摘要
目的通过细胞力学加载装置,观察推拿法对体外培养人脐静脉血管内皮细胞血管活性物质的影响,探索法的行气活血效应机制。方法培养HUVEC细胞,建立缺氧细胞模型,分别将正常细胞和缺氧细胞分为对照组、推拿组、推拿+维拉帕米组。除对照组外另两组均进行法干预,推拿方法在已建立的HUVEC细胞力学加载模型上进行,用Griess还原法分别检测各组HUVEC细胞NO的合成与释放量,采用实时荧光定量PCR技术分析推拿法对HUVEC细胞e NOSm RNA相对表达量的影响。结果缺氧可导致HUVEC合成和释放NO量显著降低(P<0.01),e NOSm RNA相对表达量显著降低(P<0.01);与对照组细胞相比,不论正常还是缺氧HUVEC细胞,推拿组NO合成和释放量显著增加(P<0.01),e NOSm RNA相对表达量显著增强(P<0.01,)推拿+Ver组NO及e NOSm RNA相对表达量并未显著增加,提示Ca2+拮抗剂Ver能显著抑制手法的这种促进作用。结论法能够激活HUVEC的分泌功能,促进内源性血管舒张因子NO的合成和释放,引发血管舒张,这是其实现行气活血效应的主要机制。但该作用可能与Ca2+有关,尚需进一步研究。
Objective To observe the effects of rolling manipulation on the vasoactive substance of HUVEC cultured in vitro through cell mechanical loading device, in order to explore the mechanism of activating the circulation of Qi and blood by rolling manipulation. Methods Firstly, HUVEC was cultivated and then part of them were cultivated in anoxic cell model. Secondly, both normal cells and anoxic cells were divided into control group, Tuina group and Tuina plus Verapamil group. Based on the establishment of mechanical loading equipment for HUVEC in vitro, both of the two groups were stimulated by rolling manipulation except the control groups. Thirdly, through Griess reduced method, the synthesis and release amount of NO of all the HUVEC groups was tested respectively. Moreover, the relative quantity of eNOSmRNA of HUVEC affected by rolling manipulation was analyzed by real - time fluorescence quantitative PCR. Results Under the condition of anoxic, the synthesis and release amount of NO of HU- VEC was remarkably decreased （P 〈 O. 01 ） , and the relative quantity of eNOSmRNA of HUVEC was also significantly decreased （P 〈 0.01 ）. Compared with the control groups ,the synthesis and release amount of NO of all the Tuina groups ,both in normal and anoxic groups ,was increased greatly（P 〈 0.01 ）. Meanwhile,the relative quantity of eNOSmRNA of the both groups was strength- ened apparently. On the other hand,both the synthesis and release amount and the relative quantity of eNOSmRNA of HUVEC in Tuina plus Verapamil group did not increase notably. This suggested that the Ca 2 ＋ antagonist, Verapamil, can apparently inhibit the promotion function of rolling manipulation. Conclusion Rolling manipulation is able to activate the secretion function of HU- VEC, and to promote the synthesis and release of NO, a kind of endogenous vascular relaxing factor. And the above effect can lead to vasodilatation. This is the main mechanism of activating the circulation of Qi and blood of rolling manipulation. The function c
出处
《时珍国医国药》
CAS
CSCD
北大核心
2015年第2期493-496,共4页
Lishizhen Medicine and Materia Medica Research
基金
国家自然基金项目(No.30300462)
关键词
推拿?法
HUVEC
细胞力学模型
行气活血
Rolling manipulation
HUVEC
Cell mechanical loading device
Activating the circulation Qi and blood
