本文選題：硫化氫　切入點：高肺血流性肺動脈高壓　出處：《吉林大學》2011年碩士論文　論文類型：學位論文

【摘要】：目的：高肺血流性肺動脈高壓是左向右分流型先天性心臟病的常見并發(fā)癥,其嚴重程度直接影響患者手術(shù)時機、手術(shù)成功率及術(shù)后長期生活質(zhì)量,嚴重威脅人類的健康和生命,但有關(guān)其發(fā)病機制尚未完全闡明。近來研究發(fā)現(xiàn)新型氣體信號分子硫化氫(hydrogen sulfide, H2S)對心血管系統(tǒng)具有重要的保護作用,能夠減輕高肺血流性肺動脈高壓的形成。內(nèi)質(zhì)網(wǎng)應激(endoplasmic reticulum stress, ERS)作為眾多心血管疾病發(fā)生發(fā)展的重要病理機制,近年來日益受到人們的廣泛關(guān)注。既往研究認為,H2S能夠通過調(diào)節(jié)內(nèi)質(zhì)網(wǎng)應激減輕組織的損傷程度,但H2S與ERS在高肺血流性肺動脈高壓的發(fā)生發(fā)展過程中有無關(guān)系及其具體機制尚不清楚。本實驗以高肺血流性肺動脈高壓大鼠為研究對象,探討H2S對高肺血流性肺動脈高壓大鼠內(nèi)質(zhì)網(wǎng)應激的影響,進而對其機制做進一步研究。 方法：選取體重140g～160g的健康雄性SD大鼠52只,隨機分為六組：對照組(n=8)、對照+硫氫化鈉(sodium hybrosulfide, NaHS)組(n=8)對照+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)。對分流組、分流+NaHS組及分流+PPG組大鼠行腹主動脈壁-下腔靜脈穿刺術(shù),造成腹主動脈與下腔靜脈瘺道建立左向右分流肺動脈高壓動物模型。分別在終止實驗前以右心導管測定肺動脈平均壓(MPAP),并取肺組織,光鏡下計算肺血管中肌性動脈(MA)、部分肌性動脈(PMA)和非肌性血管(NMA)百分比；以敏感硫電極法檢測肺組織硫化氫(H2S)含量、肺組織硫化氫生成酶(CSE)的活性；原位缺口末端標記(TUNEL)檢測平滑肌細胞的凋亡、采用免疫組織化學染色方法檢測肺動脈組織中內(nèi)質(zhì)網(wǎng)應激標志糖調(diào)節(jié)蛋白78(GRP78)、內(nèi)質(zhì)網(wǎng)相關(guān)性凋亡標志蛋白半胱天冬水解酶-12(caspasel2)和磷酸化的真核細胞蛋白質(zhì)翻譯起始復合體(p-eIF2a)的蛋白表達,并用全自動圖像分析系統(tǒng)進行半定量分析。 結(jié)果： (1)分流8周時各組肺動脈平均壓(MPAP)比較有統(tǒng)計學意義(P0.01)。分流組MPAP明顯高于對照組(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)與對照組相比,分流組肺小動脈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。與對照組相比,分流組CSE活性均明顯高于對照組(P0.05)。 (3)分流8周時,各組大鼠中、小型肺動脈平滑肌細胞中均有凋亡細胞存在。與對照組相比,分流組肺動脈平滑肌細胞凋亡率明顯減少(P0.05)；與分流組比較,分流+NaHS組肺動脈平滑肌細胞凋亡率明顯增加(P0.01),分流+PPG組肺動脈平滑肌細胞凋亡率明顯減少(P0.01)。 (4)各組大鼠肺動脈內(nèi)質(zhì)網(wǎng)應激標志蛋白GRP78、caspase12和p-eIF2a表達結(jié)果：①與對照組相比,分流組大鼠肺動脈GRP78、caspase12和p-eIF2a表達均明顯增高(P0.05)；與分流組相比,分流+NaHS組大鼠肺動脈GRP78、caspase12和p-eIF2a表達明顯降低(P0.05)；與分流組相比,分流+PPG組大鼠肺動脈caspase12和p-eIF2a蛋白表達顯著升高(P0.05)。②與對照組相比,對照+PPG組大鼠肺動脈GRP78和caspase12表達明顯升高(P0.01)；對照+NaHS組大鼠caspase12表達明顯降低(P0.05)。 結(jié)論：腹主動脈-下腔靜脈分流8周所致高肺血流性肺動脈高壓大鼠肺動脈中內(nèi)質(zhì)網(wǎng)應激顯著增強。H2S可通過抑制過度內(nèi)質(zhì)網(wǎng)應激反應拮抗高肺血流性肺動脈高壓和肺血管結(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：1600260