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Chronic effects of ketamine on gene expression changes in neurotransmitter receptors and regulators-A pcr-array study

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MOLECULAR & CELLULAR TOXICOLOGY 2015年第4期11卷 395-400页

作者： Tan, Sijie Zou, Ju Li, Meixiang Yew, David T. Univ South China Sch Med Dept Histol & Embryol Hengyang Peoples R China Chinese Univ Hong Kong Inst Chinese Med Brain Res Ctr Shatin Hong Kong Peoples R China Univ South China Sch Med Dept Parasitol Hengyang Peoples R China

of ketamine may result in dependence. We investigated the long-term effects of ketamine on anxiety like behaviors and gene expression changes in neurotransmitter receptors and regulators in mice. One month old ICR mice received sub-anesthetics dose of ketamine daily for 3 months. An Elevated Plus Maze (EPM) was employed to assess anxiety like behaviors in these mice and a pcr array was used to evaluate gene expression changes in neurotransmitter receptors and regulators in the brain. Ketamine decreased the time spent in the open arms and the visit of the open arms in the EPM. Drug dependence related genes, namely Prokr2, Npffr1 and GalR2 were differentially expressed in ketamine treated mice. The results of the present study depicted addictive potential of ketamine and may be beneficial for developing pharmacotherapies for ketamine dependence.

关键词： Ketamine Prefrontal cortex pcr-array Addiction

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Validation of commercial real-time pcr-arrays for environmental risk assessment: Application to the study of p,p'-DDE toxicity in Mus spretus mice liver

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ENVIRONMENTAL POLLUTION 2017年第Nov.期230卷 178-188页

作者： Morales-Prieto, Noelia Pueyo, Carmen Abril, Nieves Univ Cordoba Dept Biochem & Mol Biol Agrifood Campus Int Excellence CeiA3 UCO Bldg Severo Ochoa2nd FloorCampus Rabanales Cordoba Spain

Data on gene transcription profiles provide a comprehensive assessment of the toxic and defensive mechanisms that are triggered by pollutants. pcr-arrays have emerged as a reliable tool for analyzing the expression of a panel of relevant, pathway- or disease-focused genes under uniform cycling conditions. By using SYBR Green-optimized primer assays, it is possible to simultaneously amplify a sample with high specificity and amplification efficiencies. However, commercial pcr-arrays target a limited group of organisms, excluding most of those with environmental relevance, as is the case with Mus spretus mice. Our previous works with M. spretus showed a high sequence similarity between M. spretus and the model organism M. musculus allowing the use of commercial platforms with M. spretus. This work demonstrates the successful application of a commercial pcr-array designed for the model organism M. musculus to assess the biological effects caused by the organochlorine pesticide p,p'-DDE in a focused panel of stress related genes in M. spretus mice. This cost-effective, easy-to-use platform detected quantitative gene profiling differences between M. spretus hepatic RNA samples and generated data highly concordant with those obtained by absolute qRT-pcr, the most sensitive method to quantify transcripts. This platform is also suitable for use in field studies with free-living M. spretus mice for routine environmental risk assessment. Our results provide a broad impression of the biological consequences of p,p'-DDE on the hepatic health of mice. (C) 2017 Elsevier Ltd. All rights reserved.

关键词： Transcriptional profiles pcr-array p,p'-DDE Mus spretus Environmental biomonitoring

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Bisphenol-A impairs cellular function and alters DNA methylation of stress pathway genes in first trimester trophoblast cells

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REPRODUCTIVE TOXICOLOGY 2018年 82卷 72-79页

作者： Basak, Sanjay Srinivas, Vilasagaram Duttaroy, Asim K. Univ Oslo Fac Med Inst Basic Med Sci Dept Nutr POB 1046 N-0316 Oslo Norway Natl Inst Nutr Hyderabad India

Humans are exposed to Bisphenol A (BPA) from the consumer products and plastic substances. However, impacts of low levels of BPA exposure on placental developmental processes such as first trimester trophoblast cell growth, angiogenesis and epigenetic modifications are not well studied. Low concentration of BPA (1 nM) affected cell proliferation of human placental first trimester trophoblasts using a model cell, HTR8/SVneo. BPA abolished both basal- and vascular endothelial growth factor (VEGF)-stimulated tube formation in these cells. BPA significantly down regulated mRNA expression of VEGF, proliferating cell nuclear antigen, intercellular adhesion molecule 1 with concomitant upregulation of 11-beta-hydroxysteroid dehydrogenase 2 mRNA and protein expression in HTR8/SVneo cells. BPA also lowered CpG methylation of gene promoter associated with metabolic and oxidative stress. This study demonstrated that BPA at 1 nM not only affected cellular growth, development and angiogenic activities but also affected DNA methylation of stress response and down-regulation of angiogenic growth factors in first trimester trophoblast cells.

关键词： Bisphenol Tube formation Thymidine assay VEGF HTR8/SVneo First trimester placenta pcr-array DNA methylation

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Plasma levels of increased miR-195-5p correlates with the sFLT-1 levels in preeclampsia

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HYPERTENSION IN PREGNANCY 2016年第2期35卷 150-158页

作者： Sandrim, Valeria Cristina Eleuterio, Nibia Pilan, Eliane Tanus-Santos, Jose Eduardo Fernandes, Karla Cavalli, Ricardo Univ Estadual Paulista UNESP Inst Biosci Dept Pharmacol Botucatu SP Brazil Nucleo Posgrad & Pesquisa Santa Casa Belo Horizon Belo Horizonte MG Brazil Univ Sao Paulo Fac Med Ribeirao Preto Dept Pharmacol BR-14049 Ribeirao Preto SP Brazil Univ Sao Paulo Fac Med Ribeirao Preto Dept Gynecol & Obstet BR-14049 Ribeirao Preto SP Brazil

Although the role of soluble fms-like tyrosine kinase 1 (sFLT-1) in preeclampsia is well-established, the mechanism related to its synthesis remains poorly understood. We evaluated the association among the circulating microRNAs (miRs) and sFLT-1 levels in preeclampsia pregnant women. For this purpose, we measured the plasma sFLT-1 levels in 24 preeclampsia women and selected from these, three subjects with the lowest and three with the highest levels of sFLT-1 in order to screen for potential miRs associated with plasmatic sFLT-1 concentrations using a polymerase chain reaction-array (pcr-array) methodology. From screening results, we found three statistically different expressed miRs with fold change (FC-high/low levels) = 3.0: miR-195-5p (FC = 5.2 increase in group with high sFLT-1 levels), miR-16-5p (FC = 3.2;increase in group with high sFLT-1 levels), andmiR-375 (FC = -3.0;decrease in group with high sFLT-1 levels) which were later validated in all samples (n = 24). To correlate these miRs and plasma sFLT-1 levels, we used two extremes of analysis. In one part, we compared 12 preeclampsia women with the lowest sFLT-1 levels (L-50% group) to 12 with the highest levels (50% H group);and in the second analysis, 6 preeclampsia women (L-25%) from the L group to 6 preeclampsia women from the H group (H-25%). Our results showed increased expression of miR-195-5p in the H group, considering both the analyses with 50%, FC = 2.1 and 25%, FC = 4.2. Regarding other miRs, lack of correlation was found in both analyses (50% and 25%). In conclusion, our data demonstrate an association of higher levels of sFLT-1 with increased expression of miR-195-5p in preeclampsia pregnant women.

关键词： Circulating microRNA microRNA miR-195 pcr-array preeclampsia sFLT-1

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Identification of dopamine- and serotonin-related genes modulated by bisphenol A in the prefrontal cortex of male rats

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CHEMOSPHERE 2015年 139卷 235-239页

作者： Castro, Beatriz Sanchez, Pilar Miranda, Maria T. Torres, Jesus M. Ortega, Esperanza Univ Granada Dept Biochem & Mol Biol Fac Med Granada 18012 Spain Univ Granada Dept Biostat Fac Med Granada 18012 Spain Univ Granada Inst Neurosci Granada 18012 Spain

There is concern that exposure of embryos and/or infants to bisphenol A (BPA) may lead to neurological and behavioral disorders with unknown prefrontal cortex (PFC) involvement. Critical PFC functions are modulated by dopamine (DA) and serotonin (5-HT) systems, whose alterations have been associated with psychopathologies that may appear in youth and/or adulthood. This study aims to determine in the PFC of male rats exposed to a low dose of BPA (10 mu g kg(-1) d(-1)) from gestational day 12 (GD12) to postnatal day 21 (PND21): (i) DA- and 5-HT-related genes modulated by BPA at the juvenile stage (PND21);(ii) reversible and irreversible transcriptional effects;(iii) long-term consequences (effects in adult rats, PND90). In juvenile rats, BPA altered significantly the transcription of 12 out of the 84 genes analyzed using pcr-array techniques. Interestingly, transcript levels of the neurotrophic factor Gdnf were decrease by BPA in both juvenile and adult rats. At adulthood, disruptions in genes encoding rate-limiting enzymes for DA and 5-HT synthesis emerged. Overall, the results indicate that early-life exposure to BPA has consequences on DA and 5-HT systems in both juvenile- and adult-life stages. Additionally, we reveal molecular targets that could provide the foundation for future BPA neurotoxicity studies. (C) 2015 Elsevier Ltd. All rights reserved.

关键词： Bisphenol A Cortex Dopamine pcr-array Rat Serotonin

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The effect of high frequency repetitive transcranial magnetic stimulation(rTMS) on the cognitive behavior and hippocampal synaptic plasticity in aged Kunming mice

The effect of high frequency repetitive transcranial magneti...

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第六届亚太神经科学联合会学术会议暨中国神经科学学会第十一届全国学术会议

作者： Yuan GENG Bing HAN Yan-Yong WANG Pei-Zhi WANG Ming-Wei WANG Department of Neurology the First Hospital of Hebei Medical University Brain Aging and Cognitive Neuroscience Key Laboratory of Hebei

Objective To observe the effect of high frequency repetitive transcranial magnetic stimulation(rT MS) on the cognitive behavior and hippocampal synaptic plasticity in aged mice. Methods 45 Kunming mice(15-month-old) were randomly divided into three groups: sham, subthreshold and upthreshold. Two sessions of 25 Hz r TMS comprising of 1000 pulses in 10 trains were delivered every day for 14 days. Sham group were treated by the reverse side of the coil. Spontaneous motor activity, step-down test and Morris water maze(MWM) were conducted following the r TMS. The synaptic structure of hippocampal CA1 region were observed by TEM and the changes of 84 synaptic plasticity related genes were tested by pcr array. Results Compared with sham group, spontaneous motor activity were no significant difference between each test group(p >0.05). In step-down test,the latency of reaction increased and the number of errors decreased in both test groups(p<0.05). In hidden platform trials of MWM, compared with sham group, the mean escape latency were significantly shorter in subthreshold group(p<0.05), however, in spatial probe test, the time spend in and the number of crossing original platform were significantly increased in upthreshold group(p<0.05). Shown by TEM, compare with the sham group, the thickness of postsynaptic density(PSD) was increased, the synaptic cleft width and the synaptic curvature were decreased, the ratio of perforated synapse was increased(p<0.05), moreover, upthreshold group was more significant than subthreshold group. Gene expression profiles of each groups compared with sham group were identified with a >3-fold difference. In upthreshold group, Jun-β was up-regulated while Ntf5, Inhba and Fos were down-regulated. In subthreshold group, Jun-β, Egr2 and Nr4a1 were up-regulated while Ywhaq and kif17 were down-regulated. Conclusion 25 Hz high frequency r TMS has no significant effect on the emotion of aged Kunming mice. Up- and sub-threshold intensity of rT MS bot

关键词： High frequency repetitive transcranial magnetic stimulation aged Kunming mouse cognitive behavior synaptic plasticity pcr-array

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唾液酸乳糖的摄入对仔猪脑发育、认知功能、视觉发育及唾液酸表达的影响

唾液酸乳糖的摄入对仔猪脑发育、认知功能、视觉发育及唾液酸表达...

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作者：王芳厦门大学

学位级别：博士

研究背景:神经系统由中枢和周围神经系统构成,脑和脊髓组成了中枢神经系统,是人体神经系统的最主要部分。婴儿大脑发育最为迅速的时期是出生后两年内,此时是神经元细胞功能完善、突触生长和轴突髓鞘化的最重要时期。视网膜为大脑的延伸... 详细信息

研究背景:神经系统由中枢和周围神经系统构成,脑和脊髓组成了中枢神经系统,是人体神经系统的最主要部分。婴儿大脑发育最为迅速的时期是出生后两年内,此时是神经元细胞功能完善、突触生长和轴突髓鞘化的最重要时期。视网膜为大脑的延伸部分,是视觉信息形成的核心部位。虽然由遗传因素主导的视觉功能在婴儿出生时已经基本形成,但出生后在周围环境、营养和视觉习惯的调控下仍可以对视觉神经系统的功能进行进一步的完善和塑造,因此视觉系统的发育是由遗传、营养和环境因素共同塑造的。婴儿期神经系统的发育需要大量而丰富的营养物质,而在这一时期,婴儿的主要营养物质来源是母乳。母乳是婴幼儿发育早期最佳的营养来源,因为它可以提供婴儿正常生长发育需要的所有营养。此外,母乳同时也富含一些其它营养成分,可能会给婴儿提供超越传统营养的好处。母乳中富含丰富的低聚糖,母乳低聚糖(Human milk oligosaccharides,HMOs)是母乳中非常重要的一部分,它由中性及酸性低聚糖构成。酸性低聚糖,如唾液酸乳糖(Sialyllactose,SL),含有唾液酸(Sialic acid,Sia)残基;中性低聚糖,包括半乳低聚糖和低聚果糖。母乳中73.8%的唾液酸是以低聚糖结合的形式存在的,其中最主要的唾液酸化的低聚糖是3'-SL和6'-SL。6'-SL的浓度在初乳中最高(250-1300 mg/L),此后逐渐降低,然而,3'-SL的浓度则相对稳定(76-300mg/L)。SL被认为在新生儿发育中起到重要的作用,因为它们在抵抗病原体,促进肠道成熟,免疫功能及认知发育中起到重要作用。同时,唾液酸乳糖可在肠道内被分解,从而提供神经发育所必需的营养素——唾液酸。唾液酸(Sialic acid,Sia)是一类带羧基的九碳糖化合物酰基衍生物的通称。1957年,Blix等用弱酸水解方法,从唾液腺黏蛋白中分离出了与Bia's试剂反应呈紫色的物质,因此命名其为唾液酸。到目前为止已经发现了 50多种唾液酸。唾液酸一般位于糖蛋白和糖脂等大分子的末端,是动物细胞膜上的糖蛋白和糖脂的重要组成部分。唾液酸具有多种生物学功能比如:细胞的增殖,分化和识别;与病原微生物的相互作用有关,参与免疫反应,促进神经系统发育,易化记忆的形成,提高学习能力。在大脑内,唾液酸主要存在于神经节苷脂上。神经节苷脂是最复杂的鞘糖脂,以大脑灰质中含量最高。神经节苷脂在大脑的生物学活性主要体现在其对突触可塑性的调节。突触的可塑性是学习和记忆等高级的神经活动的重要机制。多聚唾液酸(polysialic acid,PSA)是一类线性、均一的,由多聚α2,8连接唾液酸的碳水化合物,它主要通过典型的N-糖苷键附着在脊椎动物神经系统神经细胞粘附分子上。多聚唾液酸通过改变神经系统神经细胞粘附分子的黏附性来调节神经细胞发育、神经导向以及突触连接的形成,从而在神经发育中起到十分重要的作用。它是存在于少数几种糖蛋白(例如:神经细胞粘附分子、鱼子糖蛋白等)中的唾液酸同聚物。基于这些潜在的好处,我们推测唾液酸乳糖和唾液酸能够作用于神经系统,是促进婴幼儿早期脑发育、视觉发育,并能提高其认知功能的非常重要关键营养素。第一部分:唾液酸乳糖摄入对新生仔猪神经发育和学习认知功能的影响研究目的:本课题以新生仔猪为动物模型,通过唾液酸乳糖的饮食干预,进行八臂迷宫行为学训练及测试,评估其学习记忆能力水平,并对其脑部相关脑区进行唾液酸表达水平和相关脑发育通路的分析,探索唾液酸乳糖对仔猪脑发育及对学习记忆的影响,并探讨其中的分子机制。研究方法:将28只3日龄的雄性仔猪随机分配到唾液酸乳糖处理组(n=14)和对照组(n=14),并连续35天在其饮食中加入一定剂量的唾液酸乳糖,其中唾液酸的摄入量为250mg/Kg/d(唾液酸乳糖组)和40mg/Kg/d(对照组)。23-36日龄之间进行八臂迷宫行为学训练及测试,评估其学习记忆能力水平,在39日龄时对其实施安乐死,取其脑部相关脑区进行唾液酸表达水平的分析,并采用pcr-array技术对海马区相关脑发育通路(包括突触可塑性通路的64个基因、神经营养因子通路的55个基因、GABA和谷氨酸盐神经递质通路的20个基因、神经形成通路的64个基因和与唾液酸代谢相关的29个基因)进行分析,通过qRCR、Western blot、ELISA等实验手段对受影响基因研究其转录、翻译及对下游基因的激活情况,探索唾液酸乳糖对仔猪脑发育及对学习记忆的影响,从而推测唾液酸乳糖影响仔猪脑发育和认知功能的分子作用机理。研究结果:膳食中添加唾液酸乳糖(1)对仔猪体重和脑重的增加无显著影响。(2)对仔猪血浆中应激激素Cortisol和ACTH的浓度无显著影响;(3)对仔猪血浆和血红细胞中唾液酸Neu5Ac和Neu5Gc的含量无显著影响;(4)对仔猪海马和前额叶中与神经

关键词：脑发育唾液酸乳糖唾液酸仔猪模型 pcr-array 视网膜发育唾液酸乳糖唾液酸仔猪模型 polySia-NCAM

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黑逍遥散对阿尔兹海默病模型大鼠海马组织Aβ代谢相关靶点影响

黑逍遥散对阿尔兹海默病模型大鼠海马组织Aβ代谢相关靶点影响

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作者：杨植媛甘肃中医学院

学位级别：硕士

目的：本实验通过建立阿尔兹海默病大鼠模型，研究黑逍遥散对阿尔兹海默病（Alzheimer’s Disease，AD）模型大鼠海马组织Aβ代谢相关靶点的影响，为黑逍遥散从肝脾论治阿尔兹海默病提供研究基础及科学依据。方法：选择Wistar大鼠120只... 详细信息

目的：本实验通过建立阿尔兹海默病大鼠模型，研究黑逍遥散对阿尔兹海默病（Alzheimer’s Disease，AD）模型大鼠海马组织Aβ代谢相关靶点的影响，为黑逍遥散从肝脾论治阿尔兹海默病提供研究基础及科学依据。方法：选择Wistar大鼠120只，雌雄各半，海马区注射Aβ25-35淀粉样蛋白建立大鼠痴呆模型，设假手术组，模型组，对照组及黑逍遥散高中低剂量组，假手术组海马注射等体积生理盐水。造模7天后灌胃给药28天（每天3ml/kg），假手术组与模型组予生理盐水，对照组予哈伯因，黑逍遥散组按组别给予不同浓度。灌胃结束后解剖分离海马，提取组织RNA及蛋白。(1)运用全组基因表达谱芯片杂交技术检测黑逍遥散对AD模型大鼠海马组织基因表达谱的影响，以qRT-pcr（quantitation Real-Time pcr）方法验证靶基因的表达。(2)运用pcr芯片（pcr-array）技术筛选AD模型大鼠Aβ代谢相关靶点基因，以qRT-pcr技术及蛋白免疫印迹法（Western Blot）检测靶基因及靶蛋白的表达。结果：（1）黑逍遥散通过调控锌指蛋白Zfp37、Znf483、ZIC4的表达影响AD发生；（2）黑逍遥散通过调控wnt信号通路相关基因Wisp-1、Crebbp、Igfbp-1、Casq2的表达而影响Aβ代谢和Tau蛋白异常磷酸化；（3）黑逍遥散通过调控Aβ代谢相关基因Clu、Map-2、AChE、Apba1的表达影响Aβ淀粉样蛋白沉积和清除；结论：黑逍遥散干预Aβ25-35诱导的大鼠阿尔兹海默病模型的作用和机制可能与调节Aβ淀粉肽在海马组织的沉积和清除等作用密切相关。此外，与涉及调节和保护多种神经元，促进神经再生方面通路相关靶点相关。

关键词：阿尔兹海默病黑逍遥散 Aβ25-35基因表达谱 pcr-array

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Early Apoptosis Signals Induced by a Low Dose of Epigallocatechin 3-Gallate Interfere with Apoptotic and Cell Death Pathways

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JOURNAL OF NANOSCIENCE AND NANOTECHNOLOGY 2012年第3期12卷 2113-2119页

作者： Berindan-Neagoe, Ioana Braicu, Cornelia Tudoran, Oana Balacescu, Ovidiu Irimie, Alexandru Univ Med & Pharm I Hatieganu Dept Surg Oncol Cluj Napoca 400427 Romania Canc Inst Ion Chiricuta Dept Funct Genom & Expt Pathol Cluj Napoca 400015 Romania Univ Med & Pharm Iuliu Hatieganu Dept Immunol Cluj Napoca 400427 Romania Univ Babes Bolyai Dept Chem Cluj Napoca 400028 Romania Ion Chiricuta Canc Inst Dept Surg Cluj Napoca 400015 Romania

Research for natural compounds with novel advantageous biological activities or antineoplastic potential is an important issue in drug discovery. An example of this type of compound is epigallocatechin 3-gallate (EGCG), which is the major polyphenol in green tea. EGCG has therapeutic effects and modulates multiple metabolic pathways, including apoptosis. The present study evaluates the effect of 10 mu M EGCG on the expression of genes that are involved in apoptosis and cell death using pcr-array technology and He La cells as an in vitro model. The specific modulatory effects of EGCG were demonstrated by significant alterations in the expression levels of the following genes that control the apoptosis pathway: the TNF receptor and its ligand family, the death effector domain family, the Bcl-2 family or TP73. The structure of EGCG suggests that it has the potential to be an anticancer agent and that EGCG serves as a promising starting point for the derivation of novel anticancer drugs. EGCC metabolite products, such as pyrogallol, gallic acid or quinones, may also play important roles in inducing apoptosis. Therefore, quantification of mRNA using pcr-array technology is a valuable tool for screening these anticarcinogenic compounds. By evaluating the action of EGCG on genes that regulate apoptosis, our results suggest that EGCG is as an activator of gene expression with direct implications in cancer therapy. In addition, the limited success in activation of the antiapoptotic gene Bcl-2 may be associated with resistance to cancer treatment.

关键词： Epigallocatechin 3-Gallate Apoptosis HeLa Cells pcr-array

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MicroRNA Modulation in Obesity and Periodontitis

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JOURNAL OF DENTAL RESEARCH 2012年第1期91卷 33-38页

作者： Perri, R. Nares, S. Zhang, S. Barros, S. P. Offenbacher, S. Univ N Carolina Dept Periodontol Chapel Hill NC USA Univ N Carolina Sch Dent Chapel Hill NC USA Univ N Carolina Ctr Oral & Syst Dis Chapel Hill NC USA

The aim of this pilot investigation was to determine if microRNA expression differed in the presence or absence of obesity, comparing gingival biopsies obtained from patients with or without periodontal disease. Total RNA was extracted from gingival biopsy samples collected from 20 patients: 10 non-obese patients (BMI < 30 kg/m(2)) and 10 obese patients (BMI > 30 kg/m(2)), each group with 5 periodontally healthy sites and 5 chronic periodontitis sites. MicroRNA expression patterns were assessed with a quantitative microRNA pcr array to survey 88 candidate microRNA species. Four microRNA databases were used to identify potential relevant mRNA target genes of differentially expressed microRNAs. Two microRNA species (miR-18a, miR-30e) were up-regulated among obese individuals with a healthy periodontium. Two microRNA species (miR-30e, miR-106b) were up-regulated in non-obese individuals with periodontal disease. In the presence of periodontal disease and obesity, 9 of 11 listed microRNAs were significantly up-regulated (miR-15a, miR-18a, miR-22, miR-30d, miR-30e, miR-103, miR-106b, miR-130a, miR-142-3p, miR-185, and miR-210). Predicted targets include 69 different mRNAs from genes that comprise cytokines, chemokines, specific collagens, and regulators of glucose and lipid metabolism. The expression of specific microRNA species in obesity, which could also target and post-transcriptionally modulate cytokine mRNA, provides new insight into possible mechanisms of how risk factors might modify periodontal inflammation and may represent novel therapeutic targets.

关键词： microRNA obesity periodontitis inflammation epigenetics pcr-array

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