TRPV1介导的艾灸血管效应的实验研究

发布时间：2018-06-25 16:33

  本文选题：艾灸 + Trpv1　； 参考：《南京中医药大学》2016年硕士论文

【摘要】：背景和目的：本课题旨在研究TRPV1(瞬态电压感受器阳离子通道,子类V,成员1)介导的艾灸血管效应。其中,以艾灸为主要干预方式,研究不同热刺激对正常大鼠局部皮肤微循环血流量的影响,艾灸激活TRPV1与局部皮肤组织肥大细胞、肠系膜动脉血管的相关性,以及血管内皮相关因子CGRP(降钙素基因相关肽)、eNOS(内皮型一氧化氮合酶)的调节作用,在此基础上进一步探讨艾灸以温促通的血管效应和作用机制。方法：包括理论研究和实验研究。理论研究部分,在系统梳理血管内皮和微循环系统研究进展的基础上,搜集并整理与局部热刺激相关的现代国内外临床与实验研究成果,对其与血管内皮功能和微循环功能相关的文献进行总结。实验研究包括：1、使用局部皮肤微循环血流检测技术,比较不同温度艾灸热刺激对正常SD大鼠(20只,随机分为两组)施灸穴位(足三里)局部皮肤微循环血流量的影响；2、免疫荧光染色方法和荧光定量PCR方法观察不同艾灸热刺激(关元)对正常SD大鼠肠系膜下动脉TRPV1蛋白和mRNA表达的差异；3、放射免疫法观察不同艾灸热刺激后正常SD大鼠血浆CGRP、eNOS含量的变化;4、通过甲苯胺蓝染色法观察艾灸正常SD大鼠关元穴对施灸局部皮肤组织肥大细胞形态和数目的影响。结果：1、在实验一中通过比较两组大鼠下肢微循环血流灌注情况可总结出,从加热方式上看,艾灸组的局部皮肤血流灌注量波动幅度更大,两组间差异不具有统计学意义(P0.05)；从温度上看,43℃血流灌注量变化幅度更大,组间差异不具有统计学意义(P0.05)；从温度后效应持续时间上看,艾灸组的温度后效应持续时间优于对照组,两组差异有统计学意义(P＜0.05)；2、在实验二中,与空白组相比,温和灸组的TRPV1蛋白水平高于空白组,差异有统计学意义(P0.05),瘢痕灸组的TRPV1蛋白水平低于空白组,差异有统计学意义(P0.05)；温和灸组和瘢痕灸组的mRNA表达均下降,差异有统计学意义(P0.05)；与空白组相比,温和灸组和瘢痕灸组大鼠血浆内CGRP含量均下降,差异有统计学意义(P0.05)；温和灸组eNOS含量与空白组相比下降,差异有统计学意义(P0.05)；4、温和灸组肥大细胞数(10.24±0.44)、瘢痕灸组肥大细胞数(12.31±0.52),较正常组(6.50±0.76)明显增多(P0.05)。5、TRPV1蛋白和mRNA水平与施灸局部皮肤肥大细胞变化量有中度正相关,差异有统计学意义(P0.05)。结论：1、不同艾灸操作方式,激活TRPV1通道可产生差异性的血管效应；2、艾灸可能通过激活皮肤肥大细胞活化TRPV1通道,引起TRPV1血管效应。
[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，

本文编号：2066761