尼古丁通过PI3K/AKT信号通路上调星形胶质细胞HSF1的表达
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摘要
目的:研究尼古丁对星形胶质细胞热休克转录因子1(HSF1)的表达的影响,并探讨其机制。方法:分离新出生24 h内乳鼠大脑皮质,进行原代培养并鉴定为星形胶质细胞,待细胞成熟后分为正常对照组、尼古丁处理组,PI3K/AKT信号通路阻断剂LY294002处理组和尼古丁+LY294002处理组,正常对照组不做任何处理;尼古丁处理组用5μmol/L尼古丁分别处理星形胶质细胞6、12、18、24 h; LY294002处理组只加10μmol/L LY294002处理;尼古丁+LY294002处理组是先加10μmol/L LY294002处理2 h后,再加入5μmol/L尼古丁共同处理18 h,免疫印迹法观察各组HSF1蛋白的表达。结果:与正常对照组相比,尼古丁处理组星形胶质细胞内HSF 1蛋白的表达上调(P <0. 05);与尼古丁处理组比较,LY294002+尼古丁处理组星形胶质细胞内HSF1表达明显受到抑制(P <0. 05)。结论:尼古丁通过PI3K/AKT信号通路上调星形胶质细胞HSF1蛋白表达。
Objective: To study the effect of nicotine on the expression of heat shock transcription factor 1( HSF1) in astrocytes and explore its mechanism. Methods: To get primary rat astrocytes,the cerebral cortexes of newborn rats within 24 h were isolated,prepared for single cell suspension,cultured and verified using GFAP immunostaining. After astrocytes matured,they were divided into four groups: control group,nicotine group,LY294002( PI3 K/AKT inhibitor) group and the nicotine +LY294002 group. The control group was not given any treatment,; the nicotine group was treated with5 μmol/L nicotine for 6 h,12 h,18 h and 24 h,respectively; LY294002 group was given 10 μmol/L LY294002; nicotine + LY294002 group was pretreated with 10 μmol/L LY294002 for 2 h and then treated with 5 μmol/L nicotine for 18 h. The expression levels of HSF1 protein were detected by immunoblotting. Results: Compared with the control group,HSF-1 level was upregulated in nicotine group( P < 0. 05). HSF1 expression level was remarkably lower in LY294002 + nicotine group than that in nicotine group( P < 0. 01). Conclusion: Nicotine upregulates HSF1 expression through PI3 K/AKT signaling pathway in astrocytes.
Objective: To study the effect of nicotine on the expression of heat shock transcription factor 1( HSF1) in astrocytes and explore its mechanism. Methods: To get primary rat astrocytes,the cerebral cortexes of newborn rats within 24 h were isolated,prepared for single cell suspension,cultured and verified using GFAP immunostaining. After astrocytes matured,they were divided into four groups: control group,nicotine group,LY294002( PI3 K/AKT inhibitor) group and the nicotine +LY294002 group. The control group was not given any treatment,; the nicotine group was treated with5 μmol/L nicotine for 6 h,12 h,18 h and 24 h,respectively; LY294002 group was given 10 μmol/L LY294002; nicotine + LY294002 group was pretreated with 10 μmol/L LY294002 for 2 h and then treated with 5 μmol/L nicotine for 18 h. The expression levels of HSF1 protein were detected by immunoblotting. Results: Compared with the control group,HSF-1 level was upregulated in nicotine group( P < 0. 05). HSF1 expression level was remarkably lower in LY294002 + nicotine group than that in nicotine group( P < 0. 01). Conclusion: Nicotine upregulates HSF1 expression through PI3 K/AKT signaling pathway in astrocytes.
引文
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[20]KAWAMATA J,SHIMOHAMA S. Stimulating nicotinic receptors trigger multiple pathways attenuating cytotoxicity in models of Alzheimer's and Parkinson's diseases[J].Journal of Alzheimers Disease Jad,2011,24(Suppl 2):95-109.
[21]任真奎,杨梅,官志忠,等. PNU上调大鼠皮质星形胶质细胞内源性Cryab抑制Aβ聚集[J].中国老年学,2016,36(22):5506-5509.
[2]BOBKOVA N V,GARBUZ D G,NESTEROVA I,et al.Therapeutic effect of exogenous hsp70 in mouse models of Alzheimer's disease[J]. Journal of Alzheimers Disease Jad,2014,38(2):425-435.
[3]TOTH M E,SZEGEDI V,VARGA E,et al. Overexpression of Hsp27 ameliorates symptoms of Alzheimer's disease in APP/PS1 mice[J]. Cell Stress&Chaperones,2013,18(6):759-771.
[4]RIVERA I,CAPONE R,CAUVI D M,et al. Modulation of Alzheimer's amyloidβpeptide oligomerization and toxicity by extracellular Hsp70[J]. Cell Stress and Chaperones,2018,23(2):269-279.
[5] GOMEZ-PASTOR R,BURCHFIEL E T,THIELE D J.Regulation of heat shock transcription factors and their roles in physiology and disease[J]. Nature Reviews Molecular Cell Biology,2017,19(1):4-19.
[6]NEEF D W,JAEGER A M,THIELE D J. Heat shock transcription factor 1 as a therapeutic target in neurodegenerative diseases[J]. Nature Reviews Drug Discovery,2011,10(12):930-944.
[7]JIANXIN S,JIE W. Nicotinic cholinergic mechanisms in Alzheimer's disease[J]. International Review of Neurobiology,2015,124(4):275-292.
[8]ZHANG B,HE P,LU Y,et al. HSF1 relieves amyloid-β-induced cardiomyocytes apoptosis[J]. Cell Biochemistry and Biophysics,2015,72(2):579-587.
[9]MCCARTHY K D,VELLIS J D. Preparation of separate astroglial and oligodendroglial cell cultures from rat cerebral tissue.[J]. Journal of Cell Biology,1980,85(3):890-902.
[10]SCHILDGE S,BOHRER C,BECK K,et al. Isolation and culture of mouse cortical astrocytes[J]. Journal of Visualized Experiments Jove,2013,71(71):e50079.
[11]CUMMINGS J,LEE G,MORTSDORF T,et al. Alzheimer's disease drug development pipeline:2017[J].Alzheimer's&Dementia:Translational Research&Clinical Interventions,2017,3(3):367-384.
[12]ASSEFA B T,GEBRE A K,ALTAYE B M. Reactive astrocytes as drug target in alzheimer’s disease[J]. Bio Med Research International,2018,2018:4160247.
[13]LIAN H,LITVINCHUK A,CHIANG A C A,et al. Astrocyte-microglia cross talk through complement activation modulates amyloid pathology in mouse models of Alzheimer's disease[J]. Journal of Neuroscience,2016,36(2):577-589.
[14]FROST G R,LI Y. The role of astrocytes in amyloid production and Alzheimer's disease[J]. Open Biology,2017,7(12):170-228.
[15]孔欣,禹文峰,官志忠.激活星形胶质细胞α7乙酰胆碱能受体抑制β-淀粉样蛋白的聚集[J].中国老年学,2016,36(13):3112-3115.
[16]MAVROUDIS I A,FOTIOU D F,ADIPEPE L F,et al.Morphological changes of the human purkinje cells and deposition of neuritic plaques and neurofibrillary tangles on the cerebellar cortex of Alzheimer's disease[J]. American Journal of Alzheimers Disease&Other Dementias,2010,25(7):585-591.
[17]JIANG Y Q,WANG X L,CAO X H,et al. Increased heat shock transcription factor 1 in the cerebellum reverses the deficiency of Purkinje cells in Alzheimer's disease[J]. Brain Research,2013,1519:105-111.
[18]ANSON P,NATALIA P,HALLORAN J J,et al. Overexpression of heat shock factor 1 phenocopies the effect of chronic inhibition of TOR by rapamycin and is sufficient to ameliorate Alzheimer's-like deficits in mice modeling the disease[J]. Journal of Neurochemistry,2013,124(6):880-893.
[19]CHONG W,JIN-TAI Y,DAN M,et al. Targeting the m TOR signaling network for Alzheimer's disease therapy[J]. Molecular Neurobiology,2014,49(1):120-135.
[20]KAWAMATA J,SHIMOHAMA S. Stimulating nicotinic receptors trigger multiple pathways attenuating cytotoxicity in models of Alzheimer's and Parkinson's diseases[J].Journal of Alzheimers Disease Jad,2011,24(Suppl 2):95-109.
[21]任真奎,杨梅,官志忠,等. PNU上调大鼠皮质星形胶质细胞内源性Cryab抑制Aβ聚集[J].中国老年学,2016,36(22):5506-5509.