IL-1β促进星形胶质细胞增殖并下调Kir4.1的表达
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摘要
目的探讨白细胞介素1β(IL-1β)对星形胶质细胞增殖及其内向整流性钾离子通道4.1(Kir4.1)表达的影响。方法体外培养新生SD大鼠大脑皮层星形胶质细胞;用IL-1β或者和白细胞介素1受体拮抗剂(IL-1Ra)处理培养的星形胶质细胞,流式细胞术检测IL-1β对星形胶质细胞细胞周期的影响;荧光定量PCR检测IL-1β对星形胶质细胞Kir4.1 mRNA表达的影响;Western blot法检测IL-1β对星形胶质细胞Kir4.1蛋白表达的影响。结果 IL-1β可以以剂量和时间依赖的方式促进星形胶质细胞的增殖。用10 ng/m L的IL-1β处理星形胶质细胞24 h后,Kir4.1 mRNA和蛋白的表达均下降,IL-1Ra可以有效对抗由IL-1β引起的Kir4.1 mRNA和蛋白的表达下降;Kir4.1 mRNA和蛋白的表达下调也能因IL-1β的去除而部分被恢复。结论 IL-1β可以促进星形胶质细胞的增殖,IL-1β是影响星形胶质细胞Kir4.1表达的关键因素。
Objective To explore the impact of IL-1β on the proliferation of astrocytes and the expression of the inwardly rectifying potassium channel 4. 1( Kir4. 1) in astrocytes. Methods Astrocytes were isolated from cerebral cortex of newborn SD rats and cultured in the presence of IL-1β or IL-1β combined with interleukin-1 receptor antagonist( IL-1Ra). The effect of IL-1β on the cell cycle of astrocytes was measured with flow cytometry; the level of Kir4. 1 mRNA was determined by quantitative real-time PCR. The level of Kir4. 1 protein was detected by Western blotting. Results IL-1β promoted astrocyte proliferation in a time-and dose-dependent manner. After astrocytes were treated with IL-1β( 10 ng/m L) for 24 hours,the levels of Kir4. 1mRNA and protein significantly decreased,but IL-1Ra significantly inhibited IL-1β-induced decrease of Kir4. 1. The downregulation of Kir4. 1 mRNA and protein expressions could be partially reverted when IL-1β was removed. Conclusion IL-1β could promote the proliferation of cultured astrocytes,and IL-1β may be a key factor influencing the expression of Kir4. 1 in astrocytes.
Objective To explore the impact of IL-1β on the proliferation of astrocytes and the expression of the inwardly rectifying potassium channel 4. 1( Kir4. 1) in astrocytes. Methods Astrocytes were isolated from cerebral cortex of newborn SD rats and cultured in the presence of IL-1β or IL-1β combined with interleukin-1 receptor antagonist( IL-1Ra). The effect of IL-1β on the cell cycle of astrocytes was measured with flow cytometry; the level of Kir4. 1 mRNA was determined by quantitative real-time PCR. The level of Kir4. 1 protein was detected by Western blotting. Results IL-1β promoted astrocyte proliferation in a time-and dose-dependent manner. After astrocytes were treated with IL-1β( 10 ng/m L) for 24 hours,the levels of Kir4. 1mRNA and protein significantly decreased,but IL-1Ra significantly inhibited IL-1β-induced decrease of Kir4. 1. The downregulation of Kir4. 1 mRNA and protein expressions could be partially reverted when IL-1β was removed. Conclusion IL-1β could promote the proliferation of cultured astrocytes,and IL-1β may be a key factor influencing the expression of Kir4. 1 in astrocytes.
引文
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[20]Bataveljic D,Nikolic L,Milosevic M,et al.Changes in the astrocytic aquaporin-4 and inwardly rectifying potassium channel expression in the brain of the amyotrophic lateral sclerosis SOD1(G93A)rat model[J].Glia,2012,60(12):1991-2003.
[21]Huang Y,Mucke L.Alzheimer mechanisms and therapeutic strategies[J].Cell,2012,148(6):1204-1222.
[22]Tong X,Ao Y,Faas G C,et al.Astrocyte Kir4.1 ion channel deficits contribute to neuronal dysfunction in Huntington’s disease model mice[J].Nat Neurosci,2014,17(5):694-703.
[23]Minkel H R,Anwer T Z,Arps K M,et al.Elevated GFAP induces astrocyte dysfunction in caudal brain regions:A potential mechanism for hindbrain involved symptoms in typeⅡAlexander disease[J].Glia,2015,63(12):2285-2297.
[24]Tan G,Sun S Q,Yuan D L.Expression of Kir 4.1 in human astrocytic tumors:correlation with pathologic grade[J].Biochem Biophys Res Commun,2008,367(4):743-747.
[25]Chen C,Chen H,Xu C,et al.Role of interleukin-1 in hypoxiainduced depression of glutamate uptake in retinal Müller cells[J].Graefes Arch Clin Exp Ophthalmol,2014,252(1):51-58.
[2]Pekny M,Pekna M,Messing A,et al.Astrocytes:a central element in neurological diseases[J].Acta Neuropathol,2016,131(3):323-345.
[3]Sudduth T L,Weekman E M,Price B R,et al.Time-course of glial changes in the hyperhomocysteinemia model of vascular cognitive impairment and dementia(VCID)[J].Neuroscience,2017,341:42-51.
[4]Larsen B R,Mac Aulay N.Kir4.1-mediated spatial buffering of K+:experimental challenges in determination of its temporal and quantitative contribution to K+clearance in the brain[J].Channels(Austin),2014,8(6):544-550.
[5]Djukic B,Casper K B,Philpot B D,et al.Conditional knock-out of Kir4.1 leads to glial membrane depolarization,inhibition of potassium and glutamate uptake,and enhanced short-term synaptic potentiation[J].J Neurosci,2007,27(42):11354-11365.
[6]Sibille J,Dao Duc K,Holcman D,et al.The neuroglial potassium cycle during neurotransmission:role of Kir4.1 channels[J/OL].PLo S Comput Biol,2015,11(3):e1004137.doi:10.1371/journal.pcbi.1004137.
[7]Nwaobi S E,Cuddapah V A,Patterson K C,et al.The role of glial-specific Kir4.1 in normal and pathological states of the CNS[J].Acta Neuropathol,2016,132(1):1-21.
[8]Zurolo E,de Groot M,Iyer A,et al.Regulation of Kir4.1 expression in astrocytes and astrocytic tumors:a role for interleukin-1β[J/OL].J Neuroinflammation,2012,9:280.doi:10.1186/1742-2094-9-280.
[9]孙美群,严海芹,邹维艳,等.脂多糖活化星形胶质细胞并下调其内向整流性钾离子通道(Kir4.1)的表达[J].细胞与分子免疫学杂志,2016,32(2):196-200.
[10]Kan M H,Yang T,Fu H Q,et al.Pyrrolidine dithiocarbamate prevents neuroinflammation and cognitive dysfunction after endotoxemia in rats[J/OL].Front Aging Neurosci,2016,8:175.doi:10.3389/fnagi.2016.00175.
[11]Saghazadeh A,Ferrari C C,Rezaei N.Deciphering variability in the role of interleukin-1βin Parkinson’s disease[J].Rev Neurosci,2016,27(6):635-650.
[12]朱晓琴,李正莉,朱长庚,等.IL-1β或IL-6对大脑皮质星形胶质细胞细胞周期的影响[J].神经解剖学杂志,2005,21(3):313-316.
[13]阮林,刘仁刚,周洁萍,等.IL-1β对星形胶质细胞的激活作用[J].中国组织化学与细胞化学杂志,2005,14(4):404-408.
[14]Revathikumar P,Bergqvist F,Gopalakrishnan S,et al.Immunomodulatory effects of nicotine on interleukin 1βactivated human astrocytes andthe role of cyclooxygenase 2 in the underlying mechanism[J/OL].J Neuroinflammation,2016,13(1):256.doi:10.1186/s12974-016-0725-1.
[15]Araque A,Carmignoto G,Haydon P G,et al.Gliotransmitters travel in time and space[J].Neuron,2014,81(4):728-739.
[16]Fields J,Ghorpade A.C/EBPβregulates multiple IL-1β-induced human astrocyte inflammatory genes[J/OL].J Neuroinflammation,2012,9:177.doi:10.1186/1742-2094-9-177.
[17]Kucheryavykh Y V,Kucheryavykh L Y,Nichols C G,et al.Downregulation of Kir4.1 inward rectifying potassium channel subunits by RNAi impairs potassium transfer and glutamate uptake by cultured cortical astrocytes[J].Glia,2007,55(3):274-281.
[18]Chever O,Djukic B,Mc Carthy K D,et al.Implication of Kir4.1channel in excess potassium clearance:an in vivo study on anesthetized glial conditional Kir4.1 knock-out mice[J].J Neurosci,2010,30(47):15769-15777.
[19]Bockenhauer D,Feather S,Stanescu H C,et al.Epilepsy,ataxia,sensorineural deafness,tubulopathy,and KCNJ10 mutations[J].N Eng J Med,2009,360(19):1960-1970.
[20]Bataveljic D,Nikolic L,Milosevic M,et al.Changes in the astrocytic aquaporin-4 and inwardly rectifying potassium channel expression in the brain of the amyotrophic lateral sclerosis SOD1(G93A)rat model[J].Glia,2012,60(12):1991-2003.
[21]Huang Y,Mucke L.Alzheimer mechanisms and therapeutic strategies[J].Cell,2012,148(6):1204-1222.
[22]Tong X,Ao Y,Faas G C,et al.Astrocyte Kir4.1 ion channel deficits contribute to neuronal dysfunction in Huntington’s disease model mice[J].Nat Neurosci,2014,17(5):694-703.
[23]Minkel H R,Anwer T Z,Arps K M,et al.Elevated GFAP induces astrocyte dysfunction in caudal brain regions:A potential mechanism for hindbrain involved symptoms in typeⅡAlexander disease[J].Glia,2015,63(12):2285-2297.
[24]Tan G,Sun S Q,Yuan D L.Expression of Kir 4.1 in human astrocytic tumors:correlation with pathologic grade[J].Biochem Biophys Res Commun,2008,367(4):743-747.
[25]Chen C,Chen H,Xu C,et al.Role of interleukin-1 in hypoxiainduced depression of glutamate uptake in retinal Müller cells[J].Graefes Arch Clin Exp Ophthalmol,2014,252(1):51-58.