專題1:心血管藥理學(xué)

專題1:心血管藥理學(xué)

T1-1

AngⅡ誘導(dǎo)兔動脈粥樣硬化下肢再狹窄磷酸化JAK/STAT信號傳導(dǎo)調(diào)控機制的實驗研究

于春利2，劉劍剛1，馬魯波2，張 童2，莊百溪2

（中國中醫(yī)科學(xué)院西苑醫(yī)院1.周圍血管科；2.心血管病中心，北京 100091）

目的觀察血管緊張素Ⅱ（AngⅡ）誘導(dǎo)兔的動脈粥樣硬化（AS）下肢動脈再狹窄模型MCP-1表達水平及對磷酸化JAK/STAT信號傳導(dǎo)的影響。方法雄性，新西蘭大耳白兔24只，隨機分為2組AngⅡ組、假手術(shù)對照組。用3%戊巴比妥鈉水溶液（1 mL·kg-1體質(zhì)量）經(jīng)耳緣靜脈注射麻醉，無菌條件下，于股深動脈血管壁逆行穿刺，股總動脈內(nèi)膜拉傷，同樣處理左側(cè)股總動脈。自右股深動脈原穿刺點注入0.5 mL的AngⅡ（100 mmol·L-1）至股總動脈球囊拉傷水平，孵育30 min，左股總動脈應(yīng)用0.5 mL生理鹽水孵育30 min，依次縫合切口。AngⅡ組高脂飼料飲食，假手術(shù)對照組為普通飼料。于1周、4周、12周、24周后經(jīng)耳緣靜脈注射麻醉，造影存留圖像，分離下肢動脈組織測定MCP1的mRNA和蛋白表達水平，檢測P-JAK2及P-Stat3蛋白表達水平的變化。結(jié)果AS兔下肢動脈組織病理結(jié)果表明，兔AS早期（1周時）并不因內(nèi)膜增殖而發(fā)生管腔縮窄，而是存在血管壁的向外擴張。但12周時，隨內(nèi)膜增殖進展，管腔出現(xiàn)明顯縮窄，殘余管腔變小，下肢動脈閉塞嚴重，尤其是經(jīng)過孵育AngⅡ的動脈，內(nèi)膜增生顯著，管腔面積明顯縮小。Western蛋白質(zhì)印跡法檢測下肢動脈蛋白的表達，和假手術(shù)組及對側(cè)支比較，動脈組織的MCP1水平和P-JAK2及P-Stat3蛋白表達顯著增加（P＜0.01）。RT-PCR結(jié)果表明，和假手術(shù)組及對側(cè)支比較，下肢動脈的MCP1mRNA水平和JAK2mRNA、Stat3mRNA的表達顯著增加（P＜0.01）。結(jié)論AngⅡ調(diào)節(jié)磷酸化JAK/STAT的信號傳導(dǎo)在動脈粥樣硬化中促進MCP-1水平表達增高導(dǎo)致STAT3磷酸化的增加，是高脂飲食促進下肢內(nèi)膜增生的關(guān)鍵因素。

血管緊張素Ⅱ；動脈粥樣硬化；下肢再狹窄；單核細胞趨化蛋白-1；蛋白磷酸化

T1-2

阿司匹林和氯吡格雷聯(lián)合應(yīng)用致急性心肌梗死后大鼠胃黏膜損傷和微血管變化的藥理機制初探

劉劍剛1，張 蕾1，張慶翔1，2，張大武1，史大卓1

（1.中國中醫(yī)科學(xué)院西苑醫(yī)院，中國中醫(yī)科學(xué)院心血管病研究所，北京 100091；2.山東省泰安市中醫(yī)醫(yī)院，山東泰安271000）

摘要：目的結(jié)扎大鼠冠狀動脈前降支造成大鼠實驗性急性心肌梗死（AMI）模型，觀察聯(lián)合應(yīng)用阿司匹林和氯吡格雷對AMI后大鼠胃黏膜微血管的影響，探討雙聯(lián)抗致大鼠胃黏膜損傷可能的藥理作用機制。方法將SD大鼠腹腔麻醉（4%水合氯醛，9 mL·kg-1體質(zhì)量）后常規(guī)處理，結(jié)扎左冠狀動脈前降支血管造成大鼠實驗性AMI模型，確定成活的大鼠隨機分為①模型（AMI）對照組：灌胃蒸餾水2 mg·kg-1·d-1；②阿司匹林組：灌胃阿司匹林9 mg·kg-1·d-1；③氯吡格雷組：灌胃氯吡格雷6.75 mg·kg-1·d-1；④雙聯(lián)抗組：灌胃阿司匹林9 mg·kg-1·d-1和氯吡格雷6.75 mg·kg-1·d-1混懸液。另取12只大鼠作為假手術(shù)組（只穿刺不結(jié)扎冠脈，其余處理方法與AMI模型組相同），每組12只，雌雄各半，連續(xù)灌胃28 d后，第29 d進行微血管形態(tài)的觀察，實驗結(jié)束后腹主動脈取血，放射免疫法測定胃泌素（GAS）、胃動素（MTL）和6-酮-前列腺素F1α（6-keto-PGF1α）和血栓素B2（TXB2）指標，大鼠胃黏膜分別按照Guth標準和Whittle方法評定損傷指數(shù)和胃黏膜損傷程度。結(jié)果經(jīng)Guth方法和Whittle標準的計分評定和假手術(shù)組比較，小劑量阿司匹林組大鼠胃黏膜損傷及程度明顯，阿司匹林聯(lián)合氯吡格雷后胃黏膜損傷程度加重，胃黏膜微血管擴張，血管壁變薄，毛細血管交叉數(shù)減少顯著（P＜0.05），大鼠血漿GAS水平顯著降低（P＜0.01），MTL水平顯著升高（P＜0.01）。阿司匹林組的GAS降低水平和MTL升高水平大于氯吡格雷組，和假手術(shù)組比較，阿司匹林組可以顯著降低TXB2水平。結(jié)論阿司匹林和氯吡格雷聯(lián)合應(yīng)用可加重AMI后大鼠胃黏膜的損傷，其機制與胃腸激素水平紊亂、干擾前列腺素合成導(dǎo)致胃黏膜微血管障礙密切相關(guān)。

關(guān)鍵詞：急性心肌梗死；氯吡格雷；阿司匹林；胃黏膜；微循環(huán)

基金項目：科技部國際科技合作與交流項目（2014DFG32700，2010DFA31690）；國家自然科學(xué)基金（81273933）

T1-3

心肌損傷修復(fù)過程中的細胞交互作用機制研究

余細勇

（廣州醫(yī)科大學(xué)藥學(xué)院，廣東廣州511436）

摘要：目的干細胞究竟如何再生和修復(fù)心肌一直是心肌重構(gòu)研究的重要課題。由于移植存活率低等原因，大多數(shù)被注射的成體干細胞并不能有效地移植到宿主心臟中。類似地，固有心臟祖細胞（CPC）和骨髓來源的間充質(zhì)干細胞（MSC）的移植存活率也很低。而與心肌注射相比，通過冠脈內(nèi)注射干細胞的移植存活率則更低。因此，亟需探討干細胞移植的新作用機制。方法磁珠分選出Scal+CPC進行培養(yǎng)，流式細胞術(shù)鑒定；分離CPC分泌的外泌體，并用Western蛋白印跡法進行表型鑒定和用Nanosight進行粒徑分析；免疫熒光法及PKH26標記技術(shù)觀察心肌細胞H9C2對外泌體的攝取作用；通過H2O2處理誘導(dǎo)H9C2細胞凋亡，觀察外泌體對心肌細胞凋亡的保護作用。結(jié)果通過熱休克模擬缺血預(yù)適應(yīng)的研究發(fā)現(xiàn)，熱休克處理后的干細胞能分泌富集了轉(zhuǎn)錄因子HSF1的外泌體并被心肌細胞吸收，使心肌細胞miR-34a啟動子區(qū)發(fā)生染色質(zhì)重構(gòu)、轉(zhuǎn)錄下調(diào)，從而使心肌細胞的凋亡顯著減少。另外，我們從缺氧預(yù)適應(yīng)MSC的條件培養(yǎng)基中提取外泌體并注射到小鼠的心梗區(qū)域，發(fā)現(xiàn)這一辦法能顯著減少心梗后的纖維化。結(jié)論外泌體介導(dǎo)的細胞間信號傳遞是干細胞修復(fù)心肌損傷的重要機制之一，外泌體有可能成為細胞治療的替代療法以及潛在的治療新靶點。

關(guān)鍵詞：干細胞；心肌修復(fù)；外泌體

基金項目：國家自然科學(xué)基金重點項目（81120108003，81330007），廣東省重大科技計劃（2014A050503047，2015B020225006）

通訊作者：余細勇，E-mail：yuxycn@aliyun.com，Tel:（020）37103261

T1-4

DRAM1通過改善自噬流抗心肌缺血損傷

覃宇燕1，陳法江2，鄭德沖1，吳曉倩1

（廣州醫(yī)科大學(xué)1.藥學(xué)院藥理教研室，2.研究生院，廣東廣州511436）

摘要：目的研究DRAM1在心肌缺血損傷中的作用及機制。方法行大鼠左前降支冠狀動脈永久性結(jié)扎術(shù)建立大鼠急性心肌梗死模型后，在結(jié)扎處附近梗死周邊區(qū)注射過表達DRAM1腺病毒或空病毒。通過免疫印跡和免疫熒光實驗檢測DRAM1的表達變化。21 d后，超聲心動圖檢測大鼠左心室功能；HE染色和Masson染色觀察心肌重構(gòu)情況；Tu?nel檢測心肌細胞凋亡；毛細血管密度測定觀察血管新生；透射電鏡觀察自噬體和自噬溶酶體；TTC染色計算心肌梗死面積；Western蛋白印跡法檢測DRAM1和自噬相關(guān)蛋白LC3和P62的表達情況。結(jié)果與空病毒組相比，超聲結(jié)果顯示DRAM1過表達明顯改善左心室功能，HE染色和Masson染色結(jié)果顯示DRAM1過表達改善心肌重構(gòu)；Tunel結(jié)果顯示，心肌細胞凋亡明顯減少；毛細血管密度測定檢測DRAM1組較空病毒組有新生血管生成；透射電鏡觀察到空病毒組自噬體堆積，自噬流障礙，過表達DRAM1自噬體堆積減少自噬溶酶體增加，自噬流改善；TTC染色結(jié)果顯示，過表達DRAM1明顯減少梗死面積；Western蛋白印跡法結(jié)果顯示，過表達DRAM1明顯逆轉(zhuǎn)長時間缺血引起的自噬蛋白LC3-Ⅱ和SQSTM1/P62堆積。結(jié)論過表達DRAM1對缺血的心肌細胞具有保護作用，可能與改善自噬流障礙有關(guān)。

關(guān)鍵詞：DRAM1；腺病毒；急性心肌梗死；心肌缺血；自噬

基金項目：國家自然科學(xué)基金（81573429）

通訊作者：吳曉倩，E-mail：wuxiaoqian2004@hotmail.com，Tel：13760686016

T1-5

川穹嗪對腎性高血壓大鼠心肌重構(gòu)及P38MAPK/AP-1信號通路的影響

呂行直1，李瑞芳1，王宏運2，李亞鵬1，汪 玲1，王建剛1

（河南科技大學(xué)1.醫(yī)學(xué)院藥理學(xué)與分子生物學(xué)重點實驗室，2.第一附屬醫(yī)院，河南洛陽 471003）

摘要：目的研究川穹嗪（TMP）對腎性高血壓大鼠心肌重構(gòu)的作用及其對P38MAPK/AP-1信號通路的影響。方法采用兩腎兩夾（2K2C）法建立腎性高血壓大鼠模型，分為假手術(shù)組、模型組、TMP 30和60 mg·kg-1及坎地沙坦（Can）10 mg·kg-1組。Can組大鼠灌胃給藥，其他各組大鼠均腹腔注射，每日1次，連續(xù)給藥2周。檢測血流動力學(xué)指標，計算左心室體重比（LVW/BW）。HE染色觀察左心室心肌細胞形態(tài)變化，Masson染色觀察管周膠原生成和心肌纖維化程度。RT-PCR檢測左心室B型心鈉素（BNP）、肌球蛋白重鏈（?-MHC）、轉(zhuǎn)化生長因子β1（TGFβ1）和結(jié)締組織生長因子（CT?GF）的mRNA表達，Western蛋白印跡法檢測信號分子P38MAPK的磷酸化表達變化和激活蛋白（AP-1）的表達。結(jié)果TMP具有明顯的降壓和改善心臟收縮和舒張功能的作用，TMP 30和60 mg·kg-1組及Can 10 mg·kg-1組高血壓大鼠主動脈收縮壓（AoSP）、主動脈舒張壓（AoDP）、左室收縮末壓（LVESP）、左室舒張末壓（LVEDP）下降而室內(nèi)壓最大上升和下降速率（±dP/dtmax）升高（P＜0.05，P＜0.01）。TMP 30和60 mg·kg-1組及Can 10 mg·kg-1組高血壓大鼠LVW/BW和管周膠原分數(shù)明顯降低（P＜0.05，P＜0.01），心肌細胞形態(tài)、排列紊亂和心肌纖維化得到明顯改善。川穹嗪能夠顯著降低高血壓大鼠左心室BNP，?-MHC，TGFβ1和CTGFmRNA的表達以及P38MAPK的磷酸化程度和AP-1的蛋白表達。結(jié)論TMP對腎性高血壓引起的左心室重構(gòu)有顯著抑制作用，其機制可能與調(diào)控P38MAPK/AP-1信號通路以及下調(diào)TGFβ1、CTGF的mRNA表達有關(guān)。

關(guān)鍵詞：川穹嗪；腎性高血壓；心肌重構(gòu)；信號傳導(dǎo)

基金項目：洛陽市科技發(fā)展計劃項目（1401085A-1）

通訊作者：李瑞芳，E-mail：ylliruifang@163.com；王宏運，whwhaust@163.com，Tel：13703499369

T1-6

硫化煙酸對血管內(nèi)皮細胞損傷模型的保護作用

李 博，陳 釧，羅麗君，歐學(xué)蘭，周春陽

（川北醫(yī)學(xué)院藥學(xué)院藥物研究所，四川南充 637007）

摘要：目的合成硫化煙酸，建立動脈粥樣硬化（AS）細胞模型，探討該縮合物是否對氧化低密度脂蛋白（ox-LDL）誘導(dǎo)的血管內(nèi)皮損傷具有保護作用。方法（1）采用酯縮合等化學(xué)合成方法，將原料藥煙酸與茴三硫縮合，應(yīng)用MS、NMR對化合物的結(jié)構(gòu)進行鑒別；（2）利用微孔濾膜吸附法對新合成的化合物進行H2S釋放檢測；（3）用濃度為80 μg·mL-1ox-LDL誘導(dǎo)人臍靜脈內(nèi)皮細胞（HUVEC）建立AS細胞模型，通過油紅O染色觀察不同階段細胞內(nèi)脂質(zhì)的活性變化規(guī)律對其生物特性進行鑒別，并采用總膽固醇、游離膽固醇試劑盒檢測不同階段細胞內(nèi)總膽固醇（TC）和游離膽固醇（Fch）含量，F(xiàn)ch占TC含量的50%，則表明造模成功；（4）采用濃度為500 μmol·L-1的硫化煙酸作用此細胞模型，設(shè)置對照組（NaSH、茴三硫、煙酸）和空白組，通過CCK8法檢測細胞增殖活力。結(jié)果（1）通過MS和NMR的檢測，表明所得產(chǎn)物合成成功。（2）微孔濾膜法H2S釋放檢測：空白組的釋放量為（18.11±1.69）nmol/（min×106cell）；對照組分別在50，100，200，500，1000 μmol·L-1時釋放量為：①NaSH（20.98±4.21，21.46±2.72，23.32±3.54，26.15±1.99，25.89±5.13）nmol/（min×106cell），②茴三硫（20.42±1.98，21.67±2.83，24.02±3.52，28.25±2.09，25.49±1.87）nmol/（min×106cell），③煙酸（18.12±1.38，17.20±3.63，18.02±4.12，18.99±3.21，19.49±2.34）nmol/（min× 106cell）；④實驗組分別在50，100，200，500，1000 μmol·L-1時釋放量為：（21.31±3.01，23.22±3.91，28.86±3.78，32.51±3.12，25.27±4.04）nmol/（min×106cell）。結(jié)果表明新合成化合物在500 μmol·L-1時，H2S釋放量最大。（3）ox-LDL誘導(dǎo)12 h開始，HUVEC細胞內(nèi)出現(xiàn)大量紅染顆粒，游離膽固醇和總膽固醇均比對照組增加，細胞游離膽固醇含量大于總膽固醇的50%，表明AS細胞模型建立成功。（4）硫化煙酸作用于AS細胞模型12 h后，通過CCK8法檢測細胞增殖活力，其中實驗組與對照組相比有明顯增加（P＜0.05）。結(jié)論利用ox-LDL誘導(dǎo)HUVEC細胞12 h-72 h后，細胞形態(tài)以及Fch/TC的比值符合AS細胞模型的生物學(xué)特征，表明成功的建立了AS細胞模型。CCK8法細胞增殖活力的檢測證明了硫化煙酸對ox-LDL誘導(dǎo)的血管內(nèi)皮損傷具有一定的保護作用，為H2S供體型藥物的研發(fā)與臨床治療提供有價值的參考。

關(guān)鍵詞：硫化氫；煙酸；茴三硫；動脈粥樣硬化；人臍靜脈內(nèi)皮細胞

通訊作者：周春陽，E-mail：chunyangzhou@hotmail.com，Tel：（0817）2242761

T1-7

白藜蘆醇通過調(diào)節(jié)PGC-1α活性與線粒體功能抑制多柔比星心肌細胞損傷

郭家彬，崔 嵐，袁海濤，張廷芬，趙 君，彭雙清

（軍事醫(yī)學(xué)科學(xué)院疾病預(yù)防控制所毒理學(xué)評價研究中心，北京 100071）

摘要：目的前期研究表明白藜蘆醇具有心肌保護作用，可抑制抗腫瘤藥物多柔比星（DOX）誘導(dǎo)的心血管毒性。過氧化物增殖體激活受體-γ共激活因子1α（PGC-1α）是一種轉(zhuǎn)錄共激活因子，在心肌細胞能量代謝、線粒體穩(wěn)態(tài)調(diào)節(jié)以及細胞損傷修復(fù)等方面具有重要作用。研究提示，白藜蘆醇的心肌保護作用可能與調(diào)節(jié)PGC-1α有關(guān)，但其具體作用機制尚不清楚。本研究旨在觀察PGC-1α以及線粒體功能在白藜蘆醇介導(dǎo)的心肌保護效應(yīng)中的作用，探討白藜蘆醇抗多柔比星心肌損傷的機制。方法人源心肌細胞（AC16）給予不同濃度的白藜蘆醇預(yù)處理，隨后再給予DOX處理。采用細胞存活率和乳酸脫氫酶漏出檢測細胞損傷，通過流式細胞儀和激光共聚焦顯微鏡觀察等檢測線粒體膜電位和超氧陰離子水平，采用Western蛋白印跡法和RT-PCR分析檢測PGC-1α及其下游靶點（NRF-1、TFAM、MnSOD和UCP2）的表達水平。采用試劑盒檢測細胞ATP水平以及沉默信息調(diào)節(jié)因子2相關(guān)酶1（SIRT1）轉(zhuǎn)錄活性，通過免疫共沉淀法檢測PGC-1α去乙?；?。應(yīng)用SiRNA敲降SIRT1，觀察SIRT1敲降后DOX誘導(dǎo)的細胞和線粒體損傷特征及對PGC-1α通路改變。結(jié)果白藜蘆醇可劑量依賴性地抑制多柔比星細胞毒性和線粒體功能紊亂，包括細胞存活率下降、LDH漏出增加、線粒體膜電位喪失、ATP水平下降和超氧離子形成增加。白藜蘆醇可增加SIRT1的活性，促進PGC-1α的去乙酰化修飾，進而激活PGC-1α通路，增強線粒體生成功能與抗氧化防御能力，減輕多柔比星誘導(dǎo)的PGC-1α及其下游分子的表達下降。SIRT1敲降能加劇DOX誘導(dǎo)的細胞損傷和線粒體功能紊亂以及氧化應(yīng)激，明顯降低白藜蘆醇對心肌細胞和線粒體功能的保護作用。結(jié)論白藜蘆醇可通過增強PGC-1α的去乙?；揎棧M而激活PGC-1α介導(dǎo)的線粒體生成和氧化應(yīng)激調(diào)節(jié)，最終抑制多柔比星誘導(dǎo)的心肌細胞與線粒體損傷。

關(guān)鍵詞：PGC-1α；多柔比星；線粒體生成；氧化應(yīng)激；心肌損傷

基金項目：國家自然科學(xué)基金項目（81470167，81430090）；國家科技重大專項（2012ZX09J12203）

T1-8

轉(zhuǎn)化生長因子 β1/Smad信號通路在肺動脈高壓中的作用

王丹姝，方蓮花，杜冠華

（中國醫(yī)學(xué)科學(xué)院藥物研究所北京市藥物靶點研究與新藥篩選重點實驗室，北京 100050）

摘要：肺動脈高壓是一種發(fā)病機制復(fù)雜的致命性疾病，其典型的病理變化是肺動脈血管重構(gòu)，主要表現(xiàn)為肺動脈平滑肌細胞增殖及細胞外基質(zhì)沉積，進而降低血管壁的靈活性增加肌束的收縮力，引起肺動脈壓力升高肺血管阻力增加，最終導(dǎo)致右心衰竭而亡。轉(zhuǎn)化生長因子β1（TGF-β1）是一個多功能分子家族，是肺動脈纖維化及血管重構(gòu)的重要調(diào)節(jié)子，并且參與到細胞增殖分化遷移及凋亡等各種細胞活動中。TGF-β1是TGF-β1細胞因子家族的一個重要成員，通過激活下游Smad2/3信號通路來調(diào)控胚胎發(fā)育、炎癥介導(dǎo)、血管增生及纖維化等多種反應(yīng)，同時參與到許多心血管疾病的發(fā)生發(fā)展中。近年來許多研究顯示，肺動脈高壓中TGF-β1顯著增加，且TGF-β1通過調(diào)控多種信號通路促進平滑肌細胞增殖及細胞外基質(zhì)沉積進而參與到肺動脈高壓的發(fā)病機制中。本文主要對TGF-β1/Smad信號通路在肺動脈高壓發(fā)生發(fā)展中發(fā)揮的作用進行綜述，從而為肺動脈高壓的發(fā)病機制及治療提供新靶點。

關(guān)鍵詞：肺動脈高壓；TGF-β1/Smad信號通路；血管重構(gòu)；肺動脈平滑肌細胞增殖；細胞外基質(zhì)沉積

基金項目：國家自然科學(xué)基金項目（81573645，81603101）；國家科技重大專項（2013ZX09103001-008）

通訊作者：方蓮花，E-mail：fanglh@imm.ac.cn，Tel：（010）63165313；杜冠華，E-mail：dugh@imm.ac.cn，Tel：（010）63165184

T1-9

甜菜堿對大鼠心肌肥厚的保護作用及機制

高 山，周延萌，王 浩，侯雪琴，趙曉民

（泰山醫(yī)學(xué)院藥學(xué)院，山東泰安 271016）

摘要：目的研究不同濃度甜菜堿對異丙腎上腺素引起的大鼠心肌肥厚的影響及作用機制。方法異丙腎上腺素3 mg· kg-1皮下注射連續(xù)給予10 d制備大鼠心肌肥厚模型，記錄大鼠狀態(tài)及體重變化，隨后分別灌胃給予甜菜堿50，100和200 mg·kg-1，持續(xù)8周，8周后測量心臟各部分重量并計算心臟重量指數(shù)，進行HE染色觀察心臟結(jié)構(gòu)變化，檢測SOD、HO-1、catalase等蛋白的含量。結(jié)果心肌肥厚模型大鼠與空白對照組比較體重明顯下降（402.2 gvs421.4 g），（LV+ SEP）/BW及（LV+SEP）/HW的值均顯著增加（2.4vs2.1，0.783vs0.762），左心室HE染色觀察到心肌細胞肥大、壞死和心肌纖維斷裂；給予甜菜堿處理后，中、高劑量組均可抑制這一變化，而低劑量組無顯著改變。模型組大鼠血漿MDA和ROS含量明顯增高，SOD和GSH-Px活力顯著降低，甜菜堿中、高劑量組大鼠血漿MDA和ROS含量顯著下降，超氧化物歧化酶和GSH-Px活力顯著提高，甜菜堿低劑量組無明顯作用。結(jié)論中、高濃度的甜菜堿對大鼠心肌肥厚具有顯著保護作用，此作用可能是通過激活抗氧化通路，抑制氧化應(yīng)激來實現(xiàn)的。

關(guān)鍵詞：甜菜堿；心肌肥厚；SOD；抗氧化；氧化應(yīng)激

基金項目：山東省醫(yī)藥衛(wèi)生科技發(fā)展計劃（2014WS0496）；山東省中青年科學(xué)家科研獎勵基金計劃（BS2014YY045）

通訊作者：高 山，E-mail：gshan84117@163.com，Tel：18864805030

T1-10

內(nèi)皮祖細胞對實驗兔動脈粥樣硬化再狹窄模型調(diào)控功能的研究

曾貴榮1，2，羅桂芳1，2，周仕達1，2，邵亞杰1，2，唐婭輝1，2，姜德建1，2

（1.新藥藥效與安全性評價湖南省重點實驗室，湖南長沙410331；湖南省藥物安全評價研究中心，湖南 長沙410331）

摘要：目的研究內(nèi)皮祖細胞對實驗兔動脈粥樣硬化再狹窄模型調(diào)控功能的影響。方法實驗兔高脂飼料連續(xù)喂養(yǎng)4周后，采用可吸收線套于右側(cè)頸動脈，隨后繼續(xù)飼養(yǎng)4周，再自右頸外動脈遠端結(jié)扎并切斷頸外動脈，由結(jié)扎近心側(cè)插入擴張球囊導(dǎo)管進行球囊擴張，繼續(xù)飼喂高脂飼料，建立動脈粥樣硬化再狹窄模型。成模后，靜脈注射不同密度的內(nèi)皮組細胞（1.0×105mL-1和2.5×105mL-1），每日1次，連續(xù)7 d，4周后再進行相關(guān)血脂相關(guān)指標檢測、血管造影及血管組織病理學(xué)檢查。結(jié)果2.5×105mL-1內(nèi)皮祖細胞血管內(nèi)移植能明顯降低實驗兔動脈粥樣硬化狹窄模型中甘油三酯（TG）、內(nèi)皮素（ET）、前列環(huán)素（PGI2）含量，降低血管內(nèi)皮細胞生長因子（VEGF）與血小板源性生長因子A（PDGF-A）表達，能明顯改善主動脈內(nèi)膜下有脂質(zhì)顆粒沉積，炎性細胞浸潤。結(jié)論內(nèi)皮祖細胞能減少實驗兔動脈粥樣硬化再狹窄模型血管內(nèi)膜炎癥浸潤，其調(diào)控作用與降低VEGF與PDGF-A表達有關(guān)。

關(guān)鍵詞：內(nèi)皮祖細胞；動脈粥樣硬化再狹窄；球囊損傷；高脂

基金項目：湖南省科技廳計劃項目（2013TT1002）

通訊作者：姜德建，E-mail：dejianjiang@yahoo.com，Tel：（0731）83285166

T1-12

Caffeoylquinic acid derivatives extract ofErigeron multiradiatusalleviated acute myocardial ischemia reperfusion injury in rats through inhibiting NF-kappaB and JNK activations

ZHANG Zhi-feng1，REN Xue-cong1，DONG Geng-ting1，LUO Pei1，ZHOU Hua1，ZHANG Hao2

（1.State Key Laboratories for Quality Research in Chi?nese Medicines，Macau University of Science and Tech?nology，Macau，China；2.Department of Medicinal Natu?ral Products，West China School of Pharmacy，Sichuan University，Chengdu 610041，China）

Abstract：Erigeron multiradiatus（Lindl.）Benth.，has been used in Tibet folk medicine to treat various inflam?matory diseases.The aim of this study was to investigate anti-myocardial ischemia and reperfusion（I/R）injury ef?fect of caffeoylquinic acids derivatives ofE.multiradiatus（AE）in vivoand to explain underling mechanism.AE was prepared using the whole plant ofE.multiradiatusand contents of 6 caffeoylquinic acid determined through HPLC analysis.Myocardial I/R were induced by left ante?rior descending coronary artery occlusion for 30 min fol?lowed by 24 h of reperfusion in rats.AE administration（10，20 and 40 mg·kg-1）inhibited I/R-induced injury as indicated by decreasing myocardial infarct size，reducing of CK and LDH activities and preventing ST-segment de?pression in dose-dependent manner.AE decreased car?diac tissue levels of pro-inflammatory factors TNF-α and IL-6 and attenuated leukocytes infiltration.AE was fur?ther demonstrated to significantly inhibit I-κB degrada?tion，nuclear translocation of p-65 and phosphorylation of JNK.Our results suggested that cardioprotective ef?fect of AE could be due to suppressing myocardial inflam?matory response and blocking NF-κB and JNK activation pathway.Thus，caffeoylquinic acids might be the active compounds inE.multiradiatuson myocardial ischemia and be a potential natural drug for treating myocardial I/R injury.

Foundation item：The project supported by the Macao Science and Technology Development Fund（052/2013/ A2）

Corresponding author：LUO Pei，E-mail：pluo@must. edu.mo

T1-13

The role of HSPB1 and ectopic F1Fo-ATPase interac?tion in hypoxia pulmonary hypertension vascular ad?ventitial vasa vasorum remodeling

LI Yu-mei1，2，WANG Xiao-yan1，CHEN Ming-gang1，ZHANG Li1，2

（1.Department of Pharmacology，Harbin Medical Univer?sity-Daqing，Daqing，China；2.Biopharmaceutical Insti?tute of the Heilongjiang Academy of Medical Sciences，Harbin，China）

Abstract：OBJECTIVETo investigate the role of adventi?tial vasa vasorum in artery remodeling during the pro?cess of pulmonary artery hypertension（PAH），we checked the small heat shock protein 27/25（HSPB1）whether involved in pathological basis of vascular remod?eling.METHODSWe explored the potential role of HSPB1 interacts with ectopic F1Fo-ATPase in the pulmo?nary vascular remodeling，investigate its effects on the endothelium cell dynamic，and further reveal its possible molecular mechanisms using hypoxic pulmonary hyper?tension rat model，transgenic mice and pulmonary ad?ventitial vasa vasorum endothelial cell culturein vitro.RE?SULTSOur studies have shown that HSPB1 improves adventitial vasa vasorum angiogenesis and remodeling. We found that hypoxia induces-HSPB1 upregulation and HSPB1 interact with ectopic F1Fo-ATPase modulate ad?ventitial vasa vasorum endothelial cell proliferation，mi?gration and tube formation.And the inhibition of HSPB1 can reverse the vascular inflammation and fibrosis amaz?ingly.CONCLUSIONAdventitial vasa vasorum plays an important role in vascular remodeling，and small heat shock protein 27/25 was involved in a variety of diseases during the development of PAH，which could an efficient therapeutic targets and prevention strategy for PAH clini?cal.

Key words：pulmonary hypertension；vascular adventi?tial remodeling；vasa vasorum；HSPB1；ectopic F1Fo-ATPase

Foundation item：The project supported by NSFC（81300036）

Corresponding author：ZHANG Li，E-mail：lizzy33991@ 163.com

T1-14

Effects of hydrogen-rich saline on platelet activation in hypertensive rats

HE Ke-gui1，3*，WANG Yun2*，MIN Xiang-dong1，HAN Ji-ju2，JIAO Peng2，CHEN Xiu-yu1，CAO Cheng-bao1，GONGMin4，ZHAO Xiao-min1

（1.Institute of Pharmacology，2.Institute of Atheroscle?rosis，4.Department of Laboratory Management，Tais?han Medical University，Taian 271000，China；3.Depart?ment of Clinical Pharmacy，Ningyang County People's Hospital，Ningyang 271400，China）

Abstract：OBJECTIVETo investigate the effect of hydro?gen-rich saline on platelet activation in hypertensive rats.METHODSThe male Wistar rats were divided into con?trol group，hypertension group，control+hydrogen-rich saline group and hypertension+hydrogen-rich saline group.Hypertension was induced by subcutaneous infusion with angiotensinⅡ0.7 mg·kg-1·d-1for 2 weeks by osmotic mini-pumps in rats.Hydrogen-rich saline（10 mL·kg-1·d-1）was administered by intraperitoneal injection for 14 d. Platelet adhesion on collagen surface was evaluated using a well-defined perfusion chamber at low shear rate（300s-1）and high shear rate（1080s-1）.The maximum aggregation rate of platelets induced by ADP was deter?mined by turbidimetry.The levels of reactive oxygen spe?cies（ROS），nitric oxide（NO）and Ca2+in platelets were measured with flow cytometry.RESULTSWhen com?pared with the control group，platelet aggregation，plate?let adhesion rate in high shear rate and low shear rate，and the level of ROS and Ca2+in platelets were elevated in hypertensive group.However，NO level in platelets decreased.Comparedwiththehypertensivegroup，hydrogen-rich saline treatment decreased platelet aggre?gation，platelet adhesion rate，the levels of ROS and Ca2+in platelets，and increased NO level in platelets of hypertensive rats.CONCLUSIONHydrogen-rich saline could inhibit platelet activation in hypertensive rats.This effect may be related to antioxidative stress.

Key words：hypertension；platelet；activation；hydro?gen；oxidative stress

Foundation item：The project supported National Natu?ral Science Foundation of China（81173061）；the Collab?orative Innovation Center for Research and Development of Traditional Chinese Medicine in Mount Tai；the Foun?dation of Overseas Distinguished Taishan Scholars of Shandong Province（FODTS）；the Natural Science Foundation of Shandong Province（ZR2014HQ007）；and the Scienceand TechnologyProjectofTaian City（201440774-25）.

Corresponding author：ZHAO Xiao-min，E-mail：zhaox?iaominty@163.com

*Co-first author.

T1-15

DDAH1-V3transcript might act as miR-21 sponge to maintain balance ofDDAH1-V1in cultured HUVECs

KUANG Da-bin1，2，3，ZHOU Ji-peng1，2，4，YULin-yu1，2，3， ZENG Wen-jing1，2，3，XIAO Jian5，ZHANG Zan-lin5，CHEN Xiao-ping1，2，3

（1.Department of Clinical Pharmacology，Xiangya Hos?pital，Central South University，Changsha 410008，Chi?na；2.Institute of Clinical Pharmacology，Central South University，Hunan Key Laboratory of Pharmacogenet?ics，Changsha 410078，China；3.Cooperative Innova?tion Center for Molecular Target New Drug Study，Uni?versity of South China，Hengyang 421001，China；4.De?partment of Cardiovascular Medicine，Xiangya Hospital，Central South University，Changsha 410008，Hunan，China；5.Department of Pharmacy，Xiangya Hospital，Central South University，Changsha 410008，China）

Abstract：OBJECTIVETo investigate whether micro RNA（miRNA）miR-21 regulates dimethylarginine dimeth?ylaminohydrolase 1 （DDAH1） expression through binding 3′-UTR regiondirectly in human umbilical venous endothelial cells（HUVECs） and to explore whetherDDAH1-V2/V3transcripts can function as microRNA sponge，thereby modulatingDDAH1-V1expression.METHODSTheDDAH13′-UTR containing miR-21 rec?ognizing sequence was cloned into PmirGLO dual-lucifer?ase miRNA target expression plasmid to construct Pmir?GLO-miR-21.The plasmid and miR-21（at concentra?tions of 25，50，100 nmol·L-1，respectively）or negative control（100 nmol·L-1）were co-transfected into HUVECs，luciferase activity was detected at 24 h.HUVECs were in?cubated with 2 μg·mL-1actinomycin D for the indicated time after miR-21（25 nmol.L-1）transfection，half-lives ofDDAH1mRNA were determined.HUVECs were trans?fected with PmirGLO-miR-21 alone or co-transfected with miR-21 for 24 h，DDAH1transcripts mRNA andDDAH1protein expression were determined.RESULTSMiR-21 decreased luciferase activity of PmirGLO-miR-21 in a dose-dependent manner（P<0.05 for 25 nmol·L-1miR-21，P<0.01 for 50 nmol·L-1and 100 nmol·L-1miR-21），and miR-21 inhibitor increased reporter activity of Pmir?GLO-miR-21 and mRNA expression of allDDAH1three transcript variants significantly（P<0.05，respectively）. The degree of increase in endogenousDDAH1mRNA expression by miR-21 inhibitor was more obvious forDDAH1-V3.Overexpression of miR-21 decreased mRNA expression and mRNA half-life time of allDDAH1tran?scripts significantly（P<0.05），andDDAH1-V2displayed significantly decreased half-life time thanDDAH1-V1and-V3 with or without miR-21 transfection（P<0.05，respectively）.MiR-21（100 nmol·L-1）decreasedDDAH1protein expression significantly（P<0.05），which was reversed by PmirGLO-miR-21 transfection（P<0.05）. Transfection of PmirGLO-miR-21 alone increased intra?cellular miR-21 expression by approximately 5.6-fold，but only showed a trend of increase inDDAH1protein expression.CONCLUSIONOur results confirmedDDAH13′-UTR as a target for miR-21，and endogenous miR-21 showed increased inhibitory effect onDDAH1-V3tran?script.DDAH13′-UTR，especially forDDAH1-V3，may function as miR-21 sponge to regulateDDAH1protein ex?pression.Modulation of miR-21-DDAH1interaction may provide a new approach for tackling cardiovascular dis?eases.

Key words：DDAH1；miR-21；microRNA sponge

Foundation item：The project supported by National NaturalScience Foundation ofChina（81170091，81373489，81422052）；Special Topic of the Major Sub?ject of National Science and Technology（2013ZX09509-107）；Provincial Natural Science Foundation of Hunan（13JJ1010）；and Funds for Hunan Education Depart?ment Program（12K006）.

Correspondingauthor：CHENXiao-ping，E-mail：chenxp 74@hotmail.com

T1-16

J24924 possesses cardiovascular and renal protec?tive effects on pristane-induced lupus through inhibi?tion of RhoA/Rho kinase pathway in mice

YAN Yu1，ZHANG Hui-fang1，ZHANG Zhi-hui1，CHEN Yu-cai1，LI Yi-huang1，F(xiàn)ANG Lian-hua1，2，DU Guan-hua1，2

（1.State Key Laboratory of Bioactive Substances and Functions of Natural Medicines，Institute of Materia Medi?ca Chinese Academy of Medical Sciences and Peking Union Medical College，Beijing 100050，China；2.Bei?jing Key Laboratory of Drug Targets Identification and Drug Screening，Institute of Materia Medica Chinese Academy of Medical Sciences and Peking Union Medical College，Beijing 100050，China）

Abstract：OBJECTIVEToexplorewhetherJ24924 could prevent the development of pristane-induced lupus in a mouse model，and whether it could protect renal and lower the cardiovascular risk.METHODSThe effect of J24924 was assessed in female BALB/c mice intraperi?toneal injected with 0.5 mL of pristane，and serum auto?antibodies were tested every month，blood pressure wasmeasured every 2 months，while serum inflammatory markers，spleen pathologic characteristics，renal injury and vascular function were observed at 6 month.RESULTSJ24924 could decrease serum autoantibodies and serum inflammatory markers in the SLE mice and improved the spleen pathologic characteristics，and at the same time improved the renal injury and decreased inflammatory responses in kidneys，reduced blood pressure and improved vascular endothelial function.Western blotting assays revealed that inhibition for the activation of NF-κB and Rho/ROCKs signaling pathways and the down?stream signaling molecules might be the potential mecha?nisms of J24924.CONCLUSIONOur findings suggest that therapy of J24924 may be a strategy to prevent SLE and ameliorate associated kidney and cardiovascular complications.

Key words：J24924；SLE；mice；ROCKs；kidney and cardiovascular complications

Foundation item：The project supported by National Sci?ence and Technology Major Project（2013ZX09103001-008，2013ZX09402203）；and the National Natural Sci?ence Foundation of China（81573645）

Corresponding authors：FANG Lian-hua，E-mail：fanglh@ imm.ac.cn；DU Guan-hua，E-mail：dugh@imm.ac.cn

T1-17

UCN enhances TGF- β-mediated mitoinhibition of VSMCs via counteracting TGF- β-induced cPLA2 ex?pression and activation

ZHU Chao，CAO Chang-chun，WANG Xiao-fei，YUAN Jie，JIN Lai，LI Sheng-nan

（Jiangsu Provincial Key Lab of Cardiovascular Diseases and Molecular Intervention，Department of Pharmacolo?gy，Nanjing Medical University，Nanjing 210029，China）

Abstract：OBJECTIVEUrocortins（UCNs）and trans?forming growth factor-β（TGF-β）have been demonstrat?ed to participate in various cardiovascular diseases，many of which involve VSMCs proliferation.And cytosol?ic phospholipase A2（cPLA2）-mediated arachidonic acid（AA）release is an important cause of vascular smooth muscle cells（VSMCs）proliferation.The work was to in?vestigate the regulation of VSMCs proliferation by UCN/ TGF-β and whether cPLA2 was a link between their sig?naling pathways.METHODSVSMC proliferation was measured by MTT assay and immunofluorescence mi?croscopy.Using cell flow cytometry，the changes in the cell cycle phases were investigated.siRNA was used to knockdown Smad2 and smad3 genes.Lentiviral Vector Particle was performed to over express cPLA2 gene.RE?SULTSBoth UCN and TGF-β inhibited VSMCs prolifera?tion and an additive effect was observed when the cells were treated with UCN plus TGF-β.TGF-β increased the percentage of cells in G1-phase while UCN increased the cell percentage in G2-phase with a concomitant de?crease in S-phase.Neither knockdown of smad2 nor smad3 reversed the role of TGF-β.Furthermore，cPLA2 expression was increased by TGF-β but decreased by UCN and UCN attenuated TGF-β-induced cPLA2 expres?sion.In primary VSMCs，TGF-β induced cPLA2 phos?phorylation，and this effect was also attenuated by UCN. Similar to UCN，the cPLA2 inhibitor，pyrrophenone（PYR），also played a role in enhancing TGF-β-mediated mitoinhibition.Inversely，over-expression of cPLA2 elimi?nated the effect of UCN on the mitoinhibition.CONCLU?SIONThe pretreatment with UCN counteracted TGF-β-mediated cPLA2 expression and activation，thereby con?tributing to TGF-β-mediated mitoinhibition of VSMCs.

Key words：urocortins；TGF-beta；cPLA2；vascular smooth muscle cells；mitoinhibition

Foundation item：The project supported by National NaturalScience FoundationofChina（81573424 & 81273510）；and Priority Academic Program Develop?ment of Jiangsu Higher Education Institutions

Corresponding author：LI Sheng-nan，Tel：（025）86863364，E-mail：snli@njmu.edu.cn

T1-18

Calpain mediated pulmonary vascular remodeling in hypoxia induced pulmonary hypertension

ZHANG Wei-fang1，ZHU Tian-tian2，GE Xiao-yue2，XIONG Ai-zhen1，HU Chang-ping2

（1.The Second Affiliated Hospital of Nanchang Universi?ty，Nanchang 330006，China；2.Department of Pharma?cology，School of Pharmaceutical Sciences，Central South University，Changsha 410078，China）

Abstract：OBJECTIVETo explore the role of calpain in in pulmonary vascular remodeling in hypoxia induced pul?monary hypertension and the underlying mechanism.METHODSSprague-Dawley rats were randomly divided into hypoxia group and normoxia control group.Right ventricular systolic pressure（RVSP）and mean pulmo?nary artery pressure（mPAP）were monitored by the method of right external jugular vein cannula.Right ven?tricular hypertrophy index was expressed as the ratio of right ventricular weight to left ventricular weight（left ven?tricle plus septum weight）.Level of calpain-1，calpain-2 and calpain-4 mRNA in pulmonary artery trunk were de?termined by real-time PCR.Expression of calpain-1，cal?pain-2 and calpain-4 protein was determined by Western Blot.Primary rat pulmonary arterial smooth muscle cells（PASMCs）were divided into 4 groups：normoxia control group，normoxia+MDL28170 group，hypoxia group and hypoxia+MDL28170 group.Cell proliferation was detect?ed by MTS and flow cytometry.Level of Ki-67 and PCNA mRNA were determined by real-time PCR.RESULTSRVSP，mPAP and right ventricular remodeling index were significantly higher in the hypoxia group than those in the normoxia group.In the hypoxia group，pulmonary vascular remodeling occurred，and the expression of calpain-1，calpain-2 and calpain-4 mRNA and protein expression wasincreased in the pulmonaryartery. MDL28170 significantly inhibited hypoxia-induced prolifer?ation of PASMCs accompanied with decreased Ki-67 and PCNA mRNA expression.CONCLUSIONCalpain mediated vascular remodeling via promoting proliferation of PASMCs in hypoxia induced pulmonary hypertension.

Key words：calpain；pulmonary hypertension；pulmo? nary vascular remodeling；pulmonary arterial smooth muscle cells；proliferation

Foundation item：The project supported by National NaturalScience Foundation ofChina（81273512，81460010）；and by Natural Science Foundation of Jiangxi province（20142BAB215035）

Corresponding author：HU Chang-ping，E-mail：34629-7715@qq.com

T1-19

Ginkgolide K protects the heart against ER stress injury by activating the IRE1α/XBP1 pathway

WANG Shou-bao1，3*，WANG Zhen-zhong2*，F(xiàn)AN Qi-ru2，4，GUO Jing1，GINA GA-LI1，DU Guan-hua3，WANG Xin1，XIAO Wei2

（1.Faculty of Life Sciences，The University of Manches?ter，Manchester M13 9NT，UK；2.State Key Laboratory of New-tech for Chinese Medicine Pharmaceutical Pro?cess，Lianyungang，China；3.Beijing Key Laboratory of Drug Targets Identification and Drug Screening，Institute of Materia Medica，Chinese Academy of Medical Sciences&Peking Union Medical College，Beijing，China；4.Faculty of Life Science and Technology，China Phar?maceutical University，Nanjing，China）

Abstract：OBJECTIVEHere we investigated the effects and the underlying mechanism of Ginkgolide K（1，10-di?hydroxy-3，14-didehydroginkgolide，GK）on cardiac ER stress.METHODSCell death，apoptosis，and ER stressrelated signalling pathwaysweremeasuredincultured neonatal rat cardiomyocytes（NRCMs），treated with the ER stress inducers tunicamycin，hydrogen peroxide，and thapsigargin.Acute myocardial infarction was estab?lished using left coronary artery occlusion in mice，and in?farct size was measured by triphenyltetrazolium chloride（TTC）staining.Echocardiography was used to assess heart function and transmission electron microscopy for evaluating ER expansion.RESULTSGK significantly de?creased ER stress-induced cell death in bothin vitroand in vivomodels.In ischemic injured mice，GK treatment re?duced infarct size，rescued heart dysfunction and amelio?rated ER dilation.Mechanistic studies revealed that the beneficial effects of GK occur through enhancement of inositol-requiring enzyme 1α（IRE1α）/X box-binding pro?tein-1（XBP1）activity，which in turn leads to increased ER-associated degradation（ERAD）-mediated clearance of misfolded proteins and autophagy.In addition，GK is also able to partially repress the pro-apoptotic action of regulated IRE1-dependent decay（RIDD）and JNK path?way.CONCLUSIONGK acts through selective activation of the IRE1α/XBP1 pathway to limit ER stress injury.GK is revealed as a promising therapeutic agent to amelio?rate ER stress for treating cardiovascular diseases.

Key words：Ginkgolide K；ER stress；IRE1α；XBP1；ER-associated degradation；autophagy

Corresponding authors：DU Guan-hua，E-mail：dugh@ imm.ac.cn），Tel：（010）63165184；WANG Xin，E-mail：xin.wang@manchester.ac.uk），Tel：+44 161 2755616，XIAO Wei，E-mail：xiaowei1959@yahoo.com

*Co-first author.

T1-20

Cerebral vasorelaxant material basis of Xiaoxuming decoction study with rat basilar artery

LI Li，ZHOU Rui，NIU Zi-ran，WANG Jin-hua，WANG Yue-hua，F(xiàn)ANG Lian-hua，DU Guan-hua

（Beijing Key Laboratory of Drug Target and Screening Research，Institute of Materia Medica，Chinese Acade?my of Medical Sciences and Peking Union Medical Col?lege，Beijing 100050，China）

Abstract：OBJECTIVETo investigate the cerebralvaso?relaxant material basis of Xiaoxuming decoction.METH?ODSAccording to the Xiaoxuming decoction herb sourc?es，we retrieved the chemical structure from the litera?tures and the Chinese Natural Product Database（http：// pharmdata.ncmi.cn）.By using microvessel tension sys?tem，we checked the vasorelaxanteffects of Xiaoxuming decoction anti-cerebral ischemia effective components group（XXMDECG）and the available composition com?pounds on pre-contracted basilar artery ring.RESULTS963 compoundsin the decoction，including 81Fangfeng，77 Mahuang，130 Shengjiang，31 Guizhi，91 Huangqin，127 Renshen，73 Chuanxiong，44 Shaoyao，39 Xin?gren，42 Fangji，62 Fuzi and 166 Gancao were collect?ed.The five largest number classes of compounds in the decoction are volatile oil（32%），flavone（32%），alka?loid（13%），saponin（7%），polyphenol and organic acid（5%）.XXMDECG at concentration from 1 to 400 μg·mL-1can dilate the KCl（60 mmol·L-1）and ET-1（0.01 μmol·L-1）pre-contracted rat basilar artery rings in a dose-depen?dent manner.There are 6 compounds with vasorelaxant ratio more than 50%at the concentration of 10 μmol·L-1.CONCLUSIONXiaoxuming decoction contains abun?dant chemical structure.It has the material basis of multi?ple ingredients and multiple targets.The XXMDECG are able to dilate the rat basilar artery rings in a dose-depen?dent manner.The network interactions between varies of chemical compounds in Xiaoxuming decoction and the vasoconstriction associated targets result in the compre?hensive regulation mechanisms of vascular function.

Key words：Xiaoxuming decoction；material basis；effec?tive compounds group；vasodilation

Foundation item：The project supported by Major Scien?tific and Technological Special Project for“Significant New Drug Creation”（2013ZX09508104，2013ZX09402203）；and by Central Public Scientific Research Institution Fun?damental Project（2014CX05）

Corresponding author：DU Guan-hua，E-mail：dugh@ imm.ac.cn

T1-21

Regulation of microRNAs in cell signaling pathwaysmediated vascular remodeling

CHEN Ying*，SUN Lan*，DU Guan-hua

（Beijing Key Laboratory of Drug Target Research and Drug Screening，State Key Laboratory for Bioactive Sub?stances and Functions of Natural Medicines，Institute of Materia Medica，Chinese Academy of Medical Science andPekingUnionMedicalCollege，Beijing100050，China）

Abstract：Vascular remodeling，which can be found in atherosclerosis，restenosis after angioplasty，hyperten?sion，and some other frequent and serious chronic dis?eases.Smooth muscle cell（SMC）phenotype change，which has been described as converting from a contrac?tile state into a synthetic phenotype，is a crucial event during vascular remodeling.Recently，microRNAs（miR?NAs）a kind of small non-coding RNA molecules，has been proven to target critical genes of cell signaling path?ways to regulate SMC phenotypic change.By searching the PubMed，Embase，reviews，and reference listsof rel?evant papers，we systematically carried out a review of the literature to provide an overview of the miRNAs and their target genes in cell signaling pathways，focus inthe pathways involving in SMC phenotype change.To be spe?cific，miRNAs that regulate genes involved in the MAPK signaling pathways（such as：miR-155，miR-92a，miR-424/503，miR-133，miR-181b，miR-31，miR-1298，miR-132，miR-200c and miR-483-3p），miRNAs target genes involved in the TGF-β signaling pathways（including miR-24，miR-17/92 cluster，miR-599，miR-21 and miR-143/ 145），miRNAs target the genes involved in the AMPK signaling pathways including miR-144/451 and miR-195，miRNAs target the genes involved in the PI3K-Akt signal?ing pathways（including miR-138，miR-34c，miR-223，miR-761，miR-10a，miR-146a），miR-199a-5ptargets the genes involved in the Wnt signaling pathways miRNAs（miR-221/222，miR-15b，miR-24/29a，miR-224）in?volved in the PDGF signaling pathways and some miR?NAs（miR-638，miR-328，miR-365，miR-663，miR-29b，miR-130，miR-142-5p，miR-424/322）which regulate SMC phenotype change by other corresponding targets were in detailed discussed in our review.Exploring the regulation of miRNAs in key cell signaling pathways-mediat?edvascular remodeling will have momentous impact on iden?tifying novel therapeutic targets for its associated disease.

Key words：microRNA；vascular remodeling；smooth muscle cell；cell signaling pathway

Foundation item：The project supported by National Nat?uralScience Foundation ofChina（81102445 and 81670456） ； Beijing Natural Science Foundation（7162132）；and the PUMC Youth Fund and the Funda?mental Research Funds for the Central Universities（33320140069）

Corresponding authors：SUN Lan，E-mail：sunhanx?ing2005@imm.ac.cn，Tel：（010）83157220；DU Guanhua，E-mail：dugh@imm.ac.cn，Tel：（010）63165184

*Co-first author.

T1-22

DL0805 derivatives protect the pulmonary arterial cells via the RhoA/ROCK pathway

YUAN Tian-yi1，ZHANG Hui-fang1，CHEN Yu-cai1，JIAO Xiao-zhen2，XIE Ping2，F(xiàn)ANG Lian-hua1，DU Guan-hua1

（1.Beijing Key Laboratory of Drug Targets Identification and Drug Screening，2.State Key Laboratory of Bioac?tive Substances and Functions of Natural Medicines，Chinese Academy of Medical Sciences and Peking Union Medical College，Beijing 100050，China）

Abstract：OBJECTIVEPulmonary artery hypertension（PAH）is a severe disease characterized by the mean pulmonary artery pressure exceeding 25 mmHg at rest. PAH could induce right heart failure and has a very high mortality rate.At present，several kinds of drugs have been used in the treatment of PAH.However，most of these drugs aim to relax pulmonary arteries and do not inhibit the injury of vessels.In other words，the drugs available for PAH treatment do not improve the survival rate of PAH patients and cannot satisfy the needs in clin?ic.To discover and develop novel candidate compounds effective on the treatment of pulmonary artery injury and remodeling will be very important.Based on these back?ground，the present study aimed to study the protective effect of two novel Rho-kinases（Rho-associated coiledcoil forming protein serine/threonine kinase，ROCK）in?hibitors，DL0805 derivatives（DL0805-1and DL0805-2），on pulmonary arterial cells and further evaluate the un?derlying mechanisms and the possibility of DL0805 deriv?atives become therapeutic drugs for PAH.METHODSThe primary cultured pulmonary arterial cells including human pulmonary artery endothelium cells（HPAECs）and human pulmonaryarterysmooth muscle cells（HPASMCs）were used in this study.HPAECs were in?jured under hypoxia environment（1%O2）and treated with or without DL0805 derivatives.After 48 h，the prolif?eration and oxidative stress were observed.CCK8 was used to detect cell viability.DCFH-DA was used as probe for reactive oxygen species（ROS）under fluores?cence imaging system.HPASMCs was stimulated by growth factors including platelet-derived growth factor-BB（PDGF-BB）and Fetal Bovine Serum（FBS）.The prolif?eration was observed in the cells treated with or without DL0805 derivatives.HPASMCs treated with or without DL0805 derivatives were further incubated with endothe?lin（ET-1），the proliferation and cytoskeleton remodeling of cells were detected by immunofluorescence assay.At last，Western blotting（WB）and immunofluorescence assay were employed to analysis the underlying mecha?nisms in the above experiments.RESULTS10 μmol·L-1DL0805-2 could inhibit the proliferation of HPAECs induced by hypoxia.Each concentration of DL0805-1 and DL0805-2 attenuated the production of ROS in HPAECs.Results from WB indicated that DL0805 derivatives decreased the injury of HPAECs induced by hypoxia through the inhibition of the expression of RhoA and the activity of ROCK.On HPASMCs，DL0805 derivatives reduced the proliferation induced by PDGF-BB and FBS and inhibited cytoskeleton remodeling induced by ET-1.Immunofluo?rescence assay showed that DL0805 derivatives inhibited ROCK activity and down regulated the phosphorylation levels of ROCK substrates.CONCLUSIONDL0805 derivatives exhibited protective effect on pulmonary arterial cells including endothelium cells and smooth muscle cells.Among the above experiments，DL0805-2 showed stronger potency than DL0805-1.These two compounds might protect the cells through the inhibition of RhoA/ ROCK pathway and they probably have the potential in the treatment of PAH and deserve further evaluation.

Key words：DL0805 derivatives；pulmonary artery endo?thelium cell；pulmonary artery smooth muscle cell；hypoxia；Rho kinases

Foundation item：The project supported by Central Public Scientific ResearchInstitution Fundamental Project（2016CX09）；and by National Natural Science Founda?tion of China（81573645）

Corresponding authors：FANG Lian-hua，Tel：（010）63165313，E-mail：fanglh@imm.ac.cn；DU Guan-hua，E-mail：dugh@imm.ac.cn，Tel：（010）63165184

T1-23

Salvianolic acid A attenuates vascular remodeling in a pulmonary arterial hypertension rat model

CHEN Yu-cai， YUAN Tian-yi， ZHANG Hui-fang，F(xiàn)ANG Lian-hua，DU Guan-hua

（Beijing Key Laboratory of Drug Targets Identification and Drug Screening，Institute of Materia Medica Chi?nese Academy of Medical Sciences and Peking Union Medical College，Beijing 100050，China）

Abstract：OBJECTIVEThe current therapeutic approaches have a limited effect on the dysregulated pulmonary vascular remodeling，which is characteristic of pulmonary arterial hypertension（PAH）.In this study we exam-ined whether salvianolic acid A（SAA）extracted from the traditional Chinese medicine′Dan Shen′attenuated vascular remodeling in a PAH rat model，and elucidated theunderlyingmechanisms.METHODSPAH was induced in rats by injecting a single dose of monocrota?line（MCT 60 mg·kg-1，sc）.The rats were orally treated with either SAA（0.3，1，3 mg·kg-1·d-1）or a positive control bosentan（30 mg·kg-1·d-1）for 4 weeks.Echocar?diography and hemodynamic measurements were performed on d 28.Then the hearts and lungs were harvested，the organ indices and pulmonary artery wall thickness were calculated， and biochemicaland histochemical analysis were conducted.The levels of apoptotic and signaling proteins in the lungswere measured using immunoblotting.RESULTSTreatment with SAA or bosentan effectively ameliorated MCT-induced pulmonary artery remodeling， pulmonary hemodynamic abnormalities and the subsequent increases of right ventricular systolic pressure（RVSP）. Furthermore， the treatments significantly attenuated MCT-inducedhypertrophicdamage ofmyocardium，parenchymal injury and collagen deposition in the lungs. Moreover，thetreatmentsattenuated MCT-induced apoptosis and fibrosis in the lungs.The treatments partiallyrestored MCT-induced reductionsofbone morphogenetic protein typeⅡ receptor（BMPRⅡ）and phosphorylated Smad1/5 in the lungs.CONCLUSIONSAA ameliorates the pulmonary arterialremodeling in MCT-induced PAH rats most likely via activating the BMPRⅡ-Smad pathway and inhibiting apoptosis.Thus，SAA may have therapeutic potential for the patients at high risk of PAH.

Keywords：salvianolic acid A； pulmonary artery hypertension；apoptosis；BMPRⅡ；Smad；vascular remolding

Foundation item：The project supported by National NaturalScience Foundation ofChina（81573645，81603101）；and the National Science and Technology Major Project（2013ZX09103001-008）

Corresponding authors：FANG Lian-hua，E-mail：fanglh@ imm.ac.cn；DU Guan-hua，E-mail：dugh@imm.ac.cn

T1-11
Salvianolic acid A attenuates isoproterenol-induced myocardial infarction in mice through PI3K/Akt sig?nal pathway

FANG Lian-hua1，NIU Zi-ran1，CHEN Yu-cai1，YUAN Tian-yi1，LI Li1，WANG Shou-bao1，WANG Yue-hua1，LYU Yang2，DU Guan-hua1

（1.Beijing Key Laboratory of Drug Targets Identification and Drug Screening，2.Beijing Key Laboratory of Poly?morphic Drugs，Institute of Materia Medica，Chinese Academy of Medical Sciences and Peking Union Medical College，Beijing 100050，China）

OBJECTIVE To investigate the protective ef?fect of salvianolic acid A（Sal A）on isoproterenol-in?duced myocardial infarction in mice and its possible mechanisms.METHODS The mice were subcutaneous?ly injected with isopropranol（ISO 8 mg·kg-1）to induce myocardial infarction and evaluated the myocardial pro?tective effect of Sal A from mortality rate，electrocardio?gram（ECG），heart function，myocardial infarction in?dex，serum myocardial enzymes and explored its possi?ble mechanisms from inflammatory，antioxidant and cells apoptosis.RESULTS Sal A can dose-dependently en?hanced the heart function of myocardial infarction mice，reduced the heart index，inhibited the myocardial en?zyme leakage，showed obvious myocardial protection ef?fects.ELISA results showed that Sal A can reduce the expression of myocardial inflammatory cytokines such as IL-6，TNF-α.Western blotting confirmed that Sal A can increase the expression of anti-apoptotic proteins Bcl-2，reduce the expression of apoptosis protein Bax，and raise the phosphorylation level of PI3K and Akt.CON?CLUSION Sal A have displayed significant protective ef?fect against isoproterenol-induced myocardial infarction and its mechanism may be related to increasing of PI3K/ Akt signal pathway and inhibition of cell apoptosis and in?flammatory reaction.

DU Guan-hua，E-mail：dugh@ imm.ac.cn，Tel：（010）63165184

Erigeron multiradiatus；caffeoylquinic acid；myocardial Ischemia reperfusion；inflammation；NF-κB；JNK

國家自然科學(xué)青年基金面上項目（81273929，81302979）

劉劍剛，E-mail：liujiangang2002@sina.com，Tel：（010）62835630

Foundation item：The project supported by National NaturalScience Foundation ofChina（81573645，81603101，81473383）

Key words：salvianolic acid A；isoproterenol；myocardi?al infarction；apoptosis；inflammation；signaling pathway