Resveratrol and Sir2 Reverse Sleep and Memory Defects Induced by Amyloid Precursor Protein / 神经科学通报·英文版

Resveratrol (RES), a natural polyphenolic phytochemical, has been suggested as a putative anti-aging molecule for the prevention and treatment of Alzheimer's disease (AD) by the activation of sirtuin 1 (Sirt1/Sir2). In this study, we tested the effects of RES and Sirt1/Sir2 on sleep and courtship memory in a Drosophila model by overexpression of amyloid precursor protein (APP), whose duplications and mutations cause familial AD. We found a mild but significant transcriptional increase of Drosophila Sir2 (dSir2) by RES supplementation for up to 17 days in APP flies, but not for 7 days. RES and dSir2 almost completely reversed the sleep and memory deficits in APP flies. We further demonstrated that dSir2 acts as a sleep promotor in Drosophila neurons. Interestingly, RES increased sleep in the absence of dSir2 in dSir2-null mutants, and RES further enhanced sleep when dSir2 was either overexpressed or knocked down in APP flies. Finally, we showed that Aβ aggregates in APP flies were reduced by RES and dSir2, probably via inhibiting Drosophila β-secretase (dBACE). Our data suggest that RES rescues the APP-induced behavioral deficits and Aβ burden largely, but not exclusively, via dSir2.

Expression, purification and micelle reconstruction of the transmembrane domain of the human amyloid precursor protein for NMR studies / 生物工程学报

The multiple-step cleavage of amyloid precursor protein (APP) generates amyloid-β peptides (Aβ), highly toxic molecules causing Alzheimer's disease (AD). The nonspecific cleavage between the transmembrane region of APP (APPTM) and γ-secretase is the key step of Aβ generation. Reconstituting APPTM under physiologically-relevant conditions is crucial to investigate how it interacts with γ-secretase and for future AD drug discovery. Although producing recombinant APPTM was reported before, the large scale purification was hindered by the use of biological protease in the presence of membrane protein. Here, we expressed recombinant APPTM in Escherichia coli using the pMM-LR6 vector and recovered the fusion protein from inclusion bodies. By combining Ni-NTA chromatography, cyanogen bromide cleavage, and reverse phase high performance liquid chromatography (RP-HPLC), isotopically-labeled APPTM was obtained in high yield and high purity. The reconstitution of APPTM into dodecylphosphocholine (DPC) micelle generated mono dispersed 2D 15N-1H HSQC spectra in high quality. We successfully established an efficient and reliable method for the expression, purification and reconstruction of APPTM, which may facilitate future investigation of APPTM and its complex in more native like membrane mimetics such as bicelle and nanodiscs.

Proteína precursora del beta-amiloide (ß-App) y daño axonal difuso tras un traumatismo craneoencefálico: un punto de vista forense / Beta-amiloid precursos protein (ß-App) and diffuse axonal damage after head injuries: a forensic point of view

Resumen La proteína precursora del β- Amiloide (β-APP) es una glicoproteína de membrana y un componente habitual de las neuronas. Tiene funciones en el crecimiento y la adhesión celular tras un traumatismo. Es transportada mediante transporte rápido axonal anterógrado y se acumula dentro de las neuronas cuando se daña citoesqueleto. Este proceso es activo, es decir consume energía. El β-APP no es específico de los traumatismos. Se acumula en cualquier circunstancia en la que se dañen los axones, tal como la hipoxia, alteraciones metabólicas, y cualquier otra causa de edema cerebral y aumento de la presión intracraneal que puedan conducir a un daño axonal difuso (DAI) En el presente estudio estudiamos la expresión de esta proteína en casos de traumatismo cráneo-encefálico con diferente evolución cronológica El daño del citoesqueleto producido por la proteólisis, junto con la alteración de las quinasas y las fosfatasas, aumentan la permeabilidad de la membrana, lo que provoca la entrada de calcio en la célula que, a su vez, activa la calmodulina que hace que los neurofilamentos se compacten, los microtúbulos desaparezcan y se rompa la espectrina. Esta disrupción del citoesqueleto tiene como consecuencia que las sustancias que se transportan a su través, se acumulen, sobre todo en las zonas afectadas por el DAI. Al final de todo este proceso, los axones se rompen, lo que se conoce como axotomía secundaria. El estudio de la acumulación del β-APP es útil para valorar la extensión del DAI y para determinar el tiempo de supervivencia tras el traumatismo o cualquier otro daño cerebral.

Abstract β-Amyloid Precursor Protein (β-APP) is a membrane glycoprotein and a common component of neurons. It is involved in adhesion and cell growth processes after traumatic events. It is carried by anterograde fast axonal transport, and it accumulates inside neurons when the cytoskeleton is damaged. This is a vital biochemical process that consumes energy. β-APP is not specific of traumatic events. It accumulates in any case of axonal damage, whatever its cause may be, like hypoxia, metabolic disorders, and any other circumstances that lead to brain swelling and intracranial pressure rising and in consequence to Diffuse Axonal Injury (DAI). In this study we review the expression of this protein in cases of traumatic brain injury with different chronological evolution. The damage of cytoskeleton due to proteolysis in addition to the disturbance of kinases and phosphatases increase the permeability of the membrane. Calcium gets into the cell and activates calmodulin, thus neurofilaments compact, microtubules disappear and spectrin breaks. This disruption of the cytoskeleton has as consequence that the transported substances accumulate in the most affected areas by DAI. At the end of this process axon breaks, which is known as secondary axotomy. The study of the accumulation of β-APP is useful to assess the extent of DAI and to determine the time elapsed after trauma or another insult to CNS.

Abdominal acupoint thread embedding therapy based on "brain-intestinal connection" for mild-to-moderate Alzheimer's disease and its effects on serum levels of APP and Aβ / 中国针灸

OBJECTIVE@#To compare the clinical efficacy of abdominal acupoint thread embedding therapy based on "brain-intestinal connection" combined with donepezil hydrochloride tablets and oral donepezil hydrochloride tablets alone for mild-to-moderate Alzheimer's disease (AD) and observe its effects on amyloid precursor protein (APP) and β-amyloid protein@*METHODS@#Sixty patients with AD were randomly divided into an observation group (30 cases, 3 cases dropped off) and a control group (30 cases, 3 cases dropped off). The patients in the control group were treated with donepezil hydrochloride tablets (5 mg per day); based on the treatment in the control group, the patients in the observation group were treated with abdominal acupoint thread embedding therapy at Zhongwan (CV 12), Xiawan (CV 10), Huaroumen (ST 24), Wailing (ST 26), Daheng (SP 15), etc., once every 10 days. Both groups were treated for 2 months. The mini-mental state examination (MMSE), Alzheimer's disease assessment scale-cognitive subscale (ADAS-Cog), activity of daily living scale (ADL), neuropsychiatric inventory questionnaire (NPI) as well as the serum levels of APP and Aβ@*RESULTS@#After treatment, the MMSE scores in the two groups were higher than those before treatment (@*CONCLUSION@#The abdominal acupoint thread embedding therapy based on the theory of "brain-intestinal connection" combined with donepezil hydrochloride tablets can improve cognitive function, self-care ability of daily life and mental behavior, and reduce the serum levels of APP and Aβ

Effect of Modified Shuyuwan on Neuroinflammation in APP/PS1 Dementia Mice Based on Nlrp3/ASC/Caspase-1 Signaling Pathway / 中国实验方剂学杂志

Objective:To observe the effect of modified Shuyuwan in amyloid precursor protein/ presenilin 1 （APP/PS1） dementia mice on cognitive and memory impairment and to explore its mechanism. Method:The 40 APP/PS1 mice were divided into model group （given Physiological saline）， low and high-dose modified Shuyuwan （14，64 g·kg^-1）group， and donepezil group （1 mg·kg^-1） and 10 wild mice were set as the blank control group （given Physiological saline）. All of the mice were administered intragastrically for 35 days. The memory and space exploration ability of mice was detected by Morris water maze， the morphology of mouse hippocampal neurons were observed by Nissl staining. The deposition of <italic>β </italic>amyloid 1-42（A<italic>β</italic>_1-42） in mouse hippocampus was detected by immunohistochemistry， and the expression of ionized calcium-binding adapter molecule 1（Iba1）， a marker of hippocampal microglia （MG） and Nitric oxide synthase（iNOS）， a marker of actived MG， were detected by immunofluorescence. The protein expression of NLR family pyrin domain containing 3（Nlrp3）， Apoptosis-associated speck-like protein containing a Caspase-recruitment domain （ASC）， cysteine protease-1（Caspase-1）and interleukin-1 beta （IL-1<italic>β</italic>） were detected by Western blot， and the expression of IL-1<italic>β</italic>， tumor necrosis factor-<italic>α</italic>（TNF-<italic>α</italic>）and interleukin-18 （IL-18） mRNA were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. Result:Compared with the blank control group， the memory and space exploration ability of the model group were significantly reduced （<italic>P</italic><0.05）， the number of hippocampal neurons decreased， the deposition of A<italic>β</italic>_1-42 increased， the markers of actived MG Iba1，iNOS increased， the protein expression of Nlrp3， ASC， Caspase-1， IL-1<italic>β</italic> increased significantly （<italic>P</italic><0.05）， and the mRNA expression of IL-1<italic>β</italic>， IL-18， and TNF-<italic>α</italic> increased significantly （<italic>P</italic><0.05）. Compared with model group， the Chinese medicine group can improve the APP/PS1 mice's space exploration ability and memory ability （<italic>P</italic><0.05）， increase the number of hippocampal neurons， reduce A<italic>β</italic>_1-42 deposition， reduce the activation of MG， and reduce the protein expression of Nlrp3， ASC， Caspase-1 and IL-1<italic>β</italic> （<italic>P</italic><0.05）， and reduced the expression of IL-1<italic>β</italic> mRNA （<italic>P</italic><0.05）. Conclusion:Modified Shuyuwan can reduce the expression of IL-1<italic>β</italic> and other inflammatory factors in the hippocampus of APP/PS1 mice by inhibiting the Nlrp3/ASC/Caspase-1 pathway， and relieve nerve inflammation and pathological injury of AD.

Roles of sodium channel-voltage-gated-beta 2B in learning and memory improvements of transgenic mice with Alzheimer's disease / 中华行为医学与脑科学杂志

Objective:To explore the potential role of amyloid precursor protein (APP) in the sodium-channel-voltage-beta 2B (SCN2B)-mediated improvement of memory and cognitive function in Alzheimer's Disease (AD).Methods:The SCN2B gene knockout mice (SCN2B -/-) were hybridized with APP gene knockout mice (APP -/-), APP gene heterozygous mice (APP +/-) and APP gene transgenic mice (APP +/+), and the tail tissue of the same mouse was genotyped by PCR gene detection.The mice were divided into SCN2B -/-APP -/- group, SCN2B -/-APP +/- group and SCN2B -/-APP +/+ group.The C57BL/6 wild-type mice were Wild type (WT) group, with 9 mice in each group.SCN2B -/-APP -/-, SCN2B -/-APP +/-, SCN2B -/-APP +/+ transgenic mice and the wild-type mice at the age of 6 months, 12 months, and 18 months were tested by Morris water Maze and Y maze test to detect the cognitive function between each group.Meanwhile, SCN2B -/-APP -/-, SCN2B -/-APP +/-, SCN2B -/-APP +/+ transgenic mice aged 6, 12, 18 months and age-match wild-type were selected to detect neuronal processes in hippocampal CA1 region, and the number of neuronal processes in basal and distal regions of hippocampal CA1 region was quantitatively analyzed.SPSS 21.0 statistical software was used for data statistics and analysis.The differences between the two groups were compared and analyzed by independent-sample t test, the comparison between multiple groups was analyzed by one way analysis of variance (ANOVA), and repeated measurement ANOVA was used to analyze behavioral deta. Results:Repeated measurement ANOVA was used to analyze the data of water maze test. The data showed that the interaction effect of escape latency group and time was significant in 18 month old mice ( Ftime×group=3.63, P<0.01). Simple effect analysis showed that compared with SCN2B -/-APP +/- group and SCN2B -/-APP -/-group, the escape latency of mice in SCN2B -/-APP +/+ group was significantly prolonged from day 4 to 6 (4th day: (47.00±2.00)s, (43.11±1.96) s, (41.89±3.06)s, t=-4.16, 1.00, both P<0.05; 5th day: (45.22±2.54) s, (36.33±2.78) s, (37.00±2.45)s, t=-7.08, -0.54, both P<0.05; 6th day: (38.11±2.03)s, (34.11±2.32)s, (33.00±2.91)s, t=-3.90, 0.90, both P<0.05). The residence time in the target quadrant was shortened((18.00±1.73)s, (25.56±1.33)s, (24.33 ±1.94)s; t=10.37, 1.56, both P<0.05). (2) Y-maze results showed that compared with SCN2B -/-APP +/- group and SCN2B -/-APP -/-group, the number of novel arm entry in 18 month old mice in SCN2B -/-APP +/+ group was decreased((50.22±3.68), (57.22±3.74), (58. 44±5.14) ; t=3.40, -0.48, both P<0.05), and the residence time of stay in the new arm was reduced((10.89±0.62)min, (14.33±0.59)min, (13.89±0.74)min; t= 8.16, 0.44, both P<0.05), and the distance of movement in the new arm was significantly reduced ((37.26±2.01)m , (45.67±2.45)m , (46.11±3.27)m ; t=7.81, 0.91, both P<0.05). (3) Golgi staining showed that SCN2B -/-APP +/- group and SCN2B -/-APP -/-group, the number of apical dendrites in hippocampal neurons of 18 month old mice in SCN2b -/-App +/+ group(number of apical dendrites: (1.78±0.37), (3.67±0.81), (3.00±1.21); t=3.36, 1.41, both P<0.05) and the number of basal dendrites (the number of basal dendrites: (1.11±0.50), (3.11±0.50), (2.56±0.69); t=4.06, 1.21, both P<0.05). Conclusion:SCN2B knockdown can improve the ability of spatial learning and memory in aged mice.Overexpression of APP can partially offset the improvement of cognitive function caused by SCN2B knockdown, and may be affected by the number of basal and distal processes of neurons in the hippocampal CA1 region of the mice.

The Essentials of Biochemistry of the Proteins as Related to Alzheimer’s Disease: A Review

Amyloid plaques and Tau tangles, constitute the pathological hallmarks of the brains of the patients suffering from Alzheimer’s disease. They are identified as far back as 1996 by Alois Alzheimer, a German psychiatrist and neuropathologist, but till this date, how they produce neuronal death remained an enigma. The amyloid cascade theory held its sway until recent times until the emphasis is shifted to the metabolites of amyloid Beta precursor protein (APP). Several metabolites of APP are formed depending on by which pathway, the APP is metabolized, either by the non - amyloidogenic pathway (forming ? - C terminal fragment - CTF? / C83 and the N - terminal fragment sAPP? / P3 and the APP intracellular domain AICD). Or amyloidogenic pathways. ( Forming extracellular A? and APP intracellular domain - AICD). The hyperphosphorylation is held responsible for the tau protein tangles. The over activity of the tau kinases or the failure of inhibition by the tau phosphatases i s implicated, in tau tangle deposits. These biochemical aspects of AD assumed importance in connection with the interventional therapeutic strategies that are developed in the years bygone, as well as those still are in the developing stage. In keeping with this fact, it is attempted to review the essentials of the biochemical aspects of the involved proteins, as related to AD, in this article

Amyloid precursor protein regulates 5-fluorouracil resistance in human hepatocellular carcinoma cells by inhibiting the mitochondrial apoptotic pathway / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Hepatocellular carcinoma (HCC) is a malignant tumor with high morbidity and mortality globally. It accounts for the majority of primary liver cancer cases. Amyloid precursor protein (APP), a cell membrane protein, plays a vital role in the pathogenesis of Alzheimer’s disease, and has been found to be implicated in tumor growth and metastasis. Therefore, to understand the relationship between APP and 5-fluorouracil (5-FU) resistance in liver cancer, Cell Counting Kit-8, apoptosis and cell cycle assays, western blotting, and reverse transcription-quantitative polymerase chain reaction (qPCR) analysis were performed. The results demonstrated that APP expression in Bel7402-5-FU cells was significantly up-regulated, as compared with that in Bel7402 cells. Through successful construction of APP-silenced (siAPP) and overexpressed (OE) Bel7402 cell lines, data revealed that the Bel7402-APP751-OE cell line was insensitive, while the Bel7402-siAPP cell line was sensitive to 5-FU in comparison to the matched control group. Furthermore, APP overexpression decreased, while APP silencing increased 5-FU-induced apoptosis in Bel7402 cells. Mechanistically, APP overexpression and silencing can regulate the mitochondrial apoptotic pathway and the expression of apoptotic suppressor genes (B-cell lymphoma-2 (Bcl-2) and B-cell lymphoma-extra large (Bcl-xl)). Taken together, these results preliminarily revealed that APP overexpression contributes to the resistance of liver cancer cells to 5-FU, providing a new perspective for drug resistance.

Effect of Huangjingwan on Antioxidant Functions and Aβ1-42 and APP Protein Expressions in Brain of Alzheimer' s Disease Rats / 中国实验方剂学杂志

Objective::To observe the effect of Huangjingwan (HW) on antioxidant functions and β-amyloid 1-42 (Aβ1-42) and amyloid precursor protein (APP) expressions in the brain of Alzheimer' s disease (AD) rats. Method::SD rats were randomly divided into normal control group, sham model control group, AD model group, and low, medium, high-dose (equivalent raw drug dose 1, 3, 9 g·kg-1·d-1) HW groups.The AD models were established through intraperitoneal injection with 1.25% D-galactose (120 mg·kg-1·d-1, 6 consecutive weeks) and then one-time right ventricular injection with Aβ1-42 (10 μg). Two weeks after modeling, the rats in each HW group received corresponding drugs through intragastric administration, once a day, while the rats in sham model control group, AD model group were given normal saline 1 mL through intragastric administration, once a day.Gastric perfusion lasted for 8 weeks.At the end of the experiment, learning and memory abilities of the rats were assessed by Platform Jumping Test.The changes of physical endurance in rats were tested by 10% weight swimming under load.The activities of superoxide dismutase (SOD), glutathione reductase (GR), glutathione peroxidase (GSH-Px) antioxidant enzymes and the contents of glutathione (GSH) and malondialdehyde (MDA) in rat brain tissue were detected by colorimetry.The changes of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), Aβ1-42 and APP protein in rat brain tissues were determined by enzyme linked immunosorbent assay (ELISA). Western blot analysis was used to measure the expression of APP protein in rat brain. Result::Compared with the normal control group, rats in AD model group showed an obvious dementia state, that is more lying and less movement, longer learning response time, significant increase in the number of learning and memory errors, significant attenuation in physical fitness, significant decrease in the activities of antioxidant enzymes (SOD, GR, GSH-Px) and anti-inflammatory factors GSH in brain, significant rise in the levels of inflammatory factors MDA, IL-1β and TNF-α and the content of Aβ1-42 protein, and significant reduction in the content of APP protein in brain (P<0.01). Low, medium and high-dose HW could ameliorate dementia symptoms in AD rats, improve the achievement of learning and memory, antagonize body weakness and increase physical fitness, promote SOD, GR, GSH-Px activities and anti-inflammatory factor GSH level in the brain, reduce the levels of MDA, IL-1β and TNF-α in the brain, decrease the level of Aβ1-42 and increase the level of APP protein in the brains of AD rats compared with the AD model group (P<0.05, P<0.01), besides, within the dose range of 1-9 g·kg-1·d-1, HW has a more obvious effect with the increase of dose. Conclusion::HW has the effects in preventing and treating AD, which is related to the HW' s mechanisms in enhancing the function of antioxidant system in brain, reducing neuroinflammatory reaction and deposition of Aβ1-42 induced by oxidative stress, and maintaining the expression level of APP protein.

Vitis vinifera acts as anti-Alzheimer's agent by modulating biochemical parameters implicated in cognition and memory

Background: Aluminum a known neuro and cholinotoxin has been implicated in the pathogenesis ofAlzheimer’s disease. Its exposure is associated with impairment of the memory and cognition.Objective: The present study was undertaken to evaluate the anti-Alzheimer’s activity of Vitis vinifera inaluminum induced Alzheimer’s disease.Materials and methods: In this study, we investigated the behavioral and biochemical effects ofaluminum in Sprague-Dawley rats. Animals were exposed to aluminum chloride (100 mg/kg/day) orallyfor a period of 8 weeks. Vitis was given in doses of 250 mg/kg and 500 mg/kg for 16 weeks and thepossible effects of Vitis vinifera on the expression of Tau and amyloid precursor protein were evaluated byPCR analysis and the possible activities of lipid peroxidation, inflammation and anti-cholinesterase activity were evaluated.Results: Aluminum intoxication was associated with significant impairment in learning and memory inMorris water maze test. A significant improvement was observed with Vitis vinifera in a dose dependentmanner.Conclusion: The findings of the present study revealed the significant neuroprotective actions of Vitisvinifera by modifying the biochemical parameters and inhibited the mRNA expression of Amyloid Precursor Protein and Tau, which are the key pathological hallmarks of Alzheimer’s disease, which wasfurther confirmed by histopathological observations.© 2017 Transdisciplinary University, Bangalore and World Ayurveda Foundation. Publishing Services byElsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Phytomolecules for Therapeutic Targets of Alzheimer’s Disease Pathology - A Virtual Analysis

Introduction: Life style of humans, with changes in diet, exercise and life style practices which play an important role in enhancing the progression of age related degenerative problems like dementia. The most common cause of dementia in the world is Alzheimer’s disease which ultimately decrease the cognitive function mainly learning and memory. The objective of the study was to find the multi target potential efficacy of the ligands, Glabridin and Diosmetin in altering the two main molecular targets of Alzheimer’s disease (AD). The target enzymes were amyloid binding alcohol dehydrogenase (ABAD) and β-site amyloid precursor protein cleaving enzyme 1(BACE1). Material and methods: In this study, we analyzed that multitarget potential of the two natural compounds on the two prior target enzymes of Alzheimer’s disease which are mainly involved in producing neurodegeneration. Drug likeness properties, absorption, digestion metabolic and toxicity profile and molecular docking were analyzed to determine therapeutic aspect by virtual methods. Results: Binding energy and Vander Waals force of Diosmetin were higher than Glabridin with the target ABAD and less than with BACE1 which showed that both drugs can be used in modulating the enzyme phosphorylation in Alzheimer’s disease. Conclusion: Glabridin and Diosmetin could be used as promising drug candidates as ABAD inhibitor and BACE1 inhibitor in Alzheimer’s disease

Effect of Electroacupuncture on Learning-memory and Expression of β-site Amyloid Precursor Protein Cleaving Enzyme 1 in APP/PS1 Mice / 中国康复理论与实践

Objective To explore the effects of electroacupuncture at Baihui (DU20) and Shenting (GV24) on Alzheimer's disease, and possible mechanism for it.Methods A total of 24 eight-month-old APP/PS1 male mice were randomly divided into model group (n = 8), electroacupuncture group (n = 8) and non-acupoint group (n = 8), and other eight wild-type mice were as wild-type group.The electroacupuncture group accepted electroacupuncture at Baihui and Shenting, while the non-acupoint group accepted electroacupuncture at bilateral subcostal non-acupoint area, and the wild-type group and the model group accepted the same grasping and fixing, for 28 days. Then they assessed with Morris water maze test. The levels of β-amyloid protein (Aβ) and β-site amyloid precursor protein cleaving enzyme 1 (BACE1) in cerebral cortex were detected with immunohistochemistry and immunofluorescence respectively, and the level of BACE1 m RNA with RT-PCR.Results Compared with the model group, the escape latency decreased in the electroacupuncture group (P ＜ 0.001), and the times crossing platforms increased (P ＜ 0.001), while the expression of BACE1 and Aβ decreased (P ＜ 0.001).Conclusion Electroacupuncture may improve the learning-memory ability by inhibiting the expression of BACE1 in the cerebral cortex of APP/PS1 mice to decreasing the level of Aβ.

Effect of Notch-Hes1 signaling blockade by a γ-secretase inhibitor on the expression of γδT17 cells in a mouse model of psoriasis-like skin inflammation / 中华皮肤科杂志

Objective@#To evaluate the effect of Notch-Hes1 signaling blockade by a γ-secretase inhibitor on the expression of γδT17 cells in a mouse model of psoriasis-like skin inflammation.@*Methods@#Forty-five healthy specific pathogen-free (SPF) male BALB/c mice were randomly divided into control group, model group and intervention group by simple random sampling. The model group and intervention group were both topically treated with imiquimod 5% cream (62.5 mg once a day) on the shaved back, the intervention group were then intraperitoneally injected with the γ-secretase inhibitor DAPT (10 mg/kg once a day) immediately after topical application of imiquimod, and the control group were topically treated with equivalent amount of vaseline once a day. After 6-day treatment, psoriasis area and severity index (PASI) was used to evaluate changes of skin lesions. On day 7, blood samples were obtained from all the mice through heart puncture after anesthetization, and spleen and skin tissues were acquired to prepare single cell suspension. Spleen index was compared among the 3 groups. Skin tissues on the mouse back were resected and subjected to hematoxylin-eosin staining to observe histopathological changes. Flow cytometry was performed to determine the percentage of γδT17 cells in the spleen and skin tissues, real-time reverse transcription (RT) -PCR to measure the mRNA expression of Hes1 in single cell suspension of the spleen, and enzyme-linked immunosorbent assay (ELISA) to determine the serum level of interleukin (IL) -17A. Statistical analysis was carried out by using one-way analysis of variance and repeated measures analysis of variance for comparison of indices among groups, and Pearson correlation analysis for evaluating the correlation between different indices.@*Results@#Twenty-four hours after the final treatment, the intervention group showed milder psoriasis-like skin inflammation, lower PASI score, and milder degree of epidermal thickening and dermal inflammatory cell infiltration compared with the model group. The model group showed significantly increased spleen index (12.534 ± 1.636) , proportions of γδT17 cells in the spleen (24.659% ± 4.603%) and skin tissues (22.127% ± 5.670%) , mRNA expression of Hes1 in the spleen (4.867 ± 0.543) , and serum level of IL-17A ([22.478 ± 2.776] ng/L) compared with the control group (all P < 0.01) . However, the above indices were significantly lower in the intervention group (9.449 ± 1.040, 14.966% ± 5.770%, 13.631% ± 5.946%, 2.541 ± 0.347, [18.639 ± 1.816] ng/L) than in the model group (all P < 0.01) . In the model group and intervention group, there were positive correlations between the proportions of γδT17 cells in the spleen and serum levels of IL-17A (r = 0.56, 0.53 respectively, both P < 0.05) , between the proportions of γδT17 cells in skin lesions and PASI scores (r = 0.56, 0.52 respectively, both P < 0.05) , as well as between the mRNA expression of Hes1 in the spleen and the proportions of γδT17 cells (r = 0.61, 0.58 respectively, both P < 0.05) or serum levels of IL-17A (r = 0.60, 0.54 respectively, both P < 0.05) .@*Conclusion@#Notch-Hes1 signaling blockade by γ-secretase inhibitor can markedly inhibit the expression of γδT17 cells, and effectively alleviate the severity of psoriasis-like skin inflammation in mouse models.

Analysis of a family with early onset familial Alzheimer′s disease caused by mutation of amyloid precursor protein gene p.V717I / 中华神经科杂志

Objective@#To analyze the clinical data of a family with early-onset familial Alzheimer′s disease and to analyze the mutation of the pathogenic gene in the family.@*Methods@#The clinical data of a proband who was clinically diagnosed as early-onset Alzheimer′s disease in the Department of Neurology, People′s Hospital of Zhengzhou University in October 2018 and her family members were collected. Moreover, whole exome sequencing was performed on blood sample from the proband, then its deleterious effects were assessed according to the Standards and guidelines for the interpretation of sequence variants, a joint consensus recommendation of the American College of Medical Genomics. Subsequently, the strong pathogenic mutation was validated by Sanger sequencing in the some members of the family and 50 sporadic Alzheimer′s disease and 50 normal individuals of the family. Apolipoprotein E (APOE) typing of 10 family members was all epsilon 3/epsilon 3.@*Results@#The proband in this family showed decreased memory, visual space disorder, verbal repetition, personality change and abnormal mental behavior. The mutation at codon 717 of exon 17 of the proband amyloid precursor protein gene was detected by gene detection. The mutation at codon 717 of exon 17 of the proband beta-amyloid precursor protein gene was also found in the other five members of the family. The mutation was not found in 50 sporadic Alzheimer′s disease patients and 50 normal individuals outside the family. The proband′s head magnetic resonance imaging (MRI) showed bilateral hippocampal atrophy on plain scan, especially on the left side. No obvious abnormality was found in the head magnetic resonance angiography. The head MRI of the proband′s sister showed brain atrophy and bilateral hippocampal atrophy.@*Conclusions@#The study identified the pathogenic mutation of the beta-amyloid precursor protein gene p.V717I in six patients of a family with early-onset familial Alzheimer′s disease, and the mutation showed a phenomenon of family segregation. This finding is of great significance to the study of early-onset Alzheimer′s disease in Chinese population.

Analysis of a family with early onset familial Alzheimer′s disease caused by mutation of amyloid precursor protein gene p.V717I / 中华神经科杂志

Objective To analyze the clinical data of a family with early?onset familial Alzheimer′s disease and to analyze the mutation of the pathogenic gene in the family. Methods The clinical data of a proband who was clinically diagnosed as early?onset Alzheimer′s disease in the Department of Neurology, People′s Hospital of Zhengzhou University in October 2018 and her family members were collected. Moreover, whole exome sequencing was performed on blood sample from the proband, then its deleterious effects were assessed according to the Standards and guidelines for the interpretation of sequence variants, a joint consensus recommendation of the American College of Medical Genomics. Subsequently, the strong pathogenic mutation was validated by Sanger sequencing in the some members of the family and 50 sporadic Alzheimer′s disease and 50 normal individuals of the family. Apolipoprotein E (APOE) typing of 10 family members was all epsilon 3/epsilon 3. Results The proband in this family showed decreased memory, visual space disorder, verbal repetition, personality change and abnormal mental behavior. The mutation at codon 717 of exon 17 of the proband amyloid precursor protein gene was detected by gene detection. The mutation at codon 717 of exon 17 of the proband beta?amyloid precursor protein gene was also found in the other five members of the family. The mutation was not found in 50 sporadic Alzheimer′s disease patients and 50 normal individuals outside the family. The proband′s head magnetic resonance imaging (MRI) showed bilateral hippocampal atrophy on plain scan, especially on the left side. No obvious abnormality was found in the head magnetic resonance angiography. The head MRI of the proband′s sister showed brain atrophy and bilateral hippocampal atrophy. Conclusions The study identified the pathogenic mutation of the beta?amyloid precursor protein gene p. V717I in six patients of a family with early?onset familial Alzheimer′s disease, and the mutation showed a phenomenon of family segregation. This finding is of great significance to the study of early?onset Alzheimer′s disease in Chinese population.

Justicidin A Reduces β-Amyloid via Inhibiting Endocytosis of β-Amyloid Precursor Protein

β-amyloid precursor protein (APP) can be cleaved by α-, and γ-secretase at plasma membrane producing soluble ectodomain fragment (sAPPα). Alternatively, following endocytosis, APP is cleaved by β-, and γ-secretase at early endosomes generating β-amyloid (Aβ), the main culprit in Alzheimer's disease (AD). Thus, APP endocytosis is critical for Aβ production. Recently, we reported that Monsonia angustifolia, the indigenous vegetables consumed in Tanzania, improved cognitive function and decreased Aβ production. In this study, we examined the underlying mechanism of justicidin A, the active compound of M. angustifolia, on Aβ production. We found that justicidin A reduced endocytosis of APP, increasing sAPPα level, while decreasing Aβ level in HeLa cells overexpressing human APP with the Swedish mutation. The effect of justicidin A on Aβ production was blocked by endocytosis inhibitors, indicating that the decreased APP endocytosis by justicidin A is the underlying mechanism. Thus, justicidin A, the active compound of M. angustifolia, may be a novel agent for AD treatment.

Herpes simplex virus type induces β-amyloid expression in human neuroblastoma cell lines sh-sy5y / 第二军医大学学报

Objective To explore the effect of herpes simplex virus type (HSV-1) infection on expression of β-amyloid (Aβ) in human neuroblastoma cell lines SH-SY5Y. Methods African green monkey kidney cell lines Vero cells were used to amplified HSV-1, and the virus titers were measured. SH-SY5Y cells were infected with HSV-1 3.2×108 plaque forming unit (PFU)/mL at a multiplicity of infection (MOI) of 10 for 12 h or 24 h, and the morphological changes of the cells were observed under microscope. PCR was used to detect the expression of HSV-1 DNA. Double-color immunofluorescence assay was performed to show the expression of Aβ and apolipoprotein E (ApoE). Western blotting was used to detect the expression levels of amyloid precursor protein (APP), melanin metabolic enzyme (MME), ApoE, glycogen synthase kinase-3β (GSK-3β) and phosphorylated glycogen synthase kinase-3β (p-GSK-3β). Results After infection with HSV-1 for 12 h, the SH-SY5Y cells had synapse reduction and neurite retraction and few neurites. And after 24 h of infection, the cells began to aggregate, and were round and shed. Compared with phosphate buffer saline (PBS) control group, the expression of Aβ was significantly increased after infection with HSV-1 for 12 h (P0.01), while the expression of ApoE protein was not significantly changed. After 24 h of infection, the expressions of Aβ and ApoE were significantly increased (P0.05), but the expression of Aβ was significantly lower than that on 12 h of post-infection (P0.01). Western blotting analysis showed that, compared with PBS control group, the expression of APP was significantly decreased on 12 h of post-infection (P0.05), and the expressions of GSK-3β, p-GSK-3β and MME were significantly increased (P0.05). However, the expression of MME was significantly decreased (P0.05) and the expression of ApoE was significantly increased (P0.05) 24 h post-infection. Conclusion HSV-1 infection induces the expression of Aβ in SH-SY5Y cells through promoting APP metabolism and Ser9 phosphorylation of GSK-3β on 12 h of post-infection, and inhibiting the degradation of Aβ on 24 h of post-infection.

Syzygium aromaticum ethanol extract reduces AlCl

Objective: To investigate the neuroprotective effects of Syzygium aromaticum (S. aromaticum) extract (500 mg/kg) on AlCl

Syzygium aromaticum ethanol extract reduces AlCl

Objective:To investigate the neuroprotective effects of Syzygium aromaticum (S. aromaticum) extract (500 mg/kg) on AlClMethods:An ethanolic extract of S. aromaticum seeds was prepared and the active compounds were identified using nuclear magnetic resonance spectroscopy. BALB/ c mice were divided into five groups (negative control, AlClResults:It was observed that AlClConclusions:The results showed that S. aromaticum is a promising antioxidant and a neuroprotective agent.

3D-QSAR Analysis and Molecular Docking Study on Biaryl Aminothiazine BACE1 Inhibitor / 中国药房

OBJECTIVE:To provide theoretic basis for the design and synthesis of novel high-activity biaryl aminothiazineβ-amyloid precursor protein cleaving enzyme 1 (BACE1) inhibitor,the research and development of new AD therapy drugs. METHODS:Totally 41 molecules of biaryl aminothiazine BACE1 inhibitors were selected. By SYBYL-X 2.0 software package, CoMFA and CoMSIA method were used to construct 3D-QSAR model of derivatized compounds. Surflex-dock molecular docking was applied to analyze binding mode of the compounds with BACE1. RESULTS:The q2 value of 3D-QSAR model established by CoMFA and CoMSIA method were all higher than 0.5,indicating good predictability. The established three dimensional contour plots could manifest the effect of substituents at different sites on activity of compounds. Surflex-dock analysis showed that biaryl aminothiazine and amino acid residues as ASP80, ASP276 and TYR246 in BACE1 had a key effect on hydrogen bonds. CONCLUSIONS:3D-QSAR model established on the basis of biaryl aminothiazine derivatized compounds show good predictability,which provides guidance for the structure optimization of the compound. TYR246 may be another potential active functional residue of biaryl aminothiazine inhibitor compound molecule combined with BACE1. Through 3D-QSAR analysis and molecular docking,new biaryl aminothiazine BACE1 inhibitor can be designed and synthesized so as to research and develop new drugs for AD.