【作者】 蒋淼；

【导师】 樊晓明；

【作者基本信息】 复旦大学 ， 消化内科学， 2011， 硕士

【摘要】 目的研究ghrelin对乙醇诱导的急性大鼠胃黏膜损伤的保护作用,并通过观察辣椒素神经拮抗剂对ghrelin保护作用的影响来进一步探讨ghrelin的作用机制。方法实验一1.64只雌性SD大鼠分成四组,空白对照组(2m1/kg 0.9%氯化钠液腹腔注射+1ml/100g 0.9%氯化钠液灌胃),乙醇模型组(2m1/kg 0.9%氯化钠液腹腔注射+1ml/10Og 75%乙醇灌胃),低剂量ghrelin组(2p,g/kg ghrelin腹腔注射+lml/100g 75%7,醇灌胃),高剂量ghrelin组(20μg/kg ghrelin腹腔注射+lml/100g 75%乙醇灌胃)2小时后四组大鼠麻醉取胃。2.计算各组大鼠胃黏膜溃疡指数。3.HE染色观察胃黏膜炎症损伤程度。4.通过荧光半定量RT-PCR法测定eNOS/iNOS和COX-1/COX-2的mRNA表达,实验二1.64只雌性SD大鼠分成四组,空白对照组(2m1/kg 0.9%氯化钠液腹腔注射+lml/100g 0.9%氯化钠液灌胃),乙醇模型组(2ml/kg 0.9%氯化钠液腹腔注射+1ml/100g 75%乙醇灌胃),ghrelin组(20μg/kg ghrelin腹腔注射+lml/100g 75%7,醇灌胃),Capsazepine组(20μg/kg ghrelin腹腔注射+1mg/kg Capsazepine腹腔注射+1ml/100g 75%乙醇灌胃),2小时后四组大鼠麻醉取胃。2.计算各组大鼠胃黏膜溃疡指数。3.HE染色观察胃黏膜炎症损伤程度。4.通过荧光半定量RT-PCR法测定eNOS(?)(?)iNOS(?)(?)COX-1/COX-2的1nRNA表达,测定CGRP的mRNA的表达。5.酶联免疫吸附试验(ELISA)法测定胃组织中CGRP含量。结果实验一1. Ghrelin组胃溃疡指数明显小于乙醇模型组,与Ghrelin呈剂量相关性。2. Ghrelin组病理炎症反应较乙醇模型组明显减轻,与Ghrelin呈剂量相关性。3.荧光半定量RT-PCR:ghrelin组与乙醇模型组相比,eNOS mRNA表达上升,COX-1 mRNA表达无变化,但iNOS mRNA和COX-2表达下降,与ghrelin呈剂量相关性。实验二1. Capsazepine组胃溃疡指数明显大于ghrelin组。2. Capsazepine组病理炎症反应明显较ghrelin组加重。3.荧光半定量RT-PCR:capsazepine组与ghrelin组相比,iNOS和COX-2mRNA表达上升；(;NOS(?)(?)CGRP mRNA表达下降,COX-1 mRNA表达无变化4.酶联免疫吸附试验(ELISA) CGRP测定：capsazepine组与ghrelin组相比,组织中CGRP含量下降。结论本实验结果提示Ghrelin具有对乙醇诱导的胃黏膜保护作用,其保护机制可能是通过参与辣椒素一敏感感觉神经保护性反射,调节CGRP的表达从而参与调控NOS和COX活性来达到胃黏膜保护的作用。更多还原

【Abstract】 ObjectiveTo investigate the gastroprotective effects of ghrelin against ethanol-induced acute gastric injury in rats and possible mechanism.MethodsPart 11.64 female Sprague-Dawley(SD) rats were divided into four groups and were intragastrically administrated as follows:Control group(2mg/kg 0.9%NaCl ip+lml/100g 0.9%NaCl ig);Ethanol model group(2ml/kg 0.9%NaCl ip+lml/100g 75%ethanol ig);Low-dose ghrelin group(2μg/kg ghrelin ip+lml/100g 75%ethanol ig);High-dose ghrelin group(20μg/kg ghrelin ip+lml/100g 75%ethanol ig).Rats were anaesthetized and the stomachs were removed after 2 hours.2. The ulcer index of gastric mucosal lesions in rats were calculated.3. Gastric mucosal inflammation were assessed by hematoxylin-eosin (H&E) staining.4. eNOS/iNOS and COX-1/COX-2 mRNA expression in these specimens were verified by semi-quantitative reverse transcriptional polymerase chain reaction (SqRT-PCR).Part 21.64 female SD rats were divided into four groups and were intragastrically administrated as follows:Control group(2mg/kg 0.9%NaCl ip+lml/100g 0.9%NaCl ig);Ethanol model group(2ml/kg 0.9%NaCl ip+lml/100g 75%ethanol ig);Ghrelin group(2μg/kg ghrelin ip+lml/100g 75%ethanol ig);Capsazepine group(20μg/kg ghrelin ip+lmg/kg Capsazepine ip+lml/100g 75%ethanol ig).Rats were anaesthetized and the stomachs were removed after 2 hours.2. The ulcer index of gastric mucosal lesions in rats were calculated.3. The inflammatory lesions of gastric mucosa were assessed by hematoxylin-eosin (H&E) staining.4. eNOS/iNOS, COX-1/COX-2 and CGRP mRNA expression in these specimens were verified by SqRT-PCR5.The levels of CGRP in these specimens were measured by Enzyme-linked immunosorbent assay (ELISA).Results Part 11. The ulcer index of ghrelin groups were significantly less than that of ethanol model group, and were positively correlated with dosages of ghrelin.2. Compared with ethanol model group,the gastric mucosal inflammation lesions of ghrelin group were significantly reduced, and were positively correlated with dosages of ghrelin.3. Compared with Ethanol model group,grhrein groups were characterized by no changes of COX-1 mRNA level, but iNOS mRNA and COX-2 mRNA decreased, eNOS mRNA increased in a dose-dependent manner.Part 21. The ulcerative index of capsazepine group were significantly greater than those of ghrelin group.2. Compared with ghrelin group,the gastric mucosal inflammation lesions of capsazepine group increased significantly.3. Compared with ghrelin group, there was no change of COX-1 mRNA level in capsazepine group,but mRNA levels of COX-2 and iNOS increased while eNOS and CGRP decreased.4. Compared with ghrelin group, the levels of CGRP in gastric tissue of capsazepine decreased.ConclusionsGhrelin had protective effects on ethanol-induced acute gastric mucosal injury in rats.The mechanism of the protective effects was related to CGRP which can be involved in regulating capsaicin-sensitive nerves and thus participated in the regulation of the activity of NOS and COX.更多还原