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学科主题: 外科学
题名:
尾悬吊模拟失重对大鼠胃窦动力影响的研究
作者: 李正鹏
答辩日期: 2013-05-20
导师: 崔彦
专业: 外科学
授予单位: 北京大学
授予地点: 北京大学解放军306医院教学医院
学位: 硕士
关键词: 模拟失重 ; 胃泌素 ; 胃动素 ; 生长抑素 ; 血管活性肠肽 ; Cajal间质细胞 ; 环氧化酶-2
其他题名: The effects of simulated weightlessness on dynamics of gastric antrum in rats
分类号: R573
摘要: 背景和目的 研究证实,失重或模拟失重对人体系统和器官产生较大影响,甚至导致病理生理性改变。胃是人体最重要的消化器官之一,失重状态下,胃的功能及其消化液的极性运动必然受到影响,探索失重对胃功能的影响具有重要意义。本实验研究尾悬吊模拟失重环境下大鼠血清GAS、MTL、SS、VIP含量以及胃窦Cajal间质细胞和胃窦肌层COX-2表达的变化,探讨模拟失重对大鼠胃窦动力的影响。     方法 健康雄性Wistar大鼠64只,随机分为8组(n=8),按模拟失重时相分别为6 h、12 h、1 d、2 d、3 d、5 d、7 d和0 h对照组。采用尾悬吊法建立模拟失重动物模型。应用放免法测定血清GAS、MTL、SS、VIP浓度,免疫组化、Western Blot和RT-PCR技术检测各组大鼠胃窦组织c-kit蛋白和mRNA以及胃窦肌层组织中COX-2蛋白和mRNA表达情况。     结果 (1) 尾悬吊6 h、12 h阶段,血清GAS浓度明显升高,与对照组比较差异有统计学意义(P<0.05)。随尾悬吊时相延长,血清GAS含量呈逐渐下降趋势,与正常对照组水平相近。尾悬吊各组大鼠血清MTL浓度均升高,12 h以后各组MTL浓度值与对照组比较,差异有统计学意义(P<0.05)。血清SS浓度在尾悬吊6 h即开始升高,12 h、2 d、3 d组明显升高,与对照组比较差异有统计学意义(P<0.05)。而1 d、5 d、7 d组血清SS浓度则与正常对照组水平相近,大体呈现先升后降的趋势。尾悬吊6 h、12 h阶段,血清VIP浓度明显升高,与对照组比较差异有统计学意义(P<0.05)。随着尾悬吊时相的延长,血清VIP含量呈逐渐下降趋势,5 d时又有一定程度上升,7 d时VIP含量明显高于正常对照组(P<0.05)。(2) 免疫组化结果显示,c-kit蛋白阳性表达为棕褐色,主要分布在Cajal间质细胞胞体和突起,正常对照组大鼠胃窦肌层ICC-MY呈连续性浓染,ICC-IM亦明晰可见;6 h - 5 d的尾悬吊过程中大鼠胃窦肌层ICC-MY连续性出现中断现象且逐渐明显,染色逐渐减弱,ICC-IM染色减弱的同时c-kit阳性ICC也明显减少。7 d组胃窦组织中c-kit蛋白表达有所恢复。COX-2蛋白阳性表达亦为棕褐色,0 h对照组胃窦平滑肌细胞胞浆染色弱阳性。悬吊6 h、12 h、1 d组大鼠胃窦平滑肌细胞浆染色变深并出现部分核周染色阳性,2 d开始逐渐恢复正常,但胞浆染色较0 h组仍为深。(3) Western Blot结果显示,悬吊6 h组胃窦肌层COX-2蛋白表达水平显著升高,12 h、1 d组持续高表达,与对照组比较,差异具有统计学意义(P<0.05),2d开始逐渐恢复正常水平。(4) 模拟失重各组及对照组大鼠胃窦组织中均有c-kit mRNA表达,尾悬吊6 h和12 h组的c-kit mRNA表达明显下降,与对照组比较,差异有统计学意义(P<0.05),1 d - 3 d组逐渐恢复,5 d组又出现明显下降(P<0.05),7 d组c-kit mRNA表达值明显复升。同时,尾悬吊6 h大鼠胃窦肌层COX-2 mRNA表达水平即出现显著增高,12 h、1 d组仍高水平表达,与对照组比较,差异具有统计学意义(P<0.05)。2 d组开始表达逐渐回落,3 d、7 d组与对照组无明显差异,5 d组轻度升高,差异不具有统计学意义(P>0.05)。     结论 (1) 尾悬吊模拟失重对大鼠血清GAS、MTL、SS、VIP和胃窦c-kit蛋白及mRNA表达造成明显影响,可能导致胃动力障碍。(2) 模拟失重使大鼠胃窦肌层组织中COX-2蛋白和mRNA表达发生明显变化,早期同步过表达,后期出现恢复趋势,提示COX-2在胃窦失重应激反应性动力障碍及恢复过程中可能发挥重要作用。
英文摘要: Background and Objective: Researches have showed that agravity or simulated weightlessness have great influence on the human body systems and organs, and even lead to pathological physiology changes. Stomach is one of the most important digestive organs of human beings, the function and digestive juices of polarity movement will be affected by the condition of weightlessness. Therefore understanding of the effects induced by the weightlessness on the structure and function of stomach is of great significance. This study investigates the alterations of serum gastrin, motilin, somatostatin, vasoactive intestinal peptide and expression of interstitial cells of Cajal in gastric antrum of rats and COX-2 in muscular layer of gastric antrum and try to evaluate the effects induced by simulated weightlessness on dynamics of gastric antrum in rats.      Methods: Sixty four adult male Wistar rats were randomly divided into 8 experimental groups (n=8) including suspension for 6 h, 12 h, 1 d, 2 d, 3 d, 5 d, 7 d and 0 h (control). Tail-suspension was used to simulate the weightlessness animal model. Serum concentrations of gastrin, motilin, somatostatin and vasoactive intestinal peptide were detected by radioimmunoassay. The c-kit protein and mRNA in rat gastric antrum and the COX-2 protein and mRNA in muscular layer of gastric antrum were assayed by immunohistochemistry, western blot and RT-PCR technique.      Results: (1) Compared with the control group, the level of serum gastrin increased significantly in the early phase of tail-suspension with a peak value at 6 h and 12 h (P<0.05) followed by declination and trend to normal range during the sustained suspension. The level of serum motilin obviously increased in all groups except that in 6 h group (P<0.05). The concentration of serum somatostatin was increased significantly in 12 h, 2 d and 3 d groups (P<0.05) and got some recovery in 5 d and 7 d groups. Similarly, the level of serum vasoactive intestinal peptide increased significantly in the early phase of tail-suspension with a peak value at 6 h and 12 h (P<0.05) followed by declination and trend to normal range during the sustained suspension and then got a sharp rise in 7 d suspension group. (2) The immunohistochemistry assay showed that c-kit protein was stained as brown particles and mainly located in cell soma and dentrites of interstitial cells of Cajal. ICC-MY and ICC-IM were deeply stained normally in rat gastric antrum. The expression of c-kit protein was obviously weakened during the period of 6 h to 5 d tail suspension, and the continuity of ICC-MY was gradually interrupted. However, the expression of c-kit protein was up-regulated in 7 d group. The COX-2 protein was also stained as brown particles. The cytoplasm of gastric smooth muscle cells was slightly stained in control group. Interestingly, the cytoplasm was deeply stained in the early hours of suspension with some positive perinuclear staining of smooth muscle cell, and had somewhat recovery in 2 d to 7 d suspension group. (3) Western blotting analysis showed that COX-2 protein expression in gastric antrum muscular layer was significantly up-regulated compared with control group in the early stage of suspension (P<0.05) and was recovered to normal ranges in the late phase of suspension. (4) The semi-quantitative RT-PCR showed that the c-kit mRNA in rats’ gastric antrum was significantly down-regulated at 6 h and 12 h of suspension compared with the control group (P<0.05), followed by a gradually recovery at 1 d - 3 d of suspension, then the c-kit mRNA was again significantly down-regulated at 5 d of suspension (P<0.05) and was markedly increased at 7 d of tai-suspension. The COX-2 mRNA in gastric antrum muscular layer was also significantly up-regulated at the early phase of suspension as compared with the control group (P<0.05), and followed by a recovery at 2 d, 3 d and 7 d of suspension. The level of COX-2 mRNA was elevated in 5 d group, but the difference was not significant compared with the control group (P>0.05).      Conclusion: (1) This study showed that the simulated weightlessness by tail-suspension could disturb the serum gastrin, motilin, somatostatin and vasoactive intestinal peptide levels and affect the c-kit expression both at protein and mRNA levels in rat gastric antrum, and thus the gastric motility may be impaired. (2) The study showed that the simulated weightlessness acted as a stressor that elevated COX-2 expression both at protein and mRNA levels in muscular layer tissues of rats’ gastric antrum, and COX-2 may played an important role in the reactive dynamics disorder of rat gastric antrum and in the process of recovery under simulated weightlessness.
语种: 中文
出处: http://xuewei.bjmu.edu.cn/simpsearch.action?keyword=尾悬吊模拟失重对大鼠胃窦动力影响的研究&dbid=72
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内容类型: 学位论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/105439
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作者单位: 北京大学解放军306医院教学医院

Recommended Citation:
李正鹏. 尾悬吊模拟失重对大鼠胃窦动力影响的研究[D]. 北京大学解放军306医院教学医院. 北京大学. 2013.
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