CAJ | 학술논문

目的应用蛋白质组学技术筛选糖尿病肾病(DN)患者尿液的相关差异蛋白,检测差异蛋白钙黏蛋白E(E-cadherin)在DN患者尿液及肾组织中的表达,以期发现新的DN相关差异蛋白.方法选择本院经肾活检确诊的1型DN(T1DN)和2型DN(T2DN)患者各12例,同时选择肾病综合征(NS)患者及健康志愿者各12例作为对照.应用2DE/MS比较蛋白质组学技术筛选DN患者及对照组人群尿中的DN相关差异蛋白.采用Western印迹法验证尿中差异蛋白的表达,免疫组织化学法检测肾组织中差异蛋白的表达.结果 2DE/MS结果显示,与NS组和健康对照组相比,T1DN组和T2DN组患者有1个差异蛋白点的蛋白丰度呈异常升高,经鉴定为E-cadherin.Western印迹结果显示,与NS和健康对照组相比,T1DN和T2DN组患者尿液中E-cadherin表达明显增高(均P＜0.01).免疫组化结果显示,E-cadherin主要表达于肾小管上皮细胞的细胞膜和细胞质,其在DN肾脏中的表达较健康对照组显著降低(P＜0.05).结论 DN患者尿液中E-cadherin蛋白特异性升高,可作为一种新的DN相关差异性蛋白.
목적 응용단백질조학기술사선당뇨병신병(DN)환자뇨액적상관차이단백,검측차이단백개점단백E(E-cadherin)재DN환자뇨액급신조직중적표체,이기발현신적DN상관차이단백.방법 선택본원경신활검학진적1형DN(T1DN)화2형DN(T2DN)환자각12례,동시선택신병종합정(NS)환자급건강지원자각12례작위대조.응용2DE/MS비교단백질조학기술사선DN환자급대조조인군뇨중적DN상관차이단백.채용Western인적법험증뇨중차이단백적표체,면역조직화학법검측신조직중차이단백적표체.결과 2DE/MS결과현시,여NS조화건강대조조상비,T1DN조화T2DN조환자유1개차이단백점적단백봉도정이상승고,경감정위E-cadherin.Western인적결과현시,여NS화건강대조조상비,T1DN화T2DN조환자뇨액중E-cadherin표체명현증고(균P＜0.01).면역조화결과현시,E-cadherin주요표체우신소관상피세포적세포막화세포질,기재DN신장중적표체교건강대조조현저강저(P＜0.05).결론 DN환자뇨액중E-cadherin단백특이성승고,가작위일충신적DN상관차이성단백.
Objective To identify novel biomarker for diabetic nephropathy (DN) by urinary proteomic methods,and to detect the expression of E-cadherin in urine and renal tissue of patients with DN.Methods Urine samples were collected from 12 cases of type 1 diabetic nephropathy patients (T1DN),12 cases of type 2 diabetic nephropathy patients (T2DN),12 cases of nephritic syndrome patients (NS),and 12 cases of healthy Controls.Comparative proteomic approach of two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS) were employed to identify DN-related biomarker in urine samples.The differential expression of the identified biomarker in urine samples and renal biopsy specimens were detected by Western blotting and immunohistochemistry method.Results E-cadherin was identified by 2DE/MS,which was significantly up-regulated in T1DN and T2DN groups (all P ＜0.01).Western blotting confirmed the expression of E-cadherin was significantly higher in T1DN and T2DN groups than in NS and Control groups (all P＜0.01).Immunohistochemical stain showed E-cadherin was mainly expressed in the membrane and cytoplasm of renal tubular epithelial cell,and its expression was markedly decreased in DN kidneys compared with healthy Controls (P ＜ 0.05).Conclusions E-cadherin is identified as a novel DN-related biomarker,which is specifically increased in urine of DN patients.