摘要
目的:改良原有的免疫印迹检测尿蛋白组分方法。分析原发性慢性肾小球疾病患者尿白蛋白聚合现象,探讨其可能的病理生理和临床意义。方法:改良原有的尿蛋白免疫印迹检测方法,利用改良法鉴定57例同步行肾活检诊断明确的原发性慢性肾小球疾病患者尿白蛋白多聚体(PAs)阳性率。结果:改良法不需要精确调整尿蛋白浓度及使用梯度凝胶电泳仪,可清晰显示PAs条带。具有肾病综合征(NS)的患者中尿PAs阳性率87.5%;无NS患者为56.1%,尿PAs阳性率与NS密切相关(P<0.05)。PAs阳性率以微小病变型肾小球肾炎(MCD)最高(91.7%),系膜增生性肾小球肾炎(MsPGN)次之(76.9%),IgA肾病(IgAN)最低(53.3%),差异具有统计学意义(P<0.05)。结论:成功改良了原有的免疫印迹检测尿蛋白组分方法。改良法较原有方法更加简便、易普及,适合国内学者研究及临床检测应用。尿PAs可能与原发性慢性肾小球疾病的某些病理生理过程有关,其作为一种无创性检测,有助于评估原发性慢性肾小球疾病的病理类型及预后。
Objective：To illustrate how to detect urinary protein components by modified western blot method. To analyze the phenomenon of urinary albumin which is one of the urinary protein components polymerization in patients with biopsy proven prima- ry glomerular diseases in order to identify its possible pathophysiologic and clinical significance. Methods： Modified the existing meth- od of western blot for urinary protein by sampling according the results of qualitative analysis of urinary protein instead of sampling based on precise concentration and electrophoresed the samples on 4% stacking gel sheet and 10% separating gel sheet instead of gra- dient 10% to 20% T polyacrylamide gel slabs. The existence of urinary PAs in 57 cases of patients with biopsy proven primary glomer- ular diseases with non - lupus giomernlonephritis, non - diabetic nephropathy, non - primary hypertension and non - cardiovascular damage disease was detected by the modified method at the same time of renal biopsy. The presence of PAs was defined as the pres- ence of at least a 100 kd band. Results ：The modified method could detect distinctly the bands of 100kd and more than 100kd of urina- ry PAs. In addition it didn＇t need to adjust the concentration of urinary protein precisely and didn＇t need gradient gel electrophoresis apparatus. PAs were present in 87.5% of 16 patients with nephrotic syndrome（ NS）, 56.1% of 41 patients without NS,the presence of PAs in NS group was significantly higher than non - NS group（P 〈0.05）. PAs were present in 91.7% minimal change disease （MCD） patients, 76.9% mesangial proliferative glomerulonephritis （ MsPGN ） patients and 53.3% IgA nephropathy （ IgAN ） patients （ P 〈 0.05 ）. Conclusion： Modified the existing method of western blot for urinary protein components successfully. The modified moth- ed is more simple and convenient, easier to popularize than the old one , is more appropriate for domestic scholars＇ study and clinical detection. Being a noninvasive detect method, PAs may be helpful to assess the pathological type and prognosis of primary glomerular diseases.
出处
《中国中西医结合肾病杂志》
2013年第1期33-35,共3页
Chinese Journal of Integrated Traditional and Western Nephrology
基金
"211工程"中山医科大学重点学科建设基金资助项目(No.98136)
