發(fā)布時(shí)間：2018-03-11 22:36

  本文選題：硫化氫　切入點(diǎn)：高肺血流性肺動(dòng)脈高壓　出處：《吉林大學(xué)》2011年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：目的：高肺血流性肺動(dòng)脈高壓是左向右分流型先天性心臟病的常見(jiàn)并發(fā)癥,其嚴(yán)重程度直接影響患者手術(shù)時(shí)機(jī)、手術(shù)成功率及術(shù)后長(zhǎng)期生活質(zhì)量,嚴(yán)重威脅人類(lèi)的健康和生命,但有關(guān)其發(fā)病機(jī)制尚未完全闡明。近來(lái)研究發(fā)現(xiàn)新型氣體信號(hào)分子硫化氫(hydrogen sulfide, H2S)對(duì)心血管系統(tǒng)具有重要的保護(hù)作用,能夠減輕高肺血流性肺動(dòng)脈高壓的形成。內(nèi)質(zhì)網(wǎng)應(yīng)激(endoplasmic reticulum stress, ERS)作為眾多心血管疾病發(fā)生發(fā)展的重要病理機(jī)制,近年來(lái)日益受到人們的廣泛關(guān)注。既往研究認(rèn)為,H2S能夠通過(guò)調(diào)節(jié)內(nèi)質(zhì)網(wǎng)應(yīng)激減輕組織的損傷程度,但H2S與ERS在高肺血流性肺動(dòng)脈高壓的發(fā)生發(fā)展過(guò)程中有無(wú)關(guān)系及其具體機(jī)制尚不清楚。本實(shí)驗(yàn)以高肺血流性肺動(dòng)脈高壓大鼠為研究對(duì)象,探討H2S對(duì)高肺血流性肺動(dòng)脈高壓大鼠內(nèi)質(zhì)網(wǎng)應(yīng)激的影響,進(jìn)而對(duì)其機(jī)制做進(jìn)一步研究。 方法：選取體重140g～160g的健康雄性SD大鼠52只,隨機(jī)分為六組：對(duì)照組(n=8)、對(duì)照+硫氫化鈉(sodium hybrosulfide, NaHS)組(n=8)對(duì)照+炔丙基甘氨酸(propargy lglycine, PPG)組(n=8)、分流組(n=10)、分流+NaHS組(n=10)、分流+PPG組(n=8)。對(duì)分流組、分流+NaHS組及分流+PPG組大鼠行腹主動(dòng)脈壁-下腔靜脈穿刺術(shù),造成腹主動(dòng)脈與下腔靜脈瘺道建立左向右分流肺動(dòng)脈高壓動(dòng)物模型。分別在終止實(shí)驗(yàn)前以右心導(dǎo)管測(cè)定肺動(dòng)脈平均壓(MPAP),并取肺組織,光鏡下計(jì)算肺血管中肌性動(dòng)脈(MA)、部分肌性動(dòng)脈(PMA)和非肌性血管(NMA)百分比；以敏感硫電極法檢測(cè)肺組織硫化氫(H2S)含量、肺組織硫化氫生成酶(CSE)的活性；原位缺口末端標(biāo)記(TUNEL)檢測(cè)平滑肌細(xì)胞的凋亡、采用免疫組織化學(xué)染色方法檢測(cè)肺動(dòng)脈組織中內(nèi)質(zhì)網(wǎng)應(yīng)激標(biāo)志糖調(diào)節(jié)蛋白78(GRP78)、內(nèi)質(zhì)網(wǎng)相關(guān)性凋亡標(biāo)志蛋白半胱天冬水解酶-12(caspasel2)和磷酸化的真核細(xì)胞蛋白質(zhì)翻譯起始復(fù)合體(p-eIF2a)的蛋白表達(dá),并用全自動(dòng)圖像分析系統(tǒng)進(jìn)行半定量分析。 結(jié)果： (1)分流8周時(shí)各組肺動(dòng)脈平均壓(MPAP)比較有統(tǒng)計(jì)學(xué)意義(P0.01)。分流組MPAP明顯高于對(duì)照組(P0.01)；與分流組比較,分流+NaHS組MPAP明顯降低(P0.01)；分流+PPG組MPAP明顯升高(P0.01)。 (2)與對(duì)照組相比,分流組肺小動(dòng)脈MA、PMA占肺小血管總數(shù)的百分比明顯升高,NMA占肺小血管總數(shù)的百分比降低。與對(duì)照組相比,分流組CSE活性均明顯高于對(duì)照組(P0.05)。 (3)分流8周時(shí),各組大鼠中、小型肺動(dòng)脈平滑肌細(xì)胞中均有凋亡細(xì)胞存在。與對(duì)照組相比,分流組肺動(dòng)脈平滑肌細(xì)胞凋亡率明顯減少(P0.05)；與分流組比較,分流+NaHS組肺動(dòng)脈平滑肌細(xì)胞凋亡率明顯增加(P0.01),分流+PPG組肺動(dòng)脈平滑肌細(xì)胞凋亡率明顯減少(P0.01)。 (4)各組大鼠肺動(dòng)脈內(nèi)質(zhì)網(wǎng)應(yīng)激標(biāo)志蛋白GRP78、caspase12和p-eIF2a表達(dá)結(jié)果：①與對(duì)照組相比,分流組大鼠肺動(dòng)脈GRP78、caspase12和p-eIF2a表達(dá)均明顯增高(P0.05)；與分流組相比,分流+NaHS組大鼠肺動(dòng)脈GRP78、caspase12和p-eIF2a表達(dá)明顯降低(P0.05)；與分流組相比,分流+PPG組大鼠肺動(dòng)脈caspase12和p-eIF2a蛋白表達(dá)顯著升高(P0.05)。②與對(duì)照組相比,對(duì)照+PPG組大鼠肺動(dòng)脈GRP78和caspase12表達(dá)明顯升高(P0.01)；對(duì)照+NaHS組大鼠caspase12表達(dá)明顯降低(P0.05)。 結(jié)論：腹主動(dòng)脈-下腔靜脈分流8周所致高肺血流性肺動(dòng)脈高壓大鼠肺動(dòng)脈中內(nèi)質(zhì)網(wǎng)應(yīng)激顯著增強(qiáng)。H2S可通過(guò)抑制過(guò)度內(nèi)質(zhì)網(wǎng)應(yīng)激反應(yīng)拮抗高肺血流性肺動(dòng)脈高壓和肺血管結(jié)構(gòu)重建形成。
[Abstract]:Objective: high flow pulmonary hypertension is a common complication of congenital heart disease with left to right shunt and the severity of patients directly affect the timing of surgery, surgical success rate and long-term life quality after the operation, a serious threat to human health and life, but its pathogenesis has not been fully elucidated. A recent study found that the new gas signal molecular hydrogen sulfide (hydrogen sulfide, H2S) has an important protective effect on the cardiovascular system, can reduce the formation of high flow pulmonary hypertension. Endoplasmic reticulum stress (endoplasmic reticulum, stress, ERS) as an important pathological mechanism of the occurrence and development of many cardiovascular diseases, in recent years has been widespread concern. Previous studies suggest that H2S can through the regulation of endoplasmic reticulum stress and tissue injury, but H2S and ERS in the occurrence and development of high flow pulmonary hypertension in No relationship and its specific mechanism are not yet clear. In this experiment, we studied the effects of H2S on endoplasmic reticulum stress in rats with pulmonary hypertension induced by high pulmonary hemodynamics, and further studied the mechanism.
Methods: the weight of 140g to 160g in 52 healthy male SD rats, were randomly divided into six groups: control group (n=8), control + sodium hydrosulfide (sodium hybrosulfide NaHS) control group (n=8) + propargylglycine (propargy, lglycine, PPG) group (n=8), shunt group (n=10) and shunt +NaHS group (n=10), +PPG group (n=8). The shunt of the shunt group and shunt +NaHS group and shunt +PPG group rats underwent abdominal aorta and inferior vena cava puncture caused by abdominal aorta and inferior vena cava by establishing animal pulmonary hypertension with left to right shunt model. Respectively before the end of experiment with right heart catheterization determination of mean pulmonary arterial pressure (MPAP), and the lung tissue, calculation of muscular artery in pulmonary arteries under light microscope (MA), part of the muscular artery (PMA) and non muscular vessels (NMA) to detect the percentage of lung tissue; sulfur sensitive electrode method of hydrogen sulfide (H2S) content in lung tissue generating enzyme (CSE) the in situ nick end activity; End labeling (TUNEL) detection of apoptosis of smooth muscle cells, immunohistochemical staining method to detect the pulmonary tissue in the endoplasmic reticulum stress marker glucose regulated protein 78 (GRP78), endoplasmic reticulum associated protein markers of apoptosis caspase -12 hydrolase (caspasel2) complex eukaryotic translation initiation protein and phosphorylated protein (p-eIF2a) the expression of semi quantitative analysis and automatic image analysis system.
Result錛，

本文編號(hào)：1600260