專題4 抑郁、癲癇：行為學(xué)與細(xì)胞機(jī)制

李云峰

（軍事醫(yī)學(xué)科學(xué)院毒物藥物研究所，北京 100850）

專題4 抑郁、癲癇：行為學(xué)與細(xì)胞機(jī)制

S4-1快速起效抗抑郁-焦慮的靶標(biāo)策略與新藥研究

李云峰

（軍事醫(yī)學(xué)科學(xué)院毒物藥物研究所，北京 100850）

李云峰，博士，研究員,2006-2008年在美國西弗吉尼亞大學(xué)醫(yī)學(xué)院作訪問學(xué)者研究，目前為軍事醫(yī)學(xué)科學(xué)院毒物藥物研究所研究室主任、博導(dǎo)、學(xué)科帶頭人。北京市“科技新星”（2005），軍隊(duì)優(yōu)秀科技人才崗位津貼獲得者（2011）。長(zhǎng)期從事抗抑郁-焦慮藥的靶標(biāo)機(jī)制和新藥創(chuàng)制研究。主持獲得國家1.1類抗抑郁化藥的臨床批件2個(gè)，其中原創(chuàng)化學(xué)結(jié)構(gòu)的SERT/5-HT1A/6多靶標(biāo)新藥羥哌吡酮具有快速起效、兼有增強(qiáng)認(rèn)知、無性功能障礙等優(yōu)勢(shì)；發(fā)現(xiàn)系列抗抑郁中藥并系統(tǒng)揭示其物質(zhì)基礎(chǔ)及其神經(jīng)調(diào)控規(guī)律，獲我國首個(gè)中藥5類抗抑郁新藥證書并上市（巴戟天寡糖膠囊，當(dāng)年入選“國內(nèi)十大重磅處方新藥”，2012），為抑郁癥治療開辟了全新的物質(zhì)類別。揭示了PDE4D剪接體與抑郁-認(rèn)知行為的密切關(guān)系及分子細(xì)胞調(diào)控機(jī)制；揭示膠質(zhì)細(xì)胞對(duì)抑郁-焦慮的調(diào)控機(jī)制，并在此基礎(chǔ)上發(fā)現(xiàn)全新機(jī)制1.1類抗焦慮-抑郁候選新藥YL-IPA08。以通訊/第一作者在J Neurosci,Neuropsychopharmacology，Br J Pharmacol，Neuropharmacology，Sci Rep等領(lǐng)域著名期刊發(fā)表SCI論文35篇；以第一完成人獲北京市科技一等獎(jiǎng)1項(xiàng)（2015），中華中醫(yī)藥學(xué)會(huì)科技一等獎(jiǎng)1項(xiàng)（2014），軍隊(duì)科技進(jìn)步二等獎(jiǎng)1項(xiàng)（2012）；累計(jì)獲國內(nèi)外授權(quán)專利23項(xiàng)。目前擔(dān)任全國和北京市神經(jīng)精神藥理專業(yè)委員會(huì)常委，北京市神經(jīng)科學(xué)學(xué)會(huì)常務(wù)理事、副秘書長(zhǎng)、青年委員會(huì)主任委員，全軍神經(jīng)內(nèi)科專委會(huì)青年委員會(huì)副主任委員，F(xiàn)ront Pharmacol等多個(gè)SCI期刊副主編或編委。

目的在成功發(fā)現(xiàn)新藥的基礎(chǔ)上歸納新藥研發(fā)的潛在靶標(biāo)策略。方法與結(jié)果基于優(yōu)化的多靶標(biāo)單胺策略，發(fā)現(xiàn)2個(gè)1.1類抗抑郁化學(xué)新藥并獲得國家CFDA批準(zhǔn)的Ⅰ～Ⅲ期臨床批件，即鹽酸羥哌吡酮（YL-0919）和鹽酸阿姆西汀(071031B)。YL-0919是原創(chuàng)化學(xué)結(jié)構(gòu)的小分子化合物，兼具5-HT重?cái)z取抑制、5-HT1A部分激動(dòng)和5-HT6激動(dòng)作用，較臨床一線藥物具有靶標(biāo)新穎、起效迅速、兼有增強(qiáng)認(rèn)知作用、無性功能障礙等優(yōu)勢(shì)；071031B是5-HT/NE重?cái)z取抑制劑度洛西汀的結(jié)構(gòu)優(yōu)化衍生化合物，具有更強(qiáng)效、肝毒性低、生物利用度高等優(yōu)點(diǎn)。基于非單胺靶標(biāo)策略，開展了膠質(zhì)細(xì)胞上18 ku轉(zhuǎn)位蛋白（TSPO）靶標(biāo)機(jī)制和新藥研究，采用TSPO敲除動(dòng)物和病毒過表達(dá)技術(shù)發(fā)現(xiàn)，TSPO-/-與TSPO+/+小鼠雖對(duì)慢性應(yīng)激具有相同的敏感性，但TSPO選擇性激活劑AC-5216在野生型小鼠上快速（2～3 d）產(chǎn)生的抗焦慮、抗抑郁、促認(rèn)知作用，且快速增強(qiáng)突觸可塑性可能是其重要機(jī)制，而在敲除小鼠上無此作用。進(jìn)而發(fā)現(xiàn)了新化學(xué)結(jié)構(gòu)和新機(jī)制的1.1類抗焦慮-抑郁、抗PTSD候選新藥YL-IPA08，并證實(shí)其具有快速抗抑郁-焦慮、促認(rèn)知行為學(xué)活性及良好的成藥性，其中TSPO及其介導(dǎo)的四氫孕酮釋放是其抗PTSD、抗焦慮-抑郁行為的核心環(huán)節(jié)。結(jié)論提出單胺能快速起效抗抑郁的多靶標(biāo)同向、多腦區(qū)協(xié)同的新藥設(shè)計(jì)思路和靶標(biāo)策略，提出介導(dǎo)快速抗抑郁效應(yīng)的“單胺-非單胺長(zhǎng)程反饋神經(jīng)環(huán)路”潛在候選假說，為研發(fā)單胺能-非單胺能協(xié)同的抗抑郁藥提供理論基礎(chǔ)。

快速起效；抗抑郁藥；羥哌吡酮；TSPO；YL-IPA08

S4-2記憶網(wǎng)絡(luò)中的N-甲基-D-天冬氨酸受體

張丹參

（河北科技大學(xué)，河北石家莊 050018）

張丹參，教授，博士生導(dǎo)師。現(xiàn)任河北科技大學(xué)副校長(zhǎng)。研究方向：神經(jīng)藥理學(xué)。學(xué)術(shù)兼職：中國藥理學(xué)會(huì)常務(wù)理事，中國麻醉藥理學(xué)專業(yè)委員會(huì)副主任委員，中國補(bǔ)益藥藥理專業(yè)委員會(huì)副主任委員，中國神經(jīng)精神藥理學(xué)專業(yè)委員會(huì)委員，河北省藥理學(xué)會(huì)、神經(jīng)科學(xué)學(xué)會(huì)、毒理學(xué)會(huì)副理事長(zhǎng)，河北省僑聯(lián)專家委員會(huì)副主任委員。全國醫(yī)藥類學(xué)術(shù)期刊《神經(jīng)藥理學(xué)報(bào)》主編。曾榮獲全國優(yōu)秀科技工作者、全國三八紅旗手、河北省模范教師等榮譽(yù)稱號(hào)。

摘要：N-甲基-D-天冬氨酸(NMDA)受體屬谷氨酸離子型受體，其與突觸的可塑性和學(xué)習(xí)記憶密切相關(guān)。中樞神經(jīng)系統(tǒng)中與學(xué)習(xí)記憶相關(guān)的，如以膽堿受體、谷氨酸受體、腺苷A1受體等為代表的諸多受體及遞質(zhì)，以谷氨酸(Glu)/GABA為代表的氨基酸能神經(jīng)通路，以長(zhǎng)時(shí)程增強(qiáng)(LTP)等為代表的腦內(nèi)神經(jīng)電活動(dòng)，以腦源性神經(jīng)營養(yǎng)因子(BDNF)等為代表的基因蛋白遺傳信息改變，甚至許多神經(jīng)退行性疾病中Glu的神經(jīng)毒性，等等，都與NMDA受體相關(guān)，通過NMDA受體功能的改變調(diào)控學(xué)習(xí)記憶功能，進(jìn)而對(duì)整個(gè)中樞神經(jīng)系統(tǒng)產(chǎn)生影響。換句話說，如果把學(xué)習(xí)記憶的信息傳遞系統(tǒng)看做一個(gè)龐大的信息網(wǎng)絡(luò)，那么NMDA受體就是學(xué)習(xí)記憶相關(guān)神經(jīng)網(wǎng)絡(luò)中一個(gè)相對(duì)中心的關(guān)鍵位點(diǎn)。因此，以探討NMDA受體在學(xué)習(xí)記憶錯(cuò)綜復(fù)雜網(wǎng)絡(luò)中的關(guān)聯(lián)為切入點(diǎn)，以微觀深入研究為基礎(chǔ)、以宏觀視野分析為引領(lǐng)，全方位評(píng)價(jià)NMDA受體在學(xué)習(xí)記憶網(wǎng)絡(luò)中的作用，或許可以引領(lǐng)未來腦功能及相關(guān)疾病系統(tǒng)的研究。

關(guān)鍵詞：N-甲基-D-天冬氨酸；受體；記憶

S4-3抑制星形膠質(zhì)細(xì)胞Hsp90-GLT1的抗癲癇機(jī)制研究

許琪

(中國醫(yī)學(xué)科學(xué)院基礎(chǔ)醫(yī)學(xué)研究所，北京協(xié)和醫(yī)學(xué)院基礎(chǔ)學(xué)院，中國醫(yī)學(xué)院神經(jīng)科學(xué)中心,北京)

許 琪，博士，中國醫(yī)學(xué)科學(xué)院基礎(chǔ)醫(yī)學(xué)研究所研究員，“協(xié)和學(xué)者”特聘教授，中國醫(yī)學(xué)科學(xué)院神經(jīng)科學(xué)中心執(zhí)行主任。國家自然科學(xué)基金“杰出青年基金”和中國青年科技獎(jiǎng)獲得者。近20年來從事精神分裂癥、抑郁癥、癲癇等重性腦疾病發(fā)生的分子機(jī)制與治療靶點(diǎn)研究，現(xiàn)已在Nature，Nature Genetics，Nature Structural&Molecular Biology，JEM，Molecular Psychiatry，American Journal of Psychiatry，Biological Psychiatry等期刊發(fā)表通訊作者或第一作者論文60余篇。

摘要：大腦內(nèi)的興奮性神經(jīng)遞質(zhì)"谷氨酸"的異常累積是導(dǎo)致癲癇發(fā)作的主要原因。星形膠質(zhì)細(xì)胞可以通過谷氨酸轉(zhuǎn)運(yùn)蛋白1（GLT-1/EAAT2）來清除胞外谷氨酸，終止神經(jīng)興奮性傳導(dǎo)。病理證據(jù)表明，GLT-1的“缺失”是導(dǎo)致谷氨酸異常累積的重要原因，但原因尚不清楚。本課題組近期的研究發(fā)現(xiàn)，致癲灶的星形膠質(zhì)細(xì)胞中過量表達(dá)的Hsp90β通過將GLT-1募集到20S蛋白酶體的方式促進(jìn)GLT-1的蛋白降解。Hsp90抑制劑能夠阻止GLT-1的過度降解，顯著提高GLT-1的蛋白水平，并增強(qiáng)星形膠質(zhì)細(xì)胞對(duì)體外谷氨酸的清除能力。我們對(duì)顳葉癲癇模型小鼠開展的治療性研究發(fā)現(xiàn)，Hsp90抑制劑17AAG具有顯著的抗癲癇效果，長(zhǎng)期用藥可大幅降低自發(fā)性癲癇發(fā)作頻率（平均抑制效果73.4%），提高無癲癇發(fā)作天數(shù)，同時(shí)能夠緩解星形膠質(zhì)細(xì)胞增生的病理現(xiàn)象。本工作拓展了癲癇發(fā)病的分子病理機(jī)制，也為顳葉癲癇等難治性癲癇的治療提供了新的思路。

關(guān)鍵詞：星形膠質(zhì)細(xì)胞；癲癇；谷氨酸轉(zhuǎn)運(yùn)蛋白1

S4-4 Disturbance of cholinergic Grb2-associated-binding protein 1 signaling participate in the pathological process of cognitive dysfunction

TAN Chao1,LU Nan-nan1,SHAO Ling-xiao1,LIU Xiu-xiu1,2,PAN Yue1,LIU Yi-jie1,YU Fang-ying1, LU Ying-mei2,HAN Feng1

(1.College of Pharmaceutical Sciences,Zhejiang University,Hangzhou,China;2.School of Medicine,Zhejiang University City College,Hangzhou,Zhejiang,China)

韓 峰，教授，博士生導(dǎo)師；藥學(xué)系主任，藥理所副所長(zhǎng)?；谀X微血管病理基礎(chǔ)的神經(jīng)精神疾病發(fā)病機(jī)制研究及藥物靶標(biāo)發(fā)現(xiàn)。已發(fā)表SCI論文50余篇，總影響因子逾260。作為通訊作者大部分工作發(fā)表在Cell Res，J Am Chem Soc，Antioxidants&Redox Signaling，J Pineal Res等雜志上。近5年論文總引用1460次，他引次數(shù)逾830次。近5年以通訊或第一作者發(fā)表SCI論文30余篇，影響因子8以上論文10余篇，單篇影響因子最高14.812。近年負(fù)責(zé)主持了國家自然科學(xué)基金重點(diǎn)國際合作研究項(xiàng)目、國家自然科學(xué)基金重大研究計(jì)劃、國家重點(diǎn)研發(fā)計(jì)劃重點(diǎn)專項(xiàng)、省部級(jí)重點(diǎn)項(xiàng)目等10多項(xiàng)科研項(xiàng)目?，F(xiàn)擔(dān)任J Pharmacol Sci副主編及多家國際刊物編委；入選“浙江省衛(wèi)生高層次人才”、“錢江人才”、“浙江省151人才”。中國毒理學(xué)會(huì)理事；中國藥理學(xué)會(huì)神經(jīng)精神藥理學(xué)專委會(huì)委員；中國藥理學(xué)會(huì)心血管藥理專委會(huì)委員；中國藥理學(xué)會(huì)生化與分子藥理學(xué)專委會(huì)委員；神經(jīng)毒理專委會(huì)委員，浙江省毒理學(xué)會(huì)副理事長(zhǎng)及秘書長(zhǎng)等。獲得中國藥學(xué)會(huì)“施慧達(dá)杯”優(yōu)秀論文一等獎(jiǎng)，浙江省高?？蒲谐晒坏泉?jiǎng)、浙江省自然科學(xué)學(xué)術(shù)獎(jiǎng)二等獎(jiǎng)及浙江省自然科學(xué)獎(jiǎng)等。

S4-5 Effects of ketamine on mouse hippocampal inflammatory cytokines IL-6, IL-1β and TNF-α levels in acute and chronic administration models

LI Yan-ning1,SHEN Rui-peng1,2,WEN Ge-hua1,DING Run-tao1,DU Ao1,ZHOU Ji-chuan3,DONG Zhi-bin1, REN Xing-hua1,YAO Hui1,ZHAO Rui1,ZHANG Guo-hua1,LU Yan3,WU Xu1

(1.Department of Forensic Pathology,School of Forensic Medicine,China Medical University, Shenyang 110122,China;2.Wujiang District Branch of Suzhou Public Security Bureau, Suzhou 215200,China;3.Key Laboratory of Health Ministry in Congenital Malformation, the Affiliated Shengjing Hospital of China Medical University,Shenyang 110004,China)

Abstract:OBJECTIVEKetamine is an injectable anesthetic and recreational drug of abuse commonly used worldwide.Many experimental studies have shown that ketamine can impair cognitive function and induce psychotic states.Neuroinflammation has been suggested to play an important role in neuro?degeneration.Meanwhile,ketamine has been showed to modulate the levels of inflammatory cytokines. Therefore,we sought to investigate whether the effects of ketamine on the central nervous system is associated with the inflammatory cytokines.METHODSWe established acute(single or multiple intra?peritoneal injection)and chronic(six months daily intraperitoneal injection)ketamine administration models in C57BL/6 mice,evaluated the spatial recognition memory and emotional response by applying the Y maze test and open field test.We analyzed the changes of inflammatory cytokines interleukin-6(IL-6),interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)levels in mouse hippocampus,employing Western blot,quantitative reverse transcriptase-polymerase chain reaction(qRT-PCR)and immunohisto?chemistry.RESULTSKetamine induced spatial recognition memory deficit,reduced anxiety-like behav?iors in mice after chronic administration,and it was dose-dependent.Moreover,we found that ketamine could increase the levels of mouse hippocampal inflammatory cytokines IL-6 and IL-1β after single,multi?ple and long-term administration in a dose-dependent manner.However,the level of TNF-α expressed differently in mouse hippocampus under different conditions.Single administration of ketamine increased the level of TNF-α,whereas multiple and long-term administration decreased it significantly.We consid?ered that TNF-α might exist bi-directional regulatory pathway,which was associated with the dose and duration of ketamine administration.CONCLUSIONOur results suggest that the alterations of inflamma?tory cytokines IL-6,IL-1β and TNF-α levels may be involved in the neurotoxicity of ketamine.

Key words:ketamine;behavior;IL-6;IL-1β;TNF-α

S4-6 A network pharmacology approach to decipher the mechanisms of anti-depression of Xiaoyaosan formula

GAO Yao,TIAN Jun-sheng,QIN Xue-mei

(Modern Research Center for Traditional Chinese Medicine of Shanxi University,Taiyuan 030006,China)

Abstract:OBJECTIVEDepression is one of the prevalent and prominent complex psychiatric diseases and the number of depressed patients has been on the rise globally during the recent decades. Xiaoyaosan,as a famous Chinese herbal formula,has been widely used in depression patients for a long time.However,the therapeutic mechanisms remain uncertain because of difficulty of depression pathophysiology and the lack of bioinformatic approach to understand the molecular connection.METHODSIn this thesis,we applied a network pharmacology approach to explain the potential mechanisms between Xiaoyaosan and depression involved in oral bioavailability screening,drug-likeness assessment, caco-2 permeability,blood-brain barrier target recognition and network analysis.RESULTS66 active compounds in Xiaoyaosan formula with favorable pharmacokinetic profiles are predicted as active compounds for anti-depression treatment.Network analyses show that these 66 compounds target 40 depression-associated proteins including especially HTR2A,NR 3C1,MAOB,XDH and CNR2.These proteins are mainly involved in the neuroactive ligand-receptor interaction,serotonergic synapse, cAMP signaling pathways and calcium signaling pathways.CONCLUSIONThe integrated network pharmacology method is an effective approach to illustrate the anti-depression mechanisms of herbs, and this in silico approach can be applied in drug discovery.

Key words:network pharmacology;Xiaoyaosan formula;depression

S4-7 Baicalin treatment regulates hyperactivity of HPA axis and alters SIRT1 related inflammation in the hypothalamus in a model of depression

YU Hai-yang1,2,ZENG Min-jie1,FU Qiang1,MA Shi-ping1

(1.Department of Pharmacology of Chinese Materia Medica,China Pharmaceutical University,Nanjing 10009,China;2.Institute of Pharmacology,Taishan Medical University,Tai′an 210046,China)

Abstract:OBJECTIVEHyperactivityof hypothalamic-pituitary-adrenal(HPA)axis is an important aetio?logical risk factor for the development of depression.Previous studies have demonstrated that the phe?nolic monomer baicalin has antidepressant-like effects and decreases serum corticosterone levels. However,the mechanism by which baicalin regulates hyperactivity of HPA axis remains unclear.This work aimed to investigate the effects of baicalin on hyperactivity of HPA axis using the olfactory bulbect?omised(OBX)rat model of depression.METHODSAnimals were anaesthetised with 10%chloral hydrate(3.3 mL·kg-1,ip).Using disinfected surgical equipment,the skull covering the olfactory bulbs was exposed by a midline incision.Two burr holes(2 mm diameter)were drilled 8 mm anterior to the bregma and 2 mm lateral to the midline.Both olfactory bulbs were aspirated and the holes filled with glass ionomer cement.The scalp was sutured closed.Sham-operated rats underwent every surgical procedure except the aspiration of the bulbs.The animals received penicillin(8×105U)intramuscularly (0.2 mL/300 g)once per day for 3 d post-surgery to prevent infection,and were subsequently housed alone in polypropylene cages.The experiments continued after 14 d of rehabilitation.The following groups were used for experiments:sham-operated(underwent surgical procedure without aspiratedolfactory bulbs and administration of vehicle only),OBX-model(underwent every surgical procedure and administration of vehicle only),OBX-amitriptyline treated(10 mg·kg-1),and OBX-baicalin treatment (20 and 40 mg·kg-1).Amitriptyline and baicalin were dissolved in physiological saline.RESULTSWe examined how baicalin altered OBX-induced changes in serum glucocorticoid level as well as inflammatory responses,sirtuin 1(SIRT1)expression,and p65 acetylation in the hypothalamus.Similar experiments were performed to analyse the effects of baicalin on lipopolysaccharide-induced inflammatory responses inhypothalamus.CONCLUSIONOur results indicate that activation of the SIRT1 in the hypothalamus contributes to hyperactivity of HPA axis,which can be alleviated by baicalin.

Key words:baicalin;SIRT1;acetylated p65;inflammation;depression

Corresponding author:MA Shi-ping,E-mail:shipingma2014@163.com

專題5 5藥物濫用：行為與分子生物學(xué)

S5-1 BDNF dependent Rac1 GTPase activation in the vmPFC contributes to aversive memory extinction by Arc-mediated GABAA receptor endocytosis

WANG Wei-sheng,JU Yun-yue,WANG Yu-jun,LIU Jing-gen

(Key Laboratory of Receptor Research,Shanghai Institute of Materia Medica,Chinese Academy of Sciences and Collaborative Innovation Center for Brain Science,Shanghai 201203,China)

劉景根，中國科學(xué)院上海藥物研究所研究員、課題組長(zhǎng)。2004年入選中國科學(xué)院“百人計(jì)劃”和獲得國家杰出青年科學(xué)基金,2007年入選“上海領(lǐng)軍人才”。2011年獲藥明康德生命化學(xué)獎(jiǎng)三等獎(jiǎng)?，F(xiàn)是中國科學(xué)院特聘研究員。主要從事阿片類藥物耐受、成癮的細(xì)胞和分子機(jī)理研究及低成癮性阿片類鎮(zhèn)痛藥和戒毒藥研究?，F(xiàn)兼任中國神經(jīng)精神藥理專業(yè)委員會(huì)常務(wù)委員、中國生化和分子藥理專業(yè)委員會(huì)委員，中國心理學(xué)會(huì)生理心理學(xué)會(huì)理事。上海市神經(jīng)藥理學(xué)專業(yè)會(huì)主任委員、上海市生理學(xué)會(huì)常務(wù)理事、上海市藥理學(xué)會(huì)理事、上海市第二屆禁毒專家委員會(huì)委員。河南省天然藥物與免疫工程重點(diǎn)實(shí)驗(yàn)室學(xué)術(shù)委員會(huì)委員。Acta Pharmacological Sinica、藥學(xué)學(xué)報(bào)(英文版)、中國藥學(xué)(英文版)、中國藥理學(xué)通報(bào)、神經(jīng)藥理學(xué)報(bào)、中國藥物依賴性雜志、藥學(xué)研究等雜志編委。

Abstract:Extinction of aversive memories has been a major concern in neuropsychiatric disorders such as anxiety disorders and drug addiction.The ventromedial prefrontal cortex(vmPFC)is important for memory extinction,but the underlying mechanisms are little known.Here,we report that extinction of conditioned place aversion(CPA),a type of aversive memory associated with drug withdrawal, required activation of Rho GTPase Rac1 in the vmPFC in a brain-derived neurotrophic factor(BDNF)-dependent manner,which triggers actin polymerization via Pak1-cofilin signaling pathway,leading to synaptic localization of activity-regulated cytoskeleton-associated protein(Arc)in the vmPFC.The synaptic Arc further determines GABAAreceptor(GABAAR)endocytosis that is necessary and sufficient for vmPFC long-term potentiation and CPA extinction.Thus,extinction of an aversive memory associated with drug withdrawal is intriguingly controlled by Rac1-dependent GABAAR endocytosis in the vmPFC, thereby suggesting therapeutic targets to promote extinction of the unwanted memory.

Key words:brain-derived neurotrophic factor;memory;GABAAreceptor

S5-2 Attenuation of opioid addiction in mice lacking aquaporin 4

LYU Yang1,2,JING Man-yi1,ZHAO Tai-yun1,PANG Chong1,WU Ning1,HU Gang2,SONG Rui1,LI Jin1

(1.Beiijing Institute of Pharmacology and Toxicology,Beijing 100850,China;2.Jiangsu Key Laboratory of Neurodegeneration,Department of Pharmacology,Nanjing Medical University,Nanjing 210029,China)

宋 睿，博士，軍事醫(yī)學(xué)科學(xué)院毒物藥物研究所，副研究員，碩士生導(dǎo)師。本科畢業(yè)于第四軍醫(yī)大學(xué)臨床醫(yī)學(xué)系；碩博畢業(yè)于軍事醫(yī)學(xué)科學(xué)院。長(zhǎng)期從事藥物成癮神經(jīng)生物學(xué)機(jī)制及防復(fù)吸候選新藥發(fā)現(xiàn)的研究。發(fā)現(xiàn)了多巴胺D3受體及二型大麻素受體調(diào)控成癮的新機(jī)制，作為藥效學(xué)負(fù)責(zé)人研制了具有自主知識(shí)產(chǎn)權(quán)的抗成癮防復(fù)吸候選新藥高選擇性多巴胺D3R拮抗劑YQA14，已獲得美國及日本等國的專利授權(quán)。作為課題負(fù)責(zé)人承擔(dān)多項(xiàng)國家自然科學(xué)基金及省部級(jí)基金項(xiàng)目，發(fā)表SCI論文15篇，授權(quán)國際、國內(nèi)專利2項(xiàng)。獲得“中央軍委后勤保障部?jī)?yōu)秀青年科技人才扶持對(duì)象”，“北京市科技新星”，“軍事醫(yī)學(xué)科學(xué)院人才戰(zhàn)略工程后備人才”?，F(xiàn)兼任中國藥物濫用協(xié)會(huì)青年委員會(huì)副主任委員，中國女醫(yī)師協(xié)會(huì)藥學(xué)委員會(huì)專業(yè)委員等職。

Abstract:OBJECTIVETo examine the effects of aquaporin 4(AQP4)on opioid addiction and underlie the mechanism behind it.METHODS①In the heroin-induced self-administration(SA)experiment,we explored the role of AQP4 on heroin-induced psychological addiction.After the mice were trained to learn heroin-induced SA under a fixedratio1(FR1)reinforcement program for 7 d,we randomly switched the heroin doses to 0.00625,0.0125,0.025,0.05 or 0.1 mg·kg-1per infusion to counterbal?ance assignment design.In the end,all mice underwent extinction training and reinstatement testing.②In oral sucrose self-administration,5%sucrose solution was used for the mice and the procedures were similar to heroin SA.③In morphine-induced hyperactivity test,mice were habituated in the test apparatus for 30 min and then were given saline(10 mL·kg-1,sc)or morphine(10 or 20 mg·kg-1,sc)to record the locomotion for 1.5 h.④ For thein vivomicrodialysis experiment,mice were surgically implanted with intracranial guide cannula into nucleus accumbens(AP+1.4 mm,ML±0.9 mm,DV-3.8 mm from bregma).After 5 d of recovery from surgery,the mice were challenged by saline(10 mL·kg-1,sc) or morphine(10 mg·kg-1,sc),and then samples were collected every 20 min.RESULTSWe found that AQP4 deletion had no effects on sucrose-seeking and sucrose-taking,but it significantly attenuated heroin-taking and heroin-seeking behaviors in heroin self-administration.Besides these,AQP4 deletion had no effects on basal level of locomotion,but dramatically decreased morphine-induced hyperactivity. Furthermore,thein vivomicrodialysis studies showed that AQP4 deficiency inhibited morphine (10mg·kg-1,sc)-induced elevation of extracellular dopamine levels in nucleus accumbens in mice.CONCLUSIONOur present findings demonstrate that AQP4 was potentially involved in the properties of opioid rewarding by inhibiting dopamine release in nucleus accumbens(NAc).

Key words:aquaporin 4;opioid addiction;hyperactivity;nucleus accumbens3.Institute of Psycopharmacology,Central Institute of Mental Health,University of Heidelberg, 68159 Mannheim,Germany;4.Departments of Behavioral Medicine&Psychiatry and Physiology &Pharmacology,West Virginia University Health Sciences Center,Morgantown,WV 26506-9137, USA;5.The Florey Institute of Neuroscience and Mental Health,University of Melbourne, Parkville,Victoria 3010,Australia)

S5-3 Brucine N-oxide reduces ethanol intake and preference in alcohol-preferring Fawn-Hooded rats

LIANG Jian-hui1,2,WEI Shou-peng2,3,LI Yu-ling2,GONG Qi2,WANG Yan-ting2,LIANG Hui2, LIU Qing2,ZHANG Han-ting4,CHEN Feng5,Andrew J LOWRENCE5

(1.Department of Molecular and Cellular Pharmacology,School of Pharmaceutical Sciences,2.Department of Pharmacology,School of Basic Medicine Sciences,Peking University,Beijing 100191,China;

LIANG Jian Jian-hui ui,MD.,PhD.,Professor of neuropsychopharmacology and Psychiatrist, Department of Molecular and Cellular Pharmacology,School of Pharmaceutical Sciences, Peking University.He served as a fellow in Toyama Pharmaceutical and Medical Univer?sity,Japan and visiting scientist in Howard Florey Institute,the University of Melbourne, Australia.At present,He has some academic positions:(1)Founder/regular member in Asian Pacific Society for Alcohol and Addiction Research(APSAAR);(2)Secretary-General of Neuropyschoharmacological Division,Chinese Pharmacological Society(CPS);(3)Vice Chair of Neuropsychopharmacological Division,Beijing Pharmacological Society(BPS); (4)Deputy Chief Editor onMetabolic Brain Disease.His laboratory has focused on neuro? psychopharmacological research,including:(1)Behavioral plasticity induced by psychoactive substance and chaperone-mediated opioid addiction in rats,mice,and zebra fishes;(2)Anti-alcoholism of extracts,fractions and compounds from traditional Chinese medicine and the mechanism underlying their pharmacological profiles in alcohol-preferring rats and mice;(3)A new model of comorbidity between depression and anxiety induced by LIANG′s contextual-stress box and the pathophysiological mechanisms in mice.

Abstract:OBJECTIVEThe alcoholism-related social problems have burdened the public health heavily. A better therapy for alcohol dependence as a chronic brain disease is highly required and interests the scientists worldwide.Our group has focused on screening the right drug with low toxicity and a sound curative effect from traditional Chinese medicine.METHODSAlcohol-preferring Fawn-Hooded(FH/ Wjd)rat was used as an animal model of alcoholism to evaluate the effects of brucine N-oxide(BNO), an alkoloid naturally existing in the seeds of Strychnos nux-vomica L,on the alcohol-drinking behaviors. Furthermore,its adverse action and toxicity were investigated.RESULTSTreatment with BNO at the doses of 30,50 and 70 mg·kg-1reduced the voluntary alcohol consumption and preference dosedependently and selectively without altering their water intake,total fluid intake,food consumption, body weight as well as sucrose preference.Remarkably,70 mg·kg-1of BNO did suppress the deprivationinduced elevation of alcohol ingestion.Moreover,BNO used at the same doses as above had no influence on locomotion in an open field test and could not result in the place preference effect.CONCLUSIONTaken together,BNO is of some significant pharmacological profiles to inhibit symptoms of alcohol dependence with high safety,and thence may be a potential pharmacotherapy.

Key words:brucine N-oxide;ethanol

Corresponding author:LIANG Jian-hui,E-mail:liangjh@bjmu.edu.cn

S5-4新型磷酸二酯酶特異性抑制劑的發(fā)現(xiàn)及識(shí)別機(jī)制

賴增偉，吳德燕，張?zhí)烊A，黃儀有，李 哲，吳一諾，羅海彬（中山大學(xué)藥學(xué)院，廣東廣州 510006）

羅海彬，博士、教授、博士生導(dǎo)師，中山大學(xué)藥學(xué)院科研副院長(zhǎng)，廣東省藥物化學(xué)“珠江學(xué)者”特聘教授，國家自然科學(xué)基金優(yōu)秀青年基金獲得者，中國藥學(xué)會(huì)－施維雅青年藥物化學(xué)獎(jiǎng)獲得者。1999年于廈門大學(xué)獲學(xué)士學(xué)位；2005年于香港浸會(huì)大學(xué)獲博士學(xué)位；2006年12月中山大學(xué)"百人計(jì)劃"引進(jìn)為藥學(xué)院副教授；2011年12月晉升為教授，2015年獲得國家自然科學(xué)基金優(yōu)秀青年基金支持，2016年入選廣東省藥物化學(xué)“珠江學(xué)者”特聘教授。羅博士主要從事磷酸二酯酶（PDE）的藥物化學(xué)和結(jié)構(gòu)生物學(xué)等方面的研究，如抗老年性癡呆、肺動(dòng)脈高壓、糖尿病、哮喘等藥物靶標(biāo)結(jié)構(gòu)生物學(xué)和發(fā)現(xiàn)研究，重點(diǎn)構(gòu)建靶標(biāo)PDE全亞型的藥物篩選體系，并進(jìn)行相關(guān)的藥物分子設(shè)計(jì)、有機(jī)合成、藥效學(xué)和作用機(jī)制研究。2000年以來,在Cell，J Med Chem，Chem Commun，Org Lett，J Chem Theory Comput，Biochem Pharm，J Chem Inf Model等國內(nèi)外雜志發(fā)表110多篇論文。已主持5項(xiàng)國家級(jí)、7項(xiàng)省部級(jí)和兩項(xiàng)市局級(jí)項(xiàng)目,6項(xiàng)發(fā)明專利授權(quán)和2項(xiàng)PCT專利。

摘要：磷酸二酯酶（PDE）通過催化水解細(xì)胞內(nèi)第二信使（環(huán)磷酸腺苷cAMP，或環(huán)磷酸鳥苷cGMP），以降低細(xì)胞內(nèi)cAMP或cGMP的濃度，從而中止這兩個(gè)第二信使所傳導(dǎo)的生理作用。PDEs作為新型的藥物作用靶點(diǎn)，參與多種疾病的治療，已成為新藥研發(fā)的重要靶點(diǎn)。截至目前，已有多個(gè)PDE抑制劑藥物上市（PDE5：5個(gè)，治療性功能障礙和肺動(dòng)脈高壓；PDE4：3個(gè)，治療慢性阻塞性肺病等），其中最出名的案例是用于治療男性勃起功能障礙和肺動(dòng)脈高壓的PDE5抑制劑西地那非。然而，低選擇性是這些PDE抑制劑的主要缺陷。近期我們針對(duì)PDE4，5，8，9和10等亞型，通過基于結(jié)構(gòu)的藥物分子設(shè)計(jì)，合成并發(fā)現(xiàn)多類高選擇性的抑制劑。如，1）合成近150個(gè)PDE9抑制劑，從中發(fā)現(xiàn)抑制效應(yīng)最強(qiáng)的高選擇性先導(dǎo)結(jié)構(gòu)3r。3r（IC50=0.6 nmol·L-1，對(duì)PDE1選擇性為788倍），活性和選擇性均明顯優(yōu)于臨床Ⅱ期的輝瑞抑制劑PF-04447943。通過晶體結(jié)構(gòu)分析，發(fā)現(xiàn)3r與PDE9特有殘基Y424形成氫鍵，是其具有高倍選擇性的結(jié)構(gòu)基礎(chǔ)，這為PDE9高選擇性抑制劑提供了新的設(shè)計(jì)策略。此外共晶結(jié)構(gòu)還揭示3r和A452形成一個(gè)氫鍵，而PF-04447943未顯示這種結(jié)合模式。通過設(shè)計(jì)與A452形成氫鍵,可提高化合物的抑制效應(yīng)。2）合成近100個(gè)多個(gè)PDE5抑制劑，從中發(fā)現(xiàn)抑制劑從中發(fā)現(xiàn)抑制效應(yīng)最強(qiáng)的高選擇性先導(dǎo)結(jié)構(gòu)1610。1610（IC50= 5.0 nmol·L-1，對(duì)PDE6選擇性為100倍）的抑制活性與輝瑞藥物西地那非相當(dāng)，但選擇性均明顯優(yōu)于西地那非（對(duì)PDE6選擇性為2～3倍），動(dòng)物實(shí)驗(yàn)也顯示該類藥物的抗動(dòng)脈高壓效果明顯優(yōu)于對(duì)照藥物西地那非。通過分子模擬分析，發(fā)現(xiàn)1610進(jìn)入靶標(biāo)結(jié)構(gòu)的Q2口袋，是1610具有100倍選擇性的結(jié)構(gòu)基礎(chǔ)，這為PDE5高選擇性抑制劑提供了新的設(shè)計(jì)策略。

關(guān)鍵詞：磷酸二酯酶；抑制劑

S5-5 Protective effects of puerarin on alcohol-induced oxidative injury in PC12 cells

LU Jie＊，WEI Zheng-jie＊，ZHU Jia-hui，LONG Yue-zhou，WANG Xiao-dan

(Institute of Pharmacology，School of Pharmaccutical Sciences，Taishan Medical University，Tai′an 271016，China)

Abstract:OBJECTIVEThe aim of this study was to investigate the protective effect of puerarin on alcoholtoxicity in rat pheochromocytoma cell line(PC12).METHODSThe PC12 cells were incubated with different concentrations of puerarin in advance.The protective effects of the puerarin on alcohol induced PC12 cell impairment were evaluated according to the following approach:the viability of PC12 cell was determined by MTT assay and the impairment level was evaluated by analysis the leakage content of the lactate dehydrogenase(LDH).The cell apoptosis degree and the pro-apoptotic p53 protein expression were measured by flow cytometry.RESULTSAlcohol significantly impaired PC12 cell viability(P＜0.05),and increased LDH leakage(P＜0.05),induced cell apoptosis and upregulated expression of p53(P＜0.05). While Puerarin significantly reversed these changes(P＜0.05).CONCLUSIONPuerarin might exert protection effect against ethanol-induced neurotoxicity via inhibition the expression of p53 protein.

Key words:puerarin;alcohol;PC12;apoptosis;p53 protein

Corresponding author:WANG Xiao-dan,E-mail:xdwang@tsmc.edu.cn

＊Co-first author.

S5-6 Protective effect of gingerol dropping pills against alcoholic liver injury in mice

LI Li,ZHANG Zhi-yuan,QI Si-jia,GAO Yong-feng,WANG Lei

(Institute of Pharmacology,Taishan Medical University,Tai′an 271016,China)

Abstract:OBJECTIVETo prepare gingerol dropping pills and to investigate its protective effect on alcoholic liver injury.METHODSThe prescription was selected by orthogonal design method and the effect of the option and ratio of ground substance,the temperature of drug.The hardness,circular degree，the tail formation and the dissolution time were studied.Totally 40 KM mice were randomly divided into control group,model group,gingerol dropping pill group(400 mg·kg-1·d-1)and positive control group(bifendate,150 mg·kg-1·d-1)of 10 mice each.The mice from the model and two drug groups were administrated with liqueur〔0.15 mL/(10 g·d)〕daily by gavage for 3 weeks,Two hours later,drug group mice were treated corresponding gingerol dropping pill and bifendate.Meanwhile,the control group were gavaged same amount of normal saline.Finally,when the model of acute alcoholic liver injury was established on the 22stday,Biochemical indicators of ocular blood in mice were observed. We also observed the change of liver morphology.RESULTSUnder optimum conditions,we can obtain dropping pills having circular shape,touching with hardness and short dissolution time．Compared with the control group,the levels of alanine transaminase(ALT),glutamic-oxaloacetic transaminase (AST)and malondialdehyde(MDA)in model group were obviously increased(P＜0.01),While the activity of Superoxide dismutase(SOD)were decreased.In addition,In model group,mice liver disorders, hepatic lobule fusion,accompanying a large number of patchy sample liver cell vacuoles,various sizes of fat vacuoles appeared in cytoplasm and inflammatory cell infiltration were visible around the central vein.On the contrary,compared with the model group,drug groups attenuated or even reversed hepatic pathological changes.Form gingerol dropping pill group,an increase in hepatic SOD activity and serum ALT and AST activities were found and a significant decrease in hepatic MDA content were also observed(P＜0.01).CONCLUSIONThe prescription of gingerol dropping pills was reasonable,and the preparation process was simple.Gingerol dropping pills can protect liver from alcoholic liver injury to some extend,and the mechanism may be related to its antioxidant effect.

Key words:gingerol dropping pills;alcoholic liver injury;antioxidation

Foundation item:The project supported by College Students Of Science and Technology Innovation Project of Tai′an City(2015D064);and the National College Students′Innovative and Entrepreneurial Training Project(201510439078)

Corresponding author：WANG Lei，E-mail：wang.lei1118@163.com，Tel：13105380208

S5-7 Overexpression of SIRT6 prevents hypoxia-induced apoptosis in osteoblast cells

WANG Jin-tao,YIN Xi-ya,GAO Shan,ZHOU Lu

(Department of Sports Medicine,Taishan Mecical University,Tai′an 271000,China)

Abstract:OBJECTIVETo investigated the role of SIRT6 in the survival of osteoblast cells against hypoxia stimulus as well as the underlying mechanism.METHODSMC3T3-E1 osteoblast cells were used.Apoptosis was induced by hypoxic treatment.MC3T3-E1 cells were transfected with adenovirus SIRT6.For SIRT6 silencing experiment,the transfection of cells were done using three siRNA duplexes directed at different regions of SIRT6 or scrambled siRNA pool.The relative levels of SIRT6 mRNA were quantifiedby using real-time PCR.Western blotting experiments were done after the specific treatment and sample collection.MTT assay was used to estimate cell viability.Caspase 3/7 activity was assayed.RESULTSThe expression of SIRT6 mRNA appeared to be markedly down-regulated in the hypoxia-treated group compared to the matched normal group.Meanwhile,western blotting analysis revealed that the expression of SIRT6 level was markedly down-regulated in hypoxia-treated group at protein level.SIRT6 level was effectively down-regulated by the transfection of SIRT6 siRNA,and significantly enhanced by SIRT6 overexpression.Compared with the control group,SIRT6 overexpression could significantly increase cell viability,and significantly decrease the percentage of apoptotic cells and the activity of caspase 3/7 in response to hypoxia treatment.By contrast,SIRT6 knockdown via treatment with SIRT6 siRNA exhibited the opposite phenotype.CONCLUSIONThese results suggest that SIRT6 plays a protective role in hypoxia-induced apoptosis in MC3T3-E1 cells,and that strategies might be beneficial for the treatment of ischemic bone disease.

Key words:SIRT6;hypoxia;apoptosis;osteoblast

Foundation item:The project supported by grants from Shandong Provincial Natural Science Foundation,China(ZR2014CQ037);National Training Programs of Innovation and Entrepreneurship for Undergraduates,China(201610439244);and National Natural Science Foundation of China(81400182)

S5-8蘆薈多糖對(duì)實(shí)驗(yàn)性口腔潰瘍大鼠NF-κB和MMP2表達(dá)的影響

王哲哲，張 雪，高永峰

（泰山醫(yī)學(xué)院藥學(xué)院，山東泰安 271016）

摘要：目的通過建立口腔潰瘍大鼠模型，探討蘆薈多糖治療口腔潰瘍的相關(guān)機(jī)制。方法SD大鼠隨機(jī)分5組，空白對(duì)照組、模型對(duì)照組、蘆薈多糖100，200和400 mg·kg-1治療組。除空白對(duì)照組外，其余各組大鼠采用50%冰醋酸造模，蘆薈多糖100,200和400 mg·kg-1治療組給予蘆薈多糖進(jìn)行干預(yù)，空白對(duì)照組和模型對(duì)照組給予生理鹽水，連續(xù)給藥7 d。計(jì)算造模后第4和第7天的潰瘍面積；Western-blot測(cè)定實(shí)驗(yàn)大鼠口腔組織中NF-κBp65和MMP2表達(dá)水平。結(jié)果蘆薈多糖各劑量組均可顯著降低造模后第4和第7天的潰瘍面積（P＜0.05）；蘆薈多糖組可降低模型大鼠口腔潰瘍組織中NF-κBp65的表達(dá)水平（P＜0.05），對(duì)MMP2表達(dá)水平有一定抑制作用但未有統(tǒng)計(jì)學(xué)意義。結(jié)論蘆薈多糖具有良好口腔潰瘍治療作用，其機(jī)制與抑制NF-κB炎癥通路有關(guān)。

關(guān)鍵詞：蘆薈多糖；口腔潰瘍；炎癥；核因子-κB；基質(zhì)金屬蛋白酶2

通訊作者：高永峰，E-mail：13583830327@163.com，Tel：13583830327

S5-9 Screening of active ingredients of Taohong Siwu Decoction on platelet membrane receptor

GUO Chun-yan,WU Xin,CHEN Shuang,GAO Yuan-yuan,WAN Ye

(Hebei North University,Zhangjiakou 075000,China)

Abstract:OBJECTIVETo screen the active ingredients preventing platelet aggregations from Taohong Siwu Decoction.METHODSFresh blood from healthy adult volunteer was collected with vacuum tube for preparation of platelet suspension.Platelet suspension was incubated with Taohong Siwu Decoction forone hour at 37℃,then centrifuged.The precipitate was washed and desorbed with buffer solutions. The differences of chemical constituents between incubated and unincubated were identified with high performance liquid chromatography-diode array detector.RESULTSAccording to the chromatograph at 254 nm,at least three kinds of different components were detected in Taohong siwu decoction before and after incubated with platelet.CONCLUSIONPlatelet suspension can be used for screening active ingredients on platelet membrane receptor from the traditional Chinese medicine.

Key words:Taohong Siwu Decoction;blood platelet;HPLC;DAD

Corresponding author：GUO Chun-yan,E-mail:guochy0311@163.com，Tel：18931316689

S5-10丹參酮ⅡA對(duì)心血管系統(tǒng)的藥理作用及其新劑型研究進(jìn)展

侯文書，張 力

(河北北方學(xué)院藥學(xué)系，河北張家口 075000)

摘要：目的本文綜述近幾年來丹參酮ⅡA對(duì)心血管系統(tǒng)的藥理作用及其開發(fā)的新劑型研究進(jìn)展。方法以近幾年來國內(nèi)外關(guān)于丹參酮ⅡA的研究為基礎(chǔ)，對(duì)丹參酮ⅡA對(duì)心血管系統(tǒng)的藥理作用及其新劑型研究進(jìn)行分析和總結(jié)。結(jié)果丹參酮ⅡA在心血管系統(tǒng)方面具有抗動(dòng)脈粥樣硬化，抗心肌缺血，抗心律失常，修復(fù)血管內(nèi)皮細(xì)胞，改善冠狀動(dòng)脈血液循環(huán)等作用，但丹參酮ⅡA有口服吸收差，生物利用度低的缺點(diǎn)，為改善丹參酮ⅡA的現(xiàn)狀，需要制備丹參酮ⅡA新劑型，主要有注射用微乳，微球，固體分散體，脂質(zhì)體，納米粒等。結(jié)論丹參酮ⅡA在心血管系統(tǒng)方面發(fā)揮重要作用，新劑型的開發(fā)使丹參酮ⅡA在治療心血管疾病發(fā)揮更大的藥用價(jià)值，為進(jìn)一步研究提供理論指導(dǎo)。

關(guān)鍵詞：丹參酮ⅡA；心血管疾??；劑型

S5-11荷葉堿在大鼠體內(nèi)的吸收、分布、代謝和排泄研究

王福剛，曹 娟，侯雪芹，曲曉蘭，張繼國

（泰山醫(yī)學(xué)院，山東泰安 271016）

摘要：目的荷葉堿為荷葉中的主要活性成分，是一種具有調(diào)節(jié)血脂和治療肥胖相關(guān)疾病的阿樸啡類生物堿。為了更好地指導(dǎo)臨床用藥，我們建立了荷葉堿在血漿及各組織勻漿中的定量測(cè)定方法，并對(duì)其在大鼠體內(nèi)的吸收、分布、代謝和排泄進(jìn)行了深入研究。方法大鼠靜脈和灌胃給予荷葉堿0.2和10 mg·kg-1，在不同時(shí)間點(diǎn)收集血漿和組織，以鹽酸巴馬汀為內(nèi)標(biāo)，HPLC-MS法測(cè)定給藥后荷葉堿在血漿和組織中的濃度。大鼠灌胃給予荷葉堿10 mg·kg-1，在不同時(shí)間點(diǎn)收集尿液和糞便，經(jīng)處理后HPLC-MS法測(cè)定給藥后荷葉堿在尿液和糞便中的濃度。結(jié)果空白血漿中荷葉堿在2～2000 ng·mL-1濃度范圍內(nèi)呈良好的線性關(guān)系，在血漿中絕對(duì)回收率為87.5%，樣品在長(zhǎng)期儲(chǔ)存過程中能保持穩(wěn)定。荷葉堿的絕對(duì)生物利用度較低為（1.9± 0.8）%。在體內(nèi)分布中腎＞脾＞肝＞心＞肺＞腦。荷葉堿在體內(nèi)主要以原形存在，尚未發(fā)現(xiàn)其代謝產(chǎn)物；荷葉堿主要通過腎排泄（80.7%）。結(jié)論本研究建立了便捷而且有效定量測(cè)定大鼠血漿中荷葉堿的方法，并對(duì)荷葉堿在大鼠體內(nèi)的吸收、分布、代謝和排泄進(jìn)行了深入研究，為臨床應(yīng)用該藥提供了理論依據(jù)。

關(guān)鍵詞：荷葉堿；藥動(dòng)學(xué)；分布；代謝；排泄

S5-12抗壞血酸插層鐵鋯復(fù)合氧化物的制備及緩釋性能評(píng)價(jià)

楊玉潔，孫凡榮，張毅偉，張忠義，張慶樂，王 浩

（泰山醫(yī)學(xué)院，山東泰安 271016）

摘要：目的抗壞血酸是一種水溶性維生素，在治療壞血病，預(yù)防癌癥、心臟病和動(dòng)脈粥硬化等方面具有較好的療效，將其制成緩釋劑型能降低臨床上頻繁給藥帶來的健康風(fēng)險(xiǎn)。鐵鋯雙金屬氧化物是一種性能優(yōu)良的新型緩釋材料，通過控制其制備工藝可提高該材料對(duì)抗壞血酸負(fù)載率和緩釋性能。方法首先，采用松針?biāo)嵋簽榫G色溶劑經(jīng)化學(xué)沉淀法合成鐵鋯復(fù)合氧化物,以抗壞血酸負(fù)載率為評(píng)價(jià)指標(biāo)，利用響應(yīng)曲面法對(duì)鐵鋯復(fù)合氧化物的制備工藝進(jìn)行優(yōu)化。其次，利用體外實(shí)驗(yàn)評(píng)價(jià)抗壞血酸插層鐵鋯復(fù)合氧化物的緩釋效果。最后，利用動(dòng)物實(shí)驗(yàn)評(píng)價(jià)了該材料的生物有效性。結(jié)果與結(jié)論首先，在100 mL松針提取液中加入0.01 mol的氯化鐵和氯氧化鋯（鐵鋯摩爾比為8∶1）、0.5 g十六烷基三甲基溴化銨，60℃持續(xù)攪拌1 h后調(diào)整溶液的pH值為9.5，105℃下烘干24 h時(shí)，制備的鐵鋯氧化物性能良好。0.1 g鐵鋯復(fù)合氧化物對(duì)50 mL濃度為50 mg·L-1抗壞血酸的最大負(fù)載率為94.10%。其次，抗壞血酸插層鐵鋯復(fù)合氧化物在模擬胃液和腸液中的最大緩釋時(shí)間為7.5 h,具有較好緩釋性能。最后，鐵鋯復(fù)合氧化物對(duì)小鼠基本沒有急性毒性，適宜作為抗壞血酸的緩釋載體材料。

關(guān)鍵詞：鐵鋯氧化物；抗壞血酸；松針提取液；緩釋

基金項(xiàng)目：國家級(jí)大學(xué)生創(chuàng)新創(chuàng)業(yè)計(jì)劃（201610439137）

S5-13芍藥苷對(duì)異丙腎上腺素誘導(dǎo)大鼠心肌重構(gòu)的干預(yù)作用

徐曉燕，王 薇，王 蕾，陳 偉，高永峰

（泰山醫(yī)學(xué)院藥學(xué)院，山東泰安 271016）

摘要：目的通研究芍藥苷（PEF）對(duì)異丙腎上腺素（ISO）誘導(dǎo)大鼠心肌重構(gòu)的干預(yù)作用，并探討可能的作用機(jī)制。方法雄性SD大鼠32只，隨機(jī)分成4組，即對(duì)照組、模型組、芍藥苷低劑量組、高劑量組（40，80 mg·kg-1），模型組和高、低劑量組連續(xù)皮下注射ISO（5 mg·kg-1）1周，對(duì)照組注射等比例生理鹽水；造模同日起，高、低劑量組灌胃芍藥苷，對(duì)照組和模型組灌胃等體積的生理鹽水，連續(xù)14 d。末次給藥后30 min，水合氯醛麻醉大鼠，腹主動(dòng)脈取血測(cè)定羥脯氨酸；處死大鼠，取出心臟分離左心室并稱重，測(cè)定左心室指數(shù)；Masson染色觀察組織中心肌纖維化程度；心肌勻漿測(cè)定組織超氧化物歧化酶（SOD）、丙二醛（MDA)及Ⅰ型膠原蛋白的含量。結(jié)果模型組血清羥脯氨酸、左心室指數(shù)及心組織MDA、I型膠原蛋白含量均高于對(duì)照組(P＜0.05)，芍藥苷給藥組均低于模型組（P＜0.05）。在Masson染色中，模型組心肌組織中的膠原明顯增加；芍藥苷低、高劑量組心肌組織中膠原纖維減少，不同劑量心肌組織膠原纖維減少不同，表現(xiàn)出對(duì)ISO引起大鼠心肌纖維化的干預(yù)作用。模型組大鼠組織SOD的活力降低；而芍藥苷高、低劑量均可明顯提高SOD活性模型組。結(jié)論芍藥苷對(duì)ISO致大鼠心肌重構(gòu)具有一定的干預(yù)作用，其作用機(jī)制可能與抗氧化作用有關(guān)。

關(guān)鍵詞：芍藥苷；心肌重構(gòu)；異丙腎上腺素

基金項(xiàng)目：山東省高等學(xué)校科技計(jì)劃（J14LM52）；山東省中醫(yī)藥科技發(fā)展計(jì)劃（2015-249）；山東省醫(yī)藥衛(wèi)生科技發(fā)展計(jì)劃（2015WS0103）

S5-14 Primary culture and identify of cerebral cortical astrocytes from Sprague-Dawley rats

ZHANG Hai-wei1,WU Chun-yang1,ZHANG Dan-shen2

(1.Department of Phrmacy,Hebei North University,Zhangjiakou 075000,China;2.Hebei University of Science and Technology,Shijiazhuang 050000,China)

Abstract:OBJECTIVETo establish a method for primary cultured and iden-tified Sprague-Dawley (SD)rats cerebral cortical astrocytes.METHODSCerebral co-rtex of SD neonatal rats within 24 h was taken with stereo microscope and was cut topieces(1 mm3),digested by Accutase and 0.1%DNAase (37℃,15 min),anddispersed cell suspension was made by mechanical method and filtered.The fibroblast cells and microglia were removed through differential adhesion and sha-ke.Passaged cells were identified by immunofluorescent with anti-Glial fibrillaryacidic protein(GFAP)antibody.RESULTSThe astrocytes of rats cerebral cortex were cultured in this method,which had a large number of cells,good activity,high purity,abundant and elongated cell processes,and were interwoven into a network,showing a typical and good growth state.The third generation of the cells comprised＞95%astrocytes.CONCLUSIONThis simple and reliable cultivation method of astrocyte from rat cerebral tissue is established with high purity,and in a good growth condition.

Key words:astrocytes;primary culture;identify

S5-15 Effect and mechanism of persimmon leaf flavonoid treatment of experimental oral ulcer

WANG Shuo,ZHANG Yong-li,HAO Mai-ling

(School of Stomatology of Taishan Medical University,Tai′an 271000,China)

Abstract:OBJECTIVETo explore the therapeutic effect and its mechanism of persimmon leaf flavonoids(PLF)on rats with oral ulcer.METHODSThe oral ulcer model was induced by acetic acid, was used for intervention of Guilin watermelon frost and different dose(20,40 and 80 mg·kg-1)of PLF. Ulcer area was calculated on the fourth and the seventh day after injury;the changes of superoxide dismutase(SOD)and malondialdehyde(MDA)in serum and ulcer tissues were observed;the level of tumor necrosis factor alpha(TNF-alpha)in ulcer tissue was measured.To observe the pathological morphological changes of H-E staining.RESULTSGuilin watermelon frost and PLF(40 and 80 mg·kg-1) can reduce the ulcer area(P＜0.05);PLF(20,40 and 80 mg·kg-1)can increase the activity of SOD and decrease the content of MDA in serum and ulcer tissue(P＜0.05);Guilin watermelon frost and PLF could significantly decrease the levels of TNF-alpha in ulcer tissue(P＜0.05),and improve the inflammatory infiltration of ulcer tissue.CONCLUSIONPLF has certain therapeutic effects on oral ulcer induced by acetic acid,and the mechanism may be related to improve oxidative damage and reduce inflammatory reaction.

Key words:persimmon leaf flavonoids;oral ulcer;oxidative stress;inflammation

Foundation item:The project supported by National Undergraduate Training Program for Innovation and Entrepreneurship（201510439015）

Corresponding author:ZHANG Yong-li

S5-16玉米須水提物對(duì)自發(fā)性高血壓大鼠血小板聚集功能的影響

秦夢(mèng)瑤，王 慧，曲曉蘭，杜寧寧，朱 超，譚 瑞，周延萌，張芳芳，趙曉民

（泰山醫(yī)學(xué)院藥理學(xué)研究所，山東泰安 271016）

摘要：目的研究玉米須水提物（AECS）對(duì)自發(fā)性高血壓大鼠血小板聚集功能的影響和機(jī)制。方法自發(fā)性高血壓大鼠隨機(jī)分為模型組、AECS 5和15 g·kg-1組，WKY大鼠作為正常對(duì)照組，各組灌胃給藥，連續(xù)25 d。第25天取大鼠肝素抗凝血制備富血小板血漿，比濁法檢測(cè)ADP和膠原誘導(dǎo)的血小板最大聚集率變化，并測(cè)定血漿總抗氧化能力、谷胱甘肽和血管緊張素Ⅱ水平。結(jié)果與WKY組比較，模型組大鼠ADP（3和10 μmol·L-1）和膠原（4 μg·mL-1）誘導(dǎo)的血小板最大聚集率增高，血漿總抗氧化能力和谷胱甘肽水平降低，血管緊張素Ⅱ水平增高；與模型組比較，AECS 5和15 g·kg-1組大鼠在上述濃度ADP和膠原誘導(dǎo)的血小板最大聚集率降低，血漿總抗氧化能力和谷胱甘肽水平增高，血管緊張素Ⅱ水平下降。結(jié)論玉米須水提物可抑制自發(fā)性高血壓大鼠的血小板聚集功能，機(jī)制與降低血管緊張素Ⅱ水平和抗氧化應(yīng)激有關(guān)。

關(guān)鍵詞：玉米須；高血壓；大鼠；血小板；聚集；氧化應(yīng)激；血管緊張素Ⅱ

基金項(xiàng)目：國家級(jí)大學(xué)生創(chuàng)新創(chuàng)業(yè)訓(xùn)練計(jì)劃項(xiàng)目（201610439111）;國家自然科學(xué)基金（81173061）;泰山地產(chǎn)中藥研發(fā)協(xié)同創(chuàng)新中心基金（zd099）;山東省自然科學(xué)基金（ZR2014HQ007）;山東省政府“泰山學(xué)者海外特聘專家”專項(xiàng)基金

通訊作者：趙曉民，E-mail：xmzhao@tsmc.edu.cn

S5-17玉米須水提物對(duì)自發(fā)性高血壓大鼠的降壓作用

王 慧，秦夢(mèng)瑤，曲曉蘭，杜寧寧，朱 超，譚 瑞，周延萌，張芳芳，趙曉民

（泰山醫(yī)學(xué)院藥理學(xué)研究所，山東泰安 271016）

摘要：目的研究玉米須水提物（AECS）對(duì)自發(fā)性高血壓大鼠的降壓作用和機(jī)制。方法自發(fā)性高血壓大鼠隨機(jī)分為模型組、AECS 5和15 g·kg-1組，WKY大鼠作為正常對(duì)照組，各組灌胃給藥，連續(xù)25 d。每5 d監(jiān)測(cè)鼠尾血壓變化，第25天取血漿測(cè)定總抗氧化能力、谷胱甘肽和血管緊張素Ⅱ水平。結(jié)果與WKY組比較，模型組大鼠收縮壓和舒張壓增高，血漿總抗氧化能力和谷胱甘肽水平降低，血管緊張素Ⅱ水平增高；與模型組比較，AECS 5和15 g·kg-1組自第5天開始收縮壓和舒張壓均下降，持續(xù)至25d，血漿總抗氧化能力和谷胱甘肽水平增高，血管緊張素Ⅱ水平下降。結(jié)論玉米須水提物對(duì)自發(fā)性高血壓大鼠有降壓作用，機(jī)制與降低血管緊張素Ⅱ水平和抗氧化應(yīng)激有關(guān)。

關(guān)鍵詞：玉米須；高血壓；大鼠；血壓；氧化應(yīng)激；血管緊張素Ⅱ

基金項(xiàng)目：國家級(jí)大學(xué)生創(chuàng)新創(chuàng)業(yè)訓(xùn)練計(jì)劃項(xiàng)目（201610439111）;國家自然科學(xué)基金（81173061）;泰山地產(chǎn)中藥研發(fā)協(xié)同創(chuàng)新中心基金（zd099）;山東省自然科學(xué)基金（ZR2014HQ007）;山東省政府“泰山學(xué)者海外特聘專家”專項(xiàng)基金

通訊作者：趙曉民，E-mail：xmzhao@tsmc.edu.cn

OBJECTIVEA causal relationshiphas been postulated between cholinergic dysfunction and the progression of cognitive decline in neurodegenerative disorders.However,the cause of the cognitive dysfunc?tion remains unclear.METHODSGab1loxP/loxPwere bred withChAT-Cremice to generateChAT-Cre;Gab1f/fmice.Excitability of cholinergic neurons wererecorded using whole-cell patch clump.A series of behavioral analyses were used to address the changes of cognitive function inChAT-Cre;Gab1f/fmice.Neurochemical changes on brain of conditional knockout mice were evaluated by using immunohistochemistry and Western blotting analysis.RESULTSGrb2-associated-binding protein 1(Gab1)is adocking/scaffolding molecule known to play an important role in cell growth and survival.Here,wereport that Gab1 is decreased in cholinergic neurons in a mousemodel of AD.We found that selective downregulation of Gab1 in the septum impaired learning and memory and hippocampal long-term potentiation,whereas overexpression of Gab1 in the same area rescued the cognitive deficitsseen inChAT-Cre;Gab1f/fandAPPswe/PS1mice.18F-FDGmicroPET imaging data indicated that Gab1 treatment had no effect on metabolic activity of glucose inAPPswe/PS1mice.More?over,we identify abnormal function of SKchannelscontributes to increased firing in cholinergic neuronsofChAT-Cre;Gab1f/fmice.CONCLUSIONGab1 signaling may serve as a potential treatment target for neuro?logical disorders involving dysfunction of central cholinergic neurons.

cholinergic neurons;cognitive dysfunction;Gab1;therapeutic target

QIN Xue-mei,E-mail:qinxm@sxu.edu.cn;Tian Jun-sheng,E-mail:jstian@sxu.edu.cn

The project supported by National Training Program of Innovation and Entrepreneur?ship for Undergraduates（201510439085）

羅海彬，E-mail:luohb77@mail.sysu.edu.cn,Tel:（020）39943031

國家級(jí)大學(xué)生創(chuàng)新創(chuàng)業(yè)訓(xùn)練項(xiàng)目（20150439074）