脱细胞处理对猪主动脉瓣与同种瓣组织构成及免疫差异的影响  

Effect of decellularization on tissue composition and immunogenicity of porcine and human aortic valves

作　　者：简旭华[1,2,3] 林秋雄[1,2,4] 

机构地区：[1]广东省心血管病研究所 [2]广东省人民医院 [3]广东省医学科学院心外科,广东广州510080 [4]广东省医学科学院医学研究中心,广东广州510080

出　　处：《南方医科大学学报》2011年第12期2039-2042,共4页Journal of Southern Medical University

基　　金：广东省自然科学基金(A-7001117)

摘　　要：目的研究脱细胞前后猪主动脉瓣瓣膜与人同种主动脉瓣的组织构成及免疫差异,探讨清除猪主动脉瓣异种抗原的方法。方法选购液氮冻存人主动脉瓣3条,猪主动脉瓣(液氮保存)4条,使用胰蛋白酶脱细胞处理,制作脱细胞瓣膜支架,分同种瓣-同种瓣脱细胞-猪主动脉瓣-猪主动脉瓣脱细胞4组,提取瓣膜蛋白,使用SDS-PAGE电泳法分析各组蛋白构成;使用各组蛋白提取物诱导U-937细胞移动,测定细胞移动量,分析瓣膜支架存在的免疫原性。结果胰蛋白酶可有效清除瓣膜细胞。SDS-PAGE蛋白电泳显示,猪主动脉瓣叶成分较同种主动脉瓣叶蛋白组成存在明显差异。脱细胞处理可显著减小猪主动脉瓣与脱细胞同种瓣的差异,但在26 000～43 000之间猪主动脉瓣叶蛋白组分仍明显较同种主动脉瓣叶条带多。U-937细胞移动实验显示脱细胞处理可明显减少细胞移动量,但比较脱细胞同种瓣,脱细胞猪主动脉瓣诱导细胞移动量较多(P<0.05)。结论胰蛋白酶脱细胞处理可降低猪主动脉瓣及同种瓣免疫原性。脱细胞处理后的瓣膜支架存在一定抗原性,其中以猪主动脉瓣脱细胞后抗原性仍较强。Objective To study the changes of tissue composition and immunogenicity of porcine and human aortic valves after decellularization.Methods Three cryopreserved human aortic valves and 4 porcine valves were decellularized with trypsin,and the leaflet tissue was homogenized for SDS-PAGE protein electrophoresis and U-937 migration assay.Results Trypsin effectively removed the cells from the valve.SDS-PAGE demonstrated an obvious difference in the tissue composition between porcine and human valves.Although decellularization significantly diminished the differences between the valves,decellularized procine aortic valve stilled contained more protein components（between 26 000 and 43 000） than human valve.U-937 migration assay showed an obvious decrease of cell migration in the valves by decellularization（from 832.7×103 to 152.4±31.1×103 for porcine valves,P0.01,and from 644.9×103 to 91.2×103 for the human valves,P0.01）.Decellularized porcine valves induced a significantly greater cell migration than decellularized human valves（P0.05）.Conclusion Decellularization with trypsin can effectively decrease the immunogenicity of human or porcine heart valve,but can not completely eliminate the antigen,and decellularized porcine valve still retain strong immunogenicity.

关 键 词：瓣膜 脱细胞处理 免疫 

分 类 号：R654.2[医药卫生—外科学]