益肾化浊方对SAMP8小鼠学习记忆能力和海马神经元及相关炎症因子的影响
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摘要
目的观察益肾化浊方对SAMP8小鼠学习记忆能力和海马CA1区神经元、肿瘤坏死因子-α(TNF-α)、核因子-κB(NF-κB)表达的影响,探讨其可能的作用机制。方法将7月龄SAMP8小鼠随机分为SAMP8组和益肾化浊方低、中、高剂量组,选择SAMR1小鼠作为对照;益肾化浊方低、中、高剂量组分别给予3.12、6.24、12.48 g/kg益肾化浊方药液灌胃,SAMR1组和SAMP8组给予等体积蒸馏水灌胃,每日1次,连续4周;采用Morris水迷宫实验评价小鼠学习记忆能力,尼氏染色观察小鼠海马CA1区神经元变化,免疫组化检测小鼠海马CA1区TNF-α和NF-κB的表达。结果水迷宫实验结果显示,各组小鼠平均游泳速度差异无统计学意义(P>0.05);与SAMR1组比较,SAMP8组小鼠逃避潜伏时间显著增加、穿越平台次数显著减少(P<0.05);与SAMP8组比较,益肾化浊方中剂量组小鼠逃避潜伏时间显著减少、穿越平台次数显著增加(P<0.05)。尼氏染色结果显示,与SAMR1组比较,SAMP8组小鼠海马CA1区神经元数量显著减少(P<0.05);与SAMP8组比较,益肾化浊方中剂量组小鼠海马CA1区神经元数量显著增加(P<0.05)。免疫组化染色结果显示,与SAMR1组比较,SAMP8组小鼠海马CA1区TNF-α和NF-κB表达显著增加(P<0.05);与SAMP8组比较,益肾化浊方中剂量组小鼠海马CA1区TNF-α和NF-κB表达显著减少(P<0.05)。结论中剂量益肾化浊方可有效改善7月龄SAMP8小鼠学习记忆能力,该作用机制可能与其减少海马CA1区神经元丢失、抑制炎症因子TNF-α、NF-κB的表达有关。
Objective To investigate the effects of Yishen Huazhuo Decoction on learning and memory ability,hippocampus neurons, tumor necrosis factor-α(TNF-α) and nuclear factor-κB(NF-κB) of senescence accelerated mouse prone 8(SAMP8) and its possible mechanism. Methods 7-month old SAMP8 mice were randomly divided into SAMP8 group, Yishen Huazhuo Decoction low-, medium-, and high-dosage groups. Senescence accelerated mouse resistant 1(SAMR1) were selected as control group. Mice in Yishen Huazhuo Decoction low-, medium-, and high-dosage groups received 3.12, 6.24, 12.48 g/kg respectively for gavage. The SAMR1 group and the SAMP8 group were given an equal volume of distilled water once a day for 4 weeks. Morris water maze test was used to evaluate the learning and memory ability of mice. Nissl staining was used to observe the changes of neurons in hippocampal CA1 region of mice, and the expressions of TNF-α and NF-κB in hippocampus of mice was detected by immunohistochemistry. Results The results of water maze test showed that there was no significant difference in the average swimming speed among the groups(P>0.05). Compared with the SAMR1 group, the escape latency of the SAMP8 group significantly increased, and the number of crossing platforms was significantly reduced(P<0.05).Compared with the SAMP8 group, the escape latency of the mice in the Yishen Huazhuo Decoction medium-dosage group was significantly reduced, and the number of crossing platforms significantly increased(P<0.05). Nissl staining showed that the number of neurons in hippocampal CA1 region of SAMP8 group significantly decreased compared with SAMR1 group(P<0.05). Compared with the SAMP8 group, the number of neurons in the hippocampal CA1 region of the Yishen Huazhuo Decoction medium-dosage group significantly increased(P<0.05).Immunohistochemical staining showed that compared with SAMR1 group, the expressions of TNF-α and NF-κB in hippocampal CA1 region of SAMP8 group significantly increased(P<0.05). Compared with the SAMP8 group, the expression of TNF-α and NF-κB in the hippocampal CA1 region of Yishen Huazhuo Decoction medium-dosage group significantly decreased(P<0.05). Conclusion Medium-dose Yishen Huazhuo Decoction can effectively improve the learning and memory ability of 7-month old SAMP8 mice, which may be related to the reduction of neuron loss in hippocampal CA1 region and the inhibition of inflammatory factors TNF-α and NF-κB.
Objective To investigate the effects of Yishen Huazhuo Decoction on learning and memory ability,hippocampus neurons, tumor necrosis factor-α(TNF-α) and nuclear factor-κB(NF-κB) of senescence accelerated mouse prone 8(SAMP8) and its possible mechanism. Methods 7-month old SAMP8 mice were randomly divided into SAMP8 group, Yishen Huazhuo Decoction low-, medium-, and high-dosage groups. Senescence accelerated mouse resistant 1(SAMR1) were selected as control group. Mice in Yishen Huazhuo Decoction low-, medium-, and high-dosage groups received 3.12, 6.24, 12.48 g/kg respectively for gavage. The SAMR1 group and the SAMP8 group were given an equal volume of distilled water once a day for 4 weeks. Morris water maze test was used to evaluate the learning and memory ability of mice. Nissl staining was used to observe the changes of neurons in hippocampal CA1 region of mice, and the expressions of TNF-α and NF-κB in hippocampus of mice was detected by immunohistochemistry. Results The results of water maze test showed that there was no significant difference in the average swimming speed among the groups(P>0.05). Compared with the SAMR1 group, the escape latency of the SAMP8 group significantly increased, and the number of crossing platforms was significantly reduced(P<0.05).Compared with the SAMP8 group, the escape latency of the mice in the Yishen Huazhuo Decoction medium-dosage group was significantly reduced, and the number of crossing platforms significantly increased(P<0.05). Nissl staining showed that the number of neurons in hippocampal CA1 region of SAMP8 group significantly decreased compared with SAMR1 group(P<0.05). Compared with the SAMP8 group, the number of neurons in the hippocampal CA1 region of the Yishen Huazhuo Decoction medium-dosage group significantly increased(P<0.05).Immunohistochemical staining showed that compared with SAMR1 group, the expressions of TNF-α and NF-κB in hippocampal CA1 region of SAMP8 group significantly increased(P<0.05). Compared with the SAMP8 group, the expression of TNF-α and NF-κB in the hippocampal CA1 region of Yishen Huazhuo Decoction medium-dosage group significantly decreased(P<0.05). Conclusion Medium-dose Yishen Huazhuo Decoction can effectively improve the learning and memory ability of 7-month old SAMP8 mice, which may be related to the reduction of neuron loss in hippocampal CA1 region and the inhibition of inflammatory factors TNF-α and NF-κB.
引文
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[24]宋宛珊,张玉莲,周震,等.益肾化浊方对快速老化小鼠认知功能与海马神经元形态学的影响及机制[J].中国老年学杂志,2016,36(11):2586-2590.
[2]田金洲.中国痴呆诊疗指南(2017年版)[M].北京:人民卫生出版社,2018:1.
[3]QUERFURTH H W,LAFERLA F M.Alzheimer's disease[J].New England Journal of Medicine,2010,362(4):329-344.
[4]SERRANOPOZO A,FROSCH M P,MASLIAH E,et al.Neuropathological alterations in Alzheimer disease.[J].Cold Spring Harbor Perspectives in Medicine,2011,1(1):a006189.
[5]NILSON A N,ENGLISH K C,GERSON J E,et al.Tau oligomers associate with inflammation in the brain and retina of tauopathy mice and in neurodegenerative diseases[J].Journal of Alzheimer's Disease,2017,55(3):1083-1099.
[6]SNOW W M,ALBENSI B C.Neuronal gene targets of NF-κB and their dysregulation in Alzheimer's disease[J].Frontiers in Molecular Neuroscience,2016,9:118.
[7]梁梅亭,朱宏,董克礼,等.补肾活血针刺法对SAMP8小鼠杏仁核蛋白质组学表达的影响[J].中国中医药信息杂志,2018,25(3):58-63.
[8]TAKEDA T,HOSOKAWA M,TAKESHITA S,et al.A new murine model of accelerated senescence[J].Mechanisms of Ageing&Development,1981,17(2):183-194.
[9]ZHANG Y,LIN C,ZHANG L,et al.Cognitive improvement during treatment for mild Alzheimer’s disease with a Chinese herbal formula:a randomized controlled trial[J].PLoS One,2015,10(6):e0130353.
[10]李强,王凯,孙伟明,等.益肾化浊方对阿尔茨海默病模型大鼠海马区Ca2+浓度及钙相关记忆蛋白表达的影响[J].时珍国医国药,2018,21(3):111-114.
[11]魏伟,吴希美,李元建.药理实验方法学:第4版[M].北京:人民卫生出版社,2010:1698.
[12]VORHEES C V,WILLIAMS M T.Morris water maze:procedures for assessing spatial and related forms of learning and memory[J].Nature Protocols,2006,1(2):848-858.
[13]MORLEY J E,ARMBRECHT H J,FARR S A,et al.The senescence accelerated mouse(SAMP8)as a model for oxidative stress and Alzheimer's disease[J].Biochim Biophys Acta,2012,1822(5):650-656.
[14]褚芹,于建春,潘建明,等.快速老化模型小鼠SAMP8行为学的增龄性变化[J].现代生物医学进展,2008,8(10):1801-1804.
[15]沈自尹,黄建华,林伟,等.从整体论到系统生物学进行肾虚和衰老的研究[J].中国中西医结合杂志,2009,29(6):548-550.
[16]JEONG Y,HUH N,LEE J,et al.Role of the hippocampal CA1 region in incrementalvaluelearning[J].Sci Rep,2018,8(1):9870.
[17]王凯,李强,孙伟明,等.不同浓度Aβ(25-35)蛋白模拟阿尔茨海默病模型学习记忆的差异[J].中国比较医学杂志,2017,27(3):14-19.
[18]TERAI K,MATSUO A,MCGEER P L.Enhancement of immunoreactivity for NF-κB in the hippocampal formation and cerebral cortex of Alzheimer's disease[J].Brain Research,1996,735(1):159-168.
[19]黄伟,张亚囡,孙蓉.何首乌不同组分单次给药对小鼠肝毒性“量-时-毒”关系研究[J].中国药物警戒,2011,8(4):193-197.
[20]MORRIS R G M.Spatial localization does not require the presence of local cues[J].Learning and Motivation,1981,12(2):239-260.
[21]LEEM Y H,CHANG H.The ameliorating effect of exercise on long-term memory impairment and dendritic retraction via the mild activation of AMP-activated protein kinase in chronically stressed hippocampal CA1 neurons[J].J Exerc Nutrition Biochem,2018,22(3):35-41.
[22]HURTADO D E,MOLINA-PORCEL L,IBA M,et al.Aβaccelerates the spatiotemporal progression of tau pathology and augments tau amyloidosis in an Alzheimer mouse model[J].American Journal of Pathology,2010,177(4):1977-1988.
[23]VALERIO A,BORONI F,BENARESE M,et al.NF-κB pathway:a target for preventingβ-amyloid(Aβ)-induced neuronal damage and Aβ42production[J].European Journal of Neuroscience,2006,23(7):1711-1720.
[24]宋宛珊,张玉莲,周震,等.益肾化浊方对快速老化小鼠认知功能与海马神经元形态学的影响及机制[J].中国老年学杂志,2016,36(11):2586-2590.