【摘要】：牡丹皮(Cortex Moutan,CM)在傳統(tǒng)中醫(yī)中常用來治療心腦血管疾病,是在亞洲和歐洲被廣泛應用的一種傳統(tǒng)中草藥。丹皮酚(Paeonol,Pae)是牡丹皮的最主要有效成分之一,是一種小分子酚類化合物(2'-羥基-4'-甲氧基苯乙酮,C9H10O3),具有抗動脈粥樣硬化,抗心律失常,抗炎癥和抗氧化等多方面的藥理作用。CM是中醫(yī)治療高血壓的降壓組方中的主要藥物之一。近年來,關(guān)于CM降壓機制的研究已經(jīng)成為相關(guān)學科的研究熱點。但是,到目前為止此類研究的結(jié)果均是在麻醉狀態(tài)下,一次或分次大量應用Pae,觀察其對血壓正常動物動脈血壓的影響。Pae對高血壓模型動物清醒狀態(tài)下動脈血壓的影響及可能機制尚不明確。一氧化氮(nitric oxide,NO)是心血管系統(tǒng)中最重要的生理調(diào)節(jié)因子之一,參與血管功能穩(wěn)態(tài)的調(diào)節(jié)。機體有三種亞型的一氧化氮合酶,參與血管功能調(diào)節(jié)的主要是內(nèi)皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)。研究證實eNOS的表達水平升高或被激活時,局部NO的產(chǎn)生量會增加,可以引起局部血管舒張、內(nèi)皮及平滑肌細胞功能變化等一系列血管機能改變。eNOS主要通過三條信號通路激活:即MEK/ERK、PI3K/Akt和cAMP/PKA途徑。Pae是否能夠通過影響內(nèi)皮細胞eNOS的表達,誘導血管舒張,從而起到降壓作用,其可能激活機制如何尚不確定。本研究旨在通過對經(jīng)典高血壓動物模型自發(fā)性高血壓大鼠(spontaneously hypertensive rat,SHR)給予Pae處理,利用功能學和分子生物學等方法,從整體動物、離體組織及蛋白表達等不同水平,系統(tǒng)動態(tài)地觀察Pae處理對SHR大鼠動脈血壓的影響,并探討其對主動脈和腎動脈舒張功能的影響和可能作用機制。研究分為以下三部分:(1)丹皮酚對自發(fā)性高血壓大鼠動脈血壓的影響;(2)丹皮酚對自發(fā)性高血壓大鼠胸主動脈舒張功能的影響;(3)丹皮酚對自發(fā)性高血壓大鼠腎動脈舒張功能的影響。第一部分丹皮酚對自發(fā)性高血壓大鼠動脈血壓的影響目的:觀察連續(xù)灌胃CM煎劑或Pae對SHR大鼠動脈血壓的影響。方法:健康雄性wky大鼠30只、shr大鼠30只,分別隨機分為wky+control、wky+cm、wky+pae、shr+control、shr+cm和shr+pae組。采用灌胃的給藥方式,用無創(chuàng)鼠尾動脈血壓測量儀測量大鼠清醒狀態(tài)下的動脈血壓。結(jié)果:1cm灌胃對wky大鼠的平均動脈壓無影響,但在給藥后0.5h,1h,2h,4h,8h均顯著降低了shr大鼠的平均動脈血壓(p0.05)。2灌胃pae對wky大鼠的平均動脈壓無影響,但在給藥后0.5h,1h,2h,4h,8h均顯著降低了shr大鼠的平均動脈血壓(p0.05)。3連續(xù)灌胃cm或pae對wky鼠的平均動脈壓無明顯影響。連續(xù)灌胃cm或pae4天后,shr大鼠的平均動脈壓降至正常水平;而且繼續(xù)應用同等劑量的cm或pae灌胃可將shr大鼠的平均動脈血壓穩(wěn)定維持在這一水平。4灌胃cm或pae對wky大鼠的收縮壓(systolicbloodpressure,sbp)和舒張壓(diastolicbloodpressure,dbp)均沒有明顯影響,但可降低shr大鼠的sbp及dbp(p0.05)。小結(jié):cm有顯著的降低shr大鼠平均動脈壓的作用,并且其主要成分pae,也能夠達到類似的降低shr大鼠平均動脈壓的作用。第二部分丹皮酚對自發(fā)性高血壓大鼠主動脈舒張功能的影響目的:觀察pae對大鼠離體主動脈環(huán)的舒張功能的影響,并分析其可能的作用機制。方法:應用離體血管環(huán)灌流裝置以及bl-420e+生物機能實驗系統(tǒng)記錄大鼠離體主動脈環(huán)的舒縮活動。westernblot法測定enos的蛋白表達量,elisa試劑盒檢測血管組織中no與血管內(nèi)皮細胞camp的表達水平。結(jié)果:1與wky組相比,cm或pae可以濃度依賴性的誘導shr大鼠離體胸主動脈環(huán)舒張(p0.05)。去內(nèi)皮或l-name預處理可以完全阻斷cm或pae的效應。2pae顯著提高了shr大鼠胸主動脈組織中enos的表達,并濃度依賴性地提高其no水平(p0.05),但對wky大鼠的胸主動脈組織中enos的表達及no水平?jīng)]有顯著影響。3 MEK阻斷劑PD98059及PI3K途徑阻斷劑LY294002預處理對Pae舒張SHR大鼠的主動脈及激活局部eNOS的作用沒有影響。4 PKA的阻斷劑H89預處理可阻斷Pae對SHR大鼠主動脈組織中eNOS的激活(P0.05)。5 Pae預處理能濃度依賴性地提高培養(yǎng)的SHR大鼠胸主動脈內(nèi)皮細胞中cAMP的表達水平(P0.05)。小結(jié):Pae通過cAMP/PKA分子通路激活eNOS,提高局部組織中NO水平,誘導SHR大鼠胸主動脈舒張。第三部分丹皮酚對自發(fā)性高血壓大鼠腎動脈舒張功能的影響目的:觀察Pae對腎動脈的舒張作用,并分析其可能的作用機制。方法:應用離體血管環(huán)灌流裝置以及BL-420E+生物機能實驗系統(tǒng)來記錄大鼠腎動脈的血管舒縮活動。內(nèi)皮型eNOS的蛋白表達量用Western blot測定,血管組織中NO與血管內(nèi)皮細胞cAMP的表達用ELISA試劑盒測定。結(jié)果:1 CM或Pae可以濃度依賴性的誘導SHR大鼠離體腎動脈環(huán)舒張(P0.05)。去內(nèi)皮或L-NAME預處理可以完全阻斷CM或Pae的效應。2 Pae顯著提高了SHR大鼠腎動脈組織中eNOS的表達,并濃度依賴性地提高其NO水平(P0.05),但對WKY大鼠的腎動脈組織中eNOS的表達及NO水平?jīng)]有顯著影響。3 MEK阻斷劑PD98059及PI3K途徑阻斷劑LY294002預處理對Pae舒張腎動脈及激活局部eNOS的作用沒有影響。4 PKA的阻斷劑H89預處理可阻斷Pae對SHR大鼠腎動脈組織eNOS的激活(P0.05)。5 Pae預處理能濃度依賴性地提高培養(yǎng)的SHR大鼠腎動脈內(nèi)皮細胞中cAMP的表達水平(P0.05)。小結(jié):丹皮酚可引起SHR大鼠離體腎動脈環(huán)舒張,其作用機制與腎動脈cAMP/PKA依賴的eNOS的激活有關(guān)。
[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
【學位授予單位】：河北醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R285.5

【參考文獻】

中國期刊全文數(shù)據(jù)庫 前3條

1 張金艷;曹永孝;翁維良;李貽奎;趙樂;;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 張廣欽,禹志領(lǐng),趙厚長;丹皮酚對大鼠反復性腦缺血的保護作用[J];中藥材;1997年12期

，

本文編號：2145874