五味子醇甲对APP/PS1小鼠学习记忆和NF-κB p65的影响

目的研究五味子醇甲(SCH)对APP/PS1双转基因痴呆小鼠行为学和NF-κB p65的影响。方法将35只APP/PS1双转基因痴呆小鼠随机分为模型(APP/PS1)组、五味子醇甲(SCH+APP/PS1)组;另以同背景同月龄的C 57 BL/6 J阴性小鼠为空白(WT)组。SCH+APP/PS1组给予SCH悬浊液灌胃(2.6 mg·kg^-1·d^-1),模型组和空白组给予等量蒸馏水灌胃。各组灌胃30 d进行Mirror水迷宫空间探索试验后取材。HE染色法观察皮层及海马CA1区神经细胞的形态结构。运用DCFH-DA法检测脑组织活性氧(ROS)含量。Western印迹检测法检验磷酸化核转录因子(NF-κB p65,pp65)的表达。结果与APP/PS 1组相比,APP/PS 1+SCH组有效区停留距离及时间明显延长,差异有统计学意义(P<0.05,P<0.01);APP/PS 1+SCH组穿越有效区次数和平台区停留距离增加,均差异有统计学意义(P<0.01)。APP/PS 1+SCH组HE染色观察皮层有部分神经细胞萎缩、减少,较APP/PS1组明显好转,海马细胞排列整齐度尚可、较均匀。与APP/PS1组相比,APP/PS1+SCH组能降低ROS含量,差异有统计学意义(P<0.05)。APP/PS 1+SCH组皮层及海马pp65蛋白相对表达量下调(P<0.05,P<0.01)。结论SCH可能通过降低ROS含量和抑制pp65保护脑组织形态结构和提高APP/PS1鼠空间探索记忆能力。     

生慧汤对APP/PS1雄性AD小鼠海马内APP代谢产物的昼夜变化影响＊

目的研究生慧汤对APP/PS1（β-amyloid precursor protein/presenilin 1246E）小鼠海马内β淀粉样前体蛋白（Amyloid precursor protein，APP）代谢产物昼夜表达的影响。方法 将56只雄性APP/PS1双转基因痴呆模型小鼠随机分为4组，分别为：痴呆模型组、褪黑素治疗组（0.78 mg·kg^-1·d^-1）、生慧汤高剂量组（27.0 g·kg^-1·d^-1）、生慧汤低剂量组（13.5 g·kg^-1·d^-1），每组14只；对照组为14只雄性C57BL/6J小鼠，连续给药30天。采用Morris水迷宫检测小鼠学习记忆能力，采用酶联免疫吸附检测（ELISA）检测不同时间段取出的海马组织sAPPα含量。对海马β淀粉样蛋白_1-40、β淀粉样蛋白_1-42表达进行免疫印迹分析。免疫组化（IHC）检测海马CA1区Aβ_1-40蛋白的表达。结果 Morris水迷宫结果表明，与对照组相比，模型组上平台潜伏期明显延长、穿越平台次数明显减少（P<0.01）；与模型组相比，生慧汤不同剂量组上平台潜伏期明显缩短（P<0.01、P<0.05），穿越平台次数明显增加（P<0.05）。ELISA结果表明，与对照组相比，模型组的sAPPα总量、各时间段（12：00、24：00）sAPPα含量明显减少（P<0.01）；与模型组相比，生慧汤不同剂量组sAPPα总量明显增多（P<0.01、P<0.05），生慧汤高剂量组仅能增加12：00 sAPPα含量（P<0.01），生慧汤低剂量组仅能增加24：00 sAPPα含量（P<0.05）。对照组sAPPα含量具有明显昼夜差异（P<0.01）。模型组sAPPα含量的昼夜差异性消失（P>0.05）。生慧汤高剂量组sAPPα含量具有昼夜差异（P<0.01），生慧汤低剂量组sAPPα含量不具有昼夜差异（P>0.05）。Western blot结果显示，与对照组相比，模型组Aβ_1-40、Aβ_1-42蛋白表达明显增加（P<0.01）；与模型组相比，生慧汤高剂量组Aβ_1-40、Aβ_1-42蛋白表达减少（P<0.05、P<0.01）；生慧汤低剂量组仅能减少Aβ_1-40蛋白的表达（P<0.05），对Aβ_1-42蛋白表达无影响（P>0.05）。免疫组化结果表明，与对照组相比，模型组Aβ_1-40表达明显增加（P<0.01）；与模型组相比，生慧汤不同剂量组Aβ_1-40表达不同程度减少（P<0.01、P<0.05）。结论 生慧汤能够改善5月龄APP/PS1阿尔茨海默病模型小鼠的学习记忆障碍，其机制可能与恢复海马内APP代谢产物sAPPα的昼夜节律性表达、从而减少Aβ的沉积有关。     

糖尿病患者移动医疗服务的使用现状及其影响因素研究

  目的  调查患者糖尿病移动医疗服务知晓情况及使用现状,分析其影响因素,旨在为其开发和推广提供参考。  方法  于2019年7—9月整群选取某市3所综合性三级医院的410名内分泌科糖尿病患者,问卷调查患者的糖尿病移动医疗服务知晓及使用情况,采用logistic回归分析患者知晓情况及使用糖尿病移动医疗服务的影响因素。  结果  患者的糖尿病移动医疗服务知晓率及使用率分别为64.1%及49.6%,回归分析结果显示,18~44岁年龄组糖尿病移动医疗知晓率是60岁及以上年龄组2.914倍(P＜0.05);初中文化程度组、高中及以上文化程度组糖尿病移动医疗服务知晓率分别是小学及以下文化程度组的2.156倍、4.265倍(均P＜0.05);城镇患者的糖尿病移动医疗服务知晓率是农村患者的1.922倍(P＜0.05);初中文化程度组、高中及以上文化程度组糖尿病移动医疗服务使用率分别是小学及以下文化程度组的2.957倍、6.856倍(均P＜0.01)。  结论  患者糖尿病移动医疗服务的知晓率及使用率呈中等水平,设计者和管理者应根据其影响因素,采取有效措施,进一步提高患者的知晓率及使用率。      

APP/PS1小鼠肠道菌群特点及补肾法干预对其的影响

目的:探究不同月龄APP/PS1双转基因模型小鼠的肠道菌群特点及补肾法对其肠道菌群失衡的影响。方法:将8只6个月龄雄性APP/PS1小鼠随机分为模型组和补肾组;4只6个月龄雄性C57BL/6小鼠作为正常对照组。补肾组小鼠给予补肾法中药灌胃3个月,模型组和正常对照组小鼠均予以等体积羧甲基纤维素(CMC)溶液灌胃。留取小鼠灌胃干预前后的粪便,运用16SrDNA技术检测各组肠道菌群的组成结构。从肠道微生态方面,探讨补肾中药对APP/PS1双转基因小鼠肠道菌群构成的影响。结果:6个月龄时模型组和正常对照组小鼠的肠道菌群差异无统计学意义;与9个月龄正常组比较,模型组小鼠肠道内芽孢杆菌纲丰度明显升高。与模型组比较,补肾组肠道内芽孢杆菌纲丰度明显下降,疣微菌门丰度呈上升趋势。结论:APP/PS1小鼠体内均存在肠道菌群失衡的情况,主要与芽孢菌纲、疣微菌门的丰度水平的变化相关。补肾法可以通过降低芽孢菌纲、升高疣微菌门的丰度来进一步改善AD小鼠肠道菌群失衡状态。     

补肾填精方防治阿尔茨海默病模型大鼠学习记忆损害的分子机制

目的探讨补肾填精方防治铝致阿尔茨海默病(AD)的分子机制。方法用长期腹腔注射AlCl3复制大鼠动物模型,用RT-PCR检测端脑皮质beta-淀粉样前体蛋白mRNA异构体-APP695mRNA水平,用Y电迷宫仪检测大鼠的学习记忆能力。结果模型组大鼠端脑皮质APP695mRNA的表达水平明显升高(P<0.05),学习记忆能力明显降低(P<0.05)。中药干预后,端脑皮质APP695mRNA表达较模型组明显降低(P<0.01),学习记忆能力明显提高(P<0.05)。结论端脑皮质APP695mRNA表达水平升高与学习记忆能力减低明显相关,通过抑制端脑皮质APP695mRNA的过量表达是补肾填精方防治AD模型大鼠学习记忆能力损害的机制之一。     

芍药苷对APP/PS1小鼠的神经细胞保护作用及机制研究

目的:探讨芍药苷(paeoniflorin,PF)通过抑制细胞凋亡通路而产生神经细胞保护作用的机制。方法:分别选用15只5月龄雄性APP/PS1非显性小鼠作为正常对照组,15只5月龄雄性APP/PS1双转基因小鼠为模型组和15只5月龄雄性APP/PS1双转基因小鼠为给药组(5 mg/kg的PF腹腔注射)。采用水迷宫实验检测各组小鼠的学习和记忆能力。采用TUNEL荧光染色法检测脑内神经细胞凋亡情况。采用Western Blot检测脑内皮层及海马区PI3K、Akt、p-PI3K、p-Akt、caspase-3、caspase-9、Bcl-2和Bax的蛋白表达水平,并用免疫组化分析caspase-3和caspase-9的蛋白表达水平及分布情况。结果:(1)与正常对照组相比,APP/PS1模型组小鼠的学习和记忆能力明显下降;与APP/PS1模型组相比,PF明显改善小鼠的学习和记忆能力。(2)与正常对照组相比,APP/PS1模型组小鼠脑内神经细胞凋亡明显增多,分布区域较广,而PF给药组小鼠凋亡细胞明显减少。(3)与APP/PS1模型组相比,PF给药组能显著下调促凋亡因子caspase-3、caspase-9和Bax的表达水平(P0.05),同时上调抑凋亡因子pPI3K、p-Akt和Bcl-2的表达水平(P0.05)。结论:PF可能通过激活PI3K/Akt通路而上调Bcl-2,下调caspase-9、caspase-3和Bax的蛋白表达水平,从而抑制神经细胞凋亡和保护神经细胞,以治疗神经退行性疾病。     

Multifunctional Activity of Polyphenolic Compounds Associated with a Potential for Alzheimer's Disease Therapy from Ecklonia cava

Five polyphenols were isolated and purified from a brown alga Ecklonia cava. These compounds showed diverse biological activities such as antioxidative, antiinflammatory, and enzyme inhibitory activities. This led us to investigate the potential of these compounds as Alzheimer's disease drugs. All of the compounds showed moderate acetylcholinesterase inhibitory activity in a micromolar range (IC₅₀ from 16.0 to 96.3 μM). For butyrylcholinesterase, a new target for the treatment of Alzheimer's disease, phlorofucofuroeckol‐A (PFF‐A), showed a particularly potent inhibitory activity (IC₅₀ 0.95 μM), which is over 100‐fold greater than for acetylcholinesterase. These compounds inhibited glycogen synthase kinase 3 beta, which is related to the formation of hyperphosphorylated tau and generation Aβ. Bieckol and PFF‐A inhibited amyloid precursor protein biosynthesis. PFF‐A also showed very strong β‐secretase inhibitory activity with IC₅₀ of submicromole. These results render these compounds as interesting potential drug candidates for Alzheimer's disease. Copyright © 2015 John Wiley & Sons, Ltd.     

Munc13 genotype regulates secretory amyloid precursor protein processing via postsynaptic glutamate receptors

The amyloid precursor protein (APP) can be proteolytically degraded via non-amyloidogenic α-secretase and amyloidogenic β-secretase pathways. Previously, we have identified the presynaptic protein Munc13-1 as a diacylglycerol/phorbolester (DAG/PE) receptor that contributes to secretory, non-amyloidogenic APP processing after PE stimulation. Here, we used organotypic brain slice cultures from wild-type mice and from Munc13-1 knock-out (KO), Munc13-2 KO and Munc13-1/2 double KO (DKO) mice for pharmacological stimulation experiments. First, we demonstrate that neuronal populations and synaptic components important for secretory APP processing develop normally in organotypic brain slice cultures of all genotypes analyzed. Blockade of voltage-gated Na⁺ channels by tetrodotoxin reduced the PE-stimulated secretory APP processing, whereas depolarization by high extracellular K⁺ concentration evoked APP secretion. Additionally, the PE-stimulated APP secretion from Munc13-1 KO brain slices was significantly lower than that from wild-type brain slices. This effect was not observed in brain slices from Munc13-2 KO mice, which is consistent with the lower abundance and subpopulation-specific distribution of Munc13-2 in presynaptic elements. In Munc13-1/2 DKO brain slices, the deficiency of Munc13-1 dominated the effect of APP processing. The Munc13-1 KO effect on APP processing could be rescued by the stimulation of postsynaptic glutamatergic receptors. This indicates that lack of postsynaptic glutamate receptor stimulation in Munc13-1 KO brain slice cultures but not presynaptic mechanisms account for compromised APP processing. We conclude that organotypic brain slices cultures are a valuable tool for studying APP processing pathways in intact neuronal circuits and that neuronal activity is important for maintenance of the non-amyloidogenic APP processing.     

Polyprenols mitigate cognitive dysfunction and neuropathology in the APP/PS1 mouse

Alzheimer's disease (AD) is a very common neurodegenerative disorder in the elderly and brings considerable financial and social problems worldwide. In this study, polyprenols were firstly evaluated the effects on the cognitive deficits and neuropathology in APP/PS1 mice model of AD. At 3 months old, the APP/PS1 mice were divided into model group; polyprenols low, middle, and high dosage group; and positive drug group. Age‐matched wild‐type mice were chosen in control group. The administration by oral gavage lasted 6 months. Polyprenols treatment significantly improved cognitive impairment of double transgenic mice compared with vehicle control treatment in behavioral tests. In addition, immunohistochemistry and enzyme‐linked immunosorbent assay showed that there were significantly reductions in neuritic plaques and the level of hyperphosphorylated tau in brain of polyprenols‐treated mice. Furthermore, we found that polyprenols treatment reduced the apoptotic cells in brain sections of 9‐month‐old APP/PS1 mice. These results reveal that polyprenols exert neuroprotective effects in APP/PS1 mice and could represent an effective treatment for AD.