【摘要】：目的：通過(guò)人為誘導(dǎo)建立大鼠急性出血壞死性胰腺炎（ANP）動(dòng)物實(shí)驗(yàn)?zāi)Ｐ停瑒?dòng)態(tài)觀察胰腺損傷的各項(xiàng)指標(biāo)，并檢測(cè)凋亡抑制蛋白Survivin的表達(dá)在ANP中的表達(dá)及作用，從而揭示Survivin蛋白在ANP發(fā)病機(jī)制中的作用，為臨床ANP診斷和治療提供新的有效的靶點(diǎn)。 方法：40只健康雄性SD大鼠隨機(jī)分為ANP造模后6h、12h、24h和假手術(shù)組(SO組)共4組，每組10只。手術(shù)前禁食12h，自由飲水。實(shí)驗(yàn)組胰膽管內(nèi)推注4.5%�；悄懰徕c1.0ml/Kg，SO組注射等量生理鹽水。取各不同時(shí)間點(diǎn)處死SD大鼠，從心臟采血測(cè)定血淀粉酶（AMY）、血脂肪酶（LPL）的濃度。各組SD大鼠處死時(shí)取胰頭部胰腺組織,應(yīng)用免疫組化及RT-PCR方法檢測(cè)Survivin蛋白在胰腺組織中的表達(dá)，TUNEL法檢測(cè)凋亡的胰腺組織細(xì)胞的凋亡指數(shù)。 結(jié)果：假手術(shù)組（SO組）大鼠胰腺組織結(jié)構(gòu)清楚，細(xì)胞形態(tài)正常，ANP實(shí)驗(yàn)組胰腺組織充血水腫明顯，胰腺局部出現(xiàn)壞死灶，少量淡紅色血性腹水，胰周脂肪組織皂化、出血。血清AMY于造模后6h即升高，12h左右達(dá)峰值，以后逐漸下降，但仍高于假手術(shù)組。SO組：(1276±213)U/L；ANP組(6343±287)U/L。兩組總體均數(shù)間差別有統(tǒng)計(jì)學(xué)意義（P0.05）。大鼠ANP建立6h血清LPL即明顯下降，隨胰腺炎癥程度的加重，血清LPL值呈逐漸下降趨勢(shì)，各實(shí)驗(yàn)組與假手術(shù)組相比較差別有統(tǒng)計(jì)學(xué)意義（P 0.01）。免疫組化：假手術(shù)組（SO組）胰腺組織細(xì)胞中未見(jiàn)有Survivin蛋白表達(dá)，造模后ANP實(shí)驗(yàn)組Survivin蛋白的表達(dá)在胰腺組織細(xì)胞中主要集中于胰腺腺泡和導(dǎo)管細(xì)胞，呈棕褐色細(xì)顆粒沙狀，ANP6h、12h、24h組Survivin的光密度值積分分別是5743±459、9041±893、13022±803，通過(guò)t-檢驗(yàn)各組間比較差別有統(tǒng)計(jì)學(xué)意義（P 0.05）。RT-PCR：假手術(shù)組胰腺組織中未見(jiàn)Survivin蛋白mRNA的表達(dá)，大鼠ANP造模后6h即可見(jiàn)到Survivin蛋白mRNA的表達(dá)，并且隨ANP病程的進(jìn)展，Survivin蛋白mRNA的表達(dá)水平呈現(xiàn)逐漸增強(qiáng)的趨勢(shì)。胰腺組織凋亡細(xì)胞呈棕黃色，假手術(shù)組胰腺組織細(xì)胞凋亡極少。ANP造模后，隨胰腺炎癥嚴(yán)重程度的加重，胰腺腺泡細(xì)胞凋亡指數(shù)逐漸降低，與假手術(shù)組相比較差別有顯著統(tǒng)計(jì)學(xué)意義（P 0.01），胰腺導(dǎo)管細(xì)胞凋亡指數(shù)逐漸升高，與假手術(shù)組相比較差別有統(tǒng)計(jì)學(xué)意義（P 0.05）。 結(jié)論：1.大鼠ANP動(dòng)物實(shí)驗(yàn)隨病程延長(zhǎng)，胰腺組織細(xì)胞損傷程度呈逐漸加重趨勢(shì)。2.血淀粉酶、血脂肪酶對(duì)ANP診斷有著重要的意義，但對(duì)于評(píng)估ANP損傷嚴(yán)重程度無(wú)重要參考意義。3.Survivin在ANP中的表達(dá)顯著增加，Survivin表達(dá)增加與ANP損傷程度有密切關(guān)系，，對(duì)于判斷ANP預(yù)后有重要意義。
[Abstract]:Aim: to establish an animal model of acute hemorrhagic necrotizing pancreatitis (ANP) in rats by artificial induction, to observe the indexes of pancreatic injury, and to detect the expression and role of apoptosis inhibitor protein Survivin in ANP, so as to reveal the role of Survivin protein in the pathogenesis of ANP and provide a new and effective target for clinical diagnosis and treatment of ANP. Methods: forty healthy male SD rats were randomly divided into 4 groups at 6 h, 12 h, 24 h and SO group (SO group) with 10 rats in each group. Fasting for 12 hours before operation, free drinking water. 4.5% sodium taurocholate 1.0 ml 路kg ~ (- 1) was injected into the pancreatic bile duct of the experimental group, and the same amount of saline was injected into the so group. SD rats were killed at different time points, and the concentration of serum amylase (AMY), blood lipase (LPL) was measured from the heart. The pancreatic tissue of pancreatic head was taken at the time of execution of SD rats in each group. The expression of Survivin protein in pancreatic tissue was detected by immunohistochemistry and RT-PCR, and the apoptosis index of apoptotic pancreatic tissue was detected by TUNEL assay. Results: in the sham operation group (SO group), the structure of pancreatic tissue was clear and the morphology of cells was normal. The hyperemia and edema of pancreatic tissue were obvious in ANP experimental group, and the local necrotic foci, a small amount of reddish hemorrhagic ascitic fluid, saponification and bleeding of peripancreatic adipose tissue were found in ANP experimental group. Serum AMY increased at 6 h after modeling, reached its peak at 12 h, and then decreased gradually, but it was still higher than that in pseudo-operation group. SO group: (1276 鹵213) U 鈮

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