脓毒症大鼠肠道TRPV1、CGRP的表达及作用

发布时间：2018-06-17 16:28

本文选题：脓毒症 + TRPV1　；参考：《皖南医学院》2014年硕士论文

【摘要】：目的：脓毒症是ICU急症，临床上发生率和死亡率都很高。近年来，人们发现脓毒症往往会造成患者胃肠功能障碍，并认为这种胃肠功能障碍在脓毒症的发生发展中起到重要作用。大量研究证明TRPV1、CGRP在胃肠道表达广泛，参与胃肠运动的调节。本次研究旨在观察脓毒症对肠动力功能的影响，并探讨TRPV1和CGRP在脓毒症引起大鼠肠动力改变中的作用。方法：雄性清洁型SD大鼠20只,随机分为2组,每组10只。A组:正常对照组;B组:脓毒症组。禁食12h后，采用腹腔内注射脂多糖(LPS)制备大鼠脓毒症模型。每只大鼠腹腔注射LPS10mg/kg，LPS用生理盐水稀释成1ml，制成脓毒症大鼠模型。正常对照组给于腹腔注射等量生理盐水作为对照。造模结束继续禁食,观察12h，记录血压心率，后给与碳素墨汁灌胃，每只大鼠2ml。观察20min后处死大鼠，剖腹取出大鼠胃及整个肠道，测定小肠墨汁推进率，每只大鼠墨汁移行距离占小肠全长的百分率即为小肠推进率。并于距盲肠15cm处快速切取一段约3cm的空肠段制备标本，备行TRPV1、CGRP免疫组织化学染色检测，镜下观察比较各指标两组间的染色结果，将阳性细胞百分率评分与染色强度评分的乘积作为最后的积分。采用SPSS18.0软件包分析各组数据。结果：腹腔注射内毒素后大鼠的疾病状态评分较正常组明显上升，，心率加快，差异均有统计学意义。脓毒症组大鼠的小肠墨汁推进率明显低于对照组，P0.05差异有统计学意义。免疫组织化学染色结果示，TRPV1、CGRP阳性染色细胞可见于粘膜层、粘膜固有层等，表达较为广泛，脓毒症组中TRPV1、CGRP的阳性细胞数与染色程度乘积的积分明显高于对照组，P0.05，差异有统计学意义。结论：1、腹腔注射内毒素10mg/kg后可以建立理想的脓毒症大鼠模型。2、脓毒症组大鼠小肠推进率较对照组显著下降，可见脓毒症大鼠发生明显的肠道动力障碍。3、脓毒症时大鼠肠道动力障碍可能与TRPV1受体表达上调有关，且CGRP的表达变化与TRPV1一致，故推测TRPV1引起肠道动力障碍可能与CGRP的表达增高有关
[Abstract]:Objective: sepsis is an emergency in ICU with high incidence and mortality. In recent years, it has been found that sepsis often causes gastrointestinal dysfunction in patients, and this gastrointestinal dysfunction plays an important role in the occurrence and development of sepsis. A large number of studies have shown that TRPV1 CGRP is widely expressed in the gastrointestinal tract, involved in the regulation of gastrointestinal motility. The purpose of this study was to observe the effect of sepsis on intestinal motility and to explore the role of TRPV1 and CGRP in intestinal motility changes induced by sepsis in rats. Methods: twenty male clean Sprague-Dawley rats were randomly divided into two groups: group A (n = 10): normal control group (n = 10): sepsis group (n = 10). After fasting for 12 hours, the sepsis model of rats was established by intraperitoneal injection of lipopolysaccharide (LPS). Each rat was injected intraperitoneally with LPS 10 mg / kg LPS and diluted into 1 ml with normal saline to make sepsis rat model. The normal control group was given intraperitoneal injection of the same amount of normal saline as the control group. At the end of the model, the rats continued to fast, observed for 12 hours, recorded the blood pressure and heart rate, and then given carbon ink to the stomach, each rat 2 ml. After observation of 20min, the rats were sacrificed, the stomach and the whole intestine were taken out by laparotomy, and the rate of small intestinal ink propelling was determined. The percentage of the migration distance of each rat's ink to the whole intestine was the small intestinal propulsion rate. A section of jejunum about 3cm was quickly cut from the cecum 15cm to prepare the specimen, and the immunohistochemical staining of TRPV1 was performed. The results of the two groups were observed and compared under the microscope. The product of positive cell percentage score and staining intensity score was used as final score. The data of each group were analyzed by SPSS 18.0 software package. Results: after intraperitoneal injection of endotoxin, the scores of disease state and heart rate in rats were significantly higher than those in normal group. The promoting rate of small intestinal ink in sepsis group was significantly lower than that in control group (P0.05). The results of immunohistochemical staining showed that the positive staining cells of TRPV1 + CGRP could be seen in the mucosal layer, lamina propria of mucosa and so on. The number of CGRP positive cells in TRPV1 in sepsis group was significantly higher than that in control group (P 0.05). Conclusion the ideal sepsis rat model can be established by intraperitoneal injection of endotoxin 10mg/kg. The intestinal propulsion rate in sepsis group is significantly lower than that in control group. It can be seen that there are obvious intestinal motility disorders in sepsis rats. The intestinal motility disorder in septic rats may be related to the up-regulation of TRPV1 receptor, and the change of CGRP expression is consistent with that of TRPV1. It is speculated that the intestinal motility disorder caused by TRPV1 may be related to the increase of CGRP expression.
【学位授予单位】：皖南医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R459.7

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