem,1998,273:4155-4162
[14]Zheng M,Zhu H,Gong Y,et al.Involvement of GMRP1,a novel mediator of Akt pathway,in brain damage after intracerebral hemorrhage[J].Int J Clin Exp Pathol,2013,6:224-229
[15]Cai X,Lytton J.Molecular cloning of a sixth member of the K+-dependent Na+/Ca2+exchanger gene family,NCKX6[J].J Biol Chem,2004,279:5867-5876
[16]Bano D,Young KW,Guerin CJ,et al.Cleavage of the plasma membrane Na+/Ca2+exchanger in excitotoxicity[J].Cell,2005,120:275-285
[17]Pignataro G,Tortiglione A,Scorziello A,et al.Evidence for a protective role played by the Na+/Ca2+exchanger in cerebral ischemia induced by middle cerebral artery occlusion in male rats[J].Neurophar macology,2004,46:439-448
[18]Pignataro G,Gala R,Cuomo O,et al.Two sodium/calcium exchanger gene products,NCX1 and NCX3,play a major role in the development of permanent focal cerebral ischemia[J].Stroke,2004,35:2566-2570
[19]Secondo A,Staiano RI,Scorziello A,et al.BHK cells transfected with NCX3 are more resistant to hypoxia followed by reoxygenation than those transfected with NCX1 and NCX2:Possible relationship with mitochondrial membrane potential[J].Cell Calcium,2007,42:521-535
[20]Cuomo O,Gala R,Pignataro G,et al.A critical role for the potassiumdependent sodium-calcium exchanger NCKX2 in protection against focal ischemic brain damage[J].J Neurosci,2008,28:2053-2063