A&beta;<sub>1~42</sub>诱导K<sub>ATP</sub>亚基SUR1/Kir6.2蛋白表达的信号转导通路研究

doi:10.3969/j.issn.1672-6731.2021.11.005

摘要/Abstract

摘要：

目的分析探讨β-淀粉样蛋白1~42（Aβ_1~42）诱导ATP敏感性钾离子通道（K_ATP）亚基SUR1/Kir6.2蛋白表达上调的潜在信号转导机制。方法采用免疫细胞化学法鉴定原代培养的大鼠皮质和海马神经元，实验组分别加入0.50 μmol/L核因子-κB（NF-κB）抑制剂SN50、2 μmol/L p38丝裂原激活蛋白激酶（MAPK）抑制剂SB203580或者2 μmol/L蛋白激酶C（PKC）抑制剂CTC，于30 min后分别加入2 μmol/L Aβ_1~42，培养72 h后采用Western blotting法检测K_ATP亚基SUR1/Kir6.2蛋白的相对表达量。结果（1）NF-κB抑制剂SN50：SN50+Aβ_1~42组SUR1/Kir6.2蛋白表达量低于对照组（t=6.237，P=0.010；t=7.136，P=0.004）和Aβ_1~42组（t=12.620，P=0.000；t=18.580，P=0.000），SN50组SUR1/Kir6.2蛋白相对表达量低于对照组（t=12.240，P=0.000；t=4.906，P=0.034）、Aβ_1~42组（t=18.620，P=0.000；t=16.350，P=0.000）和SN50+Aβ_1~42组（t=6.002，P=0.012）。（2）p38 MAPK抑制剂SB203580：SB203580+Aβ_1~42组SUR1/Kir6.2蛋白相对表达量低于Aβ_1~42组（t=13.010，P=0.000；t=8.506，P=0.001），仅Kir6.2蛋白相对表达量高于对照组（t=7.537，P=0.003）；SB203580组SUR1/Kir6.2蛋白相对表达量低于对照组（t=8.089，P=0.002；t=19.380，P=0.000）、Aβ_1~42组（t=18.870，P=0.000；t=35.430，P=0.000）和SB203580+Aβ_1~42组（t=5.869，P=0.014；t=26.920，P=0.000）。（3）PKC抑制剂CTC：CTC+Aβ_1~42组SUR1蛋白相对表达量高于对照组（t=4.756，P=0.040）、但低于Aβ_1~42组（t=15.170，P=0.000），CTC组SUR1蛋白相对表达量低于对照组（t=24.000，P=0.000）、Aβ_1~42组（t=43.930，P=0.000）和CTC+Aβ_1~42组（t=28.760，P=0.000）；CTC+Aβ_1~42组Kir6.2蛋白相对表达量低于对照组（t=15.000，P=0.000）和Aβ_1~42组（t=24.140，P=0.000），CTC组Kir6.2蛋白相对表达量低于对照组（t=9.429，P=0.001）和Aβ_1~42组（t=18.570，P=0.000）、但高于CTC+Aβ_1~42组（t=5.571，P=0.018）。结论 NF-κB、p38 MAPK和PKC信号转导通路参与Aβ_1~42上调大鼠皮质和海马神经元K_ATP亚基SUR1/Kir6.2蛋白的表达。

关键词: 淀粉样β肽类, 钾通道, NF-κB, p38丝裂原活化蛋白激酶类, 蛋白激酶C, 印迹法,蛋白质, 细胞,培养的

Abstract:

Objective To investigate the potential signal transduction mechanism of β-amyloid protein 1-42 (Aβ_1-42) induced up -regulation of ATP sensitive potassium channel K_ATP subunit SUR1/Kir6.2 proteins expression. Methods The primary cultured cortical and hippocampal neurons of rats were identified by immunocytochemistry. The experimental groups were added with 0.50 μmol/L nuclear factor-κB (NF-κB) inhibitor SN50, 2 μmol/L p38 mitogen-activated protein kinase (MAPK) inhibitor SB203580 or 2 μmol/L protein kinease C (PKC) inhibitor CTC, 2 μmol/L Aβ_1-42 (30 min later), respectively. Western blotting was used to detect the protein expression of K_ATP subunit SUR1/Kir6.2 after 72 h of culture. Results 1) NF-κB inhibitor SN50:SUR1/Kir6.2 proteins of K_ATP subunit in SN50 + Aβ_1-42 group were lower than that in control group (t=6.237, P=0.010; t=7.136, P=0.004) and Aβ_1-42 group (t=12.620, P=0.000; t=18.580, P=0.000), but the SUR1 protein expression of K_ATP subunit in SN50 + Aβ_1-42 group was higher than that in SN50 group (t=6.002, P=0.012). SUR1/Kir6.2 proteins of K_ATP subunit in SN50 group were lower than that in control group (t=12.240, P=0.000; t=4.906, P=0.034) and Aβ_1-42 group (t=18.620, P=0.000; t=16.350, P=0.000). 2) p38 MAPK inhibitor SB203580:the proteins expression of K_ATP subunit SUR1/Kir6.2 in SB203580 + Aβ_1-42 group were lower than that in Aβ_1-42 group (t=13.010, P=0.000; t=8.506, P=0.001), but higher than that in SB203580 group (t=5.869, P=0.014; t=26.920, P=0.000), but the expression of K_ATP subunit Kir6.2 protein in SB203580 + Aβ_1-42 group was higher than that in control group (t=7.537, P=0.003). The SUR1/Kir6.2 proteins expression of K_ATP subunit in SB203580 group were lower than that in control group (t=8.089, P=0.002; t=19.380, P=0.003) and Aβ_1-42 group (t=18.870, P=0.000; t=35.430, P=0.000). 3) PKC inhibitor CTC:the SUR1 protein expression of K_ATP subunit in CTC + Aβ_1-42 group was higher than that in control group (t=4.756, P=0.040) and in CTC group (t=28.760, P=0.000), but lower than that in Aβ_1-42 group (t=15.170, P=0.000). The SUR1 protein expression of K_ATP subunit in CTC group was lower than that in control group (t=24.000, P=0.000), Aβ_1-42 group (t=43.930, P=0.000) and CTC + Aβ_1-42 group. The protein expression of K_ATP subunit Kir6.2 in CTC + Aβ_1-42 group was lower than that in control group (t=15.000, P=0.000), Aβ_1-42 group (t=24.140, P=0.000) and CTC group (t=5.571, P=0.018). The expression of K_ATP subunit Kir6.2 protein in CTC group was lower than that in control group (t=9.429, P=0.001) and in Aβ_1-42 group (t=18.570, P=0.000). Conclusions NF-κB, p38 MAPK and PKC signaling pathways are involved in Aβ_1-42 up-regulation of SUR1/Kir6.2 proteins expression of K ATP subunit in cortical and hippocampal neurons of rats.

Key words: Amyloid beta-peptides, Potassium channels, NF-kappa B, p38 mitogen-activated protein kinases, Protein kinase C, Blotting, western, Cells, cultured

李艳菊, 张戈, 王晓静, 邢孝民, 马国诏. Aβ_1~42诱导K_ATP亚基SUR1/Kir6.2蛋白表达的信号转导通路研究[J]. 中国现代神经疾病杂志, 2021, 21(11): 942-950.

LI Yan-ju, ZHANG Ge, WANG Xiao-jing, XING Xiao-min, MA Guo-zhao. The effect of signal pathways on the neural K_ATP subunits SUR1/Kir6.2 expression induced by Aβ_1-42[J]. Chinese Journal of Contemporary Neurology and Neurosurgery, 2021, 21(11): 942-950.

http://journal11.magtechjournal.com/cjcnn/CN/Y2021/V21/I11/942

参考文献

[1] Kawasumi M, Hashimoto Y, Chiba T, Kanekura K, Yamagishi Y, Ishizaka M, Tajima H, Niikura T, Nishimoto I. Molecular mechanisms for neuronal cell death by Alzheimer's amyloid precursor protein-relevant insults[J]. Neurosignals, 2002, 11:236-250.
[2] Watson D, Castaño E, Kokjohn TA, Kuo YM, Lyubchenko Y, Pinsky D, Connolly ES Jr, Esh C, Luehrs DC, Stine WB, Rowse LM, Emmerling MR, Roher AE. Physicochemical characteristics of soluble oligomeric Abeta and their pathologic role in Alzheimer's disease[J]. Neurol Res, 2005, 27:869-881.
[3] Carrotta R, Di Carlo M, Manno M, Montana G, Picone P, Romancino D, San Biagio PL. Toxicity of recombinant beta-amyloid prefibrillar oligomers on the morphogenesis of the sea urchin Paracentrotus lividus[J]. FASEB J, 2006, 20:1916-1917.
[4] Ma G, Fu Q, Zhang Y, Gao J, Jiang J, Bi A, Liu K, Du Y, Chen C, Cui Y, Lu L. Effects of Abeta1-42 on the subunits of KATP expression in cultured primary rat basal forebrain neurons[J]. Neurochem Res, 2008, 33:1419-1424.
[5] Yamada S, Kane GC, Behfar A, Liu XK, Dyer RB, Faustino RS, Miki T, Seino S, Terzic A. Protection conferred by myocardial ATP-sensitive K⁺ channels in pressure overload-induced congestive heart failure revealed in KCNJ11 Kir6.2-null mutant[J]. J Physiol, 2006, 577(Pt 3):1053-1065.
[6] Lefer DJ, Nichols CG, Coetzee WA. Sulfonylurea receptor 1 subunits of ATP-sensitive potassium channels and myocardial ischemia/reperfusion injury[J]. Trends Cardiovasc Med, 2009, 19:61-67.
[7] Yu Y, Zhou L, Sun M, Zhou T, Zhong K, Wang H, Liu Y, Liu X, Xiao R, Ge J, Tu P, Fan DS, Lan Y, Hui C, Chui D. Xylocoside G reduces amyloid-β induced neurotoxicity by inhibiting NF-κB signaling pathway in neuronal cells[J]. J Alzheimers Dis, 2012, 30:263-275.
[8] Ferrer I. Stress kinases involved in tau phosphorylation in Alzheimer's disease, tauopathies and APP transgenic mice[J]. Neurotox Res, 2004, 6:469-475.
[9] Zhu X, Rottkamp CA, Hartzler A, Sun Z, Takeda A, Boux H, Shimohama S, Perry G, Smith MA. Activation of MKK6, an upstream activator of p38, in Alzheimer's disease[J]. J Neurochem, 2001, 79:311-318.
[10] Kim HJ, Kim JH, Chae SC, Park YC, Kwon KS, Hong ST. Soluble oligomeric Abeta disrupts the protein kinase C signaling pathway[J]. Neuroreport, 2004, 15:503-507.
[11] Sattiraju S, Reyes S, Kane GC, Terzic A. K(ATP) channel pharmacogenomics:from bench to bedside[J]. Clin Pharmacol Ther, 2008, 83:354-357.
[12] Selkoe DJ. Clearing the brain's amyloid cobwebs[J]. Neuron, 2001, 32:177-180.
[13] Liu J, Yin F, Zheng X, Jing J, Hu Y. Geniposide, a novel agonist for GLP-1 receptor, prevents PC12 cells from oxidative damage via MAP kinase pathway[J]. Neurochem Int, 2007, 51:361-369.
[14] Patel JR, Brewer GJ. Age-related differences in NFkappaB translocation and Bcl-2/Bax ratio caused by TNFalpha and Abeta42 promote survival in middle-age neurons and death in old neurons[J]. Exp Neurol, 2008, 213:93-100.
[15] Tickler AK, Wade JD, Separovic F. The role of Abeta peptides in Alzheimer's disease[J]. Protein Pept Lett, 2005, 12:513-519.
[16] Zeng X, Wang T, Jiang L, Ma G, Tan S, Li J, Gao J, Liu K, Zhang Y. Diazoxide and cyclosporin A protect primary cholinergic neurons against beta-amyloid (1-42)-induced cytotoxicity[J]. Neurol Res, 2013, 35:529-536.
[17] Zhi-Kun S, Hong-Qi Y, Zhi-Quan W, Jing P, Zhen H, Sheng-Di C. Erythropoietin prevents PC12 cells from beta-amyloid-induced apoptosis via PI3K/Akt pathway[J]. Transl Neurodegener, 2012, 1:7.
[18] Liu D, Slevin JR, Lu C, Chan SL, Hansson M, Elmér E, Mattson MP. Involvement of mitochondrial K⁺ release and cellular efflux in ischemic and apoptotic neuronal death[J]. J Neurochem, 2003, 86:966-979.
[19] Tan S, Ma G, Li Y, Li J, Yao W, Ren X, Liu X, Gao J. Effects of Aβ1-42 on the current of KATP channels in cultured cholinergic neurons[J]. Neurol Res, 2012, 34:707-713.
[20] Ma G, Gao J, Fu Q, Jiang L, Wang R, Zhang Y, Liu K. Diazoxide reverses the enhanced expression of KATP subunits in cholinergic neurons caused by exposure to Aβ_1-42[J]. Neurochem Res, 2009, 34:2133-2140.
[21] Elfering SL, Sarkela TM, Giulivi C. Biochemistry of mitochondria nitric -oxide synthase[J]. J Biol Chem, 2002, 277:38079-38086.
[22] Geng L, Zhang T, Liu W, Chen Y. Inhibition of miR-128 Abates Aβ-mediated cytotoxicity by targeting PPAR-γ via NF-κB inactivation in primary mouse cortical neurons and neuro2a cells[J]. Yonsei Med J, 2018, 59:1096-1106.
[23] Mattson MP, Culmsee C, Yu Z, Camandola S. Roles of nuclear factor kappaB in neuronal survival and plasticity[J]. J Neurochem, 2000, 74:443-456.
[24] Cuadrado A, Nebreda AR. Mechanisms and functions of p38 MAPK signalling[J]. Biochem J, 2010, 429:403-417.
[25] Barone FC, Irving EA, Ray AM, Lee JC, Kassis S, Kumar S, Badger AM, Legos JJ, Erhardt JA, Ohlstein EH, Hunter AJ, Harrison DC, Philpott K, Smith BR, Adams JL, Parsons AA. Inhibition of p38 mitogen-activated protein kinase provides neuroprotection in cerebral focal ischemia[J]. Med Res Rev, 2001, 21:129-145.
[26] Ossum CG, Lauritsen AN, Karottki DG, Hoffmann EK. Differential role for ERK2 in anoxia-induced activation of transcription and translation of Hsp70 in NIH 3T3 cells[J]. Cell Physiol Biochem, 2011, 27:109-120.
[27] Shukla D, Saxena S, Jayamurthy P, Sairam M, Singh M, Jain SK, Bansal A, Ilavazaghan G. Hypoxic preconditioning with cobalt attenuates hypobaric hypoxia-induced oxidative damage in rat lungs[J]. High Alt Med Biol, 2009, 10:57-69.
[28] Munoz L, Ralay Ranaivo H, Roy SM, Hu W, Craft JM, McNamara LK, Chico LW, Van Eldik LJ, Watterson DM. A novel p38 alpha MAPK inhibitor suppresses brain proinflammatory cytokine up-regulation and attenuates synaptic dysfunction and behavioral deficits in an Alzheimer's disease mouse model[J]. J Neuroinflammation, 2007, 4:21.
[29] Kim SH, Smith CJ, Van Eldik LJ. Importance of MAPK pathways for microglial pro-inflammatory cytokine IL-1 beta production[J]. Neurobiol Aging, 2004, 25:431-439.
[30] Bachstetter AD, Xing B, de Almeida L, Dimayuga, ER,Watterson DM, Van Eldik LJ. Microglial p38alpha MAPK is a key regulator of proinflammatory cytokine up-regulation induced by toll-like receptor (TLR) ligands or beta-amyloid (Abeta)[J]. J Neuroinflammation, 2011, 8:79.
[31] Bodles AM, Barger SW. Secreted beta-amyloid precursor protein activates microglia via JNK and p38-MAPK[J]. Neurobiol Aging, 2005, 26:9-16.
[32] Kikuchi M, Tenneti L, Lipton SA. Role of p38 mitogen-activated protein kinase in axotomy-induced apoptosis of retinal ganglion cells[J]. J Neurosci, 2000, 20:5037-5044.
[33] Ge B, Gram H, Di Padova F, Huang B, New L, Ulevitch Luo Y, Han J. MAPKK-independent activation of p38alpha mediated by TAB1-dependent autophosphorylation of p38alpha[J]. Science, 2002, 295:1291-1294.
[34] Li J, Miller EJ, Ninomiya-Tsuji J, Russell RR 3rd, Young LH. AMP-activated protein kinase activates p38 mitogen-activated protein kinase by increasing recruitment of p38 MAPK to TAB1 in the ischemic heart[J]. Circ Res, 2005, 97:872-879.
[35] D'Cruz BJ, Fertig KC, Filiano AJ, Hicks SD, DeFranco DB, Callaway CW. Hypothermic reperfusion after cardiac arrest augments brain-derived neurotrophic factor activation[J]. J Cereb Blood Flow Metab, 2002, 22:843-851.
[36] Xu SZ, Bullock L, Shan CJ, Cornelius K, Rajanna B. PKC isoforms were reduced by lead in the developing rat brain[J]. Int J Dev Neurosci, 2005, 23:53-64.
[37] Fu H, Dou J, Li W, Cui W, Mak S, Hu Q, Luo J, Lam CS, Pang Y, Youdim MB, Han Y. Promising multifunctional anti-Alzheimer's dimer bis(7)-cognitin acting as an activator of protein kinase C regulates activities of alpha-secretase and BACE-1 concurrently[J]. Eur J Pharmacol, 2009, 623:14-21.
[38] Lanni C, Mazzucchelli M, Porrello E, Govoni S, Racchi M. Differential involvement of protein kinase C alpha and epsilon in the regulated secretion of soluble amyloid precursor protein[J]. Eur J Biochem, 2004, 271:3068-3075.
[39] Alkon DL, Epstein H, Kuzirian A, Bennett MC, Nelson TJ. Protein synthesis required for long-term memory is induced PKC activation on days before associative learning[J]. Proc Natl Acad Sci USA, 2005, 102:16432-16437.

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2 Aβ1~42诱导KATP亚基SUR1/Kir6.2蛋白表达的信号转导通路研究

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2 Aβ_1~42诱导K_ATP亚基SUR1/Kir6.2蛋白表达的信号转导通路研究

The effect of signal pathways on the neural K_ATP subunits SUR1/Kir6.2 expression induced by Aβ_1-42