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TRPV1介導(dǎo)的艾灸血管效應(yīng)的實(shí)驗(yàn)研究

發(fā)布時(shí)間:2018-06-25 16:33

  本文選題:艾灸 + Trpv1; 參考:《南京中醫(yī)藥大學(xué)》2016年碩士論文


【摘要】:背景和目的:本課題旨在研究TRPV1(瞬態(tài)電壓感受器陽離子通道,子類V,成員1)介導(dǎo)的艾灸血管效應(yīng)。其中,以艾灸為主要干預(yù)方式,研究不同熱刺激對正常大鼠局部皮膚微循環(huán)血流量的影響,艾灸激活TRPV1與局部皮膚組織肥大細(xì)胞、腸系膜動脈血管的相關(guān)性,以及血管內(nèi)皮相關(guān)因子CGRP(降鈣素基因相關(guān)肽)、eNOS(內(nèi)皮型一氧化氮合酶)的調(diào)節(jié)作用,在此基礎(chǔ)上進(jìn)一步探討艾灸以溫促通的血管效應(yīng)和作用機(jī)制。方法:包括理論研究和實(shí)驗(yàn)研究。理論研究部分,在系統(tǒng)梳理血管內(nèi)皮和微循環(huán)系統(tǒng)研究進(jìn)展的基礎(chǔ)上,搜集并整理與局部熱刺激相關(guān)的現(xiàn)代國內(nèi)外臨床與實(shí)驗(yàn)研究成果,對其與血管內(nèi)皮功能和微循環(huán)功能相關(guān)的文獻(xiàn)進(jìn)行總結(jié)。實(shí)驗(yàn)研究包括:1、使用局部皮膚微循環(huán)血流檢測技術(shù),比較不同溫度艾灸熱刺激對正常SD大鼠(20只,隨機(jī)分為兩組)施灸穴位(足三里)局部皮膚微循環(huán)血流量的影響;2、免疫熒光染色方法和熒光定量PCR方法觀察不同艾灸熱刺激(關(guān)元)對正常SD大鼠腸系膜下動脈TRPV1蛋白和mRNA表達(dá)的差異;3、放射免疫法觀察不同艾灸熱刺激后正常SD大鼠血漿CGRP、eNOS含量的變化;4、通過甲苯胺藍(lán)染色法觀察艾灸正常SD大鼠關(guān)元穴對施灸局部皮膚組織肥大細(xì)胞形態(tài)和數(shù)目的影響。結(jié)果:1、在實(shí)驗(yàn)一中通過比較兩組大鼠下肢微循環(huán)血流灌注情況可總結(jié)出,從加熱方式上看,艾灸組的局部皮膚血流灌注量波動幅度更大,兩組間差異不具有統(tǒng)計(jì)學(xué)意義(P0.05);從溫度上看,43℃血流灌注量變化幅度更大,組間差異不具有統(tǒng)計(jì)學(xué)意義(P0.05);從溫度后效應(yīng)持續(xù)時(shí)間上看,艾灸組的溫度后效應(yīng)持續(xù)時(shí)間優(yōu)于對照組,兩組差異有統(tǒng)計(jì)學(xué)意義(P<0.05);2、在實(shí)驗(yàn)二中,與空白組相比,溫和灸組的TRPV1蛋白水平高于空白組,差異有統(tǒng)計(jì)學(xué)意義(P0.05),瘢痕灸組的TRPV1蛋白水平低于空白組,差異有統(tǒng)計(jì)學(xué)意義(P0.05);溫和灸組和瘢痕灸組的mRNA表達(dá)均下降,差異有統(tǒng)計(jì)學(xué)意義(P0.05);與空白組相比,溫和灸組和瘢痕灸組大鼠血漿內(nèi)CGRP含量均下降,差異有統(tǒng)計(jì)學(xué)意義(P0.05);溫和灸組eNOS含量與空白組相比下降,差異有統(tǒng)計(jì)學(xué)意義(P0.05);4、溫和灸組肥大細(xì)胞數(shù)(10.24±0.44)、瘢痕灸組肥大細(xì)胞數(shù)(12.31±0.52),較正常組(6.50±0.76)明顯增多(P0.05)。5、TRPV1蛋白和mRNA水平與施灸局部皮膚肥大細(xì)胞變化量有中度正相關(guān),差異有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論:1、不同艾灸操作方式,激活TRPV1通道可產(chǎn)生差異性的血管效應(yīng);2、艾灸可能通過激活皮膚肥大細(xì)胞活化TRPV1通道,引起TRPV1血管效應(yīng)。
[Abstract]:Background & AIM: the aim of this study was to investigate the vascular effect of moxibustion mediated by TRPV1 (transient voltage receptor cationic channel, subclass V, member 1). Among them, moxibustion was taken as the main intervention method to study the effect of different heat stimulation on the blood flow of local microcirculation in normal rats, and the correlation between moxibustion activation of TRPV1 and mast cells of local skin tissue and mesenteric artery vessel was studied. The regulation of calcitonin gene-related peptide (CGRP) and endothelial nitric oxide synthase (Enos) was also studied. Methods: theoretical and experimental studies were included. In the theoretical research part, on the basis of systematically combing the progress of vascular endothelial and microcirculation systems, we collect and collate the modern clinical and experimental research results related to local thermal stimulation at home and abroad. The literatures related to vascular endothelial function and microcirculation function were summarized. The experimental study included: 1. The effect of heat stimulation of moxibustion at different temperature on the blood flow of local skin microcirculation at acupoint (Zusanli) of normal SD rats (20 rats randomly divided into two groups) was compared by using the blood flow detection technique of local skin microcirculation. 2. Immunofluorescence staining and fluorescence quantitative PCR were used to observe the expression of TRPV1 protein and mRNA in the inferior mesenteric artery of SD rats with different moxibustion heat stimulation (Guanyuan). 3. Radioimmunoassay was used to observe the changes of plasma CGRP- Enos content in normal SD rats after different moxibustion heat stimulation. The effect of moxibustion on the morphology and number of mast cells in local skin tissue of normal SD rats was observed by toluidine blue staining. Results in experiment 1, by comparing the blood flow of lower extremity microcirculation between the two groups, we can conclude that the amount of local skin blood flow in moxibustion group fluctuates more from the point of view of heating mode, and the difference between the two groups is not statistically significant (P0.05). From the temperature point of view, the blood perfusion volume of 43 鈩,

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