Detection of polymorphonuclear cells, superoxide dismutase and poly C9 in glomeruli of patients with IgA nephropathy.
Detection of polymorphonuclear cells, superoxide dismutase and poly C9 in glomeruli of patients with IgA nephropathy.
复制标题
IgA肾病患者肾小球多形核细胞、超氧化物歧化酶、聚C9的检测
作者：
H. C. Chen;Y. Tomino;Y. Yaguchi;M. Fukui;H. Koide
Hong-Chun Chen，Division of Nephrology，Kaohsiung Medical College Hospital，Kaohsiung，Taiwan 80708（ROC）Dear Sir：免疫荧光法检测伊加肾病患者肾小球中的多形核细胞（PMN）、超氧化物歧化酶（SOD）和多聚C9（膜攻击复合物; MAC）。据推测，慢性肾小球肾炎的进展和/或加重可能与活性氧的活性有关。肾组织取自14例伊加肾病患者。使肾低温恒温器切片与小鼠单克隆抗人PMN抗体（Sigma，St. Louis，MO，USA，批号67 F-48161，1：10稀释），然后用FITC标记的山羊抗小鼠IG抗血清（卡佩尔实验室，Cochranville，PA，USA，1：20稀释）[1]。还用FITC标记的多克隆抗人SOD抗血清（The Binding Site，Birmingham，UK，批号G 0404，F/P摩尔比1.73，1：10稀释）对切片进行染色。将切片与小鼠单克隆抗人聚C9抗体（#MBM 10）反应，然后用FITC标记的山羊抗小鼠IG抗血清（卡佩尔实验室，宾夕法尼亚州科克伦维尔，USA，1：32稀释）。7例伊加肾病患者肾小球内均可见1 ~ 5个PMN，而对照组肾组织PMN染色基本阴性。14例伊加肾病患者中7例（50%）肾组织中可见SOD染色，而对照组肾组织中未见SOD染色。SOD在肾小球系膜区沉积，其强度从1（+）到3（+）不等。肾小管上皮细胞内可见微量SOD。Poly C9（MAC）在所有患者中均可见，位于肾小球系膜区、Bowman囊和肾小球外血管壁，其强度为1（+）~ 2（+）。在对照肾组织中未观察到聚C9染色。伊加肾病“晚期”患者肾小球内PMN、SOD、MAC的表达无统计学意义。中性粒细胞在免疫刺激下产生活性氧的能力强，其浸润可能在肾小球损伤中起重要作用，并诱导局部SOD活性异常升高，起到保护作用。此外，肾脏中存在多聚C9
Hung-Chun Chen, Division of Nephrology, Kaohsiung Medical College Hospital, Kaohsiung, Taiwan 80708 (ROC) Dear Sir, Detection of polymorphonuclear cells (PMN), super-oxide dismutase (SOD) and poly C9 (membrane attack complex; MAC) in glomeruli of patients with IgA nephropathy was examined by immunofluorescence. It is postulated that the progression and/or exacerbation of chronic glomerulonephritis might be correlated with the activities of reactive oxygen species. Renal tissues were obtained from 14 patients with IgA nephropathy. Renal cryostat sections were reacted with mouse monoclonal anti-human PMN antibody (Sigma, St. Louis, Mo., USA, Lot No. 67F-48161, 1:10 dilution) and then stained with FITC-labeled goat anti-mouse Ig antiserum (Cappel Laboratories, Cochranville, Pa., USA, 1:20 dilution) [1]. The sections were also stained with FITC-labeled polyclonal anti-human SOD antiserum (The Binding Site, Birmingham, UK, Batch No. G0404, F/P molar ratio 1.73, 1:10 dilution). The sections were reacted with mouse monoclonal anti-human poly C9 antibody (#MBM 10) and then stained with FITC-labeled goat anti-mouse Ig antiserum (Cappel Laboratories, Cochranville, Pa., USA, 1:32 dilution) as described previously [2]. One to five PMN per glomerulus was observed in all 7 patients with IgA nephropathy examined, while the PMN staining was essentially negative in the control renal tissues. Renal SOD staining was observed in 7 out of 14 patients (50%) with IgA nephropathy, but was not observed in the control tissues. SOD was deposited in the glomerular mesan-gial areas, with a variable intensity from 1( + ) to 3( + ). Trace amounts of SOD were observed in the tubular epithelial cells. Poly C9 (MAC) was observed in all patients with this disease, and was located in the glomerular mesangial areas, Bowman’s capsule, and extraglomeru-lar vascular walls with a variable intensity of 1( + ) to 2( + ). No poly C9 staining was observed in the control renal tissues. PMN, SOD and poly C9 (MAC) in the glomeruli were observed in patients with the ‘advanced stage’ of IgA nephropathy, but there was no statistical significance. It appears that the infiltration of PMN in the glomeruli, which have a high potential for production of reactive oxygen species on immunological stimulation, may play an important role in glomerular injuries, and induce an abnormally high local SOD activity for the protection. Moreover, the renal presence of poly C9