蛋白質(zhì)組學(xué)對不同透析膜吸附血清蛋白的分析
發(fā)布時間:2018-06-14 11:42
本文選題:透析膜 + 血液透析�。� 參考:《大連醫(yī)科大學(xué)》2017年碩士論文
【摘要】:目的:血液透析是尿毒癥患者的主要治療方式之一,透析膜是決定透析效果最重要的因素,對透析膜表面吸附的蛋白的研究,以便于理解透析膜表面發(fā)生的生化反應(yīng),并且為透析膜的改良提供基礎(chǔ)。方法:選我院連續(xù)兩次透析分別應(yīng)用金寶低通Polyflux14L(PAES/PVP/PA)透析器及費(fèi)森尤斯低通Fx8(PS/PVP)透析器患者2名作為材料差異組;連續(xù)兩次透析分別應(yīng)用金寶高通Polyflux140H透析器及金寶低通Polyflux14L透析器患者2名作為通量差異組。用離子液體(C12Im-Cl,氯化-1-十二烷基-3-甲基咪唑)處理入選患者使用的透析器(N=8),以洗脫透析膜上吸附的蛋白,蛋白酶解并化學(xué)標(biāo)記后混合;取患者透析前、后血液,分離血清標(biāo)本并酶解血清蛋白,化學(xué)標(biāo)記后混合。應(yīng)用LC-MS/MS的方法對血清蛋白及透析膜洗脫蛋白進(jìn)行分離和鑒定,對蛋白質(zhì)的分子量(MW)、等電點(diǎn)(p I)等分子信息和蛋白質(zhì)功能進(jìn)行對比或分析。結(jié)果:1.不同透析膜對蛋白的吸附規(guī)律不同,在吸附蛋白量方面,材料差異組:與PAES/PVP/PA合成膜相比,PS/PVP合成膜上有45種表達(dá)量升高,253種降低;通量差異組:與高通量透析膜相比,低通量透析膜上有284種表達(dá)量升高,174種表達(dá)量降低。在吸附蛋白的p I及MW分布方面,PAES/PVP/PA與PS/PVP合成膜在吸附蛋白的p I及MW無統(tǒng)計(jì)學(xué)差異(P0.05);高通量透析膜對MW40k Da,p I7.5的蛋白吸附比例高(p I:46%vs.29%,c2=11.293,P=0.001,MW:71%vs.57%,c2=8.133,P=0.004),低通量透析膜對MW 40k Da~100k Da,p I5.5~7.5的蛋白吸附比例高(p I:41%vs.30%,c2=4.906,P=0.027,MW:33%vs.22%,c2=5.978,P=0.014)。2.透析膜表面差異蛋白不同。材料差異組,甘露糖結(jié)合凝集素絲氨酸蛋白酶1,半乳糖凝集素1,特異性表達(dá)于PAES/PVP/PS合成膜上;熱休克蛋白70特異性表達(dá)于PS/PVP合成膜上。通量差異組,β2微球蛋白特異性表達(dá)于高通量透析膜上。結(jié)論:不同透析膜吸附蛋白質(zhì)的量、p I、MW不同;蛋白質(zhì)功能分析提示膜表面差異蛋白與免疫功能、應(yīng)激反應(yīng)和炎癥反應(yīng)相關(guān)。
[Abstract]:Objective: hemodialysis is one of the main treatment methods for uremic patients. Dialysis membrane is the most important factor to determine the effect of dialysis. The study of the protein adsorbed on the surface of dialysis membrane is helpful to understand the biochemical reaction on the surface of dialysis membrane. It also provides the basis for the improvement of dialysis membrane. Methods: two consecutive dialyzers were used in our hospital. Two dialyzers were used as the material difference group, one was the Jinbao low pass Polyflux14L PAES / PVP / PA dialyzer and the other was Fx8 / PVP / PVP dialyzer with Fishenyus low pass dialyzer. Two consecutive dialysis patients were treated with Jinbao Qualcomm Polyflux 140H dialyzer and Jinbao low pass Polyflux 14L dialyzer as flux difference group. C12Im-Cl and 1-dodecyl -3-methyl imidazole chloride were used to treat the dialyzer used by the selected patients to elucidate the protein adsorbed on the dialysis membrane, proteolysis and chemically labeled mixture. The blood was taken from the patients before and after dialysis. The serum samples were separated and the serum proteins were hydrolyzed by enzyme. Serum protein and dialysis membrane elution protein were isolated and identified by LC-MS / MS method. The molecular information and protein function of protein were compared and analyzed. The result is 1: 1. The adsorption of protein on different dialysis membranes was different. In terms of the amount of protein adsorbed, the material difference group: compared with PAES / PVP / PA synthetic membrane, there were 45 kinds of protein expression increased and 253 species decreased in Pae / PVP / PA synthetic membrane, and the flux difference group: compared with high flux dialysis membrane, there were 45 kinds of protein expression increased and 253 species decreased in Pas / PVP / PA synthetic membrane. On low flux dialysis membrane 284 kinds of expression increased and 174 kinds of expression decreased. There was no significant difference between pas / PVP / PA and PSP / PVP / PA / PS / PVP synthetic membrane in the protein I and MW of the adsorbed protein, and the ratio of protein adsorbed by high throughput dialysis membrane to MW40k Daapi / I7.5 was higher than that of the high throughput dialysis membrane. The ratio of protein adsorbed by high throughput dialysis membrane to MW40k Daqipi I7.5 was higher than that of the high throughput dialysis membrane C211.293P0.001MW71vs.57s. The low flux membrane was dialyzed on MW 40k Daw 100k Dap I5.57.5. The protein adsorption ratio is high (vs.30) 4.906 (P0.027MW: 33vs.22c2) 5.978U (5.978%) and 0.014% (2.23). The protein adsorption ratio is higher than that of vs.30%, and the ratio of protein adsorption is higher than that of vs.30%, and the ratio of protein adsorption is high. Dialysate membrane surface differential protein is different. Material difference group, mannose binding lectin serine protease 1, galactose agglutinin 1, specific expression in PAES- PVP / PS synthesis membrane, heat shock protein 70 specifically expressed on PS/ PVP synthetic membrane. In flux difference group, 尾 2 microglobulin was specifically expressed on high flux dialysis membrane. Conclusion: the amount of protein adsorbed by different dialysis membranes is different, and the analysis of protein function indicates that the differential protein on membrane surface is related to immune function, stress response and inflammatory reaction.
【學(xué)位授予單位】:大連醫(yī)科大學(xué)
【學(xué)位級別】:碩士
【學(xué)位授予年份】:2017
【分類號】:R692.5
【參考文獻(xiàn)】
相關(guān)期刊論文 前7條
1 賀雪明;唐麗;賀福初;;半乳糖凝集素1的免疫功能[J];生物技術(shù)通訊;2012年06期
2 謝潮鑫;孟猛;曾偉杰;鐘磊;魏春霞;羅佼;;血液透析膜材料的生物相容性[J];中國組織工程研究與臨床康復(fù);2009年08期
3 楊瑜靜;徐秀林;王新華;朱根娣;;復(fù)用血液透析器膜上蛋白黏附的種類和性質(zhì)分析[J];生物醫(yī)學(xué)工程學(xué)雜志;2009年01期
4 賈強(qiáng);;透析膜的生物相容性與遠(yuǎn)期并發(fā)癥[J];中國血液凈化;2008年02期
5 何東元;王笑云;楊俊偉;;血液透析生物相容性[J];中國血液凈化;2006年04期
6 宋艷秋,原續(xù)波,盛京;疏水性聚砜膜的共混改性研究[J];化學(xué)工業(yè)與工程;2002年01期
7 張東亮,王質(zhì)剛;血液透析膜吸附功能的研究進(jìn)展[J];國外醫(yī)學(xué).泌尿系統(tǒng)分冊;2000年05期
,本文編號:2017260
本文鏈接:http://sikaile.net/linchuangyixuelunwen/2017260.html
最近更新
教材專著