幼鼠肠炎后前扣带回皮质NR2A、NR2B表达变化

发布时间：2019-05-17 19:34

【摘要】：再发性腹痛(recurrent abdominal pain，RAP)是常见的儿童胃肠道疾病，也是儿童消化病门诊数量最多的疾病之一，因缺乏有效的诊治方法，影响患儿的生长发育及身心健康，其发病机制错综复杂，内脏高敏感(visceralhypersensitivity，VH)是RAP的重要病理生理改变之一。近年来流行病学调查研究显示，RAP症状常始于某次急性感染性疾病以后，临床上大约22％的RAP患儿之前曾患过急性胃肠炎，因此炎症后内脏敏感性升高逐渐被人们认识。有研究发现由初级传入神经纤维支配的胃肠道增敏可以引起内脏高敏，但中枢神经元可塑性的改变也同样促进了内脏高敏的形成。NMDA受体是中枢神经系统中兴奋性谷氨酸受体的一种类型，属于离子型谷氨酸受体(ionotropic glutamatereceptors，iGluRs)，参与并调控内脏伤害性信息的传递，但它们在肠道炎症后内脏高敏感中的作用尚不十分明确。研究目的：本研究使用芥子油结直肠灌注建立幼鼠肠炎症后内脏高敏感模型，分析肠道炎症对中枢NMDA受体发育的影响，并进而探讨NMDA受体发育异常与肠道炎症后内脏高敏感的关系。研究方法：以芥子油结直肠灌注建立幼鼠肠炎症后内脏高敏感模型，，并分别于生后第8、15、22、36天取大脑中枢前扣带回(anteriorcingulate cortex，ACC)区域组织，应用免疫荧光法采集不同时间节点NMDA受体的亚基NR2A和NR2B的免疫荧光图片，以IOD值反映受体亚基量的变化；以免疫印迹(Western Blot)的方法检测不同时间节点ACC处的NMDA受体的亚基NR2A和NR2B，以灰度值反映受体亚基量的变化。结果：芥子油结直肠灌注诱导幼鼠形成炎症后内脏高敏感，实验组与对照组大鼠结肠组织经HE染色，均未见明确病理学改变；免疫荧光结果显示实验组大鼠的大脑ACC处NR2A、NR2B均较对照组升高，实验组NR2A/NR2B值较对照组降低(P0.05)。Western blot结果也显示实验组大鼠的大脑ACC处NR2A、NR2B较对照组升高，但NR2A/NR2B值较对照组降低(P0.05)。结论：使用芥子油结直肠灌注可以建立幼鼠肠道炎症后内脏高敏感模型，大脑ACC处NR2A、NR2B及NR2A/NR2B值发生了变化，这表明肠道炎症后ACC处NMDA受体亚基发育异常在炎症后内脏高敏感的中发挥了一定的作用。
[Abstract]:Recurrent abdominal pain (recurrent abdominal pain,RAP) is a common gastrointestinal disease in children, and it is also one of the most numerous diseases in the clinic of digestive diseases in children. Due to the lack of effective diagnosis and treatment, it affects the growth, development and physical and mental health of children. Its pathogenesis is complex. Visceral hypersensitivity (visceralhypersensitivity,VH) is one of the important pathophysiological changes of RAP. In recent years, epidemiological studies have shown that RAP symptoms often begin after an acute infectious disease. About 22% of RAP children have suffered from acute gastroenteritis before, so the increase of visceral sensitivity after inflammation has been gradually recognized. Some studies have found that gastrointestinal tract sensitizing, dominated by primary afferent nerve fibers, can cause visceral Gao Min. However, the plasticity of central neurons also promoted the formation of visceral Gao Min. NMDA receptor is a type of excitatory glutamate receptor in central nervous system, which belongs to ionic glutamate receptor (ionotropic glutamatereceptors,iGluRs). They participate in and regulate the transmission of visceral nociceptive information, but their role in visceral hypersensitivity after intestinal inflammation is not very clear. Objective: to establish a model of visceral hypersensitivity after intestinal inflammation in young rats by colorectal perfusion of mustard oil, and to analyze the effect of intestinal inflammation on the development of central NMDA receptor. To investigate the relationship between abnormal development of NMDA receptor and visceral hypersensitivity after intestinal inflammation. Methods: the visceral hypersensitivity model of young rats with intestinal inflammation was established by colorectal perfusion of mustard oil, and the tissues of the anterior cingulate gyrus (anteriorcingulate cortex,ACC) of the brain center were taken on the 8th, 15th and 22nd day after birth, respectively. The immunofluorescence images of NR2A and NR2B of NMDA receptor subunits at different time points were collected by immunofluorescence method, and the changes of receptor subgroup quantity were reflected by Iod value. The subunits NR2A and NR2B, of NMDA receptor at ACC at different time points were detected by immunoblotting (Western Blot) to reflect the change of subbase quantity of NMDA receptor by gray value. Results: the viscera of young rats was highly sensitive after colorectal perfusion with mustard oil. No pathological changes were found in colon tissue of experimental group and control group by HE staining. The results of immunofluorescence showed that the NR2A,NR2B of ACC in the brain of the experimental group was higher than that of the control group, and the value of NR2A/ NR2B of the experimental group was lower than that of the control group (P 0.05). Western blot also showed that the NR2A,NR2B of the ACC of the brain of the experimental group was higher than that of the control group. However, the value of NR2A/ NR2B was lower than that of the control group (P 0.05). Conclusion: the visceral hypersensitivity model of young rats with intestinal inflammation can be established by colorectal perfusion of mustard oil, and the values of NR2A,NR2B and NR2A/ NR2B in ACC of the brain have changed. This suggests that the abnormal development of NMDA receptor subunits in ACC after intestinal inflammation plays a role in visceral hypersensitivity after inflammation.
【学位授予单位】：第二军医大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R725.7

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