本文選題：營養(yǎng)素　切入點：腸內(nèi)營養(yǎng)　出處：《天津醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:研究添加不同營養(yǎng)素的腸內(nèi)營養(yǎng)支持方式對大鼠全結(jié)直腸切除回腸貯袋肛管吻合術(shù)(ileal pouch anal anastomosis,IPAA)后恢復(fù)狀況及回腸貯袋黏膜屏障功能的影響,為臨床潰瘍性結(jié)腸炎(ulcerative colitis,UC)患者IPAA術(shù)后的營養(yǎng)支持提供實驗研究依據(jù)。方法:將體重在320-350g范圍內(nèi)的雄性Sprague-Dawley(SD)大鼠隨機(jī)分為4組(n=8),分別為對照組,腸內(nèi)營養(yǎng)(Enteral Nutrition,EN)組,腸內(nèi)營養(yǎng)+免疫營養(yǎng)劑組即免疫營養(yǎng)(Enteral Nutrition+Immune agents,NI)組,腸內(nèi)營養(yǎng)+益生菌組即微生態(tài)營養(yǎng)(Enteral Nutrition+Probiotics,NP)組,利用顯微外科的操作技術(shù)對大鼠進(jìn)行手術(shù),建立大鼠IPAA手術(shù)術(shù)后模型。術(shù)后前兩天,所有大鼠均給予葡萄糖氯化鈉溶液自由飲用。于術(shù)后第3天起給予對照組大鼠正常鼠糧飼養(yǎng),腸內(nèi)營養(yǎng)組大鼠給予百普素喂養(yǎng),免疫營養(yǎng)組大鼠給予百普素+谷氨酰胺喂養(yǎng),微生態(tài)營養(yǎng)組大鼠給予百普素+嗜酸乳桿菌喂養(yǎng),觀察并且記錄大鼠的一般情況(精神狀態(tài),活動度及毛發(fā),首次排便時間,糞便性狀)、體重變化等,其中采用Bristol糞便評分對大鼠糞便性狀進(jìn)行評分。于術(shù)后第30天時,留取血標(biāo)本,采用全自動生化分析儀檢測大鼠血清總蛋白(Toatal Protein,TP)、白蛋白(Albumin,ALB)、前白蛋白(Prealbumin,PA)、轉(zhuǎn)鐵蛋白(Transferrin,TF)水平的變化;留取便標(biāo)本,采用糞便快速涂片法檢測大鼠糞便菌群的變化;留取大鼠回腸貯袋標(biāo)本組織,HE染色法觀察大鼠貯袋黏膜形態(tài)的變化并進(jìn)行組織病理學(xué)評分,免疫組化方法測定各組大鼠貯袋黏膜組織中機(jī)械屏障緊密連接蛋白Occludin蛋白的表達(dá)水平。使用spss19.0軟件分析結(jié)果數(shù)據(jù),計量資料采用均數(shù)±標(biāo)準(zhǔn)差(±s)進(jìn)行描述。進(jìn)行統(tǒng)計學(xué)分析時,正態(tài)分布資料組間比較采用單因素方差分析,之后各組之間兩兩比較,采用LSD法。結(jié)果:1.大鼠的一般狀況(1)精神狀態(tài)、活動度及毛發(fā):對照組、EN組、NI組、NP組四組大鼠精神狀態(tài)均良好好,反應(yīng)較靈敏,毛發(fā)有光澤;(2)首次排便時間:四組大鼠比較,差異無統(tǒng)計學(xué)意義(P0.05);(3)糞便性狀:EN組、xNI組、NP組大鼠糞便評分高于對照組,差異有統(tǒng)計學(xué)意義(P0.01);EN組、NI組、NP組三組大鼠糞便評分比較,差異無統(tǒng)計學(xué)意義(P0.05)。2.大鼠的營養(yǎng)狀況(1)術(shù)前,4組大鼠體重比較,差異均無統(tǒng)計學(xué)意義(P0.05);術(shù)后,大鼠的體重均出現(xiàn)短期的下降,四組比較,EN、NI組、NP組大鼠體重降至最低水平所用的時間及下降的幅度均小于對照組大鼠,差異有統(tǒng)計學(xué)意義(P0.01),EN、NI組和NP組相比,后兩組所用時間及下降的幅度均小于EN組,差異有統(tǒng)計學(xué)意義(P0.05),EN、NI組相比,差異無統(tǒng)計學(xué)意義(P0.05);術(shù)后30天時,EN組、NI組和NP組大鼠體重均高于對照組,差異有統(tǒng)計學(xué)意義(F=11.657,P0.01),NI組和NP組大鼠體重均高于EN組大鼠,差異有統(tǒng)計學(xué)意義(P0.05),NI組和NP組大鼠體重比較,差異無統(tǒng)計學(xué)意義(P0.05)(2)血檢測結(jié)果顯示:術(shù)后,EN組、NI組、NP組三組大鼠血清TP水平、PA水平、ALB水平及TF水平均高于對照組大鼠,差異有統(tǒng)計學(xué)意義(P0.01),其中,NI組、NP組兩組大鼠血清TP水平、PA水平、ALB水平及TF水平高于EN組大鼠,差異有統(tǒng)計學(xué)意義(P0.05),NI組、NP組兩組大鼠之間比較,差異無統(tǒng)計學(xué)意義(P0.05)。3.大鼠的回腸貯袋粘膜屏障功能(1)糞便菌群分布檢測結(jié)果顯示:對照組大鼠糞便菌群分布:糞便細(xì)菌數(shù)較正常范圍減少,革蘭陽性桿菌顯著減少,革蘭陰性桿菌顯著增加,革蘭氏陽性球菌占比正常;EN組、NI組大鼠糞便菌群分布:糞便細(xì)菌數(shù)較正常范圍略減少,革蘭陽性桿菌略減少,革蘭陰性桿菌略增加,革蘭氏陽性球菌占比正常;NP組大鼠糞便菌群分布:細(xì)菌數(shù)及革蘭陽性桿菌、革蘭陰性桿菌在正常范圍內(nèi),革蘭陽性球菌占比正常。四組大鼠菌群分布比較,細(xì)菌數(shù)及革蘭陽性桿菌占比,EN組、NI組、NP組均高于對照組,差異均有統(tǒng)計學(xué)意義(P0.05),其中NP組高于EN組和NI組,差異有統(tǒng)計學(xué)意義(P0.05),EN組和NI組相比,差異無統(tǒng)計學(xué)意義(P0.05);革蘭陰性桿菌占比,EN組、NI組、NP組均低于對照組,差異均有統(tǒng)計學(xué)意義(P0.05)其中,NP組低于EN組及NI組,差異有統(tǒng)計學(xué)意義(P0.05),EN組及NI組比較,差異無統(tǒng)計學(xué)意義(P0.05);革蘭陽性球菌和革蘭陰性球菌占比,四組比較差異無統(tǒng)計學(xué)意義(P0.05)(2)免疫組化結(jié)果顯示:Occludin蛋白在EN組、NI組、NP組三組大鼠回腸貯袋組織表達(dá)水平高于對照組,差異有統(tǒng)計學(xué)意義(P0.05),且EN、NI組和NP組相比較,差異無統(tǒng)計學(xué)意義(P0.05)。4.病理組織學(xué)檢測結(jié)果顯示:對照組大鼠貯袋黏膜絨毛的完整性被破壞,絨毛間質(zhì)較疏松、貯袋絨毛排列不規(guī)則,絨毛出現(xiàn)中度萎縮,固有層出現(xiàn)水腫;EN組、NI組、NP組三組大鼠貯袋黏膜的基本形態(tài)正常,貯袋黏膜絨毛排列整齊,結(jié)構(gòu)完整,上皮細(xì)胞排列整齊,偶有間質(zhì)水腫;EN組、NI組、NP組病理學(xué)評分均高于對照組,差異有統(tǒng)計學(xué)意義(P0.05),EN組、NI組、NP組三組之間比較,差異無統(tǒng)計學(xué)意義(P0.05)。結(jié)論:1.腸內(nèi)營養(yǎng)、免疫營養(yǎng)劑及益生菌可以有效地改善大鼠IPAA術(shù)后的營養(yǎng)狀況,加速大鼠的術(shù)后恢復(fù),其中腸內(nèi)營養(yǎng)聯(lián)合免疫營養(yǎng)劑和腸內(nèi)營養(yǎng)聯(lián)合益生菌的作用效果優(yōu)于單純腸內(nèi)營養(yǎng)。2.腸內(nèi)營養(yǎng)聯(lián)合益生菌可以有效改善大鼠糞便的菌群分布,增加回腸貯袋屏障蛋白Occludin的表達(dá)水平,保護(hù)回腸貯袋黏膜屏障功能;腸內(nèi)營養(yǎng)和腸內(nèi)營養(yǎng)聯(lián)合免疫營養(yǎng)劑增加回腸貯袋屏障蛋白Occludin的表達(dá)水平,從而改善回腸貯袋黏膜屏障功能。3.腸內(nèi)營養(yǎng)、腸內(nèi)營養(yǎng)聯(lián)合免疫營養(yǎng)劑及腸內(nèi)營養(yǎng)聯(lián)合益生菌可以有效地改善大鼠IPAA術(shù)后的恢復(fù)狀況及回腸貯袋黏膜屏障功能,這為臨床UC患者IPAA術(shù)后的營養(yǎng)支持提供了客觀的實驗研究依據(jù)。
[Abstract]:Objective: To study the effects of different nutrition enteral nutrition on rats with total proctocolectomy with ileal pouch anal anastomosis (ileal pouch anal anastomosis, IPAA) effect after the restoration of mucosal barrier function and ileal pouch, clinical ulcerative colitis (ulcerative colitis, UC IPAA) in patients with postoperative nutritional support on the basis of experimental study. Methods: the weight in the range of 320-350g Sprague-Dawley (SD) male rats were randomly divided into 4 groups (n=8), respectively in the control group, enteral nutrition (Enteral, Nutrition, EN) group, enteral nutrition and immune nutrition group (Enteral Nutrition+Immune, agents, immune nutrition group NI) enteral nutrition and probiotics group, namely microecological nutrition (Enteral group, Nutrition+Probiotics, NP) performed surgery on rats using microsurgical technique, rat IPAA hand surgery model. The first two days after surgery, all rats were Sodium Chloride Solution dextrose free drink. In the third day after operation for the rats in the control group of normal rats fed with diets, enteral nutrition group rats were treated by peptisorb feeding, immunonutrition group rats were treated by peptisorb + glutamine feeding, microecological nutrition group rats were given by peptisorb + Lactobacillus acidophilus feeding. Observe and record the general condition of rats (mental state, activity and hair, the first defecation time, stool), body weight change, in which the Bristol stool score score on the rat stool. On the thirtieth day after operation, the blood samples, using automatic biochemical analyzer to detect the serum of rats the total protein (Toatal Protein, TP), albumin (Albumin, ALB), prealbumin (Prealbumin, PA), transferrin (Transferrin, TF) level changes; collected fecal specimens of feces smear detection by rapid changes in fecal flora in rats; the rats were Ileal pouch tissue scores observation pouch mucosa morphology in rats HE staining and pathological changes, the expression level of mechanical barrier tight junction protein Occludin protein immunohistochemistry method for determination of rat pouch mucosa. The results of data analysis using spss19.0 software, the measurement data using standard deviation (+ s) are described. By statistical analysis, normal distribution data were compared with single factor analysis of variance between the 22 groups, after comparison, using LSD method. Results: the general condition of 1. rats (1) mental state, activity and hair: control group, EN group, NI group. NP group of mental state of four groups of rats are good, sensitive reaction, hair shiny; (2) the first defecation time: comparison of four groups of rats, there was no statistically significant difference (P0.05); (3) stool: EN group, xNI group, NP group rats feces score higher than the control group, difference 寮傛湁緇熻瀛︽剰涔，

本文編號：1689438