RF/J小鼠培养的系膜细胞表型改变：一种自发性免疫复合物介导的肾小球肾炎伴进行性肾小球硬化。

Altered cultured mesangial cell phenotypes from RF/J mice: a spontaneous immune complex mediated glomerulonephritis with progressive glomerulosclerosis.

作者信息

Yashiro M, Muso E, Kamata T, Oyama A, Sasayama S, Yoshida H

机构信息

Division of Nephrology, Kyoto City Hospital, Department of Internal Medicine, Kyoto University Hospital, Kyoto, Japan.

出版信息

Exp Nephrol. 2001;9(6):420-7. doi: 10.1159/000052641.

DOI:10.1159/000052641

PMID:11702002

Abstract

BACKGROUND/AIM: RF/J mice are a model of spontaneous immune complex mediated glomerulonephritis showing massive extracellular matrix accumulation and progressive glomerulosclerosis. The aim of this study was to investigate whether there is an altered cultured mesangial cell (MC) phenotype in RF/J mice associated with these glomerular changes.

METHODS

The nature of cultured MCs from RF/J mice in the proliferative response to platelet-derived growth factor (PDGF) BB was compared with that of normal mice (BALB/c) by 3H-thymidine incorporation. The binding of PDGF-BB was examined with Scatchard analysis, and the messenger RNAs (mRNAs) of PDGF beta-receptor, collagen I, collagen IV, and fibronectin were detected using Northern blot analysis in the MCs of each mouse.

RESULTS

The 3H-thymidine incorporation of MCs from RF/J mice showed significantly lower responses to PDGF-BB stimulations with concentrations ranging from 0.5 to 10.0 ng/ml in comparison with those of BALB/c mice which exhibited a proportional dose- dependent increase of the incorporation (p < 0.05 for 0.5 ng/ml PDGF-BB, p < 0.01 for 1.0-10.0 ng/ml). According to the Scatchard analysis, MCs from BALB/c mice showed aKD of 105 pM of PDGF-BB binding to its receptors, and the density of receptors was 5.82 fmol/10(5) cells. However, no binding PDGF-BB site on the surface of MCs from RF/J mice was noted. Northern blot analysis of MCs from RF/J mice indicated negative expression of detectable PDGF-beta receptor mRNA. As for matrix protein messages, MCs from RF/J mice did not express mRNA of type I collagen, but did express a higher amount of type IV collagen and fibronectin in comparison with MCs from normal BALB/c mice.

CONCLUSIONS

An altered phenotype in MCs of RF/J mice was demonstrated, possibly contributing to the characteristic pathological glomerular changes. However, the precise association remains to be clarified.

摘要

背景/目的：RF/J小鼠是一种自发免疫复合物介导的肾小球肾炎模型，表现为大量细胞外基质积聚和进行性肾小球硬化。本研究的目的是调查RF/J小鼠中培养的系膜细胞（MC）表型是否发生改变，以及这种改变与这些肾小球变化的关系。

方法

通过³H-胸腺嘧啶核苷掺入法，比较RF/J小鼠培养的MC对血小板衍生生长因子（PDGF）BB增殖反应的性质与正常小鼠（BALB/c）的差异。用Scatchard分析检测PDGF-BB的结合情况，并使用Northern印迹分析检测每只小鼠MC中PDGFβ受体、I型胶原、IV型胶原和纤连蛋白的信使核糖核酸（mRNA）。

结果

与BALB/c小鼠相比，RF/J小鼠的MC对浓度范围为0.5至10.0 ng/ml的PDGF-BB刺激的³H-胸腺嘧啶核苷掺入反应显著降低，BALB/c小鼠的掺入量呈比例剂量依赖性增加（0.5 ng/ml PDGF-BB时p<0.05，1.0 - 10.0 ng/ml时p<0.01）。根据Scatchard分析，BALB/c小鼠的MC显示PDGF-BB与其受体结合的解离常数（KD）为105 pM，受体密度为5.82 fmol/10⁵细胞。然而，未观察到RF/J小鼠MC表面有PDGF-BB结合位点。RF/J小鼠MC的Northern印迹分析表明可检测到的PDGF-β受体mRNA呈阴性表达。至于基质蛋白信息，RF/J小鼠的MC不表达I型胶原mRNA，但与正常BALB/c小鼠的MC相比，表达更高量的IV型胶原和纤连蛋白。