本文關(guān)鍵詞：室間隔缺損潛在血清標志物鋅指蛋白41和SPARC的驗證研究　出處：《廣西醫(yī)科大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 室間隔缺損 鋅指蛋白41 血清標志物 生物信息學(xué) 室間隔缺損 SPARC 血清標志物 生物信息學(xué)

【摘要】：先天性心臟病(congenital heart disease, CHD)是出生缺陷中最常見的疾病，室間隔缺損(ventricular septal defect, VSD)作為先天性心臟病中最主要的類型，對嬰幼兒的存活及生活質(zhì)量產(chǎn)生重要影響。室間隔缺損的發(fā)病原因及機理復(fù)雜，要想全面了解，除了傳統(tǒng)相關(guān)的易感基因、環(huán)境相關(guān)因素外，更需對調(diào)控心臟異常分化的蛋白及信號通路進行研究，從而為疾病發(fā)生和診斷提供線索。 本研究針對課題組前期應(yīng)用同位素標記相對和絕對定量(iTRAQ)技術(shù)聯(lián)合液相色譜串聯(lián)基質(zhì)輔助激光解析電離飛行時間質(zhì)譜(LC-MALDI-TOF/TOF MS)方法篩選出一系列室間隔缺損相關(guān)的差異蛋白，結(jié)合實驗結(jié)果及文獻檢索分析，對其中的鋅指蛋白41(zinc finger protein41, ZNF41)和富含半胱氨酸的酸性分泌蛋白（secreted protein acidic and richin cysteine, SPARC）進行驗證性研究，應(yīng)用免疫印跡法（Western blot, WB）和酶聯(lián)免疫吸附試驗(enzyme linked immunosorbent assay, ELISA)分別檢測血清樣本中鋅指蛋白41和SPARC的表達水平，并應(yīng)用生物信息學(xué)技術(shù)對鋅指蛋白41和SPARC進行蛋白質(zhì)結(jié)構(gòu)與功能分析。研究分為兩個部分，分別探討鋅指蛋白41和SPARC是否可以作為室間隔缺損的潛在血清標志物，為室間隔缺損的早期診斷及發(fā)病機制研究提供參考依據(jù)。 第一部分室間隔缺損潛在血清標志物鋅指蛋白41的驗證研究 目的驗證鋅指蛋白41是否能作為室間隔缺損的潛在血清標志物，并應(yīng)用生物信息學(xué)進行蛋白結(jié)構(gòu)與功能分析。 方法收集0～10歲的單純性室間隔缺損、房間隔缺損(atrial septaldefect, ASD)、法洛四聯(lián)癥(tetralogy of Fallot, TOF)患者及健康對照者血清樣本各20例。應(yīng)用Western blot和ELISA檢測血清樣本中鋅指蛋白41的表達水平，應(yīng)用生物信息學(xué)技術(shù)對鋅指蛋白41的結(jié)構(gòu)功能，參與信號通路等進行分析。 結(jié)果1. VSD組、ASD組、TOF組和健康對照組間年齡、性別、民族狀況均衡可比(P0.05)。 2. Western blot檢測結(jié)果顯示，鋅指蛋白41在4組血清中均有表達，但VSD組中的相對表達水平均高于其他3組(P0.05)； 3. ELISA檢測結(jié)果顯示，VSD組患者血清中鋅指蛋白41濃度為(136.72±56.44) pg/ml， ASD組為(94.54±41.98) pg/ml， TOF組為(100.69±37.08) pg/ml，健康對照組為(82.08±42.46) pg/ml，4組間差異有統(tǒng)計學(xué)意義(P0.05)，進一步兩兩比較，VSD組與其他3組之間差異均有統(tǒng)計學(xué)意義(P0.05)，而其他3組兩兩間差異均無統(tǒng)計學(xué)意義(P0.05)。 4.生物信息學(xué)分析表明：鋅指蛋白41屬于C2H2鋅指蛋白家族，具有調(diào)節(jié)與DNA結(jié)合的活性，能調(diào)控基因表達、蛋白生成，，影響細胞分化及器官發(fā)育等功能。 結(jié)論鋅指蛋白41在室間隔缺損患者血清中表達水平較高，可能是室間隔缺損潛在的血清標志物，但仍需后續(xù)的大樣本臨床確認及鋅指蛋白41功能的研究。 第二部分室間隔缺損潛在血清標志物SPARC的驗證研究 目的驗證SPARC作為室間隔缺損潛在血清標志物的可能性并應(yīng)用生物信息學(xué)分析。 方法收集0～10歲的單純性室間隔缺損、房間隔缺損、法洛四聯(lián)癥患者及健康對照者血清樣本各20例。應(yīng)用Western blot和ELISA技術(shù)檢測SPARC在血清樣本中的表達水平，運用生物信息學(xué)進行SPARC結(jié)構(gòu)功能預(yù)測分析。 結(jié)果1.4組間年齡、性別、民族均衡可比(P0.05)。 2. Western blot檢測結(jié)果顯示，SPARC在4組血清中均有表達，但在VSD組的相對表達水平均高于其他3組(P0.05)； 3. ELISA檢測結(jié)果可知VSD組患者血清中SPARC濃度為(198.03±87.26) pg/ml， ASD組為(137.02±83.89) pg/ml， TOF組為(140.61±79.98) pg/ml，健康對照組為(91.30±55.50) pg/ml，4組間差異有統(tǒng)計學(xué)意義(P0.05)，進一步兩兩比較，VSD組與其他的3組之間差異均有統(tǒng)計學(xué)意義(P0.05)，而其他3組兩兩間差異均無統(tǒng)計學(xué)意義(P0.05)。 4.生物信息學(xué)結(jié)果，SPARC通過其特有的結(jié)構(gòu)域發(fā)揮生物學(xué)功能，影響細胞增殖和信號轉(zhuǎn)導(dǎo)，參與心臟等器官的發(fā)育。 結(jié)論SPARC在室間隔缺損的血清中表達水平較高，可能是室間隔缺損的潛在血清標志物，但仍需生物標志物第三階段的大樣本臨床確認研究，及SPARC蛋白生物學(xué)特性的研究。
[Abstract]:Congenital heart disease (congenital heart, disease, CHD) is the most common disease of birth defects, ventricular septal defect (ventricular septal defect, VSD) as the main types of congenital heart disease in infants, survival and quality of life have an important impact. The etiology and mechanism of ventricular septal defect complicated to a comprehensive understanding, in addition to the traditional related susceptibility genes, environment factors, need more protein to regulate cardiac differentiation and abnormal signal pathway were studied, providing clues for diagnosis and disease.
Based on the previous isobaric tags for relative and absolute quantitation (iTRAQ) series of matrix assisted laser desorption ionization time-of-flight mass spectrometry and liquid chromatography (LC-MALDI-TOF/TOF MS) method to screen a series of differences in ventricular septal defect associated protein, combined with the analysis of the experimental results and literature retrieval, the zinc finger protein 41 (zinc finger protein41, ZNF41) secreted protein acidic and rich in cysteine (secreted protein acidic and richin cysteine, SPARC) test, by immunoblotting (Western blot, WB) and enzyme-linked immunosorbent assay (enzyme linked immunosorbent assay, ELISA) were detected in serum samples of zinc finger protein expression levels of 41 and SPARC the application of bioinformatics, and technology of zinc finger protein 41 and SPARC analysis of protein structure and function. The research is divided into two parts, respectively. To investigate whether zinc finger protein 41 and SPARC can be used as potential serum markers for ventricular septal defect, and provide reference for early diagnosis and pathogenesis of ventricular septal defect.
Study on the validation of zinc finger protein 41, a potential serum marker of ventricular septal defect in the first part
Objective to verify whether zinc finger protein 41 can be used as a potential serum marker for ventricular septal defect and to use bioinformatics to analyze protein structure and function.
Methods isolated ventricular septal defect 0~10, atrial septal defect (atrial, septaldefect, ASD), tetralogy of Fallot (tetralogy of Fallot, TOF) patients and healthy controls serum samples of 20 cases each. The expression level of zinc finger protein 41 serum samples by Western blot and ELISA, the application of bioinformatics technology the structure and function of zinc finger protein 41, signaling pathways involved in the analysis.
Results the age, sex and national status of the 1. VSD group, the ASD group, the TOF group and the healthy control group were comparable (P0.05).
The results of 2. Western blot showed that zinc finger protein 41 was expressed in the 4 groups, but the relative expression level in the VSD group was higher than that of the other 3 groups (P0.05).
3. ELISA test results showed that the serum in patients with VSD zinc finger protein 41 concentration (136.72 + 56.44) pg/ml, ASD group (94.54 + 41.98) pg/ml, TOF group (100.69 + 37.08) pg/ml, healthy control group (82.08 + 42.46) pg/ml, there were statistically significant differences between the 4 groups (P0.05 further more, 22), VSD group and other 3 groups was statistically significant differences (P0.05), and there was no significant difference between the other 3 groups (P0.05 22).
4. bioinformatics analysis showed that zinc finger protein 41 belonged to the C2H2 zinc finger family. It had the activity of regulating DNA binding, regulating gene expression, protein production, affecting cell differentiation and organ development.
Conclusion zinc finger protein 41 is highly expressed in serum of patients with ventricular septal defect, and it may be a potential serum marker for ventricular septal defect. However, follow-up study is needed for large sample clinical confirmation and zinc finger protein 41 function.
Verification of the potential serum marker SPARC of second parts of ventricular septal defect
Objective to verify the possibility of SPARC as a potential serum marker for ventricular septal defect and to use bioinformatics analysis.
Methods the data of 0 ~ 10 years old of isolated ventricular septal defect, atrial septal defect, tetralogy of Fallot patients and healthy controls serum samples of 20 cases each. The expression level of Western blot and application of ELISA technology in detection of SPARC in serum samples, using bioinformatics of SPARC structure and function prediction analysis.
Results the age, sex, and national equilibrium of the 1.4 groups were comparable (P0.05).
The results of 2. Western blot showed that SPARC was expressed in the 4 groups of serum, but the relative expression level in the VSD group was higher than that of the other 3 groups (P0.05).
3. the results of ELISA SPARC VSD in the sera of patients was (198.03 + 87.26) pg/ml, ASD group (137.02 + 83.89) pg/ml, TOF group (140.61 + 79.98) pg/ml, healthy control group (91.30 + 55.50) pg/ml, there were statistically significant differences between the 4 groups (P0.05), a further 22 compared with the VSD group and the other 3 groups was statistically significant differences (P0.05), and there was no significant difference between the other 3 groups (P0.05 22).
4. bioinformatics results, SPARC, through its unique domain, exerts biological function, affects cell proliferation and signal transduction, and participates in the development of cardiac organs.
Conclusion the expression level of SPARC in serum of ventricular septal defect is relatively high. It may be a potential serum marker for ventricular septal defect. However, the large scale clinical confirmation study of biomarker at the third stage and the biological characteristics of SPARC protein are still needed.

【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R541;R392

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相關(guān)期刊論文 前3條

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本文編號：1420302