艾烟提取物对神经细胞凋亡的影响和机制探讨
|
摘要
目的:探讨艾烟提取物(ME)对神经细胞及神经毒性损伤模型凋亡的影响及潜在机制。方法:采用24h内新生SD大鼠乳鼠前额叶皮质及海马组织原代培养细胞,谷氨酸(GLU)诱导建立神经毒性损伤模型。运用MTT及Western Blot检测细胞存活率及目的蛋白表达,并用AKT通路抑制剂LY294002验证作用通路。结果:GLU降低神经细胞的存活率(P<0.01),ME抑制了正常神经细胞及谷氨酸诱导神经毒性损伤模型的凋亡(P<0.05);Western Blot显示,GLU降低了P-AKT、AKT、BCL-2的表达,上调active-Caspase-3的表达(P<0.01),而ME作用正好相反。结论:ME能延缓神经细胞的凋亡,其机制通过激活PI3K/AKT信号通路并调控下游的细胞凋亡蛋白BCL-2、active-Caspase-3的表达来拮抗细胞凋亡。
Objective: To investigate the effects and potential mechanisms of moxa smoke extract(ME) on apoptosis of neurons and neurotoxic injury model. Methods: Primary cultured cells were from prefrontal cortex and hippocampus of newborn SD rats within 24 hours and neurotoxicity injury model was induced by glutamate(GLU). Cell viability and protein expression were detected by MTT and Western Blot, and the pathway was verified by AKT pathway inhibitor LY294002. Results: GLU decreased the survival rate of nerve cells(P<0.01), and ME inhibited the apoptosis of normal nerve cells and GLU-induced neurotoxicinjury model(P<0.05). Western Blot showed that GLU decreased the expression of P-AKT, AKT, BCL-2, and upregulated the expression of active-Caspase-3(P<0.01), whereas ME had the opposite effect. Conclusion: ME can delay the apoptosis of nerve cells by activating the PI3 K/AKT signaling pathway and regulating the expression of downstream apoptosis proteins BCL-2 and active-Caspase-3.
Objective: To investigate the effects and potential mechanisms of moxa smoke extract(ME) on apoptosis of neurons and neurotoxic injury model. Methods: Primary cultured cells were from prefrontal cortex and hippocampus of newborn SD rats within 24 hours and neurotoxicity injury model was induced by glutamate(GLU). Cell viability and protein expression were detected by MTT and Western Blot, and the pathway was verified by AKT pathway inhibitor LY294002. Results: GLU decreased the survival rate of nerve cells(P<0.01), and ME inhibited the apoptosis of normal nerve cells and GLU-induced neurotoxicinjury model(P<0.05). Western Blot showed that GLU decreased the expression of P-AKT, AKT, BCL-2, and upregulated the expression of active-Caspase-3(P<0.01), whereas ME had the opposite effect. Conclusion: ME can delay the apoptosis of nerve cells by activating the PI3 K/AKT signaling pathway and regulating the expression of downstream apoptosis proteins BCL-2 and active-Caspase-3.
引文
[1]Johnson J L,Aprison M H.The distribution of glutamate and total free amino acids in thirteen specific regions of the cat central nervous system.Brain Research,1971,26(1):141-148
[2]Mc Donald J W,Johnston M V.Physiological and pathophysiological roles of excitatory amino acids during central nervous system developmengt.Brain Research,1990,15:41-70
[3]Olney J W,Sharpe L G.Brain lesions in an infant rhesus monkey treated with monosodium glutamate.Science,1969,166(3903):386-388
[4]Olney J W.Brain lesions,obesity,and other disturbances in mice treated with monosodium glutamate.Science,1969,164(3880):719-721
[5]Dong X,Wang Y,Qin Z.Molecular mechanisms of excitotoxicity and their relevance to pathogenesis of neurodegenerative diseases.中国药理学报(英文版),2009,30(4):379-387
[6]韩丑萍,周次利,丰晓溟,等.艾烟的研究进展.针灸推拿医学(英文版),2011,9(2):67-72
[7]Yang,Zheng,Xiaojun,et al.Effect of moxa smoke produced during combustion of Aiye(Folium Artemisiae Argyi)on behavioral changes in mice inhaling the smoke.Journal of Traditional Chinese Medicine,2016,36(6):805-811
[8]于密密,靳然,赵百孝,等.SPME-GC-MS法测定艾烟中苯酚的含量.药物分析杂志,2012(10):1870-1873
[9]Xu H,Zhao B,Cui Y,et al.Effects of moxa smoke on monoamine neurotransmitters in SAMP8 mice.Evid Based Complement Alternat Med,2013,2013:178067
[10]Huang J,Lim M Y,Zhao B,et al.PM10 mass concentration and oxidative capacity of moxa smoke.Q J M,2015,108(9):705-710
[11]Hitosugi N,Ohno R,Hatsukari I,et al.Diverse biological activity of moxa extract and smoke.In Vivo,2001,15:249-254
[12]Sakagami H,Matsumoto H,Satoh K,et al.Cytotoxicity and radical modulating activity of Moxa smoke.In Vivo,2005,19(2):391-397
[13]Matsumoto H,Shimada J,Nagasaka H,et al.Inhibition by Moxa smoke of NO production and iNOS expression in mouse macrophage-like cells Raw 264.7..In Vivo,2005,19(2):471-474
[14]孟笑男,徐焕芳,崔莹雪,等.艾燃烧生成物对快速老化模型小鼠大脑SOD、MDA和GSH-Px的影响.环球中医药,2011,4(6):413-415
[15]刘耀萌,刘钧天,黄畅,等.不同艾灸因素对阿尔茨海默小鼠PI3K/AKT通路与皮质β淀粉样蛋白沉淀的影响.世界中医药,2016,11(8):1395-1400
[16]黄文娟,万茜,徐天舒.麦粒灸对Lewis荷瘤小鼠Th1/Th2型细胞因子的影响.针刺研究,2014,39(6):477-481
[17]Wang S J,Wang Q,Ma J.Effect of moxibustion on mTOR-mediated autophagy in rotenone-induced Parkinson’s disease model rats.Neural Regeneration Research,2018,13(1):112-118
[18]黄玉海,刘平,杨必成,等.艾烟可吸入颗粒物对人肺腺癌A549细胞凋亡的影响.中国中医药信息杂志,2014,21(7):53-57
[19]蔡虹,邬继红,赵百孝.凝物对大鼠肺泡巨噬细胞NR8383活性和吞噬功能的影响.北京中医药大学学报,2013,36(7):501-504
[20]王述菊,孙国杰,马骏,等.艾灸对Alzheimer病模型大鼠行为学及海马区Bcl-2、Bax、Caspase-3影响的研究.世界科学技术-中医药现代化,2015(6):1243-1248
[21]Liu J,Zhao B,Cui Y,et al.Effects of shenque moxibustion on behavioral changes and brain oxidative state in apolipoprotein E-deficient mice.Evid Based Complement Alternat Med,2015,2015:1-8
[22]姜美驰,梁静,王静蓉,等.艾灸对AD大鼠学习记忆能力及海马Aβ、IL-1β、IL-2表达的影响.上海针灸杂志,2016,35(7):870-875
[23]王静蓉,张玉杰,姜美驰,等.艾灸对AD大鼠学习记忆、GSK-3β和磷酸化tau蛋白的影响.中国中医基础医学杂志,2015(10):1287-1290
[24]王述菊,孙国杰,马骏,等.艾灸对阿尔茨海默病模型大鼠行为学及海马超微结构的影响.中国中医药信息杂志,2015,22(11):58-61
[25]王晓兰,孙国杰,杜艳军,等.艾灸预处理对阿尔茨海默病模型大鼠海马CA1区NGF和BDNF表达的影响.中国康复,2011,26(5):323-326
[26]刘静,杜艳军,李婧,等.艾灸预刺激对AD大鼠细胞周期调节因子CyclinA,p21/cip的影响.时珍国医国药,2011,22(11):2612-2613
[27]杜艳军,宋杰,周华,等.艾灸预刺激对阿尔茨海默病大鼠Hsp70、Hsp90的影响.辽宁中医杂志,2013(3):577-579
[28]Yao R,Cooper G M.Requirement for phosphatidylinositol-3 kinase in the prevention of apoptosis by nerve growth factor.Science,1995,267(5206):2003-2006
[29]谢玮蓉,张刚.PI3K/AKT/Bcl-2凋亡信号传导通路的研究进展.中国当代医药,2015(30):22-25
[2]Mc Donald J W,Johnston M V.Physiological and pathophysiological roles of excitatory amino acids during central nervous system developmengt.Brain Research,1990,15:41-70
[3]Olney J W,Sharpe L G.Brain lesions in an infant rhesus monkey treated with monosodium glutamate.Science,1969,166(3903):386-388
[4]Olney J W.Brain lesions,obesity,and other disturbances in mice treated with monosodium glutamate.Science,1969,164(3880):719-721
[5]Dong X,Wang Y,Qin Z.Molecular mechanisms of excitotoxicity and their relevance to pathogenesis of neurodegenerative diseases.中国药理学报(英文版),2009,30(4):379-387
[6]韩丑萍,周次利,丰晓溟,等.艾烟的研究进展.针灸推拿医学(英文版),2011,9(2):67-72
[7]Yang,Zheng,Xiaojun,et al.Effect of moxa smoke produced during combustion of Aiye(Folium Artemisiae Argyi)on behavioral changes in mice inhaling the smoke.Journal of Traditional Chinese Medicine,2016,36(6):805-811
[8]于密密,靳然,赵百孝,等.SPME-GC-MS法测定艾烟中苯酚的含量.药物分析杂志,2012(10):1870-1873
[9]Xu H,Zhao B,Cui Y,et al.Effects of moxa smoke on monoamine neurotransmitters in SAMP8 mice.Evid Based Complement Alternat Med,2013,2013:178067
[10]Huang J,Lim M Y,Zhao B,et al.PM10 mass concentration and oxidative capacity of moxa smoke.Q J M,2015,108(9):705-710
[11]Hitosugi N,Ohno R,Hatsukari I,et al.Diverse biological activity of moxa extract and smoke.In Vivo,2001,15:249-254
[12]Sakagami H,Matsumoto H,Satoh K,et al.Cytotoxicity and radical modulating activity of Moxa smoke.In Vivo,2005,19(2):391-397
[13]Matsumoto H,Shimada J,Nagasaka H,et al.Inhibition by Moxa smoke of NO production and iNOS expression in mouse macrophage-like cells Raw 264.7..In Vivo,2005,19(2):471-474
[14]孟笑男,徐焕芳,崔莹雪,等.艾燃烧生成物对快速老化模型小鼠大脑SOD、MDA和GSH-Px的影响.环球中医药,2011,4(6):413-415
[15]刘耀萌,刘钧天,黄畅,等.不同艾灸因素对阿尔茨海默小鼠PI3K/AKT通路与皮质β淀粉样蛋白沉淀的影响.世界中医药,2016,11(8):1395-1400
[16]黄文娟,万茜,徐天舒.麦粒灸对Lewis荷瘤小鼠Th1/Th2型细胞因子的影响.针刺研究,2014,39(6):477-481
[17]Wang S J,Wang Q,Ma J.Effect of moxibustion on mTOR-mediated autophagy in rotenone-induced Parkinson’s disease model rats.Neural Regeneration Research,2018,13(1):112-118
[18]黄玉海,刘平,杨必成,等.艾烟可吸入颗粒物对人肺腺癌A549细胞凋亡的影响.中国中医药信息杂志,2014,21(7):53-57
[19]蔡虹,邬继红,赵百孝.凝物对大鼠肺泡巨噬细胞NR8383活性和吞噬功能的影响.北京中医药大学学报,2013,36(7):501-504
[20]王述菊,孙国杰,马骏,等.艾灸对Alzheimer病模型大鼠行为学及海马区Bcl-2、Bax、Caspase-3影响的研究.世界科学技术-中医药现代化,2015(6):1243-1248
[21]Liu J,Zhao B,Cui Y,et al.Effects of shenque moxibustion on behavioral changes and brain oxidative state in apolipoprotein E-deficient mice.Evid Based Complement Alternat Med,2015,2015:1-8
[22]姜美驰,梁静,王静蓉,等.艾灸对AD大鼠学习记忆能力及海马Aβ、IL-1β、IL-2表达的影响.上海针灸杂志,2016,35(7):870-875
[23]王静蓉,张玉杰,姜美驰,等.艾灸对AD大鼠学习记忆、GSK-3β和磷酸化tau蛋白的影响.中国中医基础医学杂志,2015(10):1287-1290
[24]王述菊,孙国杰,马骏,等.艾灸对阿尔茨海默病模型大鼠行为学及海马超微结构的影响.中国中医药信息杂志,2015,22(11):58-61
[25]王晓兰,孙国杰,杜艳军,等.艾灸预处理对阿尔茨海默病模型大鼠海马CA1区NGF和BDNF表达的影响.中国康复,2011,26(5):323-326
[26]刘静,杜艳军,李婧,等.艾灸预刺激对AD大鼠细胞周期调节因子CyclinA,p21/cip的影响.时珍国医国药,2011,22(11):2612-2613
[27]杜艳军,宋杰,周华,等.艾灸预刺激对阿尔茨海默病大鼠Hsp70、Hsp90的影响.辽宁中医杂志,2013(3):577-579
[28]Yao R,Cooper G M.Requirement for phosphatidylinositol-3 kinase in the prevention of apoptosis by nerve growth factor.Science,1995,267(5206):2003-2006
[29]谢玮蓉,张刚.PI3K/AKT/Bcl-2凋亡信号传导通路的研究进展.中国当代医药,2015(30):22-25