中华医学杂志（英文版）2001，Vol.114Issue(2)：162-166,5.
抗Fas抗体诱导人肾间质成纤维细胞凋亡
Anti-Fas antibody induces apoptosis in cultured human renal interstitial fibroblasts
程丽静 1谌贻璞2
作者信息
- 1. 北京医科大学第一医院肾内科 肾脏病研究所 北京 100034
- 2. 北京中日友好医院肾内科 北京 100029
- 折叠
摘要
Abstract
Objective To detect if Fas is expressed in human renal interstitial fibroblasts (hRIFs) and apoptosis of hRIFs can be induced by specific anti-Fas antibody.
Methods hRIFs were cultured from isolated papillae of human kidney, and identified by morphologic examination, assay of antigenic components and culture in D-valine selective medium. Fas expression in normal hRIFs was detected by RT-PCR and immunocytochemistry staining. After hRIFs were incubated with interferon gamma (γ-IFN 500 U/ml, 1000 U/ml, 1500 U/ml and 2000 U/ml, respectively) for 48 hours, Fas expression was determined by Northern blot, Western blot and flow cytometry. hRIFs pre-stimulated with γ-IFN (500 U/ml, 48 hours) were incubated with anti-Fas antibody (IgM, 0.5 μg/ml) for 12 hours. And apoptosis was identified by morphologic examination, DNA ladder assay and flow cytometry.
Results The cultured hRIFs showed a shuttle-like shape and were positively stained by labeled anti-vimentin antibody but negatively stained by anti-epithelial membrane antibody. They could not grow in the D-valine selective medium and died partly in a week. Fas mRNA and protein were expressed in normal hRIFs and markedly upregulated by stimulation with γ-IFN. Apoptosis in γ-IFN pre-stimulated hRIFs was induced by anti-Fas antibody, showing cell nuclear shrinkage and condensation in morphologic feature, internucleosomal DNA fragmentation in DNA ladder assay and a pick of hypo-diploid nuclei by flow cytometry.
Conclusion Fas is normally expressed in hRIFs and can be markedly upregulated by γ-IFN. Anti-Fas antibodies can induce apoptosis of hRIFs pre-stimulated with γ-IFN.关键词
Fas 成纤维细胞 细胞凋亡Key words
apoptosis/interstitial fibroblasts/Fas/anti-Fas antibody分类
医药卫生引用本文复制引用
程丽静,谌贻璞..抗Fas抗体诱导人肾间质成纤维细胞凋亡[J].中华医学杂志（英文版）,2001,114(2):162-166,5.
