[生物医药论文精品a]大鼠局灶性脑缺血脑组织的比较蛋白质组学研究

大鼠局灶性脑缺血脑组织的比较蛋白质组学研究王继生1，邱宗荫2，夏永鹏2，李惠芝2（1绵阳市第三人民医院绵阳621000；2重庆医科大学药学院重庆400016）A1A0A3A2建立局灶性脑缺血模型，从蛋白质整体水平探讨脑缺血/再灌注的保护作用机制。A5A4A5A4雄性、健康大鼠SD，随机分为正常组、脑缺血模型组参照KOIZUMI法并加以改进，建立MCAO大鼠脑缺血2H再灌注24H的模型；裂解液提取总蛋白质，以双向电泳技术进行蛋白质分离，考马斯亮蓝染色，获得双向电泳图谱，利用专门的软件筛选出差异蛋白质，用MALDITOFMS获得肽指纹图谱，MSFIT数据库进行搜索，获得有关的蛋白质信G5699。A7A6A7A6A7A6A9A9局灶性脑缺血模型组G994正常组脑组G13467的G8616G17751蛋白质组学G11752G12362。G994模型组G8616G17751，从正常组筛选G205223G1022差异G15932G17810蛋白G7013G9869，G1866G102513G1022G1314G15932G17810，10G1022G20652G15932G17810，G18504G4462G1866G10256G1022蛋白质，G2469G10628白G13466G14002三G9923A4水解G18250、G1431G11014G10378G14158G9620G13044G18334G6930G9620G13044G19489解G14002G3818G18250G12573G994脑缺血G11468关的蛋白质。结论G726从蛋白质组学G16294G5242G2469G10628G1114G980G1135G994脑缺血G11468关的蛋白，有G2173G1122G1182G2530进G980G8505G11752G12362脑缺血性G6451G1272的保护机制。关键词脑缺血蛋白质组学ACOMPARATIVEPROTEOMESTUDYONFOCALCEREBRALISCHEMICTISSUEOFRATBRAINWANGJISHENG1,QIUZONGYIN2,XIAYONGPENG2,LIHUIZHI21THETHIRDHOSPITALOFMIANYANG,MIANYANG6210002DEPARTMENTOFPHARMACY,CHONGQINGUNIVERSITYOFMEDICALSCIENCECHONGQING400016ABSTRACTOBJECTIVETOESTABLISHAMODELOFFOCALCEREBRALISCHEMICTISSUEOFRATBRAINANDEXPLORETHEPROTECTIVEMECHANISMOFFOCALCEREBRALISCHEMIAREPERFUSIONINWHOLELEVELOFPROTEINSMETHODMALEANDHEALTHYSDRATS,WERECLASSIFIEDTHEMINTONORMAL,MODELATRANDOMESTABLISHEDTHEMCAOMODELWITHSUTUREMETHODBYREPERFUSION24HAFTERISCHEMIC2HACCORDINGTOKOIZUMIA8SMETHOD,EXTRACTEDTOTALPROTEINSWITHLYSISBUFFER,SEPARATEDPROTEINSBYTWODIMENSIONALELECTROPHORESIS2DE,A10A11A12A13A14A15A16A17A18A19A20A21A2206S0427A23A24A25A26A27A22A28A29A30A311974A32A33A34A33A35A36A37A38A39A33A40A41A42A43A44A45A35A46A47A48A35A36A37A40A41A49A50A51A52A53A37A33EMAILA44WJS327SINACOMSTAINEDPROTEINSBYCOOMASSIEBRILLIANTBLUE,GOTTHEPATTERNS,FOUNDOUTDIFFERENTIALPROTEINS,ANDOBTAINEDPMSBYMALDITOFMS,GAINEDRELATEDTHEINFORMATIONOFPROTEINSBYMSFITDATABASERESULTACOMPARATIVEPROTEOMESTUDYOFMODELANDNORMALGROUPCOMPAREDWITHMODELGROUP,THENORMALGROUPGAINED23DIFFERENTIALPROTEINSPOTS,13SPOTSEXPRESSEDLOWLY,AND10SPOTSHIGH,IDENTIFIED6PROTEINSPOTS，FOUNDOUTTHERELATIVECEREBRALISCHEMICPROTEINSSUCHASLEUKOTRIENEA4HYDROLASE，THYROTROPINRELEASINGHORMONEDEGRADINGECTOENZYMEETCCONCLUSIONESTABLISHINGA2DETECHNOLOGYWASAPPLIEDTOPROTEINANALYSISOFBRAINTISSUE,ANDFOUNDOUTTHERELATIVEPROTEINSOFFROMPROTEOMEASPECTTHISWOULDPROFITFROMTHEPROTECTIVEMECHANISMOFCEREBRALISCHEMICTISSUEKEYWORDSCEREBRALISCHEMICPROTEOMICS；脑血G12661G11161G7171G2373G4487人G12879生G2641G994健康的常G16277G11161G2656G3822G2469G11161，以G1866G20652G2469G11161G10587、G20652G3809G2469G10587、G20652G14280G8543G10587、G20652G8527G1141G10587G1017重G2373G4487人G12879的健康。脑血G12661G11161G1025G4600以脑缺血G3822G16277，G4439G7171指脑G5502G10627血G8981G18339G1955G4581为G10317G5461的G1025G7542G12082G13475G13007G13491G11154G11161，G7171G1025G13781G5192人常G16277G11161。脑缺血G6451G1272的机制G2325分G3809G74461，G11458G2081有关脑缺血G6451G1272的机制有G19053G17241G17745，G13466G14002G1951G1141，G14270G11013G3534，G1864G3871性G17894质G12573，G17837G1135G11752G12362G4590G993G14033G4448G1852G19428G7138缺血G2462再灌注G6451G1272的机制。G3324大脑G6451G1272G1025，大G18339的局G18108缺血的脑G7787G3634，G7171G980G12193血液G1391G13485G6451G3845行为，G3324G11468关的G11161G1375G1025水G13971，G20057G1881G20652血G2399G2656G8527G1141G2356G1114802。大脑G1025G2172G14045（MIDDLECEREBRALARTERY，MCA）G1391血G2318G7171G1032G5214上缺血性脑血G12661G11161的G3822G2469G18108位，大脑G1025G2172G14045阻G3634MIDDLECEREBRALARTERYOCCLUSION,MCAO）模型被认为G7171局灶性脑缺血的标准G2172物模型，该模型制备，具有方法简便，G993改变大鼠G20057G1881G2399，缺血时间可控的优G9869，可以模拟G1032G5214上的脑缺血，适用G1122缺血/再灌注G6451G1272机制G2656保护大脑的药物G11752G12362，以G2462脑G7787G8527缺血的G11161理生理G11752G12362G12573。蛋白质G7171生物G13466G14002赖以生存的各G12193代谢G2656调控途径的主要执行者，蛋白质G993仅G7171G3822G12193G14280G11161因子对机体作用重要的靶分子，而且也成为大G3822数药物的药靶乃至直接的药物。蛋白质组学G7171近G5192来新G1864的、最具潜力的G2469G10628药靶的技术平台。蛋白质组PROTEOME源G1122蛋白质PROTEING994G3534因组GENOME两G1022英文单词的组合，G11221994G5192G11013两位澳大利亚科学家WILKINSG2656WILLIAMS提出，G4439指G13466G14002G1881G1852G18108蛋白质的存G3324G2462G1866活G2172方式，G2530引申为G980G12193G3534因组所G15932G17810的G1852套蛋白质，而G980G12193G3534因组可包括G980G12193G13466G14002乃至G980G12193生物。近G5192来，以双向电泳TWODIMENSIONALELECTROPHORESIS，2DG为核心的蛋白质分离技术G2656以质谱方法为主的蛋白质G18504G4462技术G12573体G13007已G3534本成熟3A544。本课题采用G8616G17751蛋白质组学G11752G12362策略，对局灶性脑缺血模型大鼠的脑组G13467进行G11752G12362。以双向电泳为分离手段，以G3534质辅G2173G9620光解吸飞行时间质谱（MALDITOFMS）为检测手段；利用生物信G5699学对所得实验结果进行解析，从而获取健康、G11161G10378态下脑组G13467蛋白质组G15932G17810的差异数据，以探讨脑缺血的保护机制。材料与方法一实验设备2K15G1314温离心机SIGMACOGERMANUV1601型紫G3818分光光G5242仪SHIMADZU（岛津）JAPANPHS2型酸G5242计（精G5242002PH）上海第二分析仪器厂G14270G2172双重纯水蒸馏器（SZ93）上海亚荣生化仪器厂VOYAGERDEPROG3534质辅G2173G9620光解吸离子化飞行时间质谱仪（MALDITOFMS）美国ABI公司VIRTIS型冷冻干燥机为VIRTIS公司产品。二主要试剂乙腈（ACN）为FISHER公司产品，三G8691G18271酸（TFA）为FLUKA公司产品，测G5219G13435G14020蛋白G18250（PROMEGASEPUENCINGGRADEMODIFIEDTRYPSIN）为PROMEGA公司产品，质谱G3818标G9163合液MIXTURE2包括以下G10267段G726ANGIOTENSINI129751ACTH117209446ACTH1839246672ACTH738366019INSULIN5734591,2867802、G8708G35344G13683G3534G13917G7702酸（CYANO4HYDROXYCINNAMICACID）为SIGMA公司产品。三实验方法1实验动物成G5192健康雄性SD大鼠，体重250G794300G，G11013第三G1903医大学大G3390医院实验G2172物G1025心提G1391。合G7696G16789G2507G726SCXK（G9201）2001003，G9177G8917G13435。实验G17819G12255G1025的G2172物G20294G1871G2462取G7460G3355G17993G4444实验G2172物G12661理G2656保护的有关G16280G4462。2大鼠大脑中动脉局灶脑缺血/再灌注模型制备4。G4570大鼠用35水合G8707G1828710ML/KGG14157G14120注G4568G21647G18269，G1220位G3278G4462G2530手术。G20060正G1025G2011G2487，G19283G134343CM，G19172G19057结合分离G20060G18108G12575G14192，向G3818G10313引G14028G19157G1095G12373G13920，离G7041二G14157G13920，G6183G5332G20060G2172G14045G19820，G7304G19718G2503G1403G20060总G2172G14045（CCA）、G20060G1881G2172G14045（ICA）、G20060G3818G2172G14045（ECA）；G11013CCA分G2461G3800向G3848G12483G1393G8437G9228离，电G1969ECA所有分G6915，G3324G11014G10378G14158上G2172G14045G17840G12483结G6178、G2011G7041ECA，G1363G1866主干G9228离备用；用G15533心G3853G7254时G3853G19393CCAG2656ICA，用G2110G2004G3324ECAG8543G12483作G980G10627G5430G2011G2487，G4570G20056G1820制备G3921的G3848G12483G9046有G13870G8700G18243的G4624G21869G13459G6566G1849ICA（G9046有G13937G13044），G13550G13051ECAG8543G12483G1009G13459以G19462G13459G7655G9381出G2656出血，G12239G2447G15533心G3853G4570G13459G7655G13543G5942G8851ICAG1849G20057方向G6524进，G6524进G134341720CM时G5875G2052阻力，G15932G7138G13459G7655已G13475G17902G17819大脑G1025G2172G14045的G17227G3999G18108，G4448成G980G1403大脑G1025G2172G14045的阻G3634；G7506G5332G3853G19393CCA的G15533心G3853，G9177理术G18338，G1852G4630G13553合，关G19393G2011G2487；大脑G1025G2172G14045阻G36342HG2530G6183G5332手术G2011G2487，G17743G17743G6312出G13459G7655，G5506G5875阻力时G1584G8502，实G10628再灌注，G9177理术G18338，关G19393G2011G2487。G1563手术组G19512G993G6566G13459G3818，G1866G1325G8505G20600G2528上，再灌注24H。3实验分组。WISTAR大鼠雄性12G2494，实验G2172物随机分为2组G726缺血组，对照组。4取材G3324脑缺血2H再灌注24HG9869用35水合G8707G18287G21647G18269大鼠G2530，G7041G3848取脑，G1924G11436G1025取缺血G2318脑组G13467，称重，液氮冻存。5蛋白质含量测定采用BRADFORD法进行蛋白质G4462G183395。以牛血G9177白蛋白BSA为标准蛋白，配成浓G5242为1GL1的溶液。分别取10、20、40、60、80、100L标准蛋白溶液。加G1849G993G2528试G12661，G4462容至100L。各G12661分别加G18495MLBRADFORD工作液G726001考马斯亮蓝G250、47乙醇V/V、85磷酸V/V充分G9163匀,G6930置5MIN,G1122570NMG3800测吸光G5242，20MING1881测G4448。以蛋白浓G5242为横坐标（X,G/L），吸光G5242为纵坐标（Y,OD595NM），作标准曲G13459，获得G13459性方G12255测出待测样品的595NMG3800吸光G5242值6蛋白质双向电泳分离及凝胶染色主要参考BIORADPROTEANIEFCELL型G12573电G13870焦仪操作指南进行。本实验采用G3278G11468IPGG20056制的G13459性17CM干胶条（PH310，L）。取正常大鼠脑组G13467样品G994缺血脑组G13467各3份，分别上样。蛋白质上样G18339为2MG，并G1363水化上样G13543冲液G2656样品总体积为400L，进行电泳。电泳结束G2530，G3324G3278G4462液G1025G3278G4462，G13475染色液染色，然G2530G3324脱色液G1025脱色，直至G1969胶背景变为无色。7图象分析AMERSHAMIMAGEMASTERVDSCL型G1969胶成像G13007G13491对考马斯亮蓝R250染色的2DE胶进行扫描，IMAGEMASTER2DV20021图像分析软件产生光密G5242图像（MAKEODIMAGE），随G2530进行蛋白G7013G9869检测，G1969胶匹配，根据NORMALISEDVOLUME值的变化来判G7041蛋白G7013G9869G15932G17810G18339的变化，选取G18108分差异G15932G17810候选蛋白作MALDITOF质谱分析。8蛋白质鉴定方法本课题蛋白质G18504G4462G1363用的G7171PMF法，蛋白质从二维电泳G1969胶上G2011下，G13475G14020蛋白G18250胶G1881G18250解，质谱分析得G2052肽指纹图谱，然G2530G3324互联网上进行肽指纹图谱的G8616对G18504G4462。81蛋白质的胶内酶解从2DEG1969胶上G2011下蛋白G7013G9869，G2011碎成1MM3装G1849200L的EPG12661G1025，水洗三G8437。用50ACN/25MM碳酸氢铵400L（PH80）脱色15MIN。反G3809三G8437，直至颜色脱尽。胶块浸G1849100ACN（ACNG1025G993G14033有酸性物质）G10255MIN，脱水，胶块变白，室温抽干。加10G79415LTRYPSING18250液（PROMEGASEQUENCINGGRADEMODIFIEDTRYPSIN,浓G5242为10G79415GTRYPSIN/ML，用25MMPH80的NH4HCO3溶液配制）覆盖胶块，4G6930置60MIN，胶块吸胀G2530，吸G2447G3822G1325的G18250液，37G17819夜12G79415H左G2503。吸出反应液，加G184950ACN/5TFAG9163合液30G79450L，G17743G5506振荡萃取30G79460MIN，共两G8437。萃取液（合并萃取液G2656反应液，可选）抽干G134341H，加G18493L50ACN/5TFAG9163合液溶解。进行质谱分析。82MALDITOFMS制备G3921的样品用美国ABI公司的VOYAGERDEPROMALDITOF质谱仪分析。参数设置如下G726反G4568模式，正离子谱测G4462，N2G9620光源波G19283为337NM，真空G52428108TORR，离子源加速电G2399为20KV，G14045冲宽G5242为3NS，离子延迟时间100NS，GRID75，GUIDEWIRE0003，G3534质为CHCA，G9620光强G5242为850G7941200G1881G18108单位，质谱信G2507单G8437扫描累加150G8437。G3818标采用G9163合的MIXTURE2标准液进行G11468邻校正，G1881标采用G14020蛋白G18250的G14270G2011峰（MHG726221110DA或228318或229918G265684251DA）进行两G9869或单G9869校正。9数据库搜索我们选择PROTEINPROSPECTOR检索软件，G3324MSFIT界面进行检索，检索参数为G726DATABASESWISSPROTSPECIESRATTUSNORVEGICUSTENZYMETRYPSINPEPTIDEVALUESMH,MONOISOTOPICPEPTIDEMASSTOLERANCE2PEPTIDECHARGESTATE1MAXMISSEDCLEAVAGES01POSSIBLEMODIFICATIONSMODECARBAMIDOMETHYL,C。数据库查询G2530，每G1022查询得G2052G980G1022分值2结果电泳图谱G13475G17819图像分析，模型组分辨出G13434755G1022蛋白质G7013G9869；正常组分辨出G13434780G1022蛋白质G7013G9869；模型组G994正常组的匹配G10587为79。以模型组为对照，从正常组筛选G205223G1022差异G15932G17810蛋白G7013G9869，G1866G102513G1022G1314G15932G17810，10G1022G20652G15932G17810（图1）；G18504G4462G1866G10256G1022蛋白质，G1866G10254G1022G20652G15932G17810，2G1022G1314G15932G17810，对G178376G12193蛋白质的性质G994功G14033进行总结如下G726A551A56A57A58A56A572HA59A60A61A59A6024HA62A63A64A65A66A67A64A55A68A62A63A64A65A64A55A69PH310A69A70A71A70A71A6917CMIPGA72A73A69A69A74A75A74A75A74FIG1COOMMACIEBLUESTAINED2DEIMAGENORMALANDMODELOFREPERFUSION24HAFTERISCHEMIC2HOFCEREBRALTISSUEPROTEINSINRATSIPGGELPH310L17CMAA76MODELGROUPBA76NORMALGROUPA77A77A77A7727A78A79A80A81A82A83A80A84A85A78A79A80A81A82A83A80A84A85A78A79A80A81A82A83A80A84A85A78A79A80A81A82A83A80A84A85A86A86A86A86A87A88A89A90A91A92A93A77A94A95A96A87A88A89A90A91A92A93A77A94A95A96A87A88A89A90A91A92A93A77A94A95A96A87A88A89A90A91A92A93A77A94A95A96TAB27EXPRESSIONOFRELATIVEDIFFERENTIALPROTEINSBETWEENMODELANDNORMALGROUPA97A98A97A98A97A98A97A98A99A99A99A99A100A101A100A101A100A101A100A101A102A103A104A105A102A103A104A105A102A103A104A105A102A103A104A105A106A106A106A106A102A103A107A108A102A103A107A108A102A103A107A108A102A103A107A108A109A110A111A112A109A110A111A112A109A110A111A112A109A110A111A112PI/MWA113A114A115A116A113A114A115A116A113A114A115A116A113A114A115A1161A11716A118A119A120A121A122A123A4A124A125A126A117LTA4A124A125A126A118A127A128A12969045/57A124A125A130A131A132A119A120A121A122A123A4LTA4A133A134A135A136A137A138A139A140A141A119A120A121A122A123B4LTB4A142A143A144A126A145A146A134A147A148A126AA133A149A150A1422A11722A118A151A152A153A154A155A156A157A158A155A156A159A125A121A160A126A117TRHA159A125A121A160A126A118A117TRHDEA118A117TRHA161A162A163A164A165A166A167A168A169A170A171A172117288/65TRHA173A174A175A176A177A178A161A162A163A164A179A180A1813A18215A169RAPA183A184A185A186A187A188A189A190A191A1923A182CAMPA193A194A183A184A185A186A187A188A189A190A191A192IA169A195A171A172100258/87RAPA183A184A185A186A187A188A189A190A191A192（CAMP调节鸟嘌呤核苷酸交换因子I）A196RAP1AA197RAP2AA198A199A198A200A201A202A203A204A205A206A207A208A209A210CAMPA211A212A213A2144A2157A216A217A218A219A220A221A2221A198A223A224A225A226A227A228A229A230A215A223A224A225A226A227A228A231A22919A221A222A216A232A233A23437443/51A235A236A237A238A239A240A241A242A243A244A245A2465A24713A248A249A250A251A252A253A254A255A2502A89A254GABAAA253A254A255A2502A65A0A154078/87A90A2A3A4A5A243A6A7A8A21A249A250A251A252A47A9A239A10A11A243A236A237A12A243A13A101A14A15A16A70A17A18A19GABAA20AA22A86A23A16A104A105A106A24A108A86A23A25A26A42A27A28A29A30A31A32A33A34A70A35A16A36A37A38A39A40A41A31A43A101A44A45A46A486A203A22白细胞介素前体（IL18）（干扰素诱导因子）A49A50A5122304/49A121A52A53A54A55A56A57A58A59A60A61A62A57A57A58A63A129A64A131A66A67A133A68A69TA135A71A57A58IA137A57A58A723讨论模型组与正常组的2DE图谱的差异蛋白质点，经MS分析，获得PMF图谱，经MSFIT搜索，得到6种蛋白质，现讨论如下白细胞三烯A4水解酶（LEUKOTRIENEA4HYDROLASE，LTA4H），水解环氧化白细胞三烯A4，形成白细胞三烯B4LTB4。白细胞三烯A4水解酶/氨基肽酶是一种双功能的锌金属酶，转化脂肪酸环氧化白细胞三烯A4水解白三烯B4。白细胞三烯A4水解酶的结构显示，GLU271属于一个保守的GXMEN基序，在M1家族的锌肽酶A，GLU136位于活化的位点3。白细胞三烯A4水解酶在浸润的炎症细胞中和在人的食道癌柱状细胞表达。通过大鼠食道癌模型，白细胞三烯A4水解酶抑制剂抑氨肽酶B，减少了食道腺癌的发生率大约30。根据这些研究，可以推断出LTA4水解酶过分表达出现在食道癌是一个早期事件，对预防食道癌来说可能是一个靶标。CHEN等G6265道LTA4H在人和鼠的食道癌过分表达4。白三烯，如LTA4和LTB4的G1147生，G1039G16213通过炎症细胞，如G3822形G7692细胞，G5052G3136细胞和G13945大细胞5。这些研究表G7138，白细胞三烯A4水解酶可能与G14053G13582G15892炎症G7389G1863。G1431G11014状腺G9620G13044G18334G6930G9620G13044G19489解胞G3818酶（THYROTROPINRELEASINGHORMONEDEGRADINGECTOENZYME，TRHDE），G2460G2529TRHG10317异G5627的氨基肽酶，G1039G16213分G5079在G3414G1319G2530G2506G13044G727G1039G16213在大G14053和G14053下G3414G1319表达，G13792在G13966和G13937G14051G1314表达。TRHG19489解胞G3818酶，G1328G1038一种在腺G3414G1319靶点的TRHG1461G2507G16855G14422基G3252，G1866转G5417G10301G1177在TRHG1147生的腺G11256显G14891表达。G11013G8504推断，TRHG1461G2507G1815G13044G7828G8991，通常与G13942G12483G13945大的G11161人G5353G17227的异常的GH分G8864G10301G8821G7389G11464G6521的G1863G130076。研究表G7138异常的G3414G1319G9620G13044（GH，LH和FSH的，G1134基）通过TRHG18334G69307，一G14336与通过腺G11256细胞的TRH生G10301G2524成和G1461G2507G6521G6922位点的TRHR和TRHG19489解胞G3818酶的G1039组分G7092G1863。异常的GH分G8864G10301，G993G1177G11013TRHG1147生，G13792G1000通过G1866G4439的下G1004G14053G18334G6930G3252G4388（如CRHT和LHG18334G6930G9620G13044）8。在大G3822G6980G11161G10714G451生G10714的G7477件下如G13970G15940G12481G451G12970G4627G11161G451G13937G14051G11154G11161，G14659G10301成G11282G5627G2462G3822G7053G19766G2419G3252G5353G17227的G12946G12082G19568G11873等，TRHG16837G4560GHG18334G6930。G17829G5192来的研究发现TRHG6930G1866G12879G1296G10301能G3827G1431G17839中G7542G12082经G13007G13491G6451G1272G2530功能的G5686G3809G451G1306G1867G1319G7438制G4590G993G9177G7982。G11458G2081G7389G1972种G1563说，G2265G6336G3698G2164G14053G15892G8981G18339G451G6338抗内源G5627阿片肽的G1328用G451抗白三烯和G15892小板G9620活G3252G4388的G11161G10714生G10714G1328用等。RAP鸟嘌呤G7692苷酸交换G3252G43883，G2460G2529CAMPG16855G14422鸟嘌呤G7692苷酸交换G3252G4388I，CAMPGEFI和EAPC1。G11464G6521通过CAMP活化的交换蛋白质（EXCHANGEPROTEINDIRECTLYACTIVATEDBYCAMP，EPAC）是一种RAPG10317异G5627的鸟苷酸交换G3252G4388，通过结G2524CAMP到环状的G7692苷酸一磷酸结G2524区域G13792被G9620活。G7389两种G1134型，EPAC1和EPAC2，在哺乳动G10301细胞，两者都含G7389一个G16855G14422和催化的区域，在蛋白质的N和C的末G12483。G16855G14422区域G2265含CAMP结G2524位点，在G13582乏CAMP的情况下，自动抑制催化活G5627。胞吐G3818排的EPAC1，首先出现在G10317异G5627阻滞剂G9620肽G2419酶G13582乏效应，和环G7692苷酸门控通路9。EPAC蛋白质的功能（受G1319介G4560HRASANDERKG9620活），通过RAP鸟苷三磷酸酶活G5627的抑制剂（通过RAPGAPII的表达），G13792被抑制10。环指区域蛋白1的细胞生长G16855G14422G3252G4388，G2460G2529细胞生长G16855G14422基G325219蛋白。G1039G16213存在组织纤维母细胞。G1039G16213功能能抑制G3822种细胞株的生长。组织G10317异G5627在睾丸高表达，在骨骼肌，G13937，G13966和G14053G1314表达。G16837G4560被P53G16837G456011。氨基丁酸受G1319G1134基2G2081G1319，在哺乳动G10301大G14053中氨基丁酸A型受G1319GABAARS是G1039G16213存在的抑制G5627的受G1319，通过大G3822G6980镇静剂和安眠G14659G16855制。G10317异G5627的GABAARSG1134基,就活G5627和在活G1319内的G14659G10301反应G13792言,G4590G993是十分G9177G798212。2G1134基在整个的发育和成熟的大G14053和脊索高表达,G4439大约占了整个GABAARS的6013，也是必需的GABAARS突触的聚G1287914。2G1134基是苯并二氮G12879受G1319结G2524位点的必需的结构,G1306在装配运输和插入完整的功能G5627的GABAA受G1319G12082经G1815的质膜G993是必需的15。白细胞介G1304418G2081G1319（IL18G2081G1319）G2460G2529，干扰G13044G16837G4560G3252G4388，G1039G16213分G5079于G14053，G13970上腺，G3414G1319G2081G2506等组织。CARBONEA等在人G12879的胰腺癌细胞的研究中发现，在G14659G10301治疗期间，通过T细胞16，癌细胞G1147生的IL18，可提高IFN的G1147G18339。这提示IL18充担了抗肿G11256G7438制的介G4560G1328用。G8504G3818IL18能G3827发挥与G1866G4439细胞G3252G4388的协G16855G1328用，如IL23或G3698强抗肿G11256免疫的一氧化氮G2524酶抑制剂17。在非实G1319的肿G11256模型中，这种细胞G3252G4388G8821G7389抑制G1328用，G1039G16213G1319现在人G12879的非白G15892G5627白G15892G11161细胞株中18。IL18抑制效应的G7438制归G3252于细胞分裂周期的G16855G14422G1328用，G4560致S期停止。G4439证实了在G1972种研究中，S期停止经常伴随着干扰细胞周期控制蛋白。如G13582氧G16837G4560的S期停止，在人的胸部（T47D）和颈部的（NHIK3025）癌细胞株并行向下G16855G14422细胞周期G16855G14422蛋白A19。白细胞介G13044L8还可以刺G9620白细胞介G130441G451肿G11256坏G8527G3252G4388G2462白细胞介G130448等G1866G1194炎G5627细胞G3252G4388的生成，G1431G1363炎G5627细胞向炎G5627区域聚G19610，G1861G2528G1431G1363G13582G15892G2530G14053组织中的过G5242炎症反应20。参考文献1JANARDHAN，VQURESHIAIMECHANISMSOFISCHEMICBRAININJURYJG714CURRCARDIOLREP,200462L17232NOBREMC，MONTEIROMG15G3ALBUQUERQUEAC，G72G87G15G68G79G17G39G72G70G82G80G83G85G72G86G86G76G89G72G3G70G85G68G81G76G72G70G87G82G80G92G3G73G82G85G3G87G85G72G68G87G80G72G81G87G3G82G73G3G76G81G87G85G68G70G85G68G81G76G68G79G3G75G92G83G72G85G87G72G81G86G76G82G81G3G86G72G70G82G81G71G68G85G92G3G87G82G3G79G68G85G74G72G3G76G86G70G75G72G80G76G70G3G70G72G85G72G69G85G68G79G3G76G81G73G68G85G70G87G76G82G81G29G3G68G81G68G79G92G86G76G86G3G82G73