尿液细胞外囊泡检测及其临床应用进展

doi:10.3969/j.issn.1673-8640.2017.010.022

摘要/Abstract

摘要：

细胞外囊泡（EV）是机体内细胞在向外出芽和细胞膜裂变过程中释放出的微小囊泡,内含特异性生物活性分子,如蛋白质、核酸和脂质,因此有望成为临床疾病新型标志物。EV广泛存在于人类的各种体液中,其中尿液是研究EV的良好对象,具有标本量充足、标本易得且无创等特点。尿液EV内容物是潜在的疾病标志物。文章对尿液EV的检测方法及其临床应用作了综述。

关键词: 细胞外囊泡, 尿液, 新型标志物

Abstract:

Extracelluar vesicles（EV） are vesicles which are released from most cell types during budding and cell membrane fission. EV specific molecules,such as proteins,nucleic acids and lipids,are expected as novel biomarkers. EV exists in the body fluid of human body. Urine is a good target for the research of EV,with the advantages of sufficient volume,obtaining easily and non-invasive operation. The component urine EV may be potential as biomarkers of various diseases. This review mainly focuses on the determination of urine EV and its clinical application.

Key words: Extracelluar vesicle, Urine, Novel biomarker

中图分类号:

R446.1

徐咏, 郑磊. 尿液细胞外囊泡检测及其临床应用进展[J]. 检验医学, 2017, 32(10): 933-940.

XU Yong, ZHENG Lei. Urine extracelluar vesicle determination and its research progress[J]. Laboratory Medicine, 2017, 32(10): 933-940.

图/表 2

参考文献 58

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方法	原理	优点	局限性
超高速离心^[1,11]	基于EV大小和密度不同沉淀	保持EV完整性,利于RNA、蛋白质分析	耗时长、工作量大、需特殊设备
超高速离心+蔗糖梯度^[3,8]	在超高速离心基础上,据密度不同EV悬浮于不同浓度蔗糖溶液中	去除高丰度蛋白,利于蛋白质分析,可将EV分成不同的群	耗时长、步骤复杂、不适宜大批量标本
超滤法^[13-15]	基于EV大小不同分离	简单、快速、标本量小	有蛋白污染、不适宜 RNA、蛋白质分析
离心-尺寸排阻法^[13]	基于EV大小不同分离	去除高丰度蛋白,利于蛋白质分析	耗时长、工作量大、提取效率较低
沉淀法^[8]	基于聚乙二醇分离	简单、快速、操作标准化、利于miRNA分析	有蛋白污染,不适宜蛋白质分析
免疫亲和力法^[16]	基于EV标志物与其抗体特异性结合分离	能得到某一特定类型EV	非特异性EV
微流控法^[17-18]	基于液流力学分离	简便、快速、可将EV分成不同的群	提取效率不高、不适宜大标本量标本
液压透析法^[15,19-20]	基于EV大小不同分离	可处理大量标本、保持EV完整性、无需特殊设备	需后续处理高丰度蛋白

方法	原理	优点	局限性
超高速离心^[1,11]	基于EV大小和密度不同沉淀	保持EV完整性,利于RNA、蛋白质分析	耗时长、工作量大、需特殊设备
超高速离心+蔗糖梯度^[3,8]	在超高速离心基础上,据密度不同EV悬浮于不同浓度蔗糖溶液中	去除高丰度蛋白,利于蛋白质分析,可将EV分成不同的群	耗时长、步骤复杂、不适宜大批量标本
超滤法^[13-15]	基于EV大小不同分离	简单、快速、标本量小	有蛋白污染、不适宜 RNA、蛋白质分析
离心-尺寸排阻法^[13]	基于EV大小不同分离	去除高丰度蛋白,利于蛋白质分析	耗时长、工作量大、提取效率较低
沉淀法^[8]	基于聚乙二醇分离	简单、快速、操作标准化、利于miRNA分析	有蛋白污染,不适宜蛋白质分析
免疫亲和力法^[16]	基于EV标志物与其抗体特异性结合分离	能得到某一特定类型EV	非特异性EV
微流控法^[17-18]	基于液流力学分离	简便、快速、可将EV分成不同的群	提取效率不高、不适宜大标本量标本
液压透析法^[15,19-20]	基于EV大小不同分离	可处理大量标本、保持EV完整性、无需特殊设备	需后续处理高丰度蛋白

疾病	标志物	分离方法	参考文献
急性肾损伤	Fetuin A	超高速离心法	[26]
	ATF3	超高速离心法	[27]
	NHE3	超高速离心法	[28]
肾脏缺血再灌注	水通道蛋白-1	超高速离心法	[29]
局灶节段性肾小球硬化	Wilm's tumor 1	超高速离心法	[30]
IgA肾病	α1-抗胰酶蛋白、氨肽酶N、vasorin前体、血浆铜蓝蛋白	超高速离心联合蔗糖梯度法	[31]
糖尿病肾病	miR-130、 miR-145、 miR-155 和 miR-424	超高速离心法	[32]
糖尿病肾病	明胶酶、血浆铜蓝蛋白	超高速离心法	[33]
肾小球疾病	二肽基肽酶4	滤器-离心法	[34]
	AMBP、MLL3和 VDAC1	超高速离心法	[35]
	ADAM10	超高速离心法	[36]
肾纤维化	miR-29c和miR-200	超高速离心法	[37]
肾纤维化	CD2AP	超高速离心法	[38]
慢性肾病	IL-18和 NGAL	超滤法	[39]
肾脏移植	NGAL	超高速离心法	[40]
肾脏移植	CD133	超高速离心法	[41]
多囊肾	PKD1、PKD2和PKHD1	超高速离心法	[42]
前列腺癌	ITGA3和ITGB1	超高速离心法	[24]
	miR-34a	超高速离心法	[43]
	LncRNA-p21	尿液外泌体RNA分离试剂盒	[44]
	TMPRSS2:ERG融合基因和PCA-3	超高速离心法	[45]
	TM256和LAMTOR1	超高速离心法	[46]
	钙黏蛋白3	超高速离心法	[47]
膀胱癌	EDIL-3	超高速离心联合蔗糖梯度法	[48]
	LASS2和GALNT1	超高速离心法	[49]
	TACSTD2	超高速离心法	[50]
肾癌	MMP-9、DKK4、EMMPRIN和PODXL	超高速离心法	[51]
类风湿关节炎	CD14、CD3和CD19	超高速离心法	[52]
帕金森病	Ser（P）-1292 LRRK2	超高速离心法	[53]

疾病	标志物	分离方法	参考文献
急性肾损伤	Fetuin A	超高速离心法	[26]
	ATF3	超高速离心法	[27]
	NHE3	超高速离心法	[28]
肾脏缺血再灌注	水通道蛋白-1	超高速离心法	[29]
局灶节段性肾小球硬化	Wilm's tumor 1	超高速离心法	[30]
IgA肾病	α1-抗胰酶蛋白、氨肽酶N、vasorin前体、血浆铜蓝蛋白	超高速离心联合蔗糖梯度法	[31]
糖尿病肾病	miR-130、 miR-145、 miR-155 和 miR-424	超高速离心法	[32]
糖尿病肾病	明胶酶、血浆铜蓝蛋白	超高速离心法	[33]
肾小球疾病	二肽基肽酶4	滤器-离心法	[34]
	AMBP、MLL3和 VDAC1	超高速离心法	[35]
	ADAM10	超高速离心法	[36]
肾纤维化	miR-29c和miR-200	超高速离心法	[37]
肾纤维化	CD2AP	超高速离心法	[38]
慢性肾病	IL-18和 NGAL	超滤法	[39]
肾脏移植	NGAL	超高速离心法	[40]
肾脏移植	CD133	超高速离心法	[41]
多囊肾	PKD1、PKD2和PKHD1	超高速离心法	[42]
前列腺癌	ITGA3和ITGB1	超高速离心法	[24]
	miR-34a	超高速离心法	[43]
	LncRNA-p21	尿液外泌体RNA分离试剂盒	[44]
	TMPRSS2:ERG融合基因和PCA-3	超高速离心法	[45]
	TM256和LAMTOR1	超高速离心法	[46]
	钙黏蛋白3	超高速离心法	[47]
膀胱癌	EDIL-3	超高速离心联合蔗糖梯度法	[48]
	LASS2和GALNT1	超高速离心法	[49]
	TACSTD2	超高速离心法	[50]
肾癌	MMP-9、DKK4、EMMPRIN和PODXL	超高速离心法	[51]
类风湿关节炎	CD14、CD3和CD19	超高速离心法	[52]
帕金森病	Ser（P）-1292 LRRK2	超高速离心法	[53]