丹皮酚降低自发性高血压大鼠血压的血管相关机制研究

发布时间：2018-07-26 11:32

【摘要】：牡丹皮(Cortex Moutan,CM)在传统中医中常用来治疗心脑血管疾病,是在亚洲和欧洲被广泛应用的一种传统中草药。丹皮酚(Paeonol,Pae)是牡丹皮的最主要有效成分之一,是一种小分子酚类化合物(2'-羟基-4'-甲氧基苯乙酮,C9H10O3),具有抗动脉粥样硬化,抗心律失常,抗炎症和抗氧化等多方面的药理作用。CM是中医治疗高血压的降压组方中的主要药物之一。近年来,关于CM降压机制的研究已经成为相关学科的研究热点。但是,到目前为止此类研究的结果均是在麻醉状态下,一次或分次大量应用Pae,观察其对血压正常动物动脉血压的影响。Pae对高血压模型动物清醒状态下动脉血压的影响及可能机制尚不明确。一氧化氮(nitric oxide,NO)是心血管系统中最重要的生理调节因子之一,参与血管功能稳态的调节。机体有三种亚型的一氧化氮合酶,参与血管功能调节的主要是内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)。研究证实eNOS的表达水平升高或被激活时,局部NO的产生量会增加,可以引起局部血管舒张、内皮及平滑肌细胞功能变化等一系列血管机能改变。eNOS主要通过三条信号通路激活:即MEK/ERK、PI3K/Akt和cAMP/PKA途径。Pae是否能够通过影响内皮细胞eNOS的表达,诱导血管舒张,从而起到降压作用,其可能激活机制如何尚不确定。本研究旨在通过对经典高血压动物模型自发性高血压大鼠(spontaneously hypertensive rat,SHR)给予Pae处理,利用功能学和分子生物学等方法,从整体动物、离体组织及蛋白表达等不同水平,系统动态地观察Pae处理对SHR大鼠动脉血压的影响,并探讨其对主动脉和肾动脉舒张功能的影响和可能作用机制。研究分为以下三部分:(1)丹皮酚对自发性高血压大鼠动脉血压的影响;(2)丹皮酚对自发性高血压大鼠胸主动脉舒张功能的影响;(3)丹皮酚对自发性高血压大鼠肾动脉舒张功能的影响。第一部分丹皮酚对自发性高血压大鼠动脉血压的影响目的:观察连续灌胃CM煎剂或Pae对SHR大鼠动脉血压的影响。方法:健康雄性wky大鼠30只、shr大鼠30只,分别随机分为wky+control、wky+cm、wky+pae、shr+control、shr+cm和shr+pae组。采用灌胃的给药方式,用无创鼠尾动脉血压测量仪测量大鼠清醒状态下的动脉血压。结果:1cm灌胃对wky大鼠的平均动脉压无影响,但在给药后0.5h,1h,2h,4h,8h均显著降低了shr大鼠的平均动脉血压(p0.05)。2灌胃pae对wky大鼠的平均动脉压无影响,但在给药后0.5h,1h,2h,4h,8h均显著降低了shr大鼠的平均动脉血压(p0.05)。3连续灌胃cm或pae对wky鼠的平均动脉压无明显影响。连续灌胃cm或pae4天后,shr大鼠的平均动脉压降至正常水平;而且继续应用同等剂量的cm或pae灌胃可将shr大鼠的平均动脉血压稳定维持在这一水平。4灌胃cm或pae对wky大鼠的收缩压(systolicbloodpressure,sbp)和舒张压(diastolicbloodpressure,dbp)均没有明显影响,但可降低shr大鼠的sbp及dbp(p0.05)。小结:cm有显著的降低shr大鼠平均动脉压的作用,并且其主要成分pae,也能够达到类似的降低shr大鼠平均动脉压的作用。第二部分丹皮酚对自发性高血压大鼠主动脉舒张功能的影响目的:观察pae对大鼠离体主动脉环的舒张功能的影响,并分析其可能的作用机制。方法:应用离体血管环灌流装置以及bl-420e+生物机能实验系统记录大鼠离体主动脉环的舒缩活动。westernblot法测定enos的蛋白表达量,elisa试剂盒检测血管组织中no与血管内皮细胞camp的表达水平。结果:1与wky组相比,cm或pae可以浓度依赖性的诱导shr大鼠离体胸主动脉环舒张(p0.05)。去内皮或l-name预处理可以完全阻断cm或pae的效应。2pae显著提高了shr大鼠胸主动脉组织中enos的表达,并浓度依赖性地提高其no水平(p0.05),但对wky大鼠的胸主动脉组织中enos的表达及no水平没有显著影响。3 MEK阻断剂PD98059及PI3K途径阻断剂LY294002预处理对Pae舒张SHR大鼠的主动脉及激活局部eNOS的作用没有影响。4 PKA的阻断剂H89预处理可阻断Pae对SHR大鼠主动脉组织中eNOS的激活(P0.05)。5 Pae预处理能浓度依赖性地提高培养的SHR大鼠胸主动脉内皮细胞中cAMP的表达水平(P0.05)。小结:Pae通过cAMP/PKA分子通路激活eNOS,提高局部组织中NO水平,诱导SHR大鼠胸主动脉舒张。第三部分丹皮酚对自发性高血压大鼠肾动脉舒张功能的影响目的:观察Pae对肾动脉的舒张作用,并分析其可能的作用机制。方法:应用离体血管环灌流装置以及BL-420E+生物机能实验系统来记录大鼠肾动脉的血管舒缩活动。内皮型eNOS的蛋白表达量用Western blot测定,血管组织中NO与血管内皮细胞cAMP的表达用ELISA试剂盒测定。结果:1 CM或Pae可以浓度依赖性的诱导SHR大鼠离体肾动脉环舒张(P0.05)。去内皮或L-NAME预处理可以完全阻断CM或Pae的效应。2 Pae显著提高了SHR大鼠肾动脉组织中eNOS的表达,并浓度依赖性地提高其NO水平(P0.05),但对WKY大鼠的肾动脉组织中eNOS的表达及NO水平没有显著影响。3 MEK阻断剂PD98059及PI3K途径阻断剂LY294002预处理对Pae舒张肾动脉及激活局部eNOS的作用没有影响。4 PKA的阻断剂H89预处理可阻断Pae对SHR大鼠肾动脉组织eNOS的激活(P0.05)。5 Pae预处理能浓度依赖性地提高培养的SHR大鼠肾动脉内皮细胞中cAMP的表达水平(P0.05)。小结:丹皮酚可引起SHR大鼠离体肾动脉环舒张,其作用机制与肾动脉cAMP/PKA依赖的eNOS的激活有关。
[Abstract]:Cortex Moutan (CM) is often used to treat cardiovascular and cerebrovascular diseases in traditional Chinese medicine. It is a traditional Chinese herbal medicine which is widely used in Asia and Europe. Paeonol (Pae) is one of the most important effective components of peony skin. It is a small molecular phenolic compound (2'- hydroxy -4'- methoxy Acetophenone, C9H10O3), and has the resistance to atherosclerosis. .CM is one of the major drugs in the antihypertensive group of Chinese medicine for the treatment of hypertension. In recent years, the research on the mechanism of CM has become a hot topic in the related subjects. The effects of.Pae on arterial blood pressure in normal blood pressure animals were observed by Pae. The effect and possible mechanism of.Pae on arterial blood pressure in the conscious state of hypertension model were not yet clear. Nitric oxide (nitric (NO)) is one of the most important physiological regulating factors in the cardiovascular system, and is involved in the regulation of vascular function homeostasis. The body has three The subtype of nitric oxide synthase (NOS) involved in vascular function regulation is mainly endothelial nitric oxide synthase (eNOS). Studies have shown that the increase in the expression level of eNOS can increase the production of local NO, which can lead to a series of vasodilatation, endothelial and smooth muscle cell function changes, and so on. Vasodilator.ENOS is activated mainly through three signaling pathways: whether MEK/ERK, PI3K/Akt and cAMP/PKA pathway can induce vasodilatation by affecting the expression of eNOS in endothelial cells and thus play a hypotensive effect, and how its possible activation mechanism is uncertain. This study aims to be spontaneously high in the classic hypertensive animal model. The blood pressure rats (spontaneously hypertensive rat, SHR) were treated with Pae. The effects of Pae treatment on arterial blood pressure in SHR rats were systematically investigated by means of functional and molecular biology methods. The effects of Pae treatment on arterial and renal artery diastolic function were systematically observed and the possible effects on the diastolic function of aorta and renal arteries were discussed. The study is divided into three parts: (1) the effect of Paeonol on arterial blood pressure in spontaneously hypertensive rats; (2) the effect of Paeonol on the diastolic function of thoracic aorta in spontaneously hypertensive rats; (3) the effect of Paeonol on the diastolic function of renal arteries in spontaneously hypertensive rats. Objective: To observe the effect of continuous intragastric CM decoction or Pae on arterial blood pressure in SHR rats. Methods: 30 healthy male WKY rats and 30 SHR rats were randomly divided into wky+control, wky+cm, wky+pae, shr+control, shr+cm and shr+pae groups. The state of waking state of rats was measured by a noninvasive tail artery blood pressure measuring instrument. Results: the mean arterial pressure of WKY rats was not affected by 1cm perfusion, but 0.5h, 1H, 2h, 4h, 8h significantly decreased the mean arterial pressure (P0.05).2 of SHR rats after administration, but no effect on the mean arterial pressure of rats in WKY rats, but the mean arterial pressure of rats was significantly reduced after the administration. 3 continuous perfusion of cm or PAE had no significant effect on the mean arterial pressure of WKY rats. The mean arterial pressure dropped to normal level in SHR rats after continuous gavage for cm or pae4 days, and the continued use of the same dosage of cm or PAE to maintain the average arterial pressure of SHR rats could maintain the systolic pressure of cm or PAE rats at this level. Oodpressure, SBP) and diastolic pressure (diastolicbloodpressure, DBP) had no obvious effects, but decreased SBP and DBP (P0.05) in SHR rats. Cm significantly reduced the mean arterial pressure of SHR rats, and its main component PAE, can also achieve a similar decrease in the mean arterial pressure of SHR rats. The second part of paeonol was spontaneous. The effect of PAE on diastolic function of aorta in rats with sexual hypertension: To observe the effect of PAE on the diastolic function of isolated aortic rings in rats and to analyze the possible mechanism of action. Methods: the.Westernblot method was used to determine the systolic and diastolic activity of the isolated aorta ring in rats by using the isolated vascular ring perfusion device and the bl-420e+ biological function test system. Protein expression and the expression of no in vascular tissue and expression level of camp in vascular endothelial cells by ELISA kit. Results: 1 compared with group WKY, cm or PAE can induce the concentration dependence of SHR rats' isolated thoracic aorta ring relaxation (P0.05). Endothelial or L-NAME preconditioning can completely obstruct cm or PAE effect.2pae significantly increased rat chest. The expression of eNOS in the aortic tissue and the concentration dependent enhancement of its no level (P0.05), but the expression of eNOS and the level of no in the thoracic aorta of WKY rats have no significant effect on the.3 MEK blocker PD98059 and PI3K pathway blockers. Blocking agent H89 preconditioning can block the activation of eNOS in the aortic tissue of SHR rats (P0.05).5 Pae preconditioning can increase the level of cAMP expression in the thoracic aorta endothelial cells of SHR rats (P0.05) in a concentration dependent manner. The effect of third partial paeonol on the diastolic function of renal artery in spontaneously hypertensive rats. Objective: To observe the effect of Pae on the diastolic function of the renal artery and to analyze its possible mechanism. Methods: the vasomotor activity of the renal artery in rats was recorded by using the isolated vascular loop perfusion device and the BL-420E+ biological experiment system. The protein expression of type eNOS was measured by Western blot. The expression of NO in vascular tissue and the expression of cAMP in vascular endothelial cells was determined by ELISA kit. Results: 1 CM or Pae can induce concentration dependent renal artery ring relaxation in SHR rats (P0.05). The expression of eNOS in the rat renal artery and the concentration dependent enhancement of its NO level (P0.05), but the expression of eNOS and the level of NO in the renal artery of WKY rats were not significantly affected by the.3 MEK blocker PD98059 and PI3K pathway blockers. H89 preconditioning can block the activation of eNOS in renal artery tissue of SHR rats (P0.05).5 Pae preconditioning can increase the level of cAMP expression in the renal artery endothelial cells of SHR rats (P0.05). Conclusion: paeonol can cause the relaxation of renal artery ring in SHR rats, and the mechanism of action and activation of renal artery cAMP/PKA dependent activation Of
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5

【参考文献】

中国期刊全文数据库 前3条

1 张金艳;曹永孝;翁维良;李贻奎;赵乐;;Paeonol Induces Vasodilatation in Rat Mesenteric Artery via Inhibiting Extracellular Ca~(2+) Influx and Intracellular Ca~(2+) Release[J];Chinese Journal of Integrative Medicine;2013年07期

2 ;Antiproliferation and apoptosis induction of paeonol in HepG_2 cells[J];World Journal of Gastroenterology;2007年02期

3 张广钦,禹志领,赵厚长;丹皮酚对大鼠反复性脑缺血的保护作用[J];中药材;1997年12期

，

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