發(fā)布時(shí)間：2018-04-15 11:33

  本文選題：纈沙坦 + 自發(fā)性高血壓大鼠��； 參考：《皖南醫(yī)學(xué)院》2017年碩士論文

【摘要】：晝夜節(jié)律是生物界常見(jiàn)的生物節(jié)律之一,人類許多生理生化功能和行為均呈現(xiàn)出晝夜節(jié)律變化。高血壓是最常見(jiàn)的心血管疾病之一,現(xiàn)如今,血壓晝夜節(jié)律模式已列為高血壓診斷和評(píng)估心血管事件的參考標(biāo)準(zhǔn)之一。血管緊張素Ⅱ(angiotensinⅡ,AngⅡ)與高血壓發(fā)生密切相關(guān),并參與血壓晝夜節(jié)律的調(diào)節(jié),纈沙坦是AngⅡ受體拮抗劑,能選擇性阻斷AT1受體。本課題組前期臨床研究顯示晝夜不同時(shí)間用藥,纈沙坦對(duì)非杓型高血壓患者的血壓晝夜節(jié)律有顯著影響,休息期用藥,可使高血壓患者非杓型血壓節(jié)律明顯改善,但有關(guān)其調(diào)控血壓晝夜節(jié)律的生物鐘機(jī)制尚不清楚。本實(shí)驗(yàn)以生物鐘系統(tǒng)晝夜節(jié)律調(diào)控的核心機(jī)制為理論指導(dǎo),探討纈沙坦對(duì)異常血壓晝夜節(jié)律的干預(yù)作用以及對(duì)生物鐘基因Per1、Per2表達(dá)節(jié)律的影響,從整體水平闡明生物鐘基因Per1、Per2參與纈沙坦調(diào)控血壓晝夜節(jié)律的機(jī)制。本課題研究獲得的信息將為高血壓的時(shí)辰治療提供理論基礎(chǔ)以及為尋找更理想的降壓藥物提供新思路。目的:觀察纈沙坦對(duì)自發(fā)性高血壓大鼠血壓晝夜節(jié)律的調(diào)控作用以及不同時(shí)間點(diǎn)給藥對(duì)血壓水平、血壓晝夜節(jié)律的影響,并探討其生物鐘機(jī)制。方法:10周齡雄性自發(fā)性高血壓大鼠(SHR)和10周齡雄性Wistar-Kyoto大鼠(WKY)置于時(shí)間生物學(xué)實(shí)驗(yàn)室內(nèi)適應(yīng)性喂養(yǎng)2周,參照WKY大鼠血壓晝夜節(jié)律模式作為正常對(duì)照組,確定SHR大鼠異常血壓晝夜節(jié)律模式。將其隨機(jī)分為三組:SHR模型組、纈沙坦08:00組、纈沙坦20:00組。灌胃給予纈沙坦30mg/kg,連續(xù)給藥6周。采用尾袖法,于給藥前、給藥后每2周分別測(cè)量各組大鼠連續(xù)24h尾動(dòng)脈血壓。給藥6周后,各組動(dòng)物依次分別在24h內(nèi)7個(gè)時(shí)間點(diǎn)(CT0,CT4,CT8,CT12,CT16,CT20,CT24)進(jìn)行采集樣品。采用 ELISA法檢測(cè)血漿AngⅡ、ACE、REN、ALD含量,觀察各項(xiàng)指標(biāo)24h動(dòng)態(tài)變化,分析晝夜節(jié)律;HE染色和Masson染色觀察心臟和主動(dòng)脈靶器官病理?yè)p傷程度。Real-time PCR法檢測(cè)大鼠下丘腦、主動(dòng)脈生物鐘基因Per1、Per2 mRNA表達(dá)水平,觀察24h動(dòng)態(tài)變化,分析晝夜節(jié)律;Western blot法檢測(cè)主動(dòng)脈生物鐘蛋白PER1、PER2蛋白表達(dá)水平,觀察24h動(dòng)態(tài)變化,分析晝夜節(jié)律;結(jié)果:(1)在給藥期間,與模型組相比,纈沙坦08:00組的收縮壓、舒張壓和平均動(dòng)脈壓均降低(P0.01),血壓杓型值升高(P0.01),血壓晝夜節(jié)律參數(shù)中值降低,振幅升高,峰值相位前移,其異常血壓晝夜節(jié)律得到明顯改善(P0.01);纈沙坦20:00組的收縮壓、舒張壓和平均動(dòng)脈壓均降低(P0.01),但血壓杓型值并未明顯升高。(2)給藥6周后,與模型組相比,纈沙坦08:00組的血漿AngⅡ休息期水平升高,活動(dòng)期水平降低,振幅升高(P0.01);腎素整體水平降低,血管緊張素轉(zhuǎn)化酶和醛固酮整體水平升高(P0.01),血漿中RAAS系統(tǒng)晝夜節(jié)律發(fā)生改變。(3)給藥6周后,與模型組相比,纈沙坦08:00組的臟器系數(shù)降低,心臟和主動(dòng)脈靶器官的病理?yè)p傷明顯得到緩解;纈沙坦20:00組的靶器官病理?yè)p傷緩解程度不及纈沙坦08:00組。(4)給藥6周后,與模型組相比,纈沙坦08:00組的下丘腦組織Per1 mRNA活動(dòng)期表達(dá)升高,Per2 mRNA休息期表達(dá)升高(P0.05);(5)給藥6周后,與模型組相比,纈沙坦08:00組的主動(dòng)脈組織中Per1 mRNA和Per2 mRNA整體表達(dá)降低,中值和振幅降低(P0.01);主動(dòng)脈組織中PER1蛋白活動(dòng)期表達(dá)升高,PER2蛋白整體表達(dá)升高(P0.01);生物鐘基因Per1、Per2 mRNA和蛋白表達(dá)晝夜節(jié)律發(fā)生改變。結(jié)論:相對(duì)于活動(dòng)期用藥,纈沙坦休息期用藥不僅能有效降低血壓,保護(hù)靶器官心臟和主動(dòng)脈病理?yè)p傷,還能明顯改善自發(fā)性高血壓大鼠的異常血壓晝夜節(jié)律,使其逆轉(zhuǎn)為杓型血壓晝夜節(jié)律,其機(jī)制可能與影響血漿RAAS系統(tǒng)的晝夜節(jié)律性,進(jìn)而改變下丘腦和主動(dòng)脈生物鐘基因Per1、Per2 mRNA與蛋白表達(dá)的晝夜節(jié)律性有關(guān)。
[Abstract]:Circadian rhythm is one of the common biological rhythms in biology, many human physiological and biochemical function and behavior showed circadian rhythm changes. Hypertension is one of the most common cardiovascular disease nowadays, has been listed as one of the circadian blood pressure pattern reference standard for hypertension diagnosis and assessment of cardiovascular events. Angiotensin II (angiotensin II. Ang II) is closely related to hypertension, and involved in the regulation of circadian rhythm of blood pressure. Valsartan is Ang receptor antagonist, selective blockade of AT1 receptor. Our previous clinical studies have shown that day at the same time medication, have a significant effect of valsartan on circadian rhythm of blood pressure in patients with non dipper hypertension, rest period medication, can the hypertensive patients with non dipper rhythm of blood pressure significantly improved, but the biological clock mechanism related to its regulation of circadian rhythm of blood pressure is not clear. In this experiment, the circadian clock systems The core mechanism of rhythm regulation theory, the effects of valsartan on abnormal circadian rhythm of blood pressure and the circadian clock gene Per1, Per2 expression of rhythm, from the overall level to clarify the biological clock gene Per1, Per2 participates in the regulation mechanism of valsartan on the circadian rhythm of blood pressure. Research on this topic will get information for the treatment of hypertension. To provide a theoretical basis and provide new ideas for more ideal antihypertensive drugs. Objective: To observe the effect of valsartan on regulation of circadian rhythm of blood pressure in spontaneously hypertensive rats and administration in different time on blood pressure level, affect the circadian rhythm of blood pressure, and to explore the biological clock mechanism. Methods: 10 week old male spontaneously hypertensive rats (SHR) and 10 week old male Wistar-Kyoto rats (WKY) in 2 weeks time in the biology laboratory reference adaptive feeding, circadian rhythm of blood pressure model WKY rats As the normal control group, SHR rats abnormal circadian rhythm of blood pressure. They were randomly divided into three groups: SHR model group, valsartan group 08:00, valsartan group. 20:00 intragastric administration of 30mg/kg valsartan administered continuously for 6 weeks. By using the tail cuff method, prior to drug administration, every 2 weeks after treatment respectively. Continuous 24h measuring the blood pressure of tail artery volume of rats. After 6 weeks of treatment, each animal respectively in 24h 7 time points (CT0, CT4, CT8, CT12, CT16, CT20, CT24) were collected. Plasma samples were detected with ELISA Ang II, ACE, REN, ALD in the observation 24h index analysis of dynamic change of circadian rhythm; HE staining and Masson staining were used to observe the heart and aorta were the target organs damage degree of.Real-time PCR was detected in rat hypothalamus, aortic clock gene Per1, Per2 expression level of mRNA, the dynamic changes of 24h, analysis of circadian clock protein PER; detection of aortic Western blot method 1, the expression level of PER2 protein, the dynamic changes of 24h, analysis of circadian rhythm; results: (1) during the period of delivery, compared with the model group, valsartan group 08:00 systolic pressure, diastolic pressure and mean arterial pressure (P0.01) decreased, increased blood pressure dipper value (P0.01), blood pressure circadian rhythm parameters the median decreased and the amplitude increased peak phase forward, the abnormal circadian rhythm of blood pressure was significantly improved (P0.01); valsartan group 20:00 systolic pressure, diastolic pressure and mean arterial pressure (P0.01) decreased, but the blood pressure did not increase obviously. The dipper value (2) after 6 weeks of treatment, compared with the model group. Plasma Ang II valsartan group 08:00 resting levels, the level of activity decreased and the amplitude increased (P0.01); to reduce the overall level of renin, angiotensin converting enzyme and increase the overall level of aldosterone (P0.01), change the RAAS system. The circadian rhythm of plasma (3) after 6 weeks of treatment, compared with model group valerian. Reduce the organ coefficient of 08:00 group candesartan in heart and aorta, the pathological damage of target organs has been alleviated; target organ damage relieve pathological group valsartan 20:00 less 08:00 valsartan group. (4) after 6 weeks of treatment, compared with model group, valsartan group 08:00 group Per1 active mRNA fabric hypothalamic expression increased, Per2 mRNA expression increased rest period (P0.05); (5) after 6 weeks of treatment, compared with model group, valsartan group 08:00 in aortic tissue of Per1 mRNA and Per2 mRNA expression decreased, and the median amplitude decreased (P0.01); PER1 protein activity in aorta tissue expression increased, PER2 protein expression increased overall (P0.01); clock gene Per1, Per2 mRNA and protein expression of circadian rhythm changes. Conclusion: compared with the active drug, valsartan medication rest period not only can effectively reduce blood pressure, protect the target organs of the heart and aorta pathological damage, but also Improve abnormal circadian rhythm of blood pressure in spontaneously hypertensive rats, the reversal of circadian rhythm of blood pressure, and its mechanism may be related to circadian rhythm of plasma RAAS system, and then change the hypothalamus and aortic clock gene Per1, circadian rhythm of the expression of mRNA and Per2 protein.

【學(xué)位授予單位】：皖南醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R544.1

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本文編號(hào)：1753975