大鼠急性肠粘膜损伤模型敏化点体表分布及敏化点局部CGRP、肥大细胞的变化

发布时间：2018-03-01 10:44

本文关键词： 穴位敏化肠粘膜损伤伊文思蓝渗出肥大细胞降钙基因相关肽　出处：《福建中医药大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的：观察急性肠粘膜损伤相关的体表伊文思蓝(EB)渗出点的分布及肥大细胞聚集数量和脱颗粒变化及降钙基因相关肽(CGRP)的表达。方法：(1)SD雄性大鼠36只,按芥子油浓度随机分6组,分别为5%、7.5%、10%、12.5%、15%、17.5%,每组6只。采用肠道灌注芥子油造成急性肠粘膜损伤模型,造模后观察大鼠活动情况,造模后4h全身备皮、尾静脉注射EB,造模后5h、10h、15h、20h、25h分别观察EB渗出点的分布,采用皮肤定位结合全因素聚类分析、轮廓分析的方法,分析研究渗出点随浓度及时间变化的分布规律。(2)SD雄性大鼠30只,随机分为模型组(15只)和生理盐水组(15只)。肠道灌注15%芥子油造成急性肠粘膜损伤模型,造模4h后背部备皮、尾静脉注射EB,造模后5h模型组及生理盐水组分别记录“肾俞”、“大肠俞”组织是否有蓝点,模型组有蓝点为EB渗出组,无则为非EB渗出组,之后用打孔器取下“肾俞”、“大肠俞”组织,用甲苯胺蓝染色法标记这些部位肥大细胞的聚集分布数量及脱颗粒特征,免疫组织化学方法显示局部CGRP的分布。结果：(1)造模情况：造模后25h内5%、7.5%、10%、12.5%、15%芥子油组大鼠均存活,17.5%芥子油组大鼠死亡4只。大鼠结肠组织粘膜层镜下观察可见不同程度的水肿、炎症和溃疡。(2)EB渗出点体表分布情况：不同浓度不同时间EB渗出点其规律性主要体现在腰骶段的脊柱两侧。(3)EB渗出点随浓度变化在体表分布情况：平行轮廓检验：EB渗出点行为一致(P0.05)；重合轮廓检验：EB渗出点程度相同(P0.05)；水平轮廓检验：EB渗出点与不同浓度有关(P0.05),在15%浓度渗出最明显,其次12.5%、7.5%、10%、5%。(4)EB渗出点随时间变化在体表分布情况：平行轮廓检验：EB渗出点行为一致(P0.05)；重合轮廓检验：EB渗出点程度相同(P0.05)；水平轮廓检验：EB渗出点在时间上无明显变化,造模后5hEB渗出趋于完全(P0.05)。(5)EB渗出点组肥大细胞聚集数量和脱颗粒变化：急性肠粘膜损伤后体表EB渗出点组的皮肤和皮下组织中肥大细胞呈现聚集,在肥大细胞总数上EB渗出点组与非EB渗出点组、生理盐水组无统计学差异(P0.05)；在肥大细胞脱颗粒数上EB渗出点组明显多于非EB渗出点组和生理盐水组(P0.05),脱颗粒率也显著高于非EB渗出点和生理盐水组(P0.05)。(6)EB渗出点组CGRP表达：在EB渗出点组可见CGRP蛋白的表达,水平高于非EB渗出点组和生理盐水组(P0.05)。结论：(1)急性肠粘膜损伤可促使EB在体表渗出,对于不同浓度与不同观察时间点的EB渗出现象,其规律性主要体现在腰骶段的脊柱两侧。(2)肥大细胞参与了与疾病相关的体表穴位敏化过程,可能和局部CGRP表达水平相关。
[Abstract]:Aim: to observe the distribution of exudation sites and the changes of mast cell aggregation and degranulation and the expression of calcitonin gene-related peptide CGRP in acute intestinal mucosal injury associated with EBB. Methods Thirty-six male Sprague-Dawley rats were randomly divided into 6 groups according to the concentration of mustard oil. The rats were randomly divided into 6 groups: 5 katsui 7.5g and 12.5g / 15g / L, 6 rats in each group. The acute intestinal mucosal injury model was established by intestinal perfusion of mustard oil. The activity of the rats was observed after the establishment of the model, and the skin was prepared throughout the body 4 hours after the establishment of the model. The distribution of EB exudation point was observed 5 h, 10 h, 15 h, 20 h and 25 h after injection of EB.The distribution of exudation point was studied by skin localization, whole factor cluster analysis and contour analysis. 30 SD male rats were used to study the distribution of exudation point with the change of concentration and time. Acute intestinal mucosal injury was induced by intestinal perfusion of 15% mustard oil. The rats in the model group and the saline group were respectively recorded whether there were blue spots in "Shenshu" and "Dachang Yu" tissues at 5 h after the injection of EB.The blue spots in the model group were EB exudate group, and none were non-EB exudate group. The tissues of Shenshu and Dachang Yu were removed with perforator, and the distribution of mast cells in these areas was labeled with toluidine blue staining method. The distribution of local CGRP was observed by immunohistochemical method. Results: within 25 hours after the establishment of the model, the rats in the 15% mustard oil group survived and 4 rats died. The colonic tissue mucosal layer of rats was observed under microscope to observe the edema of different degrees. Distribution of Epstein-Barr exudation points in inflammation and ulcers: the regularity of EB exudation points at different concentrations and at different times is mainly reflected in the distribution of EB exudation points along the sides of the spine of lumbosacral segment with the change of concentration: parallel contours are used to test the infiltration of EB in the body surface. The behavior of exiting point is consistent (P0.05); the superposition profile test is of the same degree of exudation point (P0.05N); the horizontal contour test of the point of exudation of EB is related to different concentration (P0.05N), and the exudation is most obvious at 15% concentration. Secondly, 12.5and 7.5and 10%, including the distribution of exudation points over time on the body surface: parallel contour test: the behavior of the exudation point of EB is the same (P0.05); the overlap contour test has the same degree of exudation point (P0.05N); the horizontal contour test has no obvious change in the time of the exudation point. 5 h after modeling, the exudation of EB tended to be complete P0. 05%. The change of mast cell aggregation and degranulation in the exudation point group: after acute intestinal mucosal injury, mast cells in skin and subcutaneous tissue showed aggregation in the group of exudation point of EB on the surface of the body. EB exudation point group and non-EB exudation point group were found in the total mast cell count. There was no significant difference in the number of mast cell degranulation in normal saline group (P 0.05), and the number of mast cell degranulation in EB exudation point group was significantly higher than that in non EB exudation point group and normal saline group, and the degranulation rate in EB exudation point group was significantly higher than that in non EB exudation point group and normal saline group. Expression: the expression of CGRP protein was observed in EB exudation point group. The level of P0. 05 was higher than that of non EB exudate point group and normal saline group. Conclusion (1) Acute intestinal mucosal injury can promote the exudation of EB on the body surface. The regularity of mast cells in lumbosacral spine is that mast cells are involved in the process of surface acupoint sensitization related to disease, which may be related to the level of local CGRP expression.
【学位授予单位】：福建中医药大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R574

【参考文献】