【摘要】：背景:從瓣膜病發(fā)生至出現(xiàn)臨床癥狀,可以經(jīng)歷20-40年的期限,在我國瓣膜性心臟病尤其是風(fēng)濕性瓣膜病仍然屬于多發(fā)病,從臨床癥狀產(chǎn)生至充血性心衰及活動耐量的明顯下降據(jù)觀察約可經(jīng)歷10-12年,二尖瓣狹窄較多見,合并心功能IV級的瓣膜病患者10年自然生存率低于15%。瓣膜置換術(shù)在目前仍將是治療瓣膜病的主要手段。而心房顫動是臨床上瓣膜病最常合并的心律失常之一,發(fā)生率高達(dá)60%左右,目前臨床上瓣膜性心臟病外科治療對患者遠(yuǎn)期生活質(zhì)量影響較大,尤其是對已經(jīng)合并心房纖顫的患者,房顫的外科治療的遠(yuǎn)期成功率較低,而對于這類患者病情變化的整個過程中往往伴隨著多種內(nèi)源性多肽組份的變化,而且這些多肽組份的變化可以給我們對患者病情、心功能狀態(tài)評估及干預(yù)治療的方式選擇上有很大意義。目的:通過對風(fēng)濕性瓣膜病患者血清多肽組學(xué)研究,篩選出表達(dá)顯著差異的多肽,探尋該差異多肽組學(xué)研究法對風(fēng)濕性瓣膜病臨床評估的意義。方法:本研究中對240例研究對象進(jìn)行分組研究,根據(jù)美國紐約心臟病學(xué)會(NYHA)1928年心功能分級標(biāo)準(zhǔn)將240例受試對象進(jìn)行分組,共分為以下三組:健康對照組即心功能正常組(不合并器質(zhì)性心臟病);患者A組即心功能I-II級組;患者B組即心功能III-IV級組,每組各80名受試對象。健康組受試對象為門診健康體檢者,符合納入排除標(biāo)準(zhǔn),體檢過程中即采集其臨床指標(biāo)和血清標(biāo)本,患者組為住院病人,病人入院后即采集各受試對象的一般臨床指標(biāo)如:年齡、體重、心率、血壓及心臟超聲檢查的相關(guān)指標(biāo)如左心房直徑、左心室舒張/收縮末期內(nèi)徑、左室射血分?jǐn)?shù)、左室縮短率、相對室壁厚度、左室后壁厚度、間隔厚度及左室質(zhì)量等指標(biāo),同時患者入院后次日抽取空腹靜脈血,檢測各研究對象的血清BNP水平,剩余血清進(jìn)行多肽組學(xué)研究,每組80個血清標(biāo)本中,將每16份血清各0.5μl混合成1個血清標(biāo)本,每組5份混合血清標(biāo)本,采用高效反向非標(biāo)記定量液相質(zhì)譜分析方法血清標(biāo)本進(jìn)行多肽組學(xué)的無標(biāo)記定量分析,每個標(biāo)本檢測重復(fù)3次后取平均值。選擇符合上述三組的5份血清樣本,進(jìn)行ELISA法驗證叢生蛋白和載脂蛋白AI。蛋白/多肽的定量分析采用基于SWATH數(shù)據(jù)的Peakview軟件進(jìn)行數(shù)據(jù)定量分析,對每個樣本采用強度549 m/z值進(jìn)行歸一化處理,數(shù)據(jù)以均數(shù)±標(biāo)準(zhǔn)差表示,統(tǒng)計分析應(yīng)用sas9.0軟件完成,兩組間比較應(yīng)用student'st檢驗分析,多肽組間差異的比較采用方差分析,p0.01為差異具有統(tǒng)計學(xué)意義。結(jié)果:各組研究對象在一般指標(biāo)如:年齡、體重、收縮壓、舒張壓等,患者兩組與健康組無明顯統(tǒng)計學(xué)差異,但患者a、b組與健康組的心率分別是88.15±8.2次/分、88.19±8.5次/分、82.91±10.64次/分,患者組明顯比健康組升高,患者組心率升高的原因有可能風(fēng)濕性瓣膜病并發(fā)心房纖顫或者陣發(fā)性心動過速等,但患者組間比較并無明顯統(tǒng)計學(xué)差異。超聲學(xué)臨床檢查實驗結(jié)果發(fā)現(xiàn),患者a/b組的左房內(nèi)徑、左室舒張末內(nèi)徑、左室收縮末內(nèi)徑、射血分?jǐn)?shù)、縮短率等指標(biāo)分別為:(57.03±8.01)/(56.59±7.05)mm、(50.2±5.24)/(53.08±5.24)mm、(34.45±4.21)/(35.48±4.47)mm、(58.32±4.65)/(55.53±6.05)%、(32.92±4.76)/(30.65±3.69)%,健康組上述指標(biāo)分別是:(31.275±2.61)mm、(44.67±6.36)mm、(29.47±3.13)mm、(68.46±1.84)%、(35.21±2.05)%,患者兩組的左房內(nèi)徑、左室舒張末內(nèi)徑、左室收縮末內(nèi)徑明顯高于健康組,但患者組間比較無明顯差異,患者兩組的射血分?jǐn)?shù)、縮短率明顯低于健康組,且患者兩組間也有顯著統(tǒng)計學(xué)差異。血清學(xué)bnp檢測實驗結(jié)果發(fā)現(xiàn),患者a組和b組與健康組的血清bnp水平結(jié)果分別是(150.96±23.91)pg/ml、(224.96±20.49)pg/ml、(52.62±12.79)pg/ml對比有顯著統(tǒng)計學(xué)差異,且患者a、b組間比較也有顯著差異。血清多肽差異組學(xué)實驗結(jié)果發(fā)現(xiàn)患者a、b組和健康對照組間均有38條蛋白質(zhì)相關(guān)多肽和95條肽具有顯著的統(tǒng)計學(xué)差異(校正后的p值0.0001),患者組間并未見明顯統(tǒng)計學(xué)差異(這可能與樣本量小、樣本均來源于西南地區(qū)等因素有關(guān)系),這些多肽包括來自包括來自組蛋白h2b、絨毛蛋白樣蛋白、補體家族c4-b、精子結(jié)構(gòu)域蛋白等來源的蛋白相關(guān)多肽。其中,補體蛋白c、c4a、c4b-2、c4b結(jié)合蛋白α鏈均被發(fā)現(xiàn)在患者組明顯下調(diào);其他蛋白如α-1-酸性糖蛋白、α-2-hs糖蛋白、抗胰蛋白酶、α-1-抗胰凝乳蛋白酶、結(jié)合珠蛋白及結(jié)合珠蛋白相關(guān)肽均下調(diào);另外,載脂蛋白ai、載脂蛋白ciii等顯著下調(diào),通過elisa檢測法進(jìn)行載脂蛋白ai和叢生蛋白血漿水平的驗證中,載脂蛋白ai血漿水平在健康組顯著降低,患者a、b組與健康組分別為(1197.82±69.57)、(1158.86±41.61)、(1414.82±98.11)μg/ml,健康組叢生蛋白血漿水平顯著降低,患者a、b組與健康組分別為(306.22±14.82)、(332.72±15.83)、(177.64±7.88)μg/ml,這些差異表達(dá)的多肽表明風(fēng)濕性瓣膜病根本的表現(xiàn)是一個動態(tài)、復(fù)雜的炎癥和血栓形成過程。結(jié)論:臨床上常用的心臟超聲檢查指標(biāo)如左室內(nèi)徑、室壁相對厚度及射血分?jǐn)?shù)、縮短分?jǐn)?shù)、bnp的檢測水平等能夠較好的從心臟結(jié)構(gòu)和功能上反映風(fēng)濕性瓣膜病患者的心臟功能狀態(tài)。我們通過血清差異多肽組學(xué)分析風(fēng)濕性瓣膜病患者與健康人群發(fā)現(xiàn)了有較顯著的多肽的差異表達(dá),而這些差異的相關(guān)蛋白/多肽大部分來源于炎癥和免疫反應(yīng)、脂質(zhì)代謝類相關(guān)蛋白,表明風(fēng)濕性瓣膜病的發(fā)病是炎癥和免疫反應(yīng)為主伴隨脂質(zhì)代謝紊亂、凝血功能失衡的復(fù)雜生物反應(yīng)過程,如果我們對這些差異的相關(guān)蛋白/多肽進(jìn)行功能及相關(guān)通路的研究,將有助于我們理解風(fēng)濕性瓣膜病的發(fā)生、發(fā)展的分子機制,同時通過差異多肽組學(xué)的研究,我們可以更深入的了解風(fēng)濕性瓣膜病的演變進(jìn)程,更有助于臨床醫(yī)師的診治和預(yù)后判斷。
[Abstract]:Background: from valvular disease to clinical symptoms, it can experience a period of 20-40 years. In our country, valvular heart disease, especially rheumatic valvular disease, still belongs to multiple diseases. It is observed from clinical symptoms to congestive heart failure and significant decrease in activity tolerance for 10-12 years. Mitral stenosis is more common, and cardiac function IV is combined. The 10 year natural survival rate of patients with valvular disease is lower than that of 15%. valve replacement, which is still the main method for the treatment of valvular disease. Atrial fibrillation is one of the most common arrhythmia in clinical valvular disease, with a rate of about 60%. The clinical treatment of valvular heart disease has a great influence on the quality of life in patients. For patients with combined atrial fibrillation, the long-term success rate of surgical treatment for atrial fibrillation is low, and the changes of endogenous polypeptide components are often accompanied by changes in the whole process of this type of patient's condition, and the changes in these polypeptide components can give us the way to the patient's disease, the assessment of heart function and the way of intervention. Objective: to search for the significance of the differential polypeptide in the clinical evaluation of rheumatic valvular disease through the study of serology of serum polypeptide in patients with rheumatic valvular disease, and to explore the significance of the differential polypeptide group in clinical evaluation of rheumatic valvular disease. Methods: in this study, 240 subjects were divided into groups, according to the New York heart disease association of the United States. NYHA) in 1928, 240 subjects were divided into three groups: the healthy control group, the normal cardiac function group (no organic heart disease), the group A, the cardiac function I-II group, the B group, the cardiac function III-IV group, and the 80 subjects in each group. The clinical indicators and serum specimens were collected during the physical examination. The patients were hospitalized and the patients were hospitalized. After admission, the general clinical indicators such as age, weight, heart rate, blood pressure, and echocardiography, such as left atrium diameter, left ventricular diastolic / end systolic diameter, left ventricular ejection score, were collected. The ratio of left ventricular shortening, relative wall thickness, left ventricular posterior wall thickness, interval thickness and left ventricular mass were measured. At the same time, the serum BNP level was detected by the patients after admission to the hospital. The residual serum was studied by polypeptides. In each group of 80 serum specimens, each of the 16 serum samples was mixed into 1 serum specimens. In each group of 5 mixed serum samples, the unmarked quantitative analysis of the serum samples was carried out by high performance reverse non labelled quantitative liquid phase mass spectrometry, and the average value of each specimen after 3 times was detected. 5 serum samples were selected in accordance with the above three groups, and the ELISA assay and apolipoprotein AI. protein / polypeptide were determined. Quantitative analysis uses Peakview software based on SWATH data to carry out quantitative analysis of data, and each sample is normalized with 549 m/z values, the data is represented by mean number of standard deviations, statistical analysis is completed by sas9.0 software, and student'st test analysis is used among the two groups, and the comparison of the differences among the polypeptides is analyzed by variance analysis, P0.01 The difference was statistically significant. Results: there was no significant difference between the two groups of the subjects in the general indexes such as age, weight, systolic pressure and diastolic pressure, but the heart rate of the two groups was 88.15 + 8.2 / min, 88.19 + 8.5 times, 82.91 + 10.64 times / scores, and the patient group was significantly higher than the health group. There was a possibility of rheumatic valvular disease complicated by atrial fibrillation or paroxysmal tachycardia in the patients with rheumatic valvular disease, but there was no significant difference between the patients. The results of the ultrasound clinical test found that the left atrium diameter, left ventricular end diastolic diameter, left ventricular end systolic diameter, ejection fraction, shortening rate, and so on were found in the a/b group. Don't be: (57.03 + 8.01) / (56.59 + 7.05) mm, (50.2 + 5.24) / (53.08 + 5.24) mm, (34.45 + 4.21) / (35.48 + 4.47) mm, (58.32 + 5.24)%. The inner diameter of the diastolic end and the left ventricular end systole was significantly higher than that of the healthy group, but there was no significant difference between the patients and the two groups. The shortening rate of the ejection fraction of the two groups was significantly lower than that of the healthy group, and there was a significant difference between the two groups. The results of serological BNP test found that the serum BNP levels of the patients and the group B and the health group were respectively, respectively. It was (150.96 + 23.91) pg/ml, (224.96 + 20.49) pg/ml and (52.62 + 12.79) pg/ml with significant statistical difference, and there was a significant difference between the patients' A and the B group. The results of serum polypeptide differential group test found that there were 38 protein related peptides and 95 peptide between the B group and the healthy control group with significant statistical differences (after correction) P value 0.0001), there was no significant statistical difference between the patients (this may be associated with the small sample size, the samples were derived from the southwest region and other factors). These peptides include protein related peptides from the sources of protein H2B, villous protein like protein, complement family c4-b, sperm domain egg white and so on. Among them, complement protein C, C4A, c4b- 2, C4B binding protein alpha chain was found to be significantly downregulated in the patient group; other proteins, such as alpha -1- acid glycoprotein, alpha -2-hs glycoprotein, anti trypsin, alpha -1- anti chymotrypsin, binding globin and globin related peptide, were downregulated, and apolipoprotein AI, apolipoprotein CIII, and so on were significantly downregulated by ELISA detection of apolipoprotein. The plasma levels of apolipoprotein AI and AI were significantly decreased in the healthy group, and in the patients with a, the group B and the healthy group were (1197.82 + 69.57), (1158.86 + 41.61), (1414.82 + 98.11) mu g/ml, the plasma level of the healthy group decreased significantly, the patients a, B group and the healthy group (332.72 + 15.83), (332.72 + 15.83), (177. 15.83), respectively, (332.72 + 15.83), (332.72 + 15.83), (332.72 + 15.83), respectively, (332.72 + 15.83), (332.72 + 15.83), respectively, (332.72 + 15.83), (332.72 + 15.83), respectively, (332.72 + 15.83), respectively, (332.72 + 15.83), (177., 15.83), respectively. 64 + 7.88) mu g/ml, these differentially expressed peptides indicate that the fundamental manifestation of rheumatic valvular disease is a dynamic, complicated process of inflammation and thrombosis. Conclusion: the commonly used echocardiographic indexes, such as the left ventricular diameter, the relative thickness of the ventricular wall and the ejection fraction, the number of shortening, the level of the detection of BNP, can be better from the heart structure. The functional state of the heart function in patients with rheumatic valvular disease. We found a significant difference in the expression of polypeptide in patients with rheumatic valvular disease and healthy people through serum differential polypeptide group, and most of these differences were derived from inflammatory and immune responses, and lipid metabolism related proteins. The pathogenesis of rheumatic valvular valvular disease is the complex bioreactive process of inflammation and immune response associated with lipid metabolism disorder and coagulation dysfunction. If we study the function and related pathways of these proteins / peptides, it will help us to understand the pathogenesis of rheumatic valvular disease and the molecular mechanism of development. Through the study of differential proteomics, we can have a better understanding of the evolution of rheumatic valvular disease and help clinicians in the diagnosis and treatment of prognosis.
【學(xué)位授予單位】：第三軍醫(yī)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：R541.2

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 任安立;李全民;司安家;董曉龍;陳新濤;;人工心臟瓣膜置換術(shù)治療風(fēng)濕性瓣膜病526例體會[J];山東醫(yī)藥;2009年23期

2 鄭文;張棣;;少年兒童風(fēng)濕性瓣膜病彩色多普勒超聲心動圖表現(xiàn)[J];醫(yī)學(xué)信息(上旬刊);2010年09期

3 鄭衍瓊,許香廣;風(fēng)濕性瓣膜病合并慢性心包縮窄2例報告[J];新醫(yī)學(xué);1993年07期

4 徐駿;林建雄;張克強;范麗娟;郭敏;;風(fēng)濕性瓣膜病繼發(fā)功能性三尖瓣關(guān)閉不全處理[J];中國實用醫(yī)藥;2006年01期

5 徐駿;林建雄;張克強;范麗娟;郭敏;;風(fēng)濕性瓣膜病繼發(fā)功能性三尖瓣關(guān)閉不全處理[J];中國實用醫(yī)藥;2006年03期

6 張喜平;;風(fēng)濕性瓣膜病合并冠心病外科治療60例[J];中國醫(yī)學(xué)工程;2014年04期

7 許曉華;風(fēng)濕性瓣膜病變彩超聲表現(xiàn)與手術(shù)方式(許曉華)選擇關(guān)系的研究(李泉水)[J];臨床超聲醫(yī)學(xué)雜志;2002年05期

8 王彪;王濤;吳樹明;張供;曹廣慶;楊洪陽;;氨基末端腦鈉肽對風(fēng)濕性瓣膜病患者圍手術(shù)期風(fēng)險性評估的意義[J];中國循環(huán)雜志;2008年01期

9 吳錫龍;;風(fēng)濕性瓣膜病變166例外科治療體會[J];貴州醫(yī)藥;2007年06期

10 馮衛(wèi)中;程兆云;張志東;趙子牛;權(quán)曉強;趙健;孫俊杰;王海波;;風(fēng)濕性瓣膜病合并冠心病的外科治療[J];醫(yī)藥論壇雜志;2011年08期

相關(guān)會議論文 前2條

1 吳錫龍;曾俊;于微;王茂;;66例風(fēng)濕性瓣膜病變的外治療[A];2012貴州省醫(yī)學(xué)會胸心血管外科年會論文集[C];2012年

2 欒波;荊全民;韓雅玲;;風(fēng)濕性瓣膜病冠狀動脈造影的資料分析[A];中華醫(yī)學(xué)會心血管病分會第八次全國心血管病學(xué)術(shù)會議匯編[C];2004年

相關(guān)博士學(xué)位論文 前1條

1 鄔曉臣;風(fēng)濕性瓣膜病患者血清差異多肽組學(xué)臨床評估意義的研究[D];第三軍醫(yī)大學(xué);2016年

相關(guān)碩士學(xué)位論文 前3條

1 李志文;紅花黃色素注射液輔助治療風(fēng)濕性瓣膜�。馓撗鲂停┋熜в^察[D];遼寧中醫(yī)藥大學(xué);2012年

2 馮衛(wèi)中;風(fēng)濕性瓣膜病合并冠心病的外科治療[D];鄭州大學(xué);2011年

3 王彪;氨基末端腦鈉肽對風(fēng)濕性瓣膜病患者圍手術(shù)期風(fēng)險性評估的意義[D];山東大學(xué);2009年

，

本文編號：2146039