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硕士学位论文海兔素对酒精性肝损伤大鼠小肠黏膜屏障的保护作用研究傅泳指导教师少儿英语系列
梁惠教授学科专业名称
心得体会网营养与食品卫生学论文答辩日期2016年5月31日硕
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订单英文博
毕业论文社区
摘要
目的研究海兔素（Aplysin ）对酒精性肝损伤大鼠血浆中D-乳酸（D-LA ）、二胺氧化酶（DAO ）、肠脂肪酸结合蛋白（FABP2）和炎性因子（TNF-α、TGF-β）及肠组织中叉头样转录因子O4（FOXO4）等的影响，以初步探讨海兔素对酒精性肝损伤大鼠肠黏膜损伤的保护作用及初步作用机制。
方法1.动物分组及模型建立：两月龄健康雄性Wistar 大鼠60只，按体重随机分为4组，每组15只。酒精模型组给予56℃白酒灌胃，第1周5.5ml/(kg•bw•d)，第2周
7.5ml/(kg•bw•d)，第3周8.0ml/(kg•bw•d)，第4周9.0ml/(kg•bw•d)，第5周至第8周11ml/(kg•bw•d)；海兔素低、高剂量组首先分别给予Aplysin100、150mg/(kg•bw•d)灌胃，1h 后再给予与模型组相同剂量的酒精灌胃；空白对照组灌胃给予10ml/(kg•bw•d)生理盐水，实验共持续8周。大鼠末次灌胃后，禁食12h ，按体重给予3%戊巴比妥钠30mg/kg 麻醉，腹主动脉取血及小肠组织。2.透射电镜观察大鼠小肠超微结构变化。3.酶联免疫吸附实验（ELISA ）检测大鼠血浆中FABP2、TNF-α、TGF-β、D-LA 及DAO 浓度。4.免疫组化实验检测小肠组织中FOXO4的表达情况，计算其阳性表达率。
结果1.透射电镜观察发现，空白对照组小肠上皮柱状细胞排列整齐、紧密，微绒毛丰富，紧密连接结构清晰完整；酒精模型组大鼠小肠上皮细胞表面微绒毛排列稀疏，细胞间紧密连接肿大，海兔素高、低剂量组小肠上皮柱状细胞及微绒毛排列情况较酒精模型组得到不同程度改善，紧密连接肿胀减轻；2.酒精模型组大鼠血浆中D-LA 、DAO 和FABP2的浓度显著增加（P <0.05），而海兔素干预后，可有效抑制酒精诱导的血浆D-LA 、DAO 和FABP2的浓度升高（P <0.05）；3.免疫组化结果显示，酒精模型组小肠组织FOXO4表达水平明显降低，较空白对照组OD 值降低约50%（P <0.05）；海兔素低、高剂量组FOXO4的表达水平显著增高，OD 值均显著高于模型组（P <0.05）。4.酒精模型组大鼠血浆中炎性因子TNF-α、TGF-β浓度均较空白对照组显著升（P <0.05），而海兔素低、高剂量组血浆中TNF-α、TGF-β浓度均较酒精模型组明显降低（P <0.05）。
结论1.通过对小肠组织超微结构的观察、血浆中D-LA 、DAO 及FABP2水平的检测，初步验证了海兔素对酒精性肝损伤大鼠肠黏膜屏障具有一定的保护作用；2.海兔素对酒精性肝损伤大鼠的小肠黏膜屏障的保护作用机制可能与海兔素调节炎性反应相关蛋白FOXO4的表达，减少炎性因子TNF-α、TGF-β的释放有关。硕博毕业论文社区
硕士研究生傅泳（营养与食品卫生学）指导教师梁惠教授关键词：海兔素；ALD ；肠黏膜屏障损伤；FOXO4硕
博youhu
毕业论文社区
Abstract
luggageObjective:To study the effects of Aplysin on rum TNF-α、FABP2、TGF-β、D-LA 、DAO concentration and FOXO4expression in intestine in rats of alcohol-induced hepatic injury.eachof
Methods:1.Experimental animals Grouping and model building:Male Wistar rats were randomly divided into the following four groups:control group,normal diet with normal saline ，ethanol-treated group,normal diet with ethanol administration ，and low-and high-do Aplysin[100,and 150mg/(kg•bw•d)]plus ethanol treatment groups.Excluding the rats in the normal control group,the other animals were initially received intragastric administration with 56%(v/v)ethanol 5.5mL kg-1day-1for the first week,7.5mL kg-1day-1for the cond week,8mL kg-1day-1for the third week,9mL kg-1day-1for the fourth week,and 11mL kg-1day-1for the rest of the 4weeks.Twelve hours after the last ethanol treatment,the rats were anesthetized,and blood was collected,the small intestine was obtained.2.the small intestine ud for ultrastructural obrvation by tansmission electron microscope. 3.plasma FABP2/TGF-β/TNF-α/D-LA/DAO concentration was measured by enzyme linked immunosorbent assay.4.The expression of FOXO4in small intestine was detected by immunohistochemistry.
Results:1.In the ethanol-treated group,the ultrastructural changes of small intestine tissue by tansmission electron microscope showed that the intestinal villi fell away,and looly arranged,tight junction were swelling,however,the control group did not show abnormal ultrastructural.In each do Aplysin,ultrastructural changes were better than the ethanol-treated group.2.In control group,the plasma concentration of D-LA and DAO were (4.78±0.67)u mol/L 、(66.65±9.07)U/L,the ethanol-treated group were (6.44±1.83)u mol/L 、(87.87±20.14)U/L,it was significantly elevated compared with control group.the plasma concentration of D-LA and DAO in alcohol+Aplysin (100mg/kg)group were (5.44±1.23)u mol/L 、(69.47±7.14)U/L,alcohol+Aplysin (150mg/kg)group were (5.06±1.32)u mol/L 、(67.92±7.68)U/L,they were significantly reduced compared with the ethanol-treated group.3.In control group,the plasma concentration of FABP2,TNF-αand TGF-βwere (2.79±1.18)ng/ml,(30.42±18.89)pg/ml and (38.67±19.22)pg/ml,the ethanol-treated group were (3.87±1.01)ng/ml,(59.31±35.01)pg/ml and (62.28±9.33)pg/ml,it was significantly elevated compared with control group.the plasma concentration of FABP2,TNF-αand TGF-βin alcohol+Aplysin (100mg/kg)group were (2.87±1.60)ng/ml,(36.32±19.58)pg/ml and 硕博毕业论文社区
