本文選題：肺動(dòng)脈高壓 + 伐地那非��； 參考：《山東大學(xué)》2014年博士論文

【摘要】：肺動(dòng)脈高壓(pulmonary arterial hypertension, PAH)是一類多病因、進(jìn)展性的疾病,其定義為靜息狀態(tài)右心導(dǎo)管(RHC)測(cè)定平均肺動(dòng)脈壓大于等于25mmHg,最終進(jìn)展為右心衰竭,直至死亡。肺動(dòng)脈高壓是多種生理途徑和細(xì)胞類型引起的多因素的病理生理改變,包括肺血管收縮、增殖,血管阻塞性重構(gòu),炎性浸潤(rùn)及原位血栓形成等機(jī)制。越來(lái)越多的研究表明,氧化應(yīng)激參與了肺動(dòng)脈高壓的發(fā)病過程。氧化應(yīng)激被定義為氧化物的增加(如過氧化氫、超氧陰離子等)伴或不伴有抗氧化物或抗氧化酶(如超氧化物歧化酶)的降低。增加的氧化物不但能夠通過氧化特定的生物分子,直接損傷組織器官,包括脂質(zhì)過氧化和DNA氧化,而且可以與一氧化氮(NO)結(jié)合,導(dǎo)致毒性分子過氧亞硝酸的產(chǎn)生和內(nèi)皮一氧化氮合成酶(endothelial nitric oxide synthase, eNOS)的脫偶聯(lián)。 在過去幾年里,肺動(dòng)脈高壓的藥物治療有了很大進(jìn)展,文獻(xiàn)報(bào)導(dǎo)顯示使用靶向藥物治療的肺動(dòng)脈高壓患者比安慰劑組的死亡率下降了43%,住院率下降了61%31。目前應(yīng)用于肺動(dòng)脈高壓治療的靶向藥物有以下3類：1)前列環(huán)素類似物；2)內(nèi)皮素受體拮抗劑；3)5-磷酸二酯酶抑制劑。其中5-磷酸二酯酶抑制劑包括西地那非、伐地那非和他達(dá)拉非,5-磷酸二酯酶抑制劑在肺血管系統(tǒng)中大量表達(dá)。5-磷酸二酯酶抑制劑通過NO/cGMP通路起到擴(kuò)張血管和抗增殖作用。5型磷酸二酯酶(phosphodiesterase-type5, PDE5)是在肺組織中廣泛表達(dá)的的磷酸二酯酶異構(gòu)酶,它能夠在鳥苷酸環(huán)化酶(guanylate cyclase)的作用下降解環(huán)磷酸鳥苷(cyclic guanosine monophosphate, cGMP),引起皿官收縮。PDE5抑制劑,包括西地那非(sildenafil)和他達(dá)拉非(tadalafil),目前已經(jīng)被批準(zhǔn)用于肺動(dòng)脈高壓的臨床治療。 伐地那非(vadenafil),屬于新型PDE5抑制劑。與傳統(tǒng)的PDE5抑制劑相比,它完全溶于水和乙醇,對(duì)PDE5的特異性作用更強(qiáng)。2011年,通過一項(xiàng)臨床雙盲、隨機(jī)、安慰劑實(shí)驗(yàn),研究者發(fā)現(xiàn)伐地那非能夠有效的改善PAH患者的運(yùn)動(dòng)能力、血流動(dòng)力學(xué)指數(shù)和心功能,但機(jī)制尚不明確。在勃起功能障礙的基礎(chǔ)和臨床研究中,研究人員發(fā)現(xiàn)除了PDE5抑制功能,PDE5抑制劑還能夠通過抑制氧自由基形成,降低NADPH氧化酶起到強(qiáng)大的抗氧化作用,從而緩解內(nèi)皮功能障礙和血管重構(gòu),但其他PAH靶向藥物,如依前列醇(epoprostenol)并沒有此功能。Ghofrani等報(bào)道給予PAH患者短期伐地那非治療后,顯示雖然伐地那非能引起肺血管收縮,但其對(duì)肺循環(huán)血管缺乏特異性,另外,伐地那非中長(zhǎng)期治療也有個(gè)案報(bào)道,顯示其對(duì)PAH的治療并沒有特殊效果。因此,伐地那非治療PAH的有效性及耐受性仍值得進(jìn)一步商榷。 2009年,荊志成教授及其課題組,通過對(duì)45名PAH患者的研究,顯示1年的伐地那非治療能夠有效地治療PAH。2011年,該課題組繼續(xù)進(jìn)行了一項(xiàng)隨機(jī)、雙盲、安慰劑-對(duì)照及多中心的研究。結(jié)果顯示與西地那非和他達(dá)拉非不同,伐地那非單藥治療即能夠有效的延緩PAH患者的臨床惡化程度,改善其血流動(dòng)力學(xué)變化。然而伐地那非治療肺動(dòng)脈高壓的具體機(jī)制仍不明確。 鑒于目前伐地那非治療PAH功能研究的缺乏,本課題收入15名特發(fā)性PAH患者,同時(shí)以野百合堿誘導(dǎo)的PAH大鼠為體內(nèi)模型,以H202誘導(dǎo)的氧化應(yīng)激內(nèi)皮細(xì)胞為體外模型,研究伐地那非治療PAH的機(jī)制研究和可能涉及的信號(hào)通路,為PAH的治療提供了依據(jù)。研究分以下三部分： 第一部分伐地那非改善氧化應(yīng)激并治療特發(fā)性肺動(dòng)脈高壓的臨床研究 本研究選擇特發(fā)性肺動(dòng)脈高壓患者作為研究對(duì)象,分析了伐地那非對(duì)特發(fā)性肺動(dòng)脈高壓患者的治療作用,及其對(duì)患者體內(nèi)氧化應(yīng)激水平的改善作用。1)我們選取了EVALUATION研究中的15名特發(fā)性肺動(dòng)脈高壓的患者,并選擇了年齡及性別與之相匹配的健康人群作為對(duì)照。入選患者給予伐地那非每次5mg,每日1次口服1個(gè)月,若患者未出現(xiàn)嚴(yán)重不良反應(yīng),則加量至每次5mg,每日2次繼續(xù)口服2個(gè)月。 2)我們于基線及3個(gè)月后評(píng)估完成以下內(nèi)容：標(biāo)準(zhǔn)病史與體格檢查,6分鐘步行距離,WHO肺動(dòng)脈高壓功能分級(jí),血生化指標(biāo),心電圖,肺功能檢查,和完整的右心導(dǎo)管檢查所測(cè)血流動(dòng)力學(xué)資料。結(jié)果顯示3個(gè)月的伐地那非治療能顯著改善特發(fā)性肺動(dòng)脈高壓患者WHO功能分級(jí)、六分鐘步行距離、心指數(shù)、肺血管阻力等指標(biāo)。盡管跟用藥前相比較,患者平均肺動(dòng)脈壓力、肺毛細(xì)血管楔壓、平均右房壓力較前有降低,但并沒有統(tǒng)計(jì)學(xué)差異。以上結(jié)果說(shuō)明伐地那非能夠改善肺動(dòng)脈高壓患者臨床癥狀及血流動(dòng)力學(xué)變化。 3)隨后我們采用商品化試劑盒測(cè)量了肺動(dòng)脈高壓患者及健康對(duì)照組血液中一氧化氮、丙二醛、8-異前列素F2α、3-硝基酪氨酸、超氧化物歧化酶等氧化應(yīng)激及氮化應(yīng)激相關(guān)產(chǎn)物的水平。發(fā)現(xiàn)肺動(dòng)脈高壓患者血漿中氧化損傷產(chǎn)物丙二醛、8-異前列素F2α、3-硝基酪氨酸明顯高于正常人,氧化應(yīng)激保護(hù)性產(chǎn)物一氧化氮、超氧化物歧化酶明顯低于正常對(duì)照者。伐地那非治療3個(gè)月后,氧化損傷產(chǎn)物8-異前列素F2α及3-硝基酪氨酸水平均較前降低,但仍高于正常人,而丙二醛的水平并沒有明顯改變。血管擴(kuò)張產(chǎn)物一氧化氮較治療前明顯升高,基本恢復(fù)正常水平�？寡趸赋趸锲缁篙^治療前升高,與正常人相比,仍處于較低水平。以上以上結(jié)果顯示,肺動(dòng)脈高壓患者體內(nèi)存在較高的氧化應(yīng)激水平,抗氧化能力明顯較正常人減弱。而伐地那非治療能夠降低患者的氧化應(yīng)激水平,改善其抗氧化能力,起到保護(hù)性作用。 第二部分伐地那非改善野百合堿誘導(dǎo)肺動(dòng)脈高壓大鼠氧化應(yīng)激水平的研究 本研究選擇了34只雄性SPF級(jí)Sprague-Dawley (SD)大鼠,體重200-220g,隨機(jī)分為正常對(duì)照組、野百合堿模型組以及伐地那非治療組。給予50mg/kg野百合堿21天后,伐地那非治療組大鼠予1mg/kg/d灌胃共21天。之后進(jìn)行如下研究： 1)應(yīng)用右心導(dǎo)管檢查術(shù)測(cè)量大鼠的血流動(dòng)力學(xué)指標(biāo),發(fā)現(xiàn)MCT組大鼠的mPAP、RVSP和PVR明顯高于正常對(duì)照組,而CO則明顯低于正常對(duì)照組。給予伐地那非治療后,伐地那非組大鼠的mPAP、RVSP和PVR較MCT組分別降低41%、25%和26%。以上結(jié)果與我們臨床試驗(yàn)結(jié)果相一致,進(jìn)一步驗(yàn)證了伐地那非對(duì)血流動(dòng)力學(xué)的改善作用。 2)為研究伐地那非對(duì)血流動(dòng)力學(xué)改善的原因,我們對(duì)大鼠肺組織進(jìn)行了組織學(xué)染色并應(yīng)用相關(guān)軟件對(duì)血管組織進(jìn)行測(cè)量。與正常對(duì)照組相比,野百合堿組大鼠的%WT和%WA明顯增高。而給予伐地那非治療后,伐地那非組的%WT和%WA明顯降低。以上結(jié)果在50μm-100μm、100-200μm直徑的肺血管中均有所體現(xiàn)。之后我們通過a-SMA抗體染色,檢測(cè)大鼠的血管肌化程度。跟正常對(duì)照組相比,野百合堿組血管肌化程度顯著增加,鏡下觀察平滑肌細(xì)胞明顯肥大增生。給予伐地那非治療后,伐地那非組肌化程度較MCT組降低了39%,平滑肌細(xì)胞增生肥大有所改善。最后我們對(duì)肺血管平滑肌細(xì)胞進(jìn)行了PCNA及TUNEL染色,發(fā)現(xiàn)伐地那非對(duì)血管厚度的改善是通過誘導(dǎo)大鼠肺動(dòng)脈平滑肌細(xì)胞的增值,同時(shí)促進(jìn)其凋亡而實(shí)現(xiàn)的。 我們應(yīng)用werstern blot的方法及商品化試劑盒,研究了伐地那非治療肺動(dòng)脈高壓的相關(guān)機(jī)制。與正常對(duì)照組相比,野百合堿組大鼠血漿及肺組織內(nèi)NO的濃度降低,.給予伐地那非治療后,跟野百合堿組相比,伐地那非組血漿及肺組織內(nèi)NO水平均較前恢復(fù)。原因是通過伐地那非上調(diào)eNOS蛋白的表達(dá)而實(shí)現(xiàn)的。之后我們發(fā)現(xiàn)伐地那非能夠改善大鼠體內(nèi)氧化應(yīng)激水平,與我們臨床試驗(yàn)結(jié)果相一致。為了進(jìn)一步研究可能的機(jī)制,我們選擇了氧化應(yīng)激過程中主要的酶,NADPH氧化酶和超氧化物歧化酶,進(jìn)行下一步研究。結(jié)果顯示與正常對(duì)照組相比,野百合堿大鼠大鼠體內(nèi)NADPH氧化酶催化亞基,NOX2和NOX4,的水平明顯上調(diào),但激活亞基RAC1水平并沒有明顯變化。伐地那非治療能顯著改善NOX2和NOX4的表達(dá)水平,卻同樣不影響RAC1亞基的表達(dá)水平。相反,MCT組大鼠體內(nèi)SOD活性較正常對(duì)照組相比明顯降低,而伐地那非治療后,SOD活性得到了顯著改善。以上結(jié)果提示,伐地那非對(duì)氧化應(yīng)激水平的改善,其中一部分是通過改善NADPH氧化酶和超氧化物歧化酶而實(shí)現(xiàn)的。 第三部分伐地那非通過BMP信號(hào)通路改善內(nèi)皮細(xì)胞氧化損傷的機(jī)制研究 通過第一部分和第二部分的體內(nèi)研究,我們明確了伐地那非能改善氧化應(yīng)激同時(shí)治療肺動(dòng)脈高壓,但具體機(jī)制仍不清楚。因此第三部分,我們應(yīng)用CRL-1730內(nèi)皮細(xì)胞系為研究模型,給予不同劑量外源性H202造成其氧化應(yīng)激損傷,進(jìn)一步研究了伐地那非對(duì)氧化應(yīng)激損傷保護(hù)性作用的具體機(jī)制。 1)通過MTT及TUNEL的方法,我們發(fā)現(xiàn)與對(duì)照組比較,H202組細(xì)胞活力顯著降低,伐地那非各劑量組與H202組比較,細(xì)胞活力顯著升高,且具有明顯的劑量依賴性。與對(duì)照組比較,H202組細(xì)胞凋亡率顯著增高,給予伐地那非處理后,細(xì)胞凋亡率較前明顯改善,且具有劑量依賴性。以上結(jié)果于體內(nèi)試驗(yàn)結(jié)果相互印證：伐地那非對(duì)內(nèi)皮細(xì)胞起到了強(qiáng)烈的保護(hù)性作用,且該作用呈劑量依賴性。 2)之后我們檢測(cè)了骨形態(tài)發(fā)生蛋白(bone morphogenetic proteins,BMPs)信號(hào)通路在各組細(xì)胞中的表達(dá),BMP信號(hào)通路廣泛參與了肺動(dòng)脈高壓、動(dòng)脈粥樣硬化等血管疾病。結(jié)果顯示,氧化應(yīng)激隨時(shí)能夠上調(diào)BMP受體1A、BMP受體1B、BMP受體2,同時(shí)上調(diào)胞漿內(nèi)p-smad水平,最后進(jìn)入細(xì)胞核內(nèi),上調(diào)轉(zhuǎn)錄因子MSX2的表達(dá)。而伐地那非能夠改善上述通路,起到保護(hù)性作用。 3)最后我們應(yīng)用BMP信號(hào)通路的抑制劑,Dorsomorphin干預(yù)細(xì)胞,并通過TUNEL檢測(cè)細(xì)胞凋亡情況。我們發(fā)現(xiàn),與對(duì)照組比較,H202組細(xì)胞內(nèi)細(xì)胞凋亡率顯著增高,伐地那非組與H202組比較,細(xì)胞凋亡率顯著下降,而與伐地那非組比較,Dorsomorphin干預(yù)組細(xì)胞凋亡率明顯上升。說(shuō)明Dorsomorphin能抑制伐地那非對(duì)內(nèi)皮細(xì)胞的保護(hù)性作用,進(jìn)一步說(shuō)明BMP信號(hào)通路參與了伐地那非對(duì)內(nèi)皮細(xì)胞氧化應(yīng)激損傷的改善。 綜上所述,我們通過臨床研究、動(dòng)物實(shí)驗(yàn)及體外細(xì)胞培養(yǎng),證明了伐地那非對(duì)PAH的治療作用以及對(duì)PAH患者體內(nèi)氧化應(yīng)激水平的改善作用,及其相關(guān)的機(jī)制。
[Abstract]:Pulmonary arterial hypertension (PAH) is a multi cause, progressive disease defined as resting right cardiac catheterization (RHC) for the determination of the mean pulmonary artery pressure greater than equal to 25mmHg, and eventually to right heart failure until death. Pulmonary hypertension is a multifactor pathogeny caused by multiple physiological pathways and cell types. A growing number of studies have shown that oxidative stress is involved in the pathogenesis of pulmonary arterial hypertension. Oxidative stress is defined as an increase in oxide (such as hydrogen peroxide, superoxide anion, etc.) with or without antioxidants or oxygen. The reduction of chemical enzymes (such as superoxide dismutase). The increased oxide can not only directly damage the tissues and organs by oxidation of specific biomolecules, including lipid peroxidation and DNA oxidation, but also can be combined with nitric oxide (NO), resulting in the production of peroxynitrite and the endothelial nitric oxid (endothelium nitric oxide synthase). Decoupling of E synthase, eNOS).
In the past few years, there has been great progress in the treatment of pulmonary hypertension. The literature has reported that the mortality of pulmonary hypertension patients using targeted drug therapy has decreased by 43% than that in the placebo group, and the hospitalization rate has decreased by 61%31., the target drugs used for the treatment of pulmonary hypertension are 3 categories: 1) prostacyclin analogues; 2) endothelium. 3) 5- phosphodiesterase inhibitors, in which 5- phosphodiesterase inhibitors, including sildenafil, dedionafil and tatabafil, 5- phosphodiesterase inhibitors in the pulmonary vascular system, expressed a large number of.5- phosphodiesterase inhibitors through the NO/cGMP pathway to expand the blood tube and anti proliferative.5 phosphodiesterase (phosphod Iesterase-type5, PDE5) is a phosphodiesterase isomerase, widely expressed in lung tissue, which can decrease cyclic guanosine monophosphate (cGMP) in the role of guanosine cyclase (guanylate cyclase), causing Petri dish officers to constriction.PDE5 inhibitors, including sildenafil (sildenafil) and tarafil (tadalafil). It has been approved for the clinical treatment of pulmonary hypertension.
Vadenafil, a new type of PDE5 inhibitor. Compared with the traditional PDE5 inhibitor, it is completely soluble in water and ethanol and is more specific to PDE5 than.2011 years. Through a double blind, randomized, placebo experiment, researchers found that FD can effectively improve the exercise ability, hemodynamic index and heart of PAH patients. In the basic and clinical studies of erectile dysfunction, the researchers found that in addition to PDE5 inhibitory function, PDE5 inhibitors can also inhibit the formation of oxygen free radicals, reduce the potent antioxidant activity of NADPH oxidase, and thus alleviate endothelial dysfunction and vascular remodeling, but other PAH targeting drugs, such as Epoprostenol, which does not have the function of.Ghofrani and other reports, gives PAH patients a short term FV treatment, showing that although FD can cause pulmonary vasoconstriction, it is not specific to the pulmonary circulatory vessels, and the medium and long term treatment of vasopunal also has a case report, showing no special effects on the treatment of PAH. The efficacy and tolerability of FD in the treatment of PAH is still open to question.
In 2009, Professor Jing Zhicheng and his team, through a study of 45 PAH patients, showed that 1 years of FDA therapy could effectively treat PAH.2011 years. The group continued to conduct a randomized, double-blind, placebo-controlled and multicenter study. It can effectively delay the clinical deterioration of PAH patients and improve their hemodynamic changes. However, the specific mechanism of the treatment of pulmonary hypertension by FD is still not clear.
In view of the current lack of PAH functional study on FV, 15 patients with idiopathic PAH were collected, and PAH rats induced by monocrotaline were in vivo model, and the oxidative stress endothelial cells induced by H202 were used as models in vitro. The mechanism of PAH treatment and the possible signaling pathways were studied, which provided for the treatment of PAH. The three parts of the study are as follows:
The first part is the clinical study of the treatment of idiopathic pulmonary hypertension by using Vastatin to improve oxidative stress.
In this study, patients with idiopathic pulmonary hypertension were selected as research subjects, and the therapeutic effect of FD on patients with idiopathic pulmonary hypertension and the improvement of oxidative stress in patients with.1) were selected. We selected 15 patients with EVALUATION in the study of idiopathic pulmonary vein hypertension and selected age and sex. The matched healthy people were given a control. The patients were given 5mg for 1 months 1 times a day. If the patient had no serious adverse reactions, the patient was given a dose of 5mg each time and 2 times a day for 2 months.
2) we evaluated the following contents at baseline and 3 months later: standard medical history and physical examination, 6 minute walking distance, WHO pulmonary hypertension function classification, blood biochemical index, electrocardiogram, lung function examination, and complete right heart catheterization test. Results showed that 3 months of FDA could significantly improve the idiopathic effect. WHO functional classification, six minute walking distance, heart index, pulmonary vascular resistance, and other indicators. Although compared with pre medication, the average pulmonary artery pressure, pulmonary capillary wedge pressure, and the mean right atrial pressure were lower than before, but there was no statistical difference. The results showed that FD could improve the pulmonary hypertension. Clinical symptoms and hemodynamic changes.
3) then we measured the levels of nitric oxide, malondialdehyde (MDA), 8- isoprost F2 a, 3- nitrotyrosine, superoxide dismutase and other oxidative stress related products in the blood of patients with pulmonary hypertension and healthy control group. Prostacyclin F2 alpha and 3- nitro tyrosine were significantly higher than normal people. Oxidative stress protective product nitric oxide and superoxide dismutase were significantly lower than normal controls. After 3 months of treatment, the levels of 8- ISO Prost F2 A and 3- nitrotyrosine were lower than those in the front, but still higher than those of normal people, while the level of malondialdehyde was not. There was an obvious change. The nitric oxide of the vasodilator increased obviously and basically recovered to the normal level. The antioxidant enzyme superoxide dismutase was higher than that before the treatment, and it was still at a lower level compared with the normal person. Above results showed that the above results showed that there was a high level of oxidative stress in the patients with pulmonary hypertension, and the antioxidant capacity was significantly higher. Normal people were weakened, while treatment with fattening could reduce oxidative stress level and improve their antioxidant capacity.
The second part is the effect of FD on improving oxidative stress in rats with pulmonary hypertension induced by monocrotaline.
In this study, 34 male SPF grade Sprague-Dawley (SD) rats and weight 200-220g were selected and randomly divided into normal control group, monocrotaline model group and FD n non treatment group. After giving 50mg/kg monocrotaline for 21 days, the rats were given a total of 21 days with 1mg/kg/d. The following study was followed:
1) the hemodynamic indexes of rats were measured by right heart catheterization, and the mPAP, RVSP and PVR in group MCT rats were significantly higher than those of normal control group, while CO was significantly lower than that of normal control group. After the treatment, the mPAP, RVSP and PVR of FD group rats decreased by 41%, 25% and more of 26%. were compared with our clinical trial. The results were consistent, which further verified the improvement of Vernon on hemodynamics.
2) in order to study the reasons for the improvement of FD's hemodynamics, we stained the lung tissue of rats and measured the vascular tissue with relevant software. Compared with the normal control group, the%WT and%WA of the rats were significantly higher than those in the normal control group. The%WT and%WA decreased significantly after treatment with FD. The results were shown in the pulmonary vessels of 50 mu M-100 mu m and 100-200 m diameter. Then we detected the degree of vascular myosis in rats by a-SMA antibody staining. Compared with the normal control group, the degree of vascular myosis in the monocrotaline group increased significantly, and the smooth muscle cells were obviously hypertrophic and proliferated under the microscope. The degree of muscle myocytes in the group was 39% lower than that in the MCT group. The proliferation and hypertrophy of smooth muscle cells was improved. Finally, we performed PCNA and TUNEL staining on the pulmonary vascular smooth muscle cells. It was found that the improvement of vascular thickness by reducing the vascular thickness was achieved by inducing the proliferation of rat pulmonary artery smooth muscle cells and promoting its apoptosis.
We used werstern blot method and commercial kits to study the mechanism of pulmonary arterial hypertension treated with FD. Compared with the normal control group, the concentration of NO in the plasma and lung tissue of the rats in the monocrotaline group decreased. After the treatment, the plasma and the NO levels in the pulmonary tissue were compared with the monocrotaline group. We found that FDA was able to increase the expression of eNOS protein. After that, we found that FD could improve the level of oxidative stress in rats, consistent with our clinical trials. In order to further study the possible mechanisms, we chose the main enzymes, NADPH oxidase, and super enzymes in the oxidative stress process. The results showed that the level of NADPH oxidase catalyzed subunits, NOX2 and NOX4 in the rats of monocrotaline increased obviously compared with the normal control group, but the level of the activated subunit RAC1 was not significantly changed. The level of NOX2 and NOX4 expression could be significantly improved, but the RAC1 was not affected by RAC1. On the contrary, the activity of SOD in the MCT group was significantly lower than that in the normal control group, and the activity of SOD was significantly improved after the treatment. The results suggested that the level of oxidative stress was improved, some of which were achieved by improving the NADPH oxidase and superoxide dismutase.
The third part is the mechanism of improving endothelial cell oxidative damage by means of BMP signaling pathway.
In the first and second parts of the body, we have identified that FD can improve oxidative stress and treat pulmonary hypertension, but the specific mechanism is still unclear. So the third part, we applied the CRL-1730 endothelial cell line as the research model, and gave different doses of exogenous H202 to cause oxidative stress damage and further study. The protective mechanism of Vastatin on oxidative stress injury is discussed.
1) through the method of MTT and TUNEL, we found that compared with the control group, the cell viability of the H202 group decreased significantly. Compared with the H202 group, the cell viability was significantly increased, and the cell viability was significantly dependent on the control group. Compared with the control group, the apoptosis rate of the H202 group was significantly higher than that in the control group. The results were proved to be dose-dependent. The results of the above results showed that FD had a strong protective effect on endothelial cells, and the effect was dose-dependent.
2) after that, we detected the expression of bone morphogenetic proteins (BMPs) signaling pathway in each cell. BMP signaling pathway is widely involved in pulmonary arterial hypertension, atherosclerosis and other vascular diseases. The results show that oxidative stress can increase the BMP receptor 1A, BMP receptor 1B, BMP receptor 2, and up - up cytoplasmic p-. Smad level finally entered the nucleus and up-regulated the expression of transcription factor MSX2, while Vastatin could improve the above pathway and play a protective role.
3) at last, we used the inhibitor of BMP signaling pathway, Dorsomorphin interfered the cells, and detected the cell apoptosis by TUNEL. We found that compared with the control group, the H202 group was within the cell.
【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類號(hào)】：R725.4

【共引文獻(xiàn)】

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本文編號(hào)：1975355