基于PEG6000富集精液来源外泌体的提取及鉴定

发布时间：2018-06-19 05:25

本文选题：精液 + 外泌体　；参考：《南方医科大学学报》2016年11期

【摘要】：目的介绍PEG6000富集精液来源外泌体的提取和鉴定方法,为精液外泌体相关研究奠定基础。方法选自南方医科大学南方医院检验中心6名正常志愿者精液,经8%PEG6000富集并多步离心结合超速离心方法获取提取物,然后采用透射电子显微镜观察其形态,纳米颗粒跟踪分析仪分析其大小,蛋白印迹法检测其蛋白组成成分。结果提取物呈类圆形、椭圆形囊泡,质膜完整,大小约为30~150 nm,内部含有低电子密度物质,均阳性表达CD63、ALIX、TSG101蛋白,且不含精子细胞内质网Calnexin蛋白。结论经过对提取物的形态、大小以及其蛋白成分分析证实所提取到的物质为精液来源的外泌体,为后续开展精液外泌体临床及基础研究提供了方法学基础。
[Abstract]:Objective to introduce the method of extraction and identification of exocrine from semen enriched by PEG6000 so as to lay a foundation for the study of semen exocrine. Methods the semen samples from 6 normal volunteers were collected from the Center of Southern Hospital of Southern Medical University. The semen was enriched by 8PEG6000 and centrifuged with ultracentrifugation. The morphology of semen was observed by transmission electron microscope (TEM). The size of nanoparticles was analyzed by tracking analyzer, and the protein components were detected by Western blotting. Results the extracts were round, oval vesicles, and the plasma membrane was intact, the size was about 30 ~ 150nm.There was a low electron density substance in the extract. All of the extracts expressed CD63 + ALIXT TSG101 protein and did not contain Calnexin protein in spermatocyte endoplasmic reticulum (ER). Conclusion the analysis of the morphology, size and protein composition of the extract confirmed that the extracted substance was the exocrine of semen, which provided a methodological basis for further clinical and basic research of semen exocrine.
【作者单位】：南方医科大学第三附属医院泌尿外科;南方医科大学南方医院检验中心;
【基金】：广东省自然科学基金(2015A030310027,2016A030310393) 广东省科技计划项目(2014A020212204,2013B021800317) 南方医科大学创新强校科技项目(QD2014N005)
【分类号】：R446.19

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