Podocytes that detach in experimental membranous nephropathy are viable
Podocytes that detach in experimental membranous nephropathy are viable
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DOI：
10.1046/j.1523-1755.2003.00217.x
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发表时间：
2003-10-01
影响因子：
19.6
通讯作者：
Shankland, SJ
中科院分区：
文献类型：
--
作者：
Petermann, AT;Krofft, R;Shankland, SJ
背景资料。足细胞丢失有助于肾小球硬化的发展。虽然在某些肾小球疾病的尿液中检测到足细胞，但分离的细胞的活性尚不清楚。收集实验性膜性肾病[被动型Heymann肾炎(PHN)模型]大鼠的尿液，离心后再悬浮于组织培养液中，将细胞接种于胶原蛋白包裹的组织培养板上。细胞在典型的细胞培养条件下生长。细胞计数，免疫组织化学染色鉴定细胞类型，光镜和电子显微镜观察细胞形态。从PHN大鼠的尿液中获得的细胞对突触素、neparin、podocin、WT-1和GLEPP1(足细胞特异性抗原)呈阳性。在细胞培养条件下体外培养时，PHN大鼠尿中获得的细胞贴附在组织培养板上，并在逆转录-聚合酶链式反应(RT-PCR)和蛋白(免疫染色)水平表达足细胞特异性蛋白。肾小球系膜细胞(OX-7)、肾小管细胞(Tamm-Horsfall蛋白)和血管内皮细胞(RecA)均未见阳性反应。电子显微镜显示足突的存在，来自PHN大鼠的足细胞C5b-9染色阳性。虽然足细胞数量在体外培养的前5天短暂增加，但此后细胞凋亡率显著增加，细胞总数减少。实验性膜性肾病大鼠将足细胞排入体外附着于组织培养板上的尿液中，并进行增殖。这些结果表明分离的足细胞是可行的。这些结果为我们理解足细胞丢失在肾小球硬化发展中的作用提供了新的视角。
Background. Podocyte loss contributes to the development of glomerulosclerosis. Although podocytes have been detected in the urine in certain glomerular diseases, the viability of detached cells is not known.Methods. Urine was collected from rats with experimental membranous nephropathy [ passive Heymann nephritis (PHN) model], centrifuged, and following resuspension in tissue culture media, cells were seeded onto collagen-coated tissue culture plates. Cells were grown under typical cell culture conditions. Cell number was measured, the cell type was identified by immunostaining with specific antibodies, and cell morphology was assessed by light and electron microscopy.Results. Cells obtained in the urine from PHN rats were positive for synaptopodin, nephrin, podocin, WT-1, and GLEPP1 (podocyte-specific antigens). When grown ex vivo under cell culture conditions, cells obtained in the urine from PHN rats adhered to tissue culture plates, and expressed podocyte-specific proteins at the mRNA [ reverse transcription-polymerase chain reaction (RT-PCR)] and protein ( immunostaining) level. Cells did not stain with antibodies to mesangial (OX-7), tubular (Tamm-Horsfall protein) and endothelial (RECA) cells. Electron microscopy showed the presence of foot processes, and podocytes from PHN rats stained positive for C5b-9. Although podocyte number increased transiently during the first 5 days ex vivo, apoptosis increased significantly thereafter, reducing overall cell number.Conclusion. Rats with experimental membranous nephropathy shed podocytes into the urine that attach to tissue culture plates ex-vivo, and proliferate. These results suggest that detached podocytes are viable. These results add new perspectives into our understanding of podocyte loss in the development of glomerulosclerosis.