本文選題：缺血再灌注 + 腦損傷　； 參考：《蚌埠醫(yī)學(xué)院》2014年碩士論文

【摘要】：目的：建立腦缺血再灌注損傷模型，觀察磷酸肌酸對(duì)腦細(xì)胞損傷的保護(hù)效果，通過對(duì)大鼠海馬組織即刻基因c-fosmRNA基因表達(dá)的檢測(cè)，探討損傷時(shí)磷酸肌酸對(duì)腦的保護(hù)作用和機(jī)制。 方法：本實(shí)驗(yàn)選用體重在300-350g之間的27只健康成年雄性Sprague-Dawley（SD）大鼠作為研究對(duì)象，隨機(jī)分組:1、假手術(shù)組(Sham組，n=9)：麻醉（10%水合氯醛，，350mg·kg-1腹腔注射)后頸部正中切口，分離出兩側(cè)頸總動(dòng)脈但不夾閉，暴露30min；2、缺血再灌注組(I/R組，n=9)：麻醉后分離并夾閉雙側(cè)頸總動(dòng)脈30min，松開動(dòng)脈夾后再灌注24h；3、藥物處理組(Drug組，n=9)：麻醉后分離并夾閉雙側(cè)頸總動(dòng)脈30min時(shí)松開動(dòng)脈夾，同時(shí)從尾靜脈緩慢注射磷酸肌酸（PCr，80mg·100g-1)，再灌注24h。術(shù)后三組大鼠均放籠飼養(yǎng)24h后直接斷頭，在冰屑上迅速開顱取海馬組織，左側(cè)海馬進(jìn)行液氮速凍后放入-80℃冰箱保存以備采用逆轉(zhuǎn)錄聚合酶鏈?zhǔn)椒磻?yīng)（RT-PCR)對(duì)c-fos mRNA的檢測(cè)，右側(cè)海馬放入4%的甲醛保存以備免疫組織化學(xué)法進(jìn)行檢測(cè)。 結(jié)果：兩只大鼠因呼吸衰竭死亡，其余均度過觀察期。Durg組的大鼠比Sham組大鼠精神狀態(tài)佳，活力強(qiáng)。1、HE染色法結(jié)果比較：Sham組的神經(jīng)細(xì)胞排列緊密、規(guī)則，形態(tài)結(jié)構(gòu)基本正常；與Sham組相比，I/R組的神經(jīng)元細(xì)胞排列散亂，細(xì)胞結(jié)構(gòu)破壞，核碎裂、核固縮；與I/R組相比，Drug組大部分神經(jīng)元細(xì)胞結(jié)構(gòu)完整，排列有序；2、免疫組化染色觀察c-fos基因表達(dá)結(jié)果比較：c-fos mRNA呈棕黃色的免疫陽性細(xì)胞在Sham組海馬組織僅有少量表達(dá)，染色淺；I/R組大鼠海馬組織中染色較深，且表達(dá)水平高；Drug組大鼠海馬組織c-fos mRNA在細(xì)胞核內(nèi)有所表達(dá)，與I/R組相比，染色減輕。3、RT-PCR技術(shù)檢測(cè)結(jié)果比較：電泳圖中三組β-actin電泳條帶寬度和亮度無明顯差別，I/R組的c-fos電泳條帶寬厚、明亮；Sham組的c-fos mRNA條帶模糊、暗淡；Drug組條帶亮度介于兩者之間。凝膠成像分析系統(tǒng)結(jié)果顯示：I/R組的c-fos/β-actin比值大于Drug組(p0.05)；Drug組的c-fos/β-actin比值大于Sham組(p0.05)。 結(jié)論：1、腦缺血再灌導(dǎo)致的腦損傷注可以增加神經(jīng)細(xì)胞c-fosmRNA的表達(dá)；2、磷酸肌酸可以降低c-fosmRNA的表達(dá)，保護(hù)腦細(xì)胞；3、C-fosmRNA可以作為腦損傷早期診斷的一個(gè)指標(biāo)。
[Abstract]:Aim: to establish a model of cerebral ischemia-reperfusion injury and to observe the protective effect of creatine phosphate on brain cell injury, and to investigate the protective effect and mechanism of creatine phosphate on brain by detecting the expression of immediate gene c-fosmRNA gene in rat hippocampal tissue.Methods: 27 healthy adult male Sprague-Dawley SD rats weighing between 300g and 350g were randomly divided into two groups: 1, sham group, 10% chloral hydrate and 350 mg kg-1 intraperitoneal injection).Separating bilateral common carotid arteries without clipping,After 30 minutes of exposure, I / R group: after anesthesia, the bilateral common carotid arteries were separated and clamped for 30 minutes, and then re-perfused for 24 hours. In the drug treatment group, when the bilateral common carotid artery 30min was separated and clipped after anesthesia, the clamp was released.At the same time, 80 mg / 100g ~ (-1) of creatine phosphate was injected slowly into the tail vein and perfused for 24 h.After 24 hours of cage feeding, the rats in the three groups were cut off directly, and the hippocampus tissue was quickly removed from the ice chip. The left hippocampus was frozen with liquid nitrogen and stored in a refrigerator at -80 鈩

本文編號(hào)：1760595