不同营养素的肠内营养配方对大鼠IPAA术后恢复的影响

发布时间：2018-03-31 06:01

  本文选题：营养素　切入点：肠内营养　出处：《天津医科大学》2017年硕士论文

【摘要】：目的:研究添加不同营养素的肠内营养支持方式对大鼠全结直肠切除回肠贮袋肛管吻合术(ileal pouch anal anastomosis,IPAA)后恢复状况及回肠贮袋黏膜屏障功能的影响,为临床溃疡性结肠炎(ulcerative colitis,UC)患者IPAA术后的营养支持提供实验研究依据。方法:将体重在320-350g范围内的雄性Sprague-Dawley(SD)大鼠随机分为4组(n=8),分别为对照组,肠内营养(Enteral Nutrition,EN)组,肠内营养+免疫营养剂组即免疫营养(Enteral Nutrition+Immune agents,NI)组,肠内营养+益生菌组即微生态营养(Enteral Nutrition+Probiotics,NP)组,利用显微外科的操作技术对大鼠进行手术,建立大鼠IPAA手术术后模型。术后前两天,所有大鼠均给予葡萄糖氯化钠溶液自由饮用。于术后第3天起给予对照组大鼠正常鼠粮饲养,肠内营养组大鼠给予百普素喂养,免疫营养组大鼠给予百普素+谷氨酰胺喂养,微生态营养组大鼠给予百普素+嗜酸乳杆菌喂养,观察并且记录大鼠的一般情况(精神状态,活动度及毛发,首次排便时间,粪便性状)、体重变化等,其中采用Bristol粪便评分对大鼠粪便性状进行评分。于术后第30天时,留取血标本,采用全自动生化分析仪检测大鼠血清总蛋白(Toatal Protein,TP)、白蛋白(Albumin,ALB)、前白蛋白(Prealbumin,PA)、转铁蛋白(Transferrin,TF)水平的变化;留取便标本,采用粪便快速涂片法检测大鼠粪便菌群的变化;留取大鼠回肠贮袋标本组织,HE染色法观察大鼠贮袋黏膜形态的变化并进行组织病理学评分,免疫组化方法测定各组大鼠贮袋黏膜组织中机械屏障紧密连接蛋白Occludin蛋白的表达水平。使用spss19.0软件分析结果数据,计量资料采用均数±标准差(±s)进行描述。进行统计学分析时,正态分布资料组间比较采用单因素方差分析,之后各组之间两两比较,采用LSD法。结果:1.大鼠的一般状况(1)精神状态、活动度及毛发:对照组、EN组、NI组、NP组四组大鼠精神状态均良好好,反应较灵敏,毛发有光泽;(2)首次排便时间:四组大鼠比较,差异无统计学意义(P0.05);(3)粪便性状:EN组、xNI组、NP组大鼠粪便评分高于对照组,差异有统计学意义(P0.01);EN组、NI组、NP组三组大鼠粪便评分比较,差异无统计学意义(P0.05)。2.大鼠的营养状况(1)术前,4组大鼠体重比较,差异均无统计学意义(P0.05);术后,大鼠的体重均出现短期的下降,四组比较,EN、NI组、NP组大鼠体重降至最低水平所用的时间及下降的幅度均小于对照组大鼠,差异有统计学意义(P0.01),EN、NI组和NP组相比,后两组所用时间及下降的幅度均小于EN组,差异有统计学意义(P0.05),EN、NI组相比,差异无统计学意义(P0.05);术后30天时,EN组、NI组和NP组大鼠体重均高于对照组,差异有统计学意义(F=11.657,P0.01),NI组和NP组大鼠体重均高于EN组大鼠,差异有统计学意义(P0.05),NI组和NP组大鼠体重比较,差异无统计学意义(P0.05)(2)血检测结果显示:术后,EN组、NI组、NP组三组大鼠血清TP水平、PA水平、ALB水平及TF水平均高于对照组大鼠,差异有统计学意义(P0.01),其中,NI组、NP组两组大鼠血清TP水平、PA水平、ALB水平及TF水平高于EN组大鼠,差异有统计学意义(P0.05),NI组、NP组两组大鼠之间比较,差异无统计学意义(P0.05)。3.大鼠的回肠贮袋粘膜屏障功能(1)粪便菌群分布检测结果显示:对照组大鼠粪便菌群分布:粪便细菌数较正常范围减少,革兰阳性杆菌显著减少,革兰阴性杆菌显著增加,革兰氏阳性球菌占比正常;EN组、NI组大鼠粪便菌群分布:粪便细菌数较正常范围略减少,革兰阳性杆菌略减少,革兰阴性杆菌略增加,革兰氏阳性球菌占比正常;NP组大鼠粪便菌群分布:细菌数及革兰阳性杆菌、革兰阴性杆菌在正常范围内,革兰阳性球菌占比正常。四组大鼠菌群分布比较,细菌数及革兰阳性杆菌占比,EN组、NI组、NP组均高于对照组,差异均有统计学意义(P0.05),其中NP组高于EN组和NI组,差异有统计学意义(P0.05),EN组和NI组相比,差异无统计学意义(P0.05);革兰阴性杆菌占比,EN组、NI组、NP组均低于对照组,差异均有统计学意义(P0.05)其中,NP组低于EN组及NI组,差异有统计学意义(P0.05),EN组及NI组比较,差异无统计学意义(P0.05);革兰阳性球菌和革兰阴性球菌占比,四组比较差异无统计学意义(P0.05)(2)免疫组化结果显示:Occludin蛋白在EN组、NI组、NP组三组大鼠回肠贮袋组织表达水平高于对照组,差异有统计学意义(P0.05),且EN、NI组和NP组相比较,差异无统计学意义(P0.05)。4.病理组织学检测结果显示:对照组大鼠贮袋黏膜绒毛的完整性被破坏,绒毛间质较疏松、贮袋绒毛排列不规则,绒毛出现中度萎缩,固有层出现水肿;EN组、NI组、NP组三组大鼠贮袋黏膜的基本形态正常,贮袋黏膜绒毛排列整齐,结构完整,上皮细胞排列整齐,偶有间质水肿;EN组、NI组、NP组病理学评分均高于对照组,差异有统计学意义(P0.05),EN组、NI组、NP组三组之间比较,差异无统计学意义(P0.05)。结论:1.肠内营养、免疫营养剂及益生菌可以有效地改善大鼠IPAA术后的营养状况,加速大鼠的术后恢复,其中肠内营养联合免疫营养剂和肠内营养联合益生菌的作用效果优于单纯肠内营养。2.肠内营养联合益生菌可以有效改善大鼠粪便的菌群分布,增加回肠贮袋屏障蛋白Occludin的表达水平,保护回肠贮袋黏膜屏障功能;肠内营养和肠内营养联合免疫营养剂增加回肠贮袋屏障蛋白Occludin的表达水平,从而改善回肠贮袋黏膜屏障功能。3.肠内营养、肠内营养联合免疫营养剂及肠内营养联合益生菌可以有效地改善大鼠IPAA术后的恢复状况及回肠贮袋黏膜屏障功能,这为临床UC患者IPAA术后的营养支持提供了客观的实验研究依据。
[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 寮傛湁缁熻�″�︽剰涔，

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