本文關鍵詞： 穴位敏化 腸粘膜損傷 伊文思藍滲出 肥大細胞 降鈣基因相關肽　出處：《福建中醫(yī)藥大學》2014年碩士論文　論文類型：學位論文

【摘要】：目的：觀察急性腸粘膜損傷相關的體表伊文思藍(EB)滲出點的分布及肥大細胞聚集數(shù)量和脫顆粒變化及降鈣基因相關肽(CGRP)的表達。 方法：(1)SD雄性大鼠36只,按芥子油濃度隨機分6組,分別為5%、7.5%、10%、12.5%、15%、17.5%,每組6只。采用腸道灌注芥子油造成急性腸粘膜損傷模型,造模后觀察大鼠活動情況,造模后4h全身備皮、尾靜脈注射EB,造模后5h、10h、15h、20h、25h分別觀察EB滲出點的分布,采用皮膚定位結合全因素聚類分析、輪廓分析的方法,分析研究滲出點隨濃度及時間變化的分布規(guī)律。(2)SD雄性大鼠30只,隨機分為模型組(15只)和生理鹽水組(15只)。腸道灌注15%芥子油造成急性腸粘膜損傷模型,造模4h后背部備皮、尾靜脈注射EB,造模后5h模型組及生理鹽水組分別記錄“腎俞”、“大腸俞”組織是否有藍點,模型組有藍點為EB滲出組,無則為非EB滲出組,之后用打孔器取下“腎俞”、“大腸俞”組織,用甲苯胺藍染色法標記這些部位肥大細胞的聚集分布數(shù)量及脫顆粒特征,免疫組織化學方法顯示局部CGRP的分布。 結果：(1)造模情況：造模后25h內5%、7.5%、10%、12.5%、15%芥子油組大鼠均存活,17.5%芥子油組大鼠死亡4只。大鼠結腸組織粘膜層鏡下觀察可見不同程度的水腫、炎癥和潰瘍。(2)EB滲出點體表分布情況：不同濃度不同時間EB滲出點其規(guī)律性主要體現(xiàn)在腰骶段的脊柱兩側。(3)EB滲出點隨濃度變化在體表分布情況：平行輪廓檢驗：EB滲出點行為一致(P0.05)；重合輪廓檢驗：EB滲出點程度相同(P0.05)；水平輪廓檢驗：EB滲出點與不同濃度有關(P0.05),在15%濃度滲出最明顯,其次12.5%、7.5%、10%、5%。(4)EB滲出點隨時間變化在體表分布情況：平行輪廓檢驗：EB滲出點行為一致(P0.05)；重合輪廓檢驗：EB滲出點程度相同(P0.05)；水平輪廓檢驗：EB滲出點在時間上無明顯變化,造模后5hEB滲出趨于完全(P0.05)。(5)EB滲出點組肥大細胞聚集數(shù)量和脫顆粒變化：急性腸粘膜損傷后體表EB滲出點組的皮膚和皮下組織中肥大細胞呈現(xiàn)聚集,在肥大細胞總數(shù)上EB滲出點組與非EB滲出點組、生理鹽水組無統(tǒng)計學差異(P0.05)；在肥大細胞脫顆粒數(shù)上EB滲出點組明顯多于非EB滲出點組和生理鹽水組(P0.05),脫顆粒率也顯著高于非EB滲出點和生理鹽水組(P0.05)。(6)EB滲出點組CGRP表達：在EB滲出點組可見CGRP蛋白的表達,水平高于非EB滲出點組和生理鹽水組(P0.05)。 結論：(1)急性腸粘膜損傷可促使EB在體表滲出,對于不同濃度與不同觀察時間點的EB滲出現(xiàn)象,其規(guī)律性主要體現(xiàn)在腰骶段的脊柱兩側。(2)肥大細胞參與了與疾病相關的體表穴位敏化過程,可能和局部CGRP表達水平相關。
[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.
【學位授予單位】：福建中醫(yī)藥大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R574

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