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探討改良線栓對(duì)局灶腦缺血模型制備的影響
【摘要】 目的： 探討改良線栓對(duì)局灶腦缺血模型制備的影響，評(píng)價(jià)該模型的穩(wěn)定性和可靠性. 方法： 雄性Wistar大鼠40只隨機(jī)分為常規(guī)線栓組和改良線栓組，每組20只. 線栓法制備右側(cè)大腦中動(dòng)脈閉塞(MCAO)致局灶腦缺血模型. 觀察改良線栓對(duì)總體手術(shù)時(shí)間、術(shù)中頸總動(dòng)脈血流阻斷時(shí)間、線栓誤入翼腭動(dòng)脈（PPA）幾率（PPA誤栓率）和模型成功率的影響，并通過神經(jīng)行為學(xué)評(píng)分、紅四氮唑(TTC)染色和磁共振檢測等方法評(píng)價(jià)該模型的穩(wěn)定性和可靠性. 結(jié)果： 改良線栓組比常規(guī)線栓組總體手術(shù)時(shí)間和術(shù)中頸總動(dòng)脈血流阻斷時(shí)間明顯縮短，線栓誤入PPA的幾率降低，模型成功率高，穩(wěn)定可靠. 結(jié)論： 改良線栓可增加模型成功率，提高局灶腦缺血模型的穩(wěn)定性和可靠性.【關(guān)鍵詞】 腦缺血；疾病模型，動(dòng)物；縫合技術(shù)；大腦中動(dòng)脈閉塞；大鼠
【Abstract】 AIM: To investigate the application of modified suture in preparation of rat model of focal cerebral ischemia and to evaluate the stability and reliability of the model. METHODS: 40 Wistar male rats were randomized into the conventional suture group and the modified suture group, with 20 in each group. The focal cerebral ischemia by MCAO was induced according to Longa?s suture method. The effects of the application of modified suture on the total operation time, the common carotid artery blood occlusion time during the course of operation, the probability that suture entered the pterygopalatine artery (PPA) and the success rate were observed. And the stability and the reliability of the model were evaluated by neurological deficit score, magnetic resonance image, triphenyltetrazolium chloride (TTC) staining in all rats. RESULTS: The total operation time and the common carotid artery blood occlusion time in the modified suture group were obviously shorter than those in the conventional suture group. And the application of modified suture in the model cut down the probability that the suture entered PPA, and increased the success rate, the stability and the reliability of the model. CONCLUSION： The application of modified suture could improve the success rate, the stability and the reliability of the rat focal cerebral ischemia induced by MCAO.
【Keywords】 brain ischemia; disease models, animal; suture techniques; middle cerebral artery occlusion; rats
0 引言
大鼠的基因與人類的基因具有98%的同源性，其腦血管結(jié)構(gòu)與人類非常相似，血管閉塞后缺血范圍恒定、重復(fù)性好. 因此，眾多有關(guān)腦缺血的研究均選用大鼠來制備模型［1-2］. 制備大鼠局灶腦缺血模型的方法較多，但以線拴法應(yīng)用最廣. 線栓法制備局灶腦缺血模型目前還存在某些不足，如線栓插入困難，栓塞成功率不高，梗塞體積不穩(wěn)定等［3］. 這與線栓直徑、弧度以及術(shù)中頸總動(dòng)脈血流阻斷時(shí)間等因素密切相關(guān).本研究重在對(duì)線栓弧度進(jìn)行改良，觀察改良線栓對(duì)局灶腦缺血模型的影響.
1 材料和方法
1.1 材料 健康雄性Wistar大鼠40只，體質(zhì)量220～250 g，8～10 wk齡，購自重慶醫(yī)科大學(xué)實(shí)驗(yàn)動(dòng)物中心，適應(yīng)性喂養(yǎng)1 wk后，隨機(jī)分為常規(guī)線栓組和改良線栓組，每組20只.
選用“貴花田”牌魚線，直徑0.23～0.26 mm，參照Ma等［4］的方法略加修改，制備改良線栓. 先將魚線剪成3 cm長的小段，60～70℃加熱2 h將魚線小段塑呈弧形，弧形外切角為30°時(shí)最佳，可確保線栓弧度與大鼠頸內(nèi)動(dòng)脈走行一致. 自然冷卻后，頭端約5 mm涂上聚胺酯，用油性記號(hào)筆在線栓頭端和距頭端約2 cm處作上標(biāo)記，以方便植入線栓時(shí)調(diào)整線栓方向，了解線栓進(jìn)入血管的長度. 制備常規(guī)線栓時(shí)，除不進(jìn)行加熱塑型外，其余步驟相同.制備好的線栓頭端向上，垂直插在海綿墊上，風(fēng)干，紫外線消毒后備用. 使用前在生理鹽水配制的肝素鈉(6250 u/mL)溶液中浸漬可用.
1.2 方法 參照Longa等［5］和羅勇等［6］報(bào)道的方法，采用經(jīng)頸內(nèi)動(dòng)脈線栓法制備大鼠右側(cè)MCAO致局灶性永久性腦缺血模型. 35 g/L水合氯醛腹腔注射麻醉大鼠，仰臥位固定，頸前區(qū)備皮、消毒. 取頸正中切口逐層切開皮膚及皮下組織，分離胸鎖乳突肌，切斷二腹肌前腹，暴露右側(cè)頸總動(dòng)脈(common carotid artery，CCA)，頸內(nèi)動(dòng)脈(internal carotid artery，ICA)和頸外動(dòng)脈(external carotid artery，ECA)，探查頸外動(dòng)脈在頸部發(fā)出第一分支（甲狀頸干）處，在其遠(yuǎn)心端（偏嘴側(cè)）約0.2 cm處結(jié)扎并切斷ECA，確保ECA殘端長度不短于0.5 cm；游離、凝閉并切斷ECA與ICA之間的交通支. 用蛙心夾暫時(shí)阻斷CCA及ICA血流，用11號(hào)手術(shù)尖刀于ECA殘端作一縱行小切口，將線栓從右側(cè)ECA殘端插入ICA；輕輕牽拉EC
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