长期负性情绪应激对大鼠学习记忆及海马Ca~(2+)/CaM-CaMKⅡ-CREB信号通路的影响
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摘要
目的探讨长期负性情绪应激对大鼠学习记忆能力的影响,并从海马钙离子(Ca2+)/钙调蛋白(Ca M)-钙调蛋白激酶(Ca MK)Ⅱ-环腺苷酸反应元件结合蛋白(CREB)信号通路探讨其可能机制。方法将大鼠分为空白对照组、D-半乳糖(D-gal)脑老化组、负性情绪应激组。通过Morris水迷宫实验检测各组大鼠学习记忆能力,采用高效液相法、荧光免疫染色法和Western印迹方法分别测定海马神经元谷氨酸(glu),Ca2+,N-甲基-D-天冬氨酸受体(NMDAR)、Ca M和磷酸化(P)-CREB的含量。结果与空白对照组相比,D-gal脑老化组和负性情绪应激组Morris水迷宫寻台时间(SPL)均显著延长(P<0. 05);与D-gal脑老化组相比,负性情绪应激组大鼠的SPL无显著差异。与空白对照组相比,D-gal脑老化组和负性情绪应激组海马神经元内glu含量、Ca2+浓度和NMDAR含量均显著增加(P<0. 05),Ca M和P-CREB的含量均显著下降(P<0. 05);与脑老化组相比,负性情绪应激组Ca M含量显著升高(P<0. 05),其他指标无明显差异。结论长期负性情绪应激是加速认知衰退进程的重要原因之一,可产生与D-gal相似的认知损伤,其部分机制可能是通过Ca M-CaMKⅡ-CREB信号通路,影响学习记忆相关蛋白表达,引起学习记忆能力下降。
Objective To investigate the effect of chronic negative emotional stress on learning and memory ability in mice,and explore the possible mechanism from Ca2+/Ca M-CaMKⅡ-CREB signaling pathway in hippocampus.Methods Forty-two mice were divided into three groups on average: blank control( BC group),D-galactose( D-gal) brain aging( D-gal) and negative emotional stress( NES) groups. The spatial memory of each group was detected by Morris water maze test. The concentrations of glutamate ( glu),calcium ion( Ca2+),N-methyl-D-aspartate receptor( NMDAR),calmodulin( Ca M) and phosphorylated cyclic adenosine monophosphate effector binding protein( P-CREB) in hippocampal neuron's were detected by high performance liquid chromatography,fluorescence immunostaining and Western blot,respectively.Results Compared with that of BC group,the searching platform latency ( SPL) in Morris water maze test was significantly prolonged( P<0.05) in both of D-gal and NES groups. No difference in SPL was found between NES group and D-gal group. The contents of glu,Ca2+and NMDAR in D-gal and NES groups were significantly higher than those in BC group( P<0.05),while the protein expression levels of Ca M and P-CREB were significantly decreased( P<0.05). Additionally,the level of Ca M was higher in NES group than that in D-gal group( P < 0. 05). Conclusions Chronic negative emotional stress is one of the important reasons of accelerating cognitive decline process,which is similar to D-gal,and the Ca M-CaMKⅡ-CREB signaling pathway might be the important mechanism through affecting the related protein's expression of learning and memory.
Objective To investigate the effect of chronic negative emotional stress on learning and memory ability in mice,and explore the possible mechanism from Ca2+/Ca M-CaMKⅡ-CREB signaling pathway in hippocampus.Methods Forty-two mice were divided into three groups on average: blank control( BC group),D-galactose( D-gal) brain aging( D-gal) and negative emotional stress( NES) groups. The spatial memory of each group was detected by Morris water maze test. The concentrations of glutamate ( glu),calcium ion( Ca2+),N-methyl-D-aspartate receptor( NMDAR),calmodulin( Ca M) and phosphorylated cyclic adenosine monophosphate effector binding protein( P-CREB) in hippocampal neuron's were detected by high performance liquid chromatography,fluorescence immunostaining and Western blot,respectively.Results Compared with that of BC group,the searching platform latency ( SPL) in Morris water maze test was significantly prolonged( P<0.05) in both of D-gal and NES groups. No difference in SPL was found between NES group and D-gal group. The contents of glu,Ca2+and NMDAR in D-gal and NES groups were significantly higher than those in BC group( P<0.05),while the protein expression levels of Ca M and P-CREB were significantly decreased( P<0.05). Additionally,the level of Ca M was higher in NES group than that in D-gal group( P < 0. 05). Conclusions Chronic negative emotional stress is one of the important reasons of accelerating cognitive decline process,which is similar to D-gal,and the Ca M-CaMKⅡ-CREB signaling pathway might be the important mechanism through affecting the related protein's expression of learning and memory.
引文
1詹向红,刘胜利,江虹,等,愤怒情志表达方式及特质对自主神经的影响[J].中华中医药杂志,2011;26(7):1475-8.
2吴帅,安书成,陈慧彬,等.慢性应激性抑郁发生中大鼠眶额叶多巴胺D1受体对谷氨酸及其NMDA受体的调节[J/OL].心理学报,2014;46(1):69-78.
3伍吉云,邓志敏.钙信号与阿尔茨海默病的关系及其作用[J].井冈山大学学报(自然科学版),2010;31(4):107-9,128.
4 Benito E,Barco A.CREB's control of intrinsic and synaptic plasticity:implications for CREB-dependent memory models[J].Trends Neurosci,2010;33(5):230-40.
5 Fu H,Li W,Lao Y,et al.Bis(7)-tacrine attenuates beta amyloid-induced neuronal apoptosis by regulating L-type calcium channels[J].JNeurochem,2006;98(5):1400-10.
6 Cross JL,Meloni BP,Bakker AJ,et al.Modes of neuronal calcium entry and homeostasis following cerebral ischemia[J].Stroke Res Treat,2010;2010:316862.
7詹向红,李伟,赵君玫,等.慢性愤怒应激对大鼠衰老进程及其神经内分泌免疫机制的影响[J].中华中医药杂志,2010;25(1):111-3.
8李文彬.D-半乳糖衰老和脑老化模型研究[A].见:陈可冀,李春生.新编抗衰老中药学[M].北京:人民卫生出版社,1998:153-62.
9王慧霞,詹向红,李伟,等.负性情绪应激大鼠模型的建立与评价[J].中国老年学杂志,2015;35(15):4140-2.
10詹向红,李伟,徐玮玮,等.慢性愤怒应激对衰老大鼠学习记忆能力的影响及其脂质过氧化机制[J].中国老年学杂志,2010;30(6):752-3.
11邢秋娟,施杞,王拥军.D-半乳糖诱导衰老动物模型的机制及其在中医药方面的应用[J].上海中医药大学学报,2010;24(3):93-8.
12许扬,吴涛,顾佳黎,等.D-半乳糖诱导衰老动物模型研究进展[J].中国老年学杂志,2009;29(13):1710-3.
13王慧霞.负性情绪积累加速D-gal衰老大鼠认知衰退的神经内分泌机制研究[D].郑州:河南中医学院,2014.
14 Van der Jeugd A,Ahmed T,Burnouf S,et al.Hippocampal tauopathy in tau transgenic mice coincides with impaired hippocampus-dependent learning and memory,and attenuated late-phase long-term depression of synaptic transmission[J].Neurobiol Learn Mem,2011;95(3):296-304.
15 Bratt AM,Kelley SP,Knowles JP,et al.Long term modulation of the HPA axis by the hippocampus:Behavioral,biochemical and immunologi-cal endpoints in rats exposed to chronic mild stress[J].Psychoneuroendocrinology,2001;26(2):121-45.
16 Al-Hullaq RA,Conrads TP,Veenstra TD,et al.NMDA diheteromeric receptor populations and associated proteins in rathippocampus[J].JNeurosci,2007;27(31):8334-43.
17余瑞元,王燕峰,徐长法.CREB研究进展[J].中国生物工程杂志,2003;23(1):39-42,47.
18 Leissring MA,Murphy MP,Mead TR,et al.A physiologic signalling role for the gama-secretase-derived intracellular fragent of APP[J].Proc Natl Acad Sci USA,2002;99(7):4697-702.
19韩冬云.钙调蛋白对豚鼠心室肌细胞Cav1.2钙通道的剂量依赖性和钙依赖性作用[D].沈阳:中国医科大学,2007.
2吴帅,安书成,陈慧彬,等.慢性应激性抑郁发生中大鼠眶额叶多巴胺D1受体对谷氨酸及其NMDA受体的调节[J/OL].心理学报,2014;46(1):69-78.
3伍吉云,邓志敏.钙信号与阿尔茨海默病的关系及其作用[J].井冈山大学学报(自然科学版),2010;31(4):107-9,128.
4 Benito E,Barco A.CREB's control of intrinsic and synaptic plasticity:implications for CREB-dependent memory models[J].Trends Neurosci,2010;33(5):230-40.
5 Fu H,Li W,Lao Y,et al.Bis(7)-tacrine attenuates beta amyloid-induced neuronal apoptosis by regulating L-type calcium channels[J].JNeurochem,2006;98(5):1400-10.
6 Cross JL,Meloni BP,Bakker AJ,et al.Modes of neuronal calcium entry and homeostasis following cerebral ischemia[J].Stroke Res Treat,2010;2010:316862.
7詹向红,李伟,赵君玫,等.慢性愤怒应激对大鼠衰老进程及其神经内分泌免疫机制的影响[J].中华中医药杂志,2010;25(1):111-3.
8李文彬.D-半乳糖衰老和脑老化模型研究[A].见:陈可冀,李春生.新编抗衰老中药学[M].北京:人民卫生出版社,1998:153-62.
9王慧霞,詹向红,李伟,等.负性情绪应激大鼠模型的建立与评价[J].中国老年学杂志,2015;35(15):4140-2.
10詹向红,李伟,徐玮玮,等.慢性愤怒应激对衰老大鼠学习记忆能力的影响及其脂质过氧化机制[J].中国老年学杂志,2010;30(6):752-3.
11邢秋娟,施杞,王拥军.D-半乳糖诱导衰老动物模型的机制及其在中医药方面的应用[J].上海中医药大学学报,2010;24(3):93-8.
12许扬,吴涛,顾佳黎,等.D-半乳糖诱导衰老动物模型研究进展[J].中国老年学杂志,2009;29(13):1710-3.
13王慧霞.负性情绪积累加速D-gal衰老大鼠认知衰退的神经内分泌机制研究[D].郑州:河南中医学院,2014.
14 Van der Jeugd A,Ahmed T,Burnouf S,et al.Hippocampal tauopathy in tau transgenic mice coincides with impaired hippocampus-dependent learning and memory,and attenuated late-phase long-term depression of synaptic transmission[J].Neurobiol Learn Mem,2011;95(3):296-304.
15 Bratt AM,Kelley SP,Knowles JP,et al.Long term modulation of the HPA axis by the hippocampus:Behavioral,biochemical and immunologi-cal endpoints in rats exposed to chronic mild stress[J].Psychoneuroendocrinology,2001;26(2):121-45.
16 Al-Hullaq RA,Conrads TP,Veenstra TD,et al.NMDA diheteromeric receptor populations and associated proteins in rathippocampus[J].JNeurosci,2007;27(31):8334-43.
17余瑞元,王燕峰,徐长法.CREB研究进展[J].中国生物工程杂志,2003;23(1):39-42,47.
18 Leissring MA,Murphy MP,Mead TR,et al.A physiologic signalling role for the gama-secretase-derived intracellular fragent of APP[J].Proc Natl Acad Sci USA,2002;99(7):4697-702.
19韩冬云.钙调蛋白对豚鼠心室肌细胞Cav1.2钙通道的剂量依赖性和钙依赖性作用[D].沈阳:中国医科大学,2007.