连翘酯苷对东莨菪碱模型小鼠学习记忆的影响及其机制研究

目的:研究连翘酯苷对东莨菪碱模型小鼠学习记忆的影响,并探讨其作用机制,为连翘酯苷抗阿尔茨海默病研究提供数据支撑。方法:昆明种小鼠,随机分为4组,分别为正常组,东莨菪碱模型组,多奈哌齐组(3 mg·kg-1)和连翘酯苷(200 mg·kg-1)组,每组12只。各组连续给药14 d。给药第14天,东莨菪碱模型组、多奈哌齐组和连翘酯苷组给予东莨菪碱(3 mg·kg-1),腹腔注射。20 min后,进行跳台实验。同时,观察连翘酯苷对乙酰胆碱酯酶(Ach E)和环磷酸腺苷-细胞外信号调节激酶通路的影响。另设一批实验,昆明种小鼠分为4组,分别为正常组,东莨菪碱模型组,维生素E组(100 mg·kg-1)和连翘酯苷(200 mg·kg-1)组,给药14 d后,断头处死动物,检测连翘酯苷对东莨菪碱模型超氧化物歧化酶(SOD),丙二醛(MDA),单胺氧化酶(MAO)的影响。结果:跳台实验获得期和巩固期,东莨菪碱模型组与正常组比较,安全期时间比率显著下降(P0.05),连翘酯苷显著增加安全期时间比率(P0.05)。连翘酯苷显著降低东莨菪碱模型小鼠大脑皮层和海马Ach E活性(P0.05)。连翘酯苷显著升高东莨菪碱模型小鼠海马P-ERK含量,与东莨菪碱组比较,具有显著性差异(P0.05)。同时,东莨菪碱组与正常组比较,显著降低小鼠大脑皮层和海马SOD活性、升高MDA含量、升高MAO活性(P0.05)。连翘酯苷能显著升高小鼠大脑皮层和海马SOD活性、降低MDA含量和MAO活性,与东莨菪碱组比较,具有显著性差异(P0.05)。结论:连翘酯苷可提高东莨菪碱模型小鼠的学习记忆能力,其机制可能与抑制模型小鼠大脑皮层Ach E活性,促进环磷酸腺苷(c AMP)表达,活化细胞外信号调节激酶(ERK)及抗氧化有关。     

Anti‐depressant Effects of Aqueous Extract from Acanthopanax senticosus in Mice

In this paper, the anti‐depressant effects of Acanthopanax senticosus extract (ASE) were studied using animal models of depression including the forced swimming and tail suspension tests. The anti‐depressive mechanism of ASE was explored by monitoring the levels of monoamine neurotransmitters including 5‐hydroxytrylamine (5‐HT), norepinephrine (NE), and dopamine (DA), as well as cAMP response element‐binding (CREB) protein expression in the whole brain of mice following the tail suspension test. Our results showed that intragastric administration of ASE at a dose of 2000 mg/kg for seven days significantly reduced the duration of immobility in both the forced swimming test and the tail suspension test. These results indicate that ASE possesses antidepressant‐like properties. Pre‐treatment with 2000 mg/kg of ASE for seven days significantly elevated the levels of 5‐HT, NE, and DA in the whole brain of mice. Moreover, ASE at doses of 1000 and 2000 mg/kg significantly up‐regulated the level of CREB protein. Taken together, these findings suggest that the anti‐depressive mechanism of ASE may be mediated via the central monoaminergic neurotransmitter system and CREB protein expression. Therefore, administration of ASE may be beneficial for patients with depressive disorders. Copyright © 2013 John Wiley & Sons, Ltd.     

Ginseng Protein Reverses Amyloid Beta Peptide and H2O2 Cytotoxicity in Neurons,and Ameliorates Cognitive Impairment in AD Rats Induced by a Combination of D‐Galactose and AlCl3

Ginseng (Panax ginseng C.A. Meyer) is one of the most widely used herbal medicines worldwide. The present study evaluated the neuroprotective effects of ginseng protein (GP) and its possible mechanisms in a cellular and animal model of AD. The results demonstrated that GP (10–100 µg/mL) significantly improved the survival rate of neurons and reduced the cells' apoptosis and the mRNA expression of caspase‐3 and Bax/Bcl‐2. In addition, GP (0.1 g/kg) significantly shortened the escape latency, prolonged the crossing times and the percentage of residence time; reduced the level of Aβ_1–42 and p‐tau, the activity of T‐NOS and iNOS, and the content of MDA and NO, improved the activity of SOD, the concentration of cAMP and the protein expression of p‐PKA/PKA and ‐CREB/CREB. The results demonstrated that GP significantly inhibited Alzheimer‐like pathophysiological changes induced by Aβ_25–35 or H₂O₂ in cells or those induced by D‐gal/ Al in animals. These neuroprotective effects of GP may be associated with the cAMP/PKA/CREB pathway. Also, in combination with our previous studies, these results indicate that the anti‐AD mechanism of GP was likely to activate the CREB pathway through multiple channels. Copyright © 2016 John Wiley & Sons, Ltd.     

Inhibitory Effect of Ethanol Extract of Magnolia officinalis on Memory Impairment and Amyloidogenesis in a Transgenic Mouse Model of Alzheimer's Disease via Regulating β‐Secretase Activity

Alzheimer's disease (AD) is the most common form of dementia and is characterized by deposition of amyloid beta (Aβ) in the brain. The components of the herb Magnolia officinalis are known to have antiinflammatory, antioxidative and neuroprotective activities. In this study we investigated the effects of ethanol extract of M. officinalis on memory dysfunction and amyloidogenesis in a transgenic mouse model of AD. Oral pretreatment of ethanol extract of M. officinalis (10 mg/kg in 0.05% ethanol) into drinking water for 3 months inhibited memory impairment and Aβ deposition in the brain of Tg2576 mice. Ethanol extract of M. officinalis also decreased activity of β‐secretase, cleaving Aβ from amyloid precursor protein (APP), and expression of β‐site APP cleaving enzyme 1 (BACE1), APP and its product, C99. Our results showed that ethanol extract of M. officinalis effectively prevented memory impairment via down‐regulating β‐secretase activity. Copyright © 2012 John Wiley & Sons, Ltd.     

White Ginseng Protects Mouse Hippocampal Cells Against Amyloid‐Beta Oligomer Toxicity

Amyloid‐beta oligomer (AβO) is a soluble oligomer form of the Aβ peptide and the most potent amyloid‐beta form that induces neuronal damage in Alzheimer's disease. We investigated the effect of dried white ginseng extract (WGE) on neuronal cell damage and memory impairment in intrahippocampal AβO (10 μM)‐injected mice. Mice were treated with WGE (100 and 500 mg/kg/day, p.o.) for 12 days after surgery. WGE improved memory impairment by inhibiting hippocampal cell death caused by AβO. In addition, AβO‐injected mice treated with WGE showed restoration of reduced synaptophysin and choline acetyltransferase intensity and lower levels of ionized calcium‐binding adaptor molecule 1 in the hippocampus compared with those of vehicle‐treated controls. These results suggest that WGE reverses memory impairment in Alzheimer's disease by attenuating neuronal damage and neuroinflammation in the AβO‐injected mouse hippocampus. Copyright © 2017 John Wiley & Sons, Ltd.     

桂皮醛通过阻止海马神经炎症反应改善阿尔茨海默病老年小鼠记忆障碍的作用研究＊

目的 探讨桂皮醛（trans-cinnamaldehyde，TCA）对老年具有阿尔茨海默病（Alzheimer''s disease，AD）样表型的早老素1/2条件性双基因敲除（Presenilin1/2 conditional double knockout，PS cDKO）小鼠海马神经炎症反应和突触破坏导致的记忆障碍的改善作用及其机制研究。方法 选用9月龄PS cDKO小鼠和其同窝野生型对照小鼠随机分为四组：野生型组（wildtype，WT）、野生型 + TCA（WT + TCA）、PS cDKO组（cDKO）和PS cDKO + TCA组（cDKO + TCA），每组10只。WT + TCA和cDKO + TCA组的小鼠给予含有TCA（240 ppm）的饲料治疗，WT和cDKO组的小鼠给予普通饲料，共治疗90 d，治疗60 d后进行行为学测试，测试期间继续进行治疗，测试结束后取小鼠海马进行分子生物学实验。其中，Morris水迷宫实验观察TCA治疗后对PS cDKO小鼠空间参考记忆的影响，恐惧实验检测TCA对PS cDKO小鼠联合记忆能力的影响；Western Blot检测TCA对PS cDKO小鼠海马突触蛋白表达的影响；RT-PCR检测TCA对PS cDKO小鼠海马促炎症因子mRNA水平的影响。结果 TCA明显改善PS cDKO小鼠的空间参考记忆障碍（P < 0.05）和联合记忆损伤（P < 0.05）。另外，TCA上调PS cDKO小鼠海马中N-甲基-D-天冬氨酸受体（N-methyl-D-aspartate receptor，NMDAR）的亚基NR1和NR2A、突触小泡蛋白（Synaptophysin， SYP）的蛋白表达，下调Tau蛋白的过度磷酸化；同时，TCA降低海马促炎症因子白介素-1β（interleukin-1β，IL-1β）、诱导性一氧化碳酶（inducible nitric oxide synthase，iNOS）、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、环氧合酶-2（cyclooxygenase-2，COX-2）的mRNA水平（P < 0.05）。结论 TCA改善老年PS cDKO小鼠记忆障碍可能是通过抑制海马的神经炎症反应并提高突触蛋白的表达来发挥神经保护作用。     

Microarray Expression Analysis for the Paradoxical Roles of Acanthopanax senticosus Harms in Treating α‐Synucleinopathies

α‐Synuclein is a key player in the pathogenesis of neurodegenerative disorders with Lewy bodies. Our previous studies have also showed that Acanthopanax senticosus harms (AS) could significantly suppress α‐synuclein overexpression and toxicity. Identifying the RNAs related to α‐synucleinopathies may facilitate understanding the pathogenesis of the diseases and the safe application of AS in the clinic. Microarray expression profiling of long non‐coding RNAs (lncRNAs) and mRNAs was undertaken in control non‐transgenic and human α‐synuclein transgenic mice. The effects of AS on central nervous system (CNS) in pathology and physiology were investigated based on the lncRNA/mRNA targets analysis. In total, 341 lncRNAs and 279 mRNAs were differentially expressed by α‐synuclein stimulus, among which 29 lncRNAs and 25 mRNAs were involved in the anti‐α‐synucleinopathies mechanism of AS. However, the levels of 19/29 lncRNAs and 12/25 mRNAs in AS group were similar to those in α‐synuclein group, which may cause potential neurotoxicity analogous to α‐synuclein. This study demonstrated that some of lncRNAs/mRNAs were involved in α‐synuclein related pathophysiology, and AS produced the bidirectional effects on CNS under pathological and physiological conditions. Copyright © 2015 John Wiley & Sons, Ltd.     

苦参碱对LPS诱导的阿尔茨海默病小鼠模型学习记忆功能和脑内神经炎症的影响

目的:研究苦参碱对阿尔茨海默病(Alzheimer's disease,AD)模型小鼠学习记忆损伤的治疗作用并探讨其可能的作用机制。方法:ICR小鼠随机分为正常组、模型组、苦参碱高、中、低剂量组和阳性药组。除正常组之外其余各组小鼠侧脑室注射脂多糖(lipopolysaccharide,LPS)建立AD小鼠模型,正常组小鼠注射等量生理盐水。之后连续灌胃给药21 d,每天1次,其中苦参碱各给药组高、中、低剂量分别为40,20,10 mg·kg~(-1),阳性药多奈哌齐剂量为5 mg·kg~(-1)。采用新物体识别(novel object recognition,NOR)实验,Y迷宫实验评价各组动物的学习记忆能力;酶联免疫吸附测定(ELISA)检测各组小鼠脑内海马组织中炎症因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α),白细胞介素-1β(interleukin-1β,IL~(-1)β)和活性氧(reactive oxygen species,ROS)的含量;蛋白免疫印迹法(Western blot)检测各组小鼠脑内海马区烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶亚基gp91phox,p47phox蛋白的含量。结果:与正常组比较,模型组小鼠的新物体偏爱指数和自发交替反应率明显降低(P 0. 01),海马组织中TNF-α,IL~(-1)β和ROS的含量明显升高(P 0. 01),NADPH氧化酶亚基gp91phox,p47phox蛋白的表达明显升高(P 0. 01)。与模型组比较,苦参碱高、中、低剂量组均可使小鼠的新物体偏爱指数和自发交替反应率明显升高(P 0. 05,P 0. 01),明显改善AD小鼠的学习记忆能力;同时,苦参碱各剂量组可显著降低AD模型小鼠海马组织中TNF-α,IL~(-1)β和ROS的含量(P 0. 05,P 0. 01),并下调NADPH氧化酶亚基gp91phox,p47phox蛋白的表达(P 0. 05,P 0. 01)。结论:苦参碱可缓解LPS诱导的学习记忆功能损伤和脑内神经炎症,这些作用可能是通过抑制NADPH氧化酶亚基gp91phox和p47phox的蛋白表达来发挥的。     

Chinese Herbal Medicine for Mild Cognitive Impairment: A Systematic Review and Meta‐Analysis of Cognitive Outcomes

Mild cognitive impairment (MCI) is a condition that may be prodromal to the development of dementia. There remain, as yet, no approved pharmaceutical interventions for MCI. Chinese herbal medicines (CHMs) have a long history of use for cognitive impairments and some plant ingredients have shown neuroprotective actions in experimental studies. This review assesses the current clinical evidence from controlled clinical trials for the effects of CHMs on cognitive outcomes as measured by Mini‐mental state examination (MMSE) or Alzheimer's Disease Assessment Scale‐Cognitive subscale (ADAS‐Cog). Fifty one studies (4026 participants) were included. These compared CHM with placebo, supportive care, pharmaceutical treatment or combined CHM with a pharmaceutical in an integrative setting. For the eight randomised controlled trials (RCTs) of comparisons with placebo, MMSE was significantly higher in the CHM groups (MD 1.56 [0.78, 2.34] I² = 85%, n = 503), similarly for eight RCTs of comparisons with supportive care (MD 1.77 [1.33, 2.21] I² = 0%, n = 555). Benefits were also evident in comparisons with some pharmaceuticals and with integrative treatment. The small size of most studies and methodological weaknesses mean that these results should be interpreted with caution. Further studies employing rigorous methods are required to investigate the potential benefits of these CHMs for MCI. Copyright © 2016 John Wiley & Sons, Ltd.     

四君子丸对SAMP8小鼠海马CA3区神经元Lon蛋白表达的影响

目的：研究SAMP8小鼠海马Lon蛋白及线粒体动力学相关蛋白表达变化,为健脾益气法治疗阿尔茨海默病提供理论依据。方法：3月龄SAMR1小鼠8只作为正常组,3月龄SAPM8小鼠32只分为模型组、多奈哌齐组(0.013 g·kg^-1)、四君子丸低、高剂量组(3.24、12.56 g·kg^-1),每组8只,多奈哌齐组灌胃多奈哌齐,四君子丸低、高剂量组灌胃四君子丸溶液,灌胃30 d;第25天开始水迷宫定位航行实验,第30天开始水迷宫空间探索实验;第30天取材后,免疫组化检测线粒体融合蛋白2(MFN2)蛋白表达变化,酶联免疫吸附测定法(ELISA)检测腺苷酸活化蛋白激酶(AMPK)表达,比色法检测三磷酸腺苷(ATP)含量,电镜检测神经元线粒体微观结构变化,蛋白免疫印迹法(Western blot)检测β淀粉样蛋白(Aβ)、Lon蛋白、线粒体动力相关蛋白1(DRP1)蛋白、MFN1蛋白表达变化。结果：与正常组比较,模型组小鼠逃避潜伏期时间增加、穿越次数减少、AMPK表达上调,ATP含量降低,Aβ蛋白表达升高(P<0.01),DRP1蛋白表达显著升高...     

Neurotherapeutic Effects of Pueraria mirifica Extract in Early‐ and Late‐Stage Cognitive Impaired Rats

We determined the neurotherapeutic effects of Pueraria mirifica extract (PME) and pure puerarin (PU) in comparison with 17β‐estradiol (E₂) in early‐ and late‐stage cognitive impaired rats. Rats were ovariectomized (OVX), kept for 2 and 4 months to induce early‐ and late‐stage cognitive impairment, respectively, and divided into four groups that were treated daily with (i) distilled water, (ii) 100 mg/kg of PME, (iii) 7 mg/kg of PU, and (iv) 80 µg/kg of E₂ for 4 months. The estrogen deficiency symptoms of OVX rats were abrogated by treatment with E₂ or PME, but not by treatment with PU. The mRNA level of genes associated with amyloid production (App and Bace1) and hyperphosphorylated Tau (Tau4) were upregulated together with the level of impaired cognition in the 2‐ and 4‐month OVX rats. Treatment with E₂ reduced the level of cognitive impairment more than that with PME and PU, and 2‐month OVX rats were more responsive than 4‐month OVX rats. All treatments down‐regulated the Bace1 mRNA level in 2‐month OVX rats, while PU and PME also decreased the App mRNA level in 2‐ and 4‐month OVX rats, respectively. Only PU suppressed Tau4 expression in 2‐month OVX rats. Thus, PME and PU elicit neurotherapeutic effects in different pathways, and earlier treatment is optimal. Copyright © 2016 John Wiley & Sons, Ltd.     

生慧汤通过调节神经递质改善阿尔茨海默病模型小鼠认知损伤和昼夜节律紊乱

目的 观察生慧汤对阿尔茨海默病（AD）小鼠血清中神经递质含量的影响,并探讨其改善AD认知损伤和昼夜节律紊乱的机制。方法 将27只APP/PS1小鼠随机分为模型组、多奈哌齐组和生慧汤组,另将9只野生型C57BL/6JNju正常小鼠设为空白组。多奈哌齐组（0.92×10^-4 g·kg^-1·d^-1）和生慧汤组（13.5 g·kg^-1·d^-1）分别灌服多奈哌齐和生慧汤,空白组和模型组灌服等体积的纯水,各组连续灌胃4周。采用Morris水迷宫实验和自主活动实验评估小鼠的认知功能和昼夜节律。采用液相色谱-质谱联用（LC-MS/MS）技术检测小鼠血清中乙酰胆碱（ACh）、胆碱乙酰转移酶（ChAT）、去甲肾上腺素（NE）、肾上腺素（E）、谷氨酸（Glu）、5-羟基吲哚乙酸（5-HIAA）和多巴胺（DA）的表达水平。结果 与空白组比较,模型组小鼠的平台潜伏期、游泳距离、初次抵达平台时间、光照活动时间、黑暗活动时间和总活动时间显著增加（P<0.01）,穿越平台次数和目标象限游泳时间则显著减少（P<0.01）;与模型组比较,多奈哌齐组和生慧汤组小鼠的平台潜伏期、游泳距离、初次抵达平台时间、光照活动时间、黑暗活动时间和总活动时间减少（P<0.05,P<0.01）,穿越平台次数和目标象限游泳时间增加（P<0.05,P<0.01）。与空白组比较,模型组小鼠血清中ACh、ChAT的表达水平显著减少（P<0.01）;与模型组比较,多奈哌齐组和生慧汤组小鼠血清中ACh、ChAT的表达水平增加（P<0.05,P<0.01）。与空白组比较,模型组小鼠血清中Glu的表达水平显著增加（P<0.01）,NE、5-HIAA和DA的表达水平显著减少（P<0.01）;与模型组比较,生慧汤组小鼠血清中Glu的表达水平降低（P<0.05）,NE、5-HIAA和DA的表达水平增加（P<0.05,P<0.01）;多奈哌齐组小鼠血清中NE、Glu、5-HIAA、DA的表达水平较模型组变化不明显,且差异没有统计学意义。各组小鼠血清中E的表达水平变化不明显,且差异没有统计学意义。结论 生慧汤可改善AD小鼠的认知损伤和昼夜节律紊乱,其机制可能与其调节神经递质有关。     

基于16S rDNA测序研究当归芍药散调控肠道菌群改善SAMP8小鼠认知能力的作用机制

目的 基于16S rDNA测序研究当归芍药散（DSS）对阿尔茨海默病（AD）模型小鼠SAMP8肠道菌群的影响。方法 按随机数字表法,将24只7月龄SAMP8小鼠分为DSS低、中、高剂量组（14.4、28.8、57.6 g·kg^-1·d^-1）及模型组,另取6只同月龄SAMR1小鼠,为正常组。连续给药8周后,采用16S rDNA高通量测序技术检测小鼠粪便肠道菌群的变化;Morris水迷宫实验评价小鼠定位航行与空间探索能力;尼氏染色观察海马CA1区神经元病理变化;酶联免疫吸附测定法（ELISA）检测海马β淀粉样蛋白（Aβ）和Tau蛋白过度磷酸化（p-Tau）蛋白含量。结果 与正常组比较,模型组小鼠肠道菌群α多样性降低、β多样性明显改变;逃避潜伏期延长（P<0.05）,穿越平台象限次数和平台象限停留时间明显减少（P<0.05）;CA1区神经元尼氏小体数量明显减少（P<0.05）;Aβ和p-Tau蛋白水平明显升高（P<0.05）。DSS干预后,各组小鼠肠道菌群α多样性均增加,中、高剂量组β多样性与正常组相似、以中剂量组效果最佳;在门水平上,厚壁菌门丰度明显增加（P<0.05）,拟杆菌门、变形菌门丰度明显降低（P<0.05）;在属水平上,乳酸杆菌等菌属丰度明显增加（P<0.05）,拟杆菌、幽门螺杆菌、理研菌、副杆状菌、萨特氏菌、黏液性菌等菌属丰度明显降低（P<0.05）;小鼠逃避潜伏期明显缩短（P<0.05）、穿越平台象限次数和平台象限停留时间明显增加（P<0.05）;尼氏小体数量明显增加（P<0.05）,Aβ和p-Tau蛋白水平明显降低（P<0.05）;Pearson相关性分析表明黏液性菌、拟杆菌和萨特氏菌菌属丰度与SAMP8小鼠认知能力呈负相关,乳酸杆菌和Butyricimonas菌属丰度与SAMP8小鼠认知能力呈正相关。结论 DSS可改善SAMP8小鼠认知能力,其作用机制可能与调节肠道菌群多样性、群落组成有关。     

补阳还五汤对阿尔茨海默病小鼠海马凋亡因子及学习记忆能力的影响

目的:探讨补阳还五汤(BYHWT)对阿尔茨海默病小鼠海马组织凋亡因子及学习记忆能力的影响。方法:40只雄性APP/PS1双转基因小鼠随机分为模型组,多奈哌齐组(0.001 g·kg~(-1)),BYHWT高、中、低剂量组(分别为37.06,18.53,9.26 g·kg~(-1))。C57小鼠作为空白组。连续灌胃BYHWT 30 d,进行水迷宫行为学测试后,取材。用蛋白免疫印迹法(Western blot)检测小鼠海马中凋亡因子B细胞淋巴瘤/白血病-2(Bcl-2),Bcl-2相关X蛋白(Bax)和半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)的变化。结果:与空白组比较,模型组小鼠学习记忆能力明显降低,海马组织中凋亡因子Bcl-2明显减少(P0.01),Bax和Caspase-3明显增加(P0.01)。与模型组比较,BYHWT高、中剂量组能明显改善小鼠学习记忆能力,凋亡因子Bcl-2明显增加(P0.01),Bax和Caspase-3明显减少(P0.01)。结论:补阳还五汤能够明显改善阿尔茨海默病小鼠的学习记忆能力,降低海马组织中Bax和Caspase-3的表达,增加Bcl-2的表达。     

补肾填精法对肾虚阿尔茨海默小鼠海马自噬的干预作用

目的:通过研究六味地黄丸对肾虚阿尔茨海默病(AD)小鼠海马神经元自噬水平的改善作用及部分机制,探讨补肾填精法对AD的部分治疗机制。方法:健康雄性C57-B6小鼠分为正常组,AD组,肾虚AD组,六味地黄丸组(1. 08 g·kg~(-1))。正常组与AD组每日皮下注射生理盐水(15 m L·kg~(-1)),肾虚AD组和六味地黄丸组每日皮下注射氢化可的松注射液(15 m L·kg~(-1)),连续注射20 d。第21天除正常组侧脑室注射无菌生理盐水外,其余3组侧脑室注射β淀粉样蛋白25-35(Aβ_(25-35)),6μg/只。采用酶联免疫吸附测定(ELISA)检测各组小鼠血清皮质醇、睾酮水平;透射电镜观察海马神经元细胞形态变化;免疫荧光检测微管相关蛋白1轻链3(LC3)的表达;蛋白免疫印迹法(Western blot)检测选择性自噬接头蛋白(p62)的蛋白表达。结果:与正常组比较,AD组和肾虚AD组小鼠的血清皮质醇、睾酮水平显著降低(P 0. 01),小鼠海马LC3表达显著减少(P 0. 01),小鼠海马p62蛋白表达显著增加(P 0. 01);与AD组比较,肾虚AD组小鼠的血清皮质醇、睾酮水平明显降低(P 0. 05,P 0. 01),小鼠海马p62蛋白表达显著增加(P 0. 01);与AD组和肾虚AD组比较,六味地黄丸组小鼠的血清皮质醇、睾酮水平显著升高(P 0. 01),小鼠海马LC3表达显著增多(P 0. 01),p62蛋白表达显著减少(P 0. 01)。正常组和六味地黄丸组小鼠神经元细胞内可见自噬小体,AD组和肾虚AD组小鼠神经元细胞内自噬小体鲜见。结论:补肾填精法能保护海马神经元细胞,增加海马神经元中LC3表达,降低p62表达水平,提高海马神经元自噬水平,对肾虚阿尔茨海默病具有一定的治疗作用。     

Genistein protects against amyloid‐beta‐induced toxicity in SH‐SY5Y cells by regulation of Akt and Tau phosphorylation

Alzheimer's disease is a neurodegenerative disorder characterized by extracellular deposition of amyloid‐β (Aβ) peptide and hyperphosphorylation of Tau protein, which ultimately leads to the formation of intracellular neurofibrillary tangles and cell death. Increasing evidence indicates that genistein, a soy isoflavone, has neuroprotective effects against Aβ‐induced toxicity. However, the molecular mechanisms involved in its neuroprotection are not well understood. In this study, we have established a neuronal damage model using retinoic‐acid differentiated SH‐SY5Y cells treated with different concentrations of Aβ_25–35 to investigate the effect of genistein against Aβ‐induced cell death and the possible involvement of protein kinase B (PKB, also termed Akt), glycogen synthase kinase 3β (GSK‐3β), and Tau as an underlying mechanism to this neuroprotection. Differentiated SH‐SY5Y cells were pre‐treated for 24 hr with genistein (1 and 10 nM) and exposed to Aβ_25–35 (25 μM), and we found that genistein partially inhibited Aβ induced cell death, primarily apoptosis. Furthermore, the protective effect of genistein was associated with the inhibition of Aβ‐induced Akt inactivation and Tau hyperphosphorylation. These findings reinforce the neuroprotective effects of genistein against Aβ toxicity and provide evidence that its mechanism may involve regulation of Akt and Tau proteins.     

针刺对比西药改善阿尔茨海默病患者认知功能障碍和精神行为症状的临床研究现状与分析＊

阿尔茨海默病（Alzheimer''s disease，AD）是一种起病隐匿且呈进行性发展的神经系统退行性疾病。临床上AD患者主要表现为认知功能障碍（Cognitive Impairment，CI）和精神行为症状（Behavioral and Psychological Symptoms，BPS），并可能伴有日常功能活动障碍等症状。目前，西药疗法为AD患者的主要治疗方法，但此法只可延缓患者病情，且副作用较多。最新研究认为，针刺疗法对于AD患者CI和BPS等症状具有改善作用。因此，本文对近二十五年针刺对比西药改善AD患者CI和BPS的文献进行梳理，将从针刺方法、针刺选穴、治疗时间以及针刺效果等方面进行分析，发现针刺对轻中度AD患者的CI和BPS治疗效果显著，并探讨分析了相应的治疗方案，为针刺治疗AD提供理论指导。     

镇心省睡益智方及其精油对AD模型小鼠学习记忆的影响

目的:观察镇心省睡益智方水提液及其精油对β淀粉样前体蛋白基因/早老素基因(APP/PS1)双转基因小鼠学习记忆的影响并探讨其可能机制。方法:采用APP/PS1双转基因痴呆小鼠及同月龄相同遗传背景C57BL/6JNju小鼠2种小鼠。C57BL/6JNju小鼠作为正常组,APP/PS1双转基因痴呆小鼠随机分为模型组,镇心省睡益智方精油低、高质量浓度组(12.13,48.50 mg·L~(-1)),镇心省睡益智方水提液组(0.46 g·kg~(-1)),每组12只。每天给药1次,连续给药22 d。给药结束后采用跳台实验、Morris水迷宫实验对小鼠行为学能力进行检测,采用尼氏染色观察海马CA1区神经元变化,采用硫磺素(Th S)染色观察海马DG区老年斑(SP)沉积,采用免疫组化法检测小鼠脑组织中葡萄糖转运蛋白1(GLUT1),胰岛素受体底物-1(IRS-1)的表达,采用酶联免疫吸附测定(ELISA)检测小鼠海马组织中乙酰胆碱(ACH),γ-氨基丁酸(GABA),谷氨酸(GLU)含量的变化。结果:与正常组比较,模型组小鼠跳台实验潜伏期显著缩短,错误次数显著增加(P0.01),Morris水迷宫实验定位航行逃避潜伏期明显延长(P0.05,P0.01),海马CA1区神经元出现缺失,DG区出现明显的老年斑沉积(P0.05),ACH,GLUT1含量均显著下降(P0.01),GABA,GLU水平及IRS-1的表达均显著升高(P0.01);与模型组比较,各给药组均可显著延长小鼠跳台实验潜伏期、减少跳台错误次数(P0.01),明显降低定位航行小鼠逃避潜伏期(P0.05,P0.01),可一定程度保护小鼠海马CA1区神经元,减少DG区老年斑沉积(P0.05,P0.01),明显增加小鼠脑组织中ACH,GLUT1表达(P0.05,P0.01),显著降低GABA,GLU水平及IRS-1的表达(P0.01)。结论:镇心省睡益智方水提液及其精油能够改善APP/PS1小鼠的学习记忆行为,保护神经元,增加脑组织GLUT1的表达,减少脑组织IRS-1的表达,减少老年斑沉积,升高ACH含量,降低GABA,GLU含量,可能是其防治阿尔茨海默症的机制。     

大黄酚对铅中毒小鼠学习记忆能力的影响

目的:研究大黄酚对铅中毒小鼠学习记忆能力及组织内铅水平的改善作用,并探讨其可能的作用机制.方法:采用连续8 dip7 mg?kg-1醋酸铅造成铅中毒小鼠模型,采用跳台实验,观察ip大黄酚(10.0,1.0,0.1 mg? kg-1)对铅中毒模型小鼠记忆能力的改善作用,并于大黄酚治疗14 d后测定小鼠心、肝、脾、肾内铅水平,同时测定小鼠肝、肾组织内丙二醛(MDA)含量及超氧化酶歧化酶(SOD),谷胱甘肽过氧化物酶(GSH-Px)活力.结果:连续8 dip7 mg?kg-1醋酸铅造成铅中毒小鼠模型学习记忆障碍,使小鼠心、肝、脾、肾内铅水平显著升高,小鼠肝、肾内SOD和GSH-Px活性显著降低,使MDA含量增加;而连续ip大黄酚14 d治疗后,可不同程度提高小鼠铅中毒后学习记忆能力,与模型组相比大黄酚治疗组均可显著降低心、肝、脾、肾铅含量(P＜0.01);与模型组相比大黄酚10.0,1.0 mg?kg-1治疗组可显著升高肝、肾内SOD和GSH -Px的活性,降低MDA含量(P＜0.01),而大黄酚0.1 mg?kg-1治疗组可显著升高肾内SOD和GSH-Px的活性,降低MDA含量(P＜0.01),对肝内SOD,GSH-Px的活性及MDA含量无显著影响.结论:大黄酚可显著改善铅中毒小鼠学习记忆能力,降低铅中毒小鼠各组织的铅水平,提高小鼠肝、肾组织内抗氧化酶的活性,改善铅中毒造成的脂质过氧化.     

加减薯蓣丸介导线粒体自噬改善APP/PS1小鼠氧化应激损伤及学习记忆能力

目的:探讨加减薯蓣丸对阿尔茨海默病(AD)小鼠学习记忆能力减退的作用及相关机制。方法:将40只5月龄SPF级APP/PS1小鼠随机分为模型组、多奈哌齐组、加减薯蓣丸组、加减薯蓣丸+氯喹(CQ)组,每组10只,同背景野生型C57BL/6J小鼠10只设定为正常组。其中加减薯蓣丸组予以加减薯蓣丸煎剂(10 g·kg^-1)灌胃,多奈哌齐组予以盐酸多奈哌齐溶液(0.45 mg·kg^-1)灌胃,加减薯蓣丸+CQ组则在加减薯蓣丸组基础上予以CQ(10 mg·kg^-1)腹腔注射1次,与灌胃同时进行,正常组与模型组予以等量生理盐水灌胃,1次/d,共35 d。给药结束后,Morris水迷宫实验和新物体识别实验检测小鼠空间记忆能力;原位末端标记法(TUNEL)染色检测小鼠海马CA1神经元凋亡水平;生化检测小鼠海马神经元活性氧(ROS)、超氧化物歧化酶(SOD)水平;透射电镜观察小鼠海马CA1区神经元线粒体超微结构;蛋白免疫印迹法(Western blot)检测小鼠海马线粒体自噬接头蛋白(p62)、微管相关蛋白1轻链3Ⅱ(LC3Ⅱ)、PTEN诱...