摘要
目的探究尿液细胞外囊泡中足细胞表面标志蛋白podocalyxin(PCX)水平在糖尿病肾病诊断中的应用价值。方法收集2017年3月~2017年9月在本院确诊收治的糖尿病患者共57例,其中34例单纯糖尿病患者,23例糖尿病肾病患者,并收集21例其他类型肾病患者,11例健康体检者作为对照。采用透射电镜(TEM)和纳米示踪技术(NTA)验证尿液中细胞外囊泡的分离效果,Western blot法检测分离出的各组细胞外囊泡的分子标记物(TSG101、PCX),ELISA法检测细胞外囊泡中PCX水平。结果TEM结果发现尿液中细胞外囊泡含量丰富、形态完整、大小不等,NTA结果发现绝大多数颗粒直径小于300 nm。Western blot法显示4组样本中均可以检测出TSG101表达,且糖尿病肾病患者样本中PCX均呈阳性表达,而在健康体检组、单纯糖尿病患者以及其他肾病患者样本中呈阴性表达。ELISA法显示糖尿病肾病组24 h尿液细胞外囊泡中PCX水平(3.27±2.30 ng/μmol)显著高于健康体检组(1.22±0.36 ng/μmol)、单纯糖尿病组(2.22±1.29 ng/μmol)以及其他肾病组(1.24±0.45 ng/μmol),有统计学意义(P<0.05)。结论糖尿病肾病患者尿液细胞外囊泡中PCX水平明显升高,可以作为临床检测糖尿病肾病的潜在标志物之一,具有重要的临床意义。
Objective To explore the value of detecting podocalyxin（PCX） level in urinary extracellular vesicles for the diagnosis of diabetic nephropathy. Methods This study was conducted among 57 diabetic patients admitted during the period from March to September, 2017, including 34 with uncomplicated diabetics and 23 with diabetic nephropathy; 21 patients with other types of nephropathy and 11 healthy individuals were also included to serve as the controls. Transmission electron microscopy（TEM） and nanoparticle tracking analysis（NTA） were used to verify the separation of urinary extracellular vesicles. The molecular markers of extracellular vesicles（TSG101 and podocalyxin [PCX]） were detected using Western blotting. PCX levels in extracellular vesicles were also detected using ELISA. Results TEM reveal the presence of numerous extracellular vesicles in the urine with intact morphology and different sizes, and most of them were below 300 nm in diameter as shown by NTA.TSG101 expression was detected in the samples from all the 4 groups. Positive expression of PCX was detected in the samples from patients with diabetic nephropathy but not in the other groups. In patients with diabetic nephropathy, the mean PCX levels（3.27±2.30 ng/μmol）was significantly higher than those in the healthy control group（1.22±0.36 ng/μmol）, uncomplicated diabetes group（2.22±1.29 ng/μmol） and nephropathy group（1.24±0.45 ng/μmol）. Conclusion PCX level in urinary extracellular vesicles is significantly increased in patients with diabetic nephropathy, suggesting the value of PCX as a potential marker for clinical diagnosis of diabetic nephropathy.
作者
吴凡
陈韵颖
肖花
邹子良
宁靖
陈海珊
邹和群
WU Fan;CHEN Yunyin;XIAO Hua;ZOU Ziliang;NING Jing;CHEN Haishan;ZOU Hequn(Department of Nephrology,Affiliated Hospital of Putian University,Putian 351100,China;Department of Nephrolog Third Affiliated Hospital of Southern Medical University,Guangzhou 510630,China;Shunde Nanfang Medical University Science Park,Shunde 528000,China)
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2018年第9期1126-1130,共5页
Journal of Southern Medical University
基金
广东省自然科学基金(2016A030313559)
广东省科技厅科技计划项目(2014A020212196)
南方医科大学科研启动计划项目(PY2014N094)
中国南方智谷引进创新团队和项目(CXTD-04)
广州市健康医疗协同创新重大专项(201604020015)
欧盟第七框架计划资助项目(Uro Sense-286386)
