The Study for Clinical Pathophysiolongy of Feline Membranous Nephropathy and Diagnostic Significance of Urinary Micro-Protein Analyses.

采用阳离子化牛血清白蛋白（BSA）静脉注射，每周3次，连续3个月，建立大鼠膜性肾病模型。5例中4例肾小球基底膜增生、棘突形成、IgG沉积。血液生化检查中，TP、Alb降低，F-Cho、Cho-E、PL升高，NEFA、TG无变化。血清脂蛋白分析显示乳糜微粒和极低密度脂蛋白无变化，但低密度脂蛋白和高密度脂蛋白升高。结果表明，本病为Ⅱ a型高脂血症。即使Cre、UN和内源性Cre清除率无变化，尿蛋白浓度、尿蛋白Cre指数和24小时尿蛋白排泄量也增加。结果表明，尿蛋白分析对膜性肾病的诊断是有价值的。尿蛋白SDS-PAGE电泳结果显示，在分子量为160.5kDa、113.2kDa和113.2kDa的蛋白质条带中， ...更多信息 在分子量为131.4kDa、77.8kDa、42.5kDa、39.8kDa、31.5kDa和24.6kDa的蛋白质条带中检测到了新的蛋白质条带。从这9条蛋白带中筛选出160.5kDa蛋白（160 UP）和113.2kDa蛋白（113 UP），并进行纯化。此外，这些蛋白质通过氨基酸序列分析鉴定。采用丙烯酰胺凝胶电洗脱与高效液相色谱相结合的方法，可以对113 UP进行纯化。其氨基酸序列与猫血清白蛋白（FSA）一致，但分子量与FSA不一致。经二硫苏糖醇（DTT）处理后，SDS-PAGE显示5条蛋白带，分子量分别为113.2 kDa、69.7 kDa、68.3 kDa、65. 1 kDa和60.3kDa。这5条蛋白带的氨基酸序列与FSA的氨基酸序列一致。这些结果表明，113 UP可能是FSA代谢产物通过二硫键形成的二聚体。160 UP可通过电洗脱从丙烯酰胺凝胶中纯化。160 UP经DTT处理后，经SDS-PAGE电泳分析，160 UP由分子量为50.5kDa、47.8kDa和25.0kDa的片段组成。这些片段的氨基酸序列与其他物种的IgG同源。此外，用抗猫IgG进行的Western blotting表明，160 UP是免疫猫IgG。本研究证明猫尿白蛋白和尿IgG可作为膜性肾病的诊断工具。少

The membranous nephropathy model producted by cationized-BSA administration in vein three times a week for three months. Four of them from 5 showed hyperplasia of glomerular basement membrane, formation of spike, deposition of IgG. In blood biochemistry examination, the plasma TP and Alb decreased, and plasma F-Cho, Cho-E and PL increased, but NEFA and TG showed no change. The serum lipoprotein analyses revealed that chylomicron and VLDL showed no change but LDL and HDL increased. This results demonstrated that this disease state was Iia type of hyperlipidemia. Even if the Cre, UN and Endogenous Cre clearance did not change, urinary protein concentration, Cre index of urinary protein and excretion of urinary protein for 24hrs increased. This results demonstrated that urinary protein analyses is useful for diagnosis of the membranous nephropathy. In a result of SDS-PAGE of urinary protein, the relative concentration of protein bands increased on the molecular weight 160.5kDa, 113.2kDa a … More nd 67.7kDa, and new the protein bands were able to detect on the molecular weight 131.4kDa, 77.8kDa, 42.5kDa, 39.8kDa, 31.5kDa and 24.6kDa. The 160.5 kDa protein (160UP) and the 113.2 kDa protein (113UP) among these 9 protein bands were selected and purified. Furthermore, these proteins were identified by amino acid sequence analyses. 113UP was able to purify by the combination method of electric elution form acryramide gels and HPLC. Its amino acid sequence agreed with feline serum albumin (FSA), but its molecular weight did not agree with FSA. In the result of SDS-PAGE which the protein incubated with dithiothreitol (DTT), 5 protein bands of molecular weight 113.2 kDa, 69.7 kDa, 68.3 kDa, 65. 1 kDa and 60.3kDa were detected. The amino acid sequence of these 5 protein bands were homologous with that of FSA. These results demonstrated that 113UP would be a dimer of FSA metabolite formed by disulfide linkage. Further 160UP was able to purify by the electric elution form acryramide gels. The SDS-PAGE of 160UP incubated with DTT revealed that 160UP consist of the some fragment of molecular weight 50.5 kDa, 47.8 kDa and 25.0kDa. The amino acid sequences of these fragments were homologous with IgG of other species, Furthermore, Western blotting used Anti-feline IgG revealed that 160UP is immunological feline IgG. This research was proved that feline urinary albumin and feline urinary IgG would be a tool for diagnosis of membranous nephropathy. Less