發(fā)布時間：2018-03-04 23:34

  本文選題：SPA　切入點：EEN　出處：《瀘州醫(yī)學(xué)院》2013年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：通過檢測早期腸內(nèi)營養(yǎng)對牛黃膽酸鈉誘導(dǎo)大鼠重癥急性胰腺炎模型的血清淀粉酶(AMY)、血糖(GLU)、靜脈血中內(nèi)毒素的含量，末段回腸和結(jié)腸peyer結(jié)MAdcAM-1分子(mucosal addressin celladhesion molecule-1)CD4+、CD8+含量。和大鼠門靜脈血、胰腺、肺臟、腸系膜淋巴結(jié)的細(xì)菌培養(yǎng)。以及大鼠腸道的TLR4表達(dá)，觀察早期腸內(nèi)營養(yǎng)在大鼠重癥急性胰腺炎腸道免疫屏障里的功能和研究EEN功效原理。方法：把成年身體狀況良好的60只雄性大鼠體重約為300克左右，隨機(jī)分成三組，每組20只大鼠。即假手術(shù)組(SO組)：開腹后僅輕輕翻動十二指腸及胰腺組織后關(guān)腹，不作處理。將3.8％�；悄懰徕clml從大鼠胰腺被膜下各點徐徐勻速注入胰腺從而完成動物模型建立。造模成功后按組分別給予腸外營養(yǎng)和腸內(nèi)營養(yǎng)。腸外營養(yǎng)組(TPN組)，造模后12小時，經(jīng)頸外靜脈置管輸注腸外營養(yǎng)液。腸內(nèi)營養(yǎng)組(EEN組)，造模12小時后，經(jīng)胃，空腸做空腸造瘺管，予以腸內(nèi)營養(yǎng)。各組于造模后一天，經(jīng)SD大鼠頸靜脈取血，檢測血清淀粉酶(AMY)、血糖(GLU)水平。腸外營養(yǎng)、腸內(nèi)營養(yǎng)持續(xù)給予五天，第六天兩組分別統(tǒng)計SD大鼠死亡率，隨即處死大鼠并收集標(biāo)本，胰腺組織HE染色后行光鏡病理學(xué)檢查�？漳c、回腸、結(jié)腸HE染色后觀察腸粘膜形態(tài)學(xué)。免疫組化法測定末端回腸和Peyer結(jié)腸道的MAdcAM-1、CD4+、CD8+的含量。Westen blot法測定SD大鼠腸道組織TLR4的表達(dá)。SD大鼠靜脈血檢測內(nèi)毒素含量。取SD大鼠門靜脈血、胰腺、肺臟、腸系膜淋巴結(jié)做細(xì)菌培養(yǎng)，檢測細(xì)菌移位率。此課題為四川省綿陽市中心醫(yī)院趙平武教授申請的四川省應(yīng)用基礎(chǔ)研究的課題。結(jié)果：(1)各組SD大鼠5天的生存率：假手術(shù)組(SO組)SD大鼠全部存活生存率是100％，死亡率為零；腸外營養(yǎng)組(TPN組)5天死亡率為50%，早期腸內(nèi)營養(yǎng)組(EEN組)5天死亡率為25%，S組和其它實驗組的死亡率的對照存在明顯差異(P0.01)。TPN和EEN組的死亡率對照無明顯差異無統(tǒng)計學(xué)意義(P=0.0847)。證明EEN是安全可行的。(2)SAP下SD大鼠血清淀粉酶改變：TPN、EEN組血淀粉酶含量在建模24小時后都明顯上升，與SO組對照有明顯差異(P0.01)。在建模24小時后，EEN組和TPN對照，血清淀粉酶含量降低，有統(tǒng)計學(xué)意義(P0.01)。(3)血糖變化：造模后TPN、EEN組血糖水平在建模24小時后均顯著升高，與SO組比較有顯著性統(tǒng)計學(xué)差異(P0.01)；在建模24小時后，EEN組與TPN組比，血糖含量無明顯變化，但有顯著統(tǒng)計學(xué)差異(P0.01)。(5)胰腺組織學(xué)評分：TPN、EEN組建模5天后所采集標(biāo)本組織學(xué)評分與SO組相比，均明顯升高。與S組比較有顯著差異性(P0.01)。而TPN、EEN組相比，TPN組評分高于EEN組而且有顯著性統(tǒng)計學(xué)差異(P0.01)。(6)各組SD大鼠靜脈血內(nèi)毒素含量測定：本實驗中EEN、TPN兩組動物在建立SAP模型5天后血內(nèi)毒素水平明顯比SO組升高，說明EEN、TPN兩組均有不同程度的腸道屏障損傷，而EEN組血漿內(nèi)毒素含量與TPN組對比明顯偏低，具有統(tǒng)計學(xué)意義，(P0.05)。說明SAP早期腸內(nèi)營養(yǎng)有效降低了SD大鼠SAP時腸粘膜的通透性，有效保護(hù)了其腸粘膜屏障。(7)胰腺、肺臟、腸系膜淋巴結(jié)細(xì)菌培養(yǎng)和細(xì)菌易位率測定：細(xì)菌類型主要為革蘭陰性菌，其中大腸桿菌占85.7％(12/14)。各種細(xì)菌易位率EEN組遠(yuǎn)低于TPN組，具有統(tǒng)計學(xué)意義(P0.01)。而EEN、TPN組與SO組相比細(xì)菌移位率遠(yuǎn)遠(yuǎn)高于后者，具有明顯統(tǒng)計學(xué)意義P0.01.(8)末端回腸黏膜和Peyer結(jié)CD4+、CD8+、MadcAM-1，免疫組化測定：CD4+、CD8+、MadcAM-1在EEN、TPN組表達(dá)均低于SO組(P0.05)，其中TPN組低于EEN組(p0.05）(9)回腸黏膜光鏡下改變：在第5d時，TPN組腸粘膜萎縮程度包括黏膜厚度、絨毛長度與EEN組對照，腸粘膜萎縮程度明顯加重，有顯著性差異(p0.01)。(10)Westen blot法測定SD大鼠腸道組織TLR4的表達(dá)：光密度掃描分析顯示，EEN、TPN組與SO組相比腸組織TLR4表達(dá)增強(qiáng)(P0.01）。EEN組與TPN組相比，，腸組織TLR4表達(dá)減弱(P0.01)。結(jié)論：(1)將3．8％�；悄懰徕clml緩慢勻速注入SD大鼠胰腺被膜下，能建立SAP動物模型。它具有重復(fù)性好，操作簡便，價格便宜的優(yōu)點。而且SD大鼠SAP動物模型病情呈漸進(jìn)性發(fā)展。和患者SAP的疾病演變過程十分相像。5d死亡率穩(wěn)定。完全能夠滿足SAP的早期腸內(nèi)營養(yǎng)治療研究。(2)早期腸內(nèi)營養(yǎng)不加重胰腺炎病情，是安全、可行的，而且早期腸內(nèi)營養(yǎng)在維護(hù)重癥胰腺炎SD大鼠腸道屏障方面的作用效果顯著，對腸道屏障的化學(xué)、免疫、生物、機(jī)械的幾方面的功能提高幾乎均有效。尤其是EEN能增加MAdcAM-1、CD4+、CD8+表達(dá)，提高SAP時腸免疫屏障，減少茵群易位，減少局部及全身感染。可以提高急性重癥胰腺炎SD大鼠5d生存率。(3)研究成果預(yù)測，TLR4通過和它主要識別G—細(xì)菌細(xì)胞壁成分脂多糖（LPS）相結(jié)合后其復(fù)合物在SAP時導(dǎo)致的腸道屏障受損中起到關(guān)鍵作用，TLR4表達(dá)強(qiáng)度與SAP時炎癥的強(qiáng)弱有一定的正相關(guān)性。
[Abstract]:Objective: through the detection of serum amylase of early enteral nutrition on yellow taurocholate induced severe acute pancreatitis model in rats (AMY), blood glucose (GLU), the content of endotoxin in blood, the terminal ileum and colon Peyer MAdcAM-1 (mucosal addressin celladhesion molecule-1) molecules CD4+, CD8+ content. And the rat portal vein blood. The pancreas, lung, bacterial culture of mesenteric lymph nodes. And rat intestinal TLR4 expression, observation function and study of EEN effect principle of early enteral nutrition on intestinal barrier in rats with severe acute pancreatitis. Methods: 60 male adult rats in good physical condition is about 300 grams, random divided into three groups, 20 rats in each group. Sham operation group (SO group): abdominal laparotomy only gently flip the duodenum and pancreas after no treatment. 3.8% sodium taurocholate LML from rat pancreatic capsule each The point slowly injected into the pancreas to complete the animal model. After the success of the model group were given parenteral nutrition and enteral nutrition Parenteral Nutrition Group (TPN group), 12 hours after modeling, through external jugular venous catheter infusion of parenteral nutrition. Enteral nutrition group (EEN group). Modeling after 12 hours, the stomach, jejunum do jejunal fistula, given enteral nutrition. Each day after modeling, jugular vein in SD rats blood serum amylase (AMY), blood glucose (GLU) levels. Parenteral nutrition, enteral nutrition for five days, sixth the two day mortality statistics were SD rats, then the rats were sacrificed and samples were collected, pancreatic tissue HE staining under light microscope. The pathological examination of jejunum, ileum, colon mucosal morphology were observed after HE staining. Immunohistochemical determination of terminal ileum and colon Peyer by MAdcAM-1, CD4+, TLR4 determination of intestinal tissue SD the content of.Westen blot CD8+ in the To detect the expression of endotoxin in blood of.SD rats. SD rats of portal vein blood, pancreas, lung, mesenteric lymph nodes were cultured for bacteria, the detection rate of bacterial translocation. Subject to the basic application research of Sichuan Province, Sichuan province Mianyang Central Hospital professor Zhao Pingwu for the study. Results: (1) the 5 day survival rate in each group SD rats: sham operation group (group SO) SD rats survived survival rate was 100%, the mortality rate is zero; parenteral nutrition group (TPN group) the 5 day mortality rate was 50%, the early enteral nutrition group (EEN group) the 5 day mortality rate was 25%, there are obvious differences in S control group and the other groups the mortality rate (P0.01) and.TPN EEN group were no significant difference between the mortality rate was not statistically significant (P=0.0847). EEN is safe and feasible. (2) serum amylase in SD rats under SAP change: TPN, amylase content of blood EEN 24 hours after modeling were significantly increased, with control group and SO group 鏄庢樉宸紓(P0.01).鍦ㄥ緩妯

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