蛋白质组学对不同透析膜吸附血清蛋白的分析

发布时间：2018-06-14 11:42

本文选题：透析膜 + 血液透析　；参考：《大连医科大学》2017年硕士论文

【摘要】：目的:血液透析是尿毒症患者的主要治疗方式之一,透析膜是决定透析效果最重要的因素,对透析膜表面吸附的蛋白的研究,以便于理解透析膜表面发生的生化反应,并且为透析膜的改良提供基础。方法:选我院连续两次透析分别应用金宝低通Polyflux14L(PAES/PVP/PA)透析器及费森尤斯低通Fx8(PS/PVP)透析器患者2名作为材料差异组;连续两次透析分别应用金宝高通Polyflux140H透析器及金宝低通Polyflux14L透析器患者2名作为通量差异组。用离子液体(C12Im-Cl,氯化-1-十二烷基-3-甲基咪唑)处理入选患者使用的透析器(N=8),以洗脱透析膜上吸附的蛋白,蛋白酶解并化学标记后混合;取患者透析前、后血液,分离血清标本并酶解血清蛋白,化学标记后混合。应用LC-MS/MS的方法对血清蛋白及透析膜洗脱蛋白进行分离和鉴定,对蛋白质的分子量(MW)、等电点(p I)等分子信息和蛋白质功能进行对比或分析。结果:1.不同透析膜对蛋白的吸附规律不同,在吸附蛋白量方面,材料差异组:与PAES/PVP/PA合成膜相比,PS/PVP合成膜上有45种表达量升高,253种降低;通量差异组:与高通量透析膜相比,低通量透析膜上有284种表达量升高,174种表达量降低。在吸附蛋白的p I及MW分布方面,PAES/PVP/PA与PS/PVP合成膜在吸附蛋白的p I及MW无统计学差异(P0.05);高通量透析膜对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),低通量透析膜对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.透析膜表面差异蛋白不同。材料差异组,甘露糖结合凝集素丝氨酸蛋白酶1,半乳糖凝集素1,特异性表达于PAES/PVP/PS合成膜上;热休克蛋白70特异性表达于PS/PVP合成膜上。通量差异组,β2微球蛋白特异性表达于高通量透析膜上。结论:不同透析膜吸附蛋白质的量、p I、MW不同;蛋白质功能分析提示膜表面差异蛋白与免疫功能、应激反应和炎症反应相关。
[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.
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R692.5

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