氯胺酮與乙醇對小鼠海馬PKA-CREB 信號通路的影響

南通大學(xué)學(xué)報(醫學(xué)版Journal of Nantong University(Medical Sciences) 2013: 33(4283氯胺酮與乙醇對小鼠海馬PKA-CREB信號通路的影響楊美玉13”,丁飛2,蔣小崗3,顧振綸3,卞士中”(蘇州大學(xué)醫學(xué)部法醫學(xué)系,蘇州215123;2上海市公安局寶山分局刑事科學(xué)研究所;3蘇州中藥研究所)摘要]目的:研究氯胺酮與乙醇對小鼠海馬ⅣKA-CREB信號通路的影響,探討氯胺酮與乙醇致學(xué)習記憶障礙的分子學(xué)機制。方法:40只ICR小鼠隨機分為4組:對照組、氯胺酮組、乙醇組、氯胺酮乙醇合用組,每組10只。氯胺酮和乙醇分別釆取腹腔注射和灌胃的給藥方式,1次/d,連續14d。通過(guò)RT-PCR檢測海馬中c-AMP反應單元結合蛋白(c- AMP response element binding protein,CREB)蛋白激酶A( protein kinase A,FKA)及腺苷酸環(huán)化酶( adenylate cyclase,AC)mRNA的表達; Western blot檢測 PCREB/CREB的蛋白表達。結果:RT-PCR檢測結果:氯胺酮乙醇合用組可顯著(zhù)降低海馬區CREB、PKA及 AC mRNA的表達(P<0.05), Western Blot檢測顯示 PCREB/CREB的蛋白表達顯著(zhù)降低(P<0.05)。結論:氯胺酮與乙醇可協(xié)同抑制小鼠的學(xué)習記憶行為,其機制可能與大腦海馬區CREB信號通路相關(guān)[關(guān)鍵詞]氯胺酮;乙醇;學(xué)習記憶;環(huán)磷酸腺苷反應結合蛋白;小鼠中圖分類(lèi)號]R971文獻標志碼]A[文章編號]1674-7887(2013)04-0283-05Effects of ketamine on ethanol-induced learning and memory impairment in miceYANG Meiyu., DING Fei JIA NG Xiaogang, GU Zhenlun, BlA N Shizhong (Department of Forensic MedicineMedical College of Soochow University, Institute of Forensic Sciences of Soochow University, Suzhou 215123; Institute ofCriminal Sciences, Baoshan Branch of Shanghai Public Security Bureau; Suzhou Institute of Chinese Materia Medica)[Abstract] Objective: To study the effects of ketamine and ethanol on PKA-CREB pathway of hippocampus -dependentlearning and memory in mice and its possible mechanism. Methods: A total of 40 male mice were divided into 4 groups: anormal control group, a ketamine group, an ethanol group and an ethanol plus ketamine group. Ketamine and ethanol wererespectively taken by intraperitoneal injection and oral administration, once per day for 14 d. The expressions of CreBPKA and AC gene in hippocampus were detected by RT-PCR. The expressions of pCREB/CREB were detected by WesternBlot. Results: After the combined treatment with ketamin and ethanol. CREB. PKA and AC mRNA were down regulated byRT-PCR(P<0. 05), and p CREB/CREB were down regulated by Westem Blot(P<0. 05). Conclusion: The data indicatethat combination of ketamine and ethanol is synergic in the inhibition of learning and memory in mice, its mechanism maybe related to the CreB signal transduction pathway.[Key words] ketamine; ethanol; learning and memory; C-AMP-response element binding protein; mouse氯胺酮是N-甲基D-天(門(mén))冬氨酸(N- methy-人細胞核磷酸化CREB,然后激活TEGs(C-os)等,其D- -aspartate,NMDA)受體的非特異性阻斷劑,其作激活產(chǎn)物進(jìn)一步編碼LTP所需要的蛋白質(zhì)。用機制復雜。目前研究認為,麻醉劑量的氯胺酮本研究以小鼠為實(shí)驗對象,觀(guān)察氯胺酮與酒精可通過(guò)阻斷NMDA受體抑制學(xué)習記憶,該受體對神聯(lián)合給藥對小鼠學(xué)習記憶的影響。采用RT-PCR技經(jīng)元突觸長(cháng)時(shí)程增強(ong- -term potentiation,TP)的術(shù)檢測小鼠海馬中CREB、PKA及腺甘酸環(huán)化酶產(chǎn)生、維持和學(xué)習記憶功能至關(guān)重要。adenylate cyclase, AC)mRNA的表達; Western blot環(huán)磷腺苷 (cyclin adenosine monophosphate,c-AMP)檢測 pCREB/CREB的蛋白表達情況,以探討氯胺酮反應元件結合蛋白(c- AMP response element binding與乙醇對小鼠學(xué)習記憶行為學(xué)影響的分子機制。protein,CREB)是一種依賴(lài)c-AMP的轉錄因子,可將細胞內的c-AMP的量的變化轉錄為基因表達的1材料與方法形式。CREB在記憶的形成過(guò)程中發(fā)揮了及其重要1.1實(shí)驗動(dòng)物ICR小鼠40只,雄性,體質(zhì)量25的作用,其過(guò)程可能是:神經(jīng)遞質(zhì)等作用神經(jīng)元細30g。蘇州大學(xué)實(shí)驗動(dòng)物中心提供。實(shí)驗前小鼠適應胞,導致胞質(zhì)內c-AMP濃度升高,使蛋白激酶A性飼養3d,維持室溫(22±1)℃,在整個(gè)實(shí)驗過(guò)程中protein kinase A,PKA)激活,PKA的催化亞單位進(jìn)小鼠自由飲水和進(jìn)食。*作者簡(jiǎn)介楊美玉,女,漢族,生于1984年3月,山東省濟寧市人,碩土在讀,研究方向:法醫毒理病理學(xué)通信作者卞士中,電話(huà):0512-67580893,E- mail. bianshizhongt@ suda. edu. en南通大學(xué)學(xué)報(醫學(xué)版)2013:33(4)1.2主要試劑及儀器鹽酸氯胺酮注射液(國藥準表2內參和目的基因擴增條件字H32022820,批號:KH071004,2ml:0.1g)購自江基因擴增條件蘇恒瑞醫藥公司;56%v/紅星二鍋頭酒購自北京紅 GAPDH94℃4min94℃30s-55℃30s72℃星股份有限公司;PCR擴增試劑盒(批號:DR0A)63lmin+72℃10min+4℃購自 Takara公司;RNA逆轉錄引物:由上海生物工℃5min→94℃1min→+57℃30s-72℃程有限公司合成;兔抗鼠 phospho-CREB-1(Ser13345s72℃10min*4℃抗體(一抗)(1:100)購自 slabs;抗小鼠CREB抗體94℃4min→→94℃30s57℃30s+72℃30 cycles)lmin+72℃10min-+4℃(一抗)(1:100購自武漢博士德生物技術(shù)有限公司CREB94℃2min+94℃30s→55℃45s72℃actin(1:1000)購自 Santa cruze;所用抗鼠和抗兔熒(30 cyeles)45s+72℃7min-+4℃光二抗均為 Odyssey公司產(chǎn)品1.5 Western blot1.3模型制備與給藥40只小鼠隨機分成4組,分1.5.1蛋白樣品制備取液氮內凍存的小鼠海馬組別為正常對照組,氯胺酮組,乙醇組,聯(lián)合給藥組,每織約30mg,加入組織裂解液05m,超聲勻漿后于組10只。給藥方式:對照組以生理鹽水腹腔注射+生4℃,12000 r/min離心10min,吸取上清,分裝,另理鹽水灌胃,乙醇組以生理鹽水腹腔注射+乙醇灌外留少許待測蛋白溶液(于-40℃冷藏。胃,氯胺酮組以氯胺酮腹腔注射+生理鹽水灌胃,聯(lián)1.5.2BCA試劑盒檢測蛋白濃度(1)蛋白標準曲合給藥組以氯胺酮腹腔注射+乙醇灌胃。生理鹽水、線(xiàn)繪制取標準蛋白液,用PBS梯度稀釋成4、8、12、氯胺酮溶液(50mgkg)腹腔注射1次l,連續14d;生16、20、24μg/μL蛋白溶液,每組40μL。取BCA試理鹽水、乙醇紅星二鍋頭稀釋至20%灌胃1次,劑盒A液和B液按50:1配成工作液。稀釋后的標連續14d。各組小鼠每日9:00給藥。準蛋白液與BCA工作液按1:8混合均勻后,37℃水1.4PCR檢測浴30min,于酶標儀上測定OD值,每組設3復孔。1.4.1總RNA提取取液氮內凍存的小鼠海馬組由OD值,繪制蛋白標準曲線(xiàn);(2)待測蛋白濃度的測織約30mg,加人Tiol試劑0.5mL,按說(shuō)明書(shū)進(jìn)行定取待測蛋白液每組10μL。將待測蛋白液與BCARNA提取,所用RNA的D/D2均在1.8~2.0。工作液按1:8混勻后,37℃水浴30min,于酶標儀上1.4.2模板cDNA合成提取的總RNA逆轉錄合測其OD值,每組設3復孔。根據OD值,于蛋白標成cDNA。準曲線(xiàn)上計算相關(guān)蛋白濃度。根據蛋白濃度測定值1.4.3 RT-PCR調整樣本上樣量,使各組蛋白量為60μg,按4:1與14.3.1內參和目的基因的引物序列及產(chǎn)物長(cháng)度5× padding Buffer混勻,水浴100℃變性5min后,見(jiàn)表1。冰浴備用4.3.2內參和目的基因擴增條件均為25μL1.5.3SDS-PAGE電泳先將實(shí)驗用玻片洗凈,再反應體系,應用 GAPDH作為參照。內參和目的基用無(wú)水乙醇脫脂,晾干后組裝制膠板。配12%的分離因擴增條件見(jiàn)表2。PCR產(chǎn)物用1.5%瓊脂糖凝膠膠,注入制膠板中,加適量無(wú)水乙醇封閉。待膠凝固,進(jìn)行電泳,結束后于全自動(dòng)凝膠成像分析系統進(jìn)倒去乙醇,用蒸餾水清洗3次。配制濃縮膠,兩端交行灰度掃描。以 GAPDH擴増產(chǎn)物校正作光密度相替加入制膠板,插入梳子后放置約1h待膠凝固后對分析。使用。安裝電泳裝置,上樣品,進(jìn)行電泳,濃縮膠時(shí)設表1內參和目的基因的引物序列及產(chǎn)物長(cháng)度置電壓90V,染料條帶至分離膠時(shí)電壓設置至基因引物110V,染料接近分離膠底部,停止電泳。F5'-CCGGAACATGGACCTCTACTAC-31.54轉膜將丙烯酰胺凝膠從玻璃板上剝離,置(284 bp) R5'-ATAGGTGGGAGGAGATGGACTT-3入轉移緩沖液中,將海綿、濾紙、凝膠、NC膜、濾紙F5 '-GGAGAGCGTGAAAGAGTTCCTA-3海綿按順序疊加,用玻璃棒小心趕走氣泡,放置于轉(339 bp) R5'-CCAGCTACATACTCCATGACCA-3移電泳槽中恒流400mA,轉移120mi。轉膜后,CREBF5=AGTATGCACAGACCACTGATGG-3用1×TBST洗膜。(392 bp) R5'-TCCACCGTAACAGGAGAGAACT-31.5.5封閉1×TBST洗膜3次,每次10min,然后GAPDH F5 -GCCATCAACGACCCCTTCATT-3將膜浸于5%脫脂牛奶中封閉1h(413 bp) R5'-CGCCTGCTTCACCACCTTCTT-31.5.6抗體孵育封閉結束后再用1×TBST洗膜3楊美玉,等氯胺酮與乙醇對小鼠海馬PKA-CREB信號通路的影響285次,每次l0mins將NC膜放入小塑料袋內,加入5%的降低用統計學(xué)意義(P<0.05,圖2)。提示:氯胺酮脫脂牛奶按比例稀釋后的_抗,膜。去除袋內飲酒、氯胺酮+飲酒聯(lián)合給藥均可下調 pCREB氣泡,用封膜機封口,置于搖床上室溫孵育3h過(guò)CREB蛋白表達,且聯(lián)合用藥組 pCREB/CREB降低夜。取膜,用1×TBST洗滌3次,每次10min后,用更明顯5%脫脂牛奶按1:10000的比例稀釋的二抗封閉,置對照組乙醇氯胺酮氯胺酮+乙醇于搖床上,室溫避光孵育1h。取膜,用1×TBST在暗pCREB-盒中洗滌3次,每次10min。15.7顯影近紅外雙色激光成像系統顯影CREB-42k1.6圖像處理及數據分析RT-PCR, Western blot圖像掃描后均采用 SigmaScan pro5軟件系統分析,圖2氯胺酮與乙醇對小鼠海馬CREB, PCREB蛋白表達的實(shí)驗數據以x±s表示。顯著(zhù)性檢驗采用 One way影響ANOVA,P<0.05為差異有統計學(xué)意義。3討論2結果CREB是堿性氨基酸亮氨酸(Leu)拉鏈轉錄因子2.lRT-PCR檢測結果在連續給藥14d,小鼠海家族,受細胞內多條信號通路的調節,與生長(cháng)抑素馬組織RT-PCR檢測結果分別以PKA、AC、CREB基因的cAMP反應元件 CAMP response element,CRE與內參基因 GAPDH的密度比值表示組織中PKA、有高度親和力。CREB家族在結構上具有DNA結AC、 CREB MRNA的相對表達強度。實(shí)驗結果表明,合結構域和轉錄激活結構域,因而具有DNA結合活與正常對照組相比,飲酒、氯胺酮、氯胺酮+飲酒聯(lián)合性和轉錄調節活性。但只有當Ser133被PKA、蛋白給藥可誘導小鼠海馬組織中的RKA、AC、CREB激酶 C(protein kinase C,ⅨKC或鈣/鈣調蛋白激酶Ca27mRNA表達下調均P0.05。與單用藥組相比,聯(lián)合 calmodulin- dependent protein kinase, Ca+/CaMK等磷給藥組小鼠海馬組織pKA、AC、 CREB mRNA表達酸化后才能啟動(dòng)轉錄,磷酸化的CREB形成二聚體顯著(zhù)下降(均P<0.05圖1)。提示:飲酒、氯胺酮、氯后活化,并與靶基因CRE序列結合,調節其轉錄,發(fā)胺酮+飲酒聯(lián)合給藥可使小鼠海馬組織中PKA、AC、揮多種生物學(xué)效應。CREB及其相應的基因表達下調,其中氯胺酮+飲酒在中樞神經(jīng)系統中,CREB調節神經(jīng)細胞的生聯(lián)合給藥組作用更明顯。長(cháng)發(fā)育及營(yíng)養,參與神經(jīng)細胞突觸可塑性和長(cháng)時(shí)程Marker對照組乙醇氯胺酮氯胺酮+乙醇記憶的形成,是長(cháng)時(shí)程記憶和LTP所必需的基因開(kāi)關(guān)閃。CREB通過(guò)調控目的基因的表達,影響晚時(shí)(413bp相的LTP和長(cháng)時(shí)程記憶實(shí)驗證實(shí)CREB是學(xué)習記憶中一個(gè)關(guān)鍵因子參與海馬的空間記憶調節。研究認為迷宮實(shí)驗(284bp練可使大鼠海馬和紋狀體內 PCREB免疫活性顯著(zhù)500bp升高。海馬CREB功能降低,可導致空間記憶功能的400bp減退。譚蕾等研究結果表明氯胺酮通過(guò)下調大鼠海馬神經(jīng)元 pCREB水平,誘導新生大鼠海馬神經(jīng)元凋亡,突觸形成受損。 S C Pandey等研究發(fā)現大鼠(392bp)圖1氯胺酮與乙醇對小鼠海馬PKA、AC、CREB基因表達的過(guò)度飲酒,可導致杏仁核CRE-DNA結合力下降,海影響馬內總CREB及 pCREB表達下調,進(jìn)而影響大鼠的2.2 Western blot檢測結果CREB的磷酸化水平學(xué)習記憶功能。與學(xué)習記憶密切有關(guān)。連續給予氯胺酮、飲酒、聯(lián)合氯胺酮為苯環(huán)己哌啶(N-1- phenyeyc lohexy-給藥14d后,與正常對照組相比,各用藥組小鼠海 piperidine,PCP)的衍生物,是NMDA受體的非特異馬組織 PCREB、CREB蛋白表達量均降低(P<0.05),性阻斷劑。氯胺酮拮抗NMDA受體,可通過(guò)阻斷各且磷酸化水平 pCREB/CREB的降低有統計學(xué)意義刺激產(chǎn)生的長(cháng)時(shí)程增強效應,干擾學(xué)習記憶過(guò)程。有P005與單用藥組相比聯(lián)合用藥組 PCREB/CREB報道稱(chēng)氯胺酮可致其濫用者記憶的長(cháng)久損害作用。南通大學(xué)學(xué)報(醫學(xué)版)2013:33(4)經(jīng)多次氯胺酮麻醉的大鼠其空間學(xué)習記憶能力明顯小鼠學(xué)習記憶的影響[中國藥理學(xué)通報,199,15(3)下降,且海馬神經(jīng)元超微結構也有異常改變,損害程90-93度與使用氯胺酮的劑量水平有關(guān)叫。本實(shí)驗室研究結5 Molkentin JI, Dorn gW. Cytoplasmic signaling pathways在海馬內,乙醇通過(guò)抑制NMDA受體亞型2001,636330小 hypertrophy. Annu rey physiol果表明,氯胺酮多次給藥可致小鼠空間學(xué)習記憶能that regulate cardiac力障礙。Wu GY, Deisseroth K, Tsien RW. Activity -dependentNR2B的活性來(lái)抑制NMDA受體。早期研究表明CREB Phospheof a fast. sensitivecalmodulin kinase pathway and a slow, less sensitive mite乙醇在較低濃度時(shí)就可對NMDA受體的一個(gè)亞型gen-activated protein kinase pathway J. Proe Natl Acad起作用,以某種方式改變該受體亞型之間的相互配Seius a.2001,985)2808-2813合,從而使谷氨酸利用受體進(jìn)行的信號傳遞受到破7] Setlow b, Roozendaal b. McGaugh JI. Involvement of a壞,進(jìn)而影響包括LTP在內的分子信號傳導通路受basolateral amygdala complex-nucleus accumbens pathway阻,導致學(xué)習記憶功能障礙。體外研究證明,乙醇in glucocorticoid-induced modulation of memory consoli可明顯降低CREB的活性。dation. Eur J Neurosci, 2000, 12(1): 367-375本研究分子生物學(xué)實(shí)驗硏究結果表明:氯胺酮、18] Puzo d.iolo. Trinchese h,etal. Amyloid- -beta pep乙醇、氯胺酮聯(lián)合乙醇作用小鼠14d后,小鼠海馬ide inhibits activation of the nitric oxidelegMPicAMP-re組織AC、PKA、 CREB MRNA的表達減少,且兩兩比onsive element-binding protein pathwaypocampal synaptic plasticity [J). J Neurosci較顯示聯(lián)合用藥組與單用藥差異均有統計學(xué)意義6887-6897提示氯胺酮和乙醇可能在基因轉錄水平上對CREBLiu ry fioravante d. shah S. et al. cAMP信號傳導通路協(xié)同抑制。 Western blot檢測結果表ment-binding protein 1 feedback loop is necessary for明:用藥14d后,小鼠海馬組織中, pCREB/CREB蛋onsolidation of long-term synaptic facilitation in aplysiaJI白表達也有所下降,提示氯胺酮和乙醇在蛋白質(zhì)水J Neurosci.,2008,28(8):1970-1976平上影響CREB信號傳導通路。與本實(shí)驗室行為學(xué)10 Colombo PJ, Brightwell JJ. Countryman RA. Cognitive str結果一致,提示氯胺酮、乙醇可加強抑制小鼠海馬組ategy -specific increases in phosphorylated cAMP re-織CREB信號傳導通路。sponse element-binding protein and c-Fos in the hip.CREB參與成癮和學(xué)習記憶的形成,但還不能and dorsal striatum[J]. J Neurosci, 2003, 23(8)3547-3554僅用CREB解釋行為改變的長(cháng)久性。藥物抑制和經(jīng)m1譚蕾,羅愛(ài)林,王金韜,等氯胺酮對大鼠海馬神經(jīng)元驗誘導的CREB活性相對短暫,且注射pCRE只能CREB磷酸化水平的影響J中華麻醉學(xué)雜志,2008部分促進(jìn)突觸的刺激。這表明成癮和記憶的長(cháng)時(shí)轉7637-639變可能啟動(dòng)更加穩定調節機制,有其他因子的參與。12] Pandey so., Mittal n. Lumeng L, et al. Involvement of the對于精神活性物質(zhì)的成癮仍需要進(jìn)一步的實(shí)驗研cyclic AMP-responsive element binding protein gene tran-究,為臨床戒斷和脫毒提供理論數據支持。scription factor in genetic preference for alcohol drinkingbehavior[J]. Alcohol Clin Exp Res, 1999, 23(9): 1425[參考文獻][1] Morgan CJ, Riccelli M, Maitland CH, et al. Long-term ef- [13] Goulart BK, de Lima MN, de Farias CB, et al. Ketaminfects of ketamine: evidence for a persisting impairment ofimpairs recognition memory consolidation and preventssource memory in recreational users[J. Drug Alcohol Delearning-induced increase in hippocampal brain-derivedpend,2004,75(3):301-308eurotrophic factor levels[J]. Neuroscience, 2010, 167(4)[2 Wang JH. Fu Y, Wilson FA, et al. 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J Neural Transm, 2005, 112(8)南通大學(xué)學(xué)報(醫學(xué)版Journal of Nantong University(Medical Sciences) 2013: 33(4Wnt5a在嚴重燙傷大鼠早期腸黏膜組織中的表達及胰島素的調控作用*俊”,胡克蘇,張云,張逸(南通大學(xué)附屬醫院燒傷整形科,南通226001)摘要]目的:研究在嚴重燙傷大鼠腸黏膜組織中Wn5a的動(dòng)態(tài)表達變化及其意義。方法:將48只大鼠背部浸人恒溫水浴盆(92℃中20s達Ⅲ度燙傷,制作燙傷創(chuàng )面,采用隨機數字法將大鼠分為對照組、單純燙傷組燙傷組)、燙傷后胰島素治療組(治療組)。傷后取腸組織觀(guān)察大體變化,取腸道黏膜H染色后,行組織形態(tài)學(xué)觀(guān)察,分別于燙傷后6、1224、48h觀(guān)察,取腸黏膜組織以免疫組織化學(xué)的方法,檢測Wnt5a的表達情況。結果:在燙傷組和治療組Wnt5a表達均有顯著(zhù)表達,對照組有少量表達(P<0.05),傷后6-12hWn5a的表達持續升高,燙傷組12-24h開(kāi)始表達逐漸減少,24h后表達又重新增多,各組在48h表達達到最高峰;與燙傷組相比,治療組wnt5a的表達水平明顯降低(P<0.05,病理形態(tài)學(xué)變化顯示燙傷早期大鼠腸黏膜組織損傷明顯,治療組大鼠腸黏膜組織損傷較燙傷組明顯減輕。結論:嚴重燙傷早期SD大鼠腸黏膜組織損傷明顯,wnt5a可能參與了燒傷膿毒癥的發(fā)生發(fā)展過(guò)程。胰島素能減少其表達,可能是其保護腸黏膜機制的因素之一鍵詞]燒傷;腸黏膜;Wnt5a;大鼠「中圖分類(lèi)號1R644「文獻標志碼]A[文章編號]1674-7887(2013)04-0287-04Expression of Wnt5a in intestinal mucosa of rats during the early stage after severe burninjury and the effect of the insulinQI Jun, HU Kesu, ZHANG Yun, ZHANG Yi(Department of Plastic Surgery, the Affiliated Hospital of Nantong Uniity, Nantong 226001)[Abstract] Objective: To investigate the changes of the expressions of Wnt5a in intestinal mucosa of rats during the earlyage after severe burn Injury and its sigficance. Methods: 48 healthy SD rats were set upscalded models, which wererandomly divided into control group, scald group and scald +insulin group(treatment group). The intestinal mucosa tissuesample separately after 6, 12, 24, 48 hours were posted scald injury. In the treatment group, subcutaneous injection of 3 U/kgsulin after scald injury. The expression of Wnt5a in the intestinal mucosa was detected with immunohistochemistry methodsnd analyzed by a micro-image analysis system, and at the same time we observed the pathological changes of the intestinalmucosa tissue of each groups. Results: In the scalded group, the expression of Wnt5a was higher than that in control group atthe same time(P<0.05),6-12 hours post scald injury the expression was increased. Following the expression of Wnt5adecreased obviously 24 hours post scald injury, 48 hours post scald injury the expression was increased. The expression ofWnt5a in the treatment group was significantly smaller than that in scalded group at most time points at the same time(P<0.05). Conclusions: In the early phase of severe burns injury, the intestinal mucosa tissue injury is very severe. Wnt5amay take part in the procession of burn sepsis. Insulin can deduce its expression, which may be one of factors protecting theIKey words burn; intestinal mucosa; Wnt5a;rat臨床很早就發(fā)現嚴重的燒傷可導致胃、十二指量的細菌和內毒素,主要是依賴(lài)于其腸道屏障的功腸潰瘍,甚至岀現消化道大岀血或穿孔等并發(fā)癥。能。對其屏障的功能行深人地研究,有可能使燒傷的其影響因素有多種,目前尚未完全明了,胃腸黏膜缺救治水平再上1個(gè)臺階。Wnt5a為分泌性信號轉導血和胃液氫離子反滲是主要的發(fā)病原因腸道是人體分子Wnt家族成員。近來(lái),不少研究資料②顯示內最大的“內毒素庫”。正常的腸道之所以能抵抗大Wnt5a在單核巨噬細胞的激活中發(fā)揮著(zhù)重要作用。*[基金項目]南通大學(xué)自然科學(xué)類(lèi)科研課題(117017)*[作者簡(jiǎn)介]祁俊,男,漢族,生于1980年8月,江蘇省南通市人,碩士,主治醫師,研究方向:燒傷整形等*[通信作者]張逸,電話(huà):0513-81168901,E- mail: qijunsky@126cm1005-1013for addiction[J]. Brain Res, 2011, 1406: 59-64[17 Kumar D, Deb L, Chakraborty J, et al. A polymorphism of收稿日期]2013-03-10the CreB binding protein(CrEBBP) gene is a risk fac