Morphometry of interstitial fibrosis
Morphometry of interstitial fibrosis
复制标题
DOI：
10.1093/ndt/15.suppl_6.72
复制
发表时间：
2000-01-01
影响因子：
6.1
通讯作者：
van Es, LA
中科院分区：
文献类型：
--
作者：
De Heer, E;Sijpkens, YWJ;van Es, LA
几项临床研究已经证实，肾小管间质室的组织形态学变化包含预测进行性肾功能不全发展的最佳相关参数。间质纤维化的过程伴随着炎性细胞的流入，纤维化细胞因子如转化生长因子-β和碱性成纤维细胞生长因子的上调，其拮抗剂的瞬时下调，肌成纤维细胞的产生和增殖，以及最后，间质胶原和蛋白聚糖的积累。间质纤维化的仔细形态计量学分析需要敏感的参数，通过这些参数可以量化严重程度，并可以预测肾功能不全的进展。我们已经解决了这些问题的形态分析人类活检和完善现有的大鼠实验模型。使用配备有来自Zeiss-Kontron的全色3CCD照相机和KS-400图像分析软件的Zeiss显微镜进行形态测定分析。对于人体材料的研究，检查了各种肾脏疾病患者的活检，包括慢性同种异体移植功能障碍患者。间质纤维化的发展与临床参数相关。在实验模型中，我们分析了牛血清白蛋白（BSA）诱导的蛋白超载肾病和人IgG诱导的慢性血清病肾炎大鼠的间质组成和最终肾小球的变化。最后，我们改进并完善了大鼠输尿管梗阻性间质纤维化模型。为此，将定制的钛夹（S & T，Neuhaus，Switzerland）植入大鼠腹部的输尿管周围以阻塞输尿管而不引起坏死。在输尿管梗阻后的不同时间点（1 - 14天）取出夹子。此后研究了肾小管的随后重塑，以确定尿毒症诱导的间质纤维化是否在任何时候都保持可逆。在大鼠模型中，我们发现蛋白质超载诱导的和血清疾病诱导的间质纤维化都伴随着局灶性和节段性肾小球硬化的发展。只有在输尿管梗阻模型中才发生选择性间质纤维化，并且在研究的所有时间都保持可逆。为了可靠地评估间质纤维化，我们发现间质纤维化与肾功能的最佳相关参数是：（i）TGF-β-1及其拮抗剂核心蛋白聚糖的蛋白质积累比率;（ii）平滑肌α-肌动蛋白的间质表达;和（iii）间质胶原蛋白的积累（通过免疫过氧化物酶和天狼星红染色确定）。
Several clinical studies have confirmed that histomorphometric changes in the tubulointerstitial compartment contain the best correlating parameters to predict the development of progressive renal insufficiency. The process of interstitial fibrosis is accompanied by an influx of inflammatory cells, up-regulation of fibrogenic cytokines such as transforming growth factor-beta and basic fibroblast growth factor, transient down-modulation of their antagonists, generation and proliferation of myofibroblasts, and, finally, by accumulation of interstitial collagens and proteoglycans. A careful morphometric analysis of interstitial fibrosis requires sensitive parameters through which the severity can be quantified and by which the progression into renal insufficiency can be predicted. We have addressed these issues by morphometric analysis of both human biopsies and by refining existing experimental models in the rat. Morphometric analysis was performed using a Zeiss microscope equipped with a full colour 3CCD camera and KS-400 image analysis software from Zeiss-Kontron. For studies with human material, biopsies were examined from patients with various renal diseases including patients with chronic allotransplant dysfunction. The development of interstitial fibrosis was correlated with clinical parameters. In experimental models, we analysed the interstitial composition and eventual glomerular alterations in rats with bovine serum albumin (BSA)-induced protein overload nephropathy and with human IgG-induced chronic serum sickness nephritis. Finally, we adapted and refined the model of ureter obstruction-induced interstitial fibrosis in the rat. For this purpose, custom-made titanium clips (S&T, Neuhaus, Switzerland) were implanted around the ureter in the abdomen of rats to obstruct the ureter without causing necrosis. The clips were removed at various time points after obstruction of the ureter (1-14 days). The subsequent remodelling of the interstitium was studied thereafter, in order to establish whether uraemia-induced interstitial fibrosis remains reversible at all times. In rat models, we have found that both protein overload-induced and serum sickness-induced interstitial fibrosis are accompanied by the development of focal and segmental glomerulosclerosis. Only in the ureter obstruction model did selective interstitial fibrosis develop, and remained reversible at all times studied. For the reliable assessment of interstitial fibrosis we have found that the best correlating parameters of interstitial fibrosis with renal function were: (i) the ratio of protein accumulation of TGF-beta -1 and its antagonist decorin; (ii) interstitial expression of smooth muscle alpha -actin; and (iii) accumulation of interstitial collagens (as determined by immunoperoxidase and by Sirius red staining).