CYR61 Expression and Function in rapidly progressive glomerulonephritis

为了证实CYR61蛋白的表达，我们用抗CYR61抗体对174份人肾活检样本进行了免疫染色。其中19例为快速进展性肾小球肾炎。19例快速进行性肾小球肾炎中抗cyr61抗体阳性6例（30%），细胞月牙形阳性4例，管状阳性2例。155例慢性肾小球肾炎标本中抗cyr61抗体阳性25例（16.1%）。慢性肾小球肾炎标本肾小球未见反应。25个阳性样品在试管中出现反应。“新月阳性RPGN”组血清肌酐为7.2mg/dl，“新月阴性RPGN”组血清肌酐为4.3mg/dl。两组之间没有差异。在慢性肾小球样本中，CYR61阳性组间质损伤较CYR61阴性组更为严重。但两组之间没有任何临床差异。由于与多种抗体有交叉反应性，采用RT-PCR检测CYR61的表达。从冷冻肾活检样本中提取mRNA。少量肾活检标本在肾活检时快速冷冻，进行免疫荧光染色。将冰冻组织切成5μm厚度，用70%乙醇快速固定，去除OCT化合物。5分钟后，快速风干组织切片，用4M GTC溶液提取RNA。用该cDNA进行RT反应和PCR。通过常规35环PCR检测CYR61 mRNA产物。实时PCR结果显示，临床预后与CYR表达无相关性。mRNA质量、样本数量，尤其是快速进展性肾小球肾炎较少是导致阴性结果的几个原因。为了确定CYR61在尿中的表达，Western Blot的敏感性不足以观察到CYR61。改变样品采集和浓缩方法。CYR61在严重肾小球肾炎、快速进展肾小球肾炎中均有表达。特别是在快速进展的肾小球肾炎样本中，这种反应在新月形中可见。因此CYR61在人类快速进行性肾小球肾炎中会发挥一定的作用。少

To certify the expression of CYR61 protein, 174 human renal biopsy samples were immuno-stained with anti-CYR61 antibody. 19 of 174 cases were rapidly progressive glomerulonephritis. 6 (30%) of 19 rapidly progressive glomerulonephritis showed positive reaction with anti-CYR61 Antibody. 4 cases showed positive reaction in cellular Crescents. 2 cases showed positive reaction in tubles. 25 (16.1%) of 155 chronic glomerulonephritis samples showed positive reaction with anti-CYR61 antibody. No reaction was seen in glomeruli of chronic glomerulonephritis samples. 25 positive samples showed the reaction in tubles. Serum creatinine was 7.2mg/dl in the patients of "crescent positive stained RPGN", and 4.3mg/dl in "crescent negative stained RPGN". There were no differences between both groups. In chronic Glomerular samples, CYR61 positive group had more severe interstitial damages compare to CYR61 negative group. But there were any clinical differences in both groups.Because of cross reactivity o … More f antibodies, CYR61 expression was checked by another method, RT-PCR. mRNA was extracted from frozen renal biopsy samples. Small amount of renal biopsy samples were quickly frozen at the time of renal biopsy for immuno-fluorescent staining. Several frozen tissue were cut in 5μm thickness and quickly fixed with 70% ethanol to remove OCT compound. After 5 minutes, tissue sections were quickly air dried and RNA was extracted by 4M GTC solution. RT reaction was performed and PCR was performed with this cDNA. By regular 35cycle PCR, CYR61 mRNA products were seen. By real time PCR, there was no relation between the clinical future and CYR expression. Several reason of this negative result were the quality of mRNA, number of samples, especially less number of rapidly progressive glomerulonephritis.To determine the urinaly expression of CYR61, sensitivity of Western Blot was not enough to see CYR61. Sample collection and concentration method should be changed.The Expression of CYR61 was seen in severe glomerulonephritis, rapidly progressive glomerulonephritis. Especially the reaction was seen in crescents in rapidly progressive glomerulonephritis samples. So CYR61 will perform some reaction in human rapidly progressive glomerulonephritis. Less