Firestein G S, Yeo M, Zvaifler N J
University of California, San Diego Medical Center, La Jolla, California 92093, USA.
J Clin Invest. 1995 Sep;96(3):1631-8. doi: 10.1172/JCI118202.
RA synovial tissue (ST) was studied to determine if and where apoptosis occurs in situ. Genomic DNA was extracted from 5 RA and 1 osteoarthritis ST samples. Agarose gel electrophoresis demonstrated DNA ladders characteristic for apoptosis from each tissue. In situ and labeling (ISEL) was used to identify DNA strand breaks consistent with apoptosis in frozen sections. 12 RA and 4 osteoarthritis ST were studied by ISEL and all were positive, but only 2 of 4 normal tissues were positive. The primary location of apopotic cells was the synovial lining. Some sublining cells were also positive, but lymphoid aggregate staining was conspicuously absent. Immunohistochemistry and ISEL were combined and showed that the lining cells with DNA strand breaks were mainly macrophages, although some fibroblastlike cells were also labeled. Sublining cells with fragmented DNA included macrophages and fibroblasts, but T cells in lymphoid aggregates, which expressed large amounts of bcl-2, were spared. DNA strand breaks in cultured fibroblastlike synoviocytes was assessed using ISEL. Apoptosis could be induced by actinomycin D, anti-fas antibody, IL-1, and TNF-alpha but not by IFN-gamma. Fas expression was also detected on fibroblast-like synoviocytes using flow cytometry. Therefore, DNA strand breaks occur in synovium of patients with arthritis. Cytokines regulate this process, and the cytokine profile in RA (high IL-1/TNF; low IFN-gamma) along with local oxidant injury might favor induction of apoptosis.
研究类风湿性关节炎(RA)滑膜组织(ST)以确定凋亡是否以及在何处原位发生。从5个RA和1个骨关节炎ST样本中提取基因组DNA。琼脂糖凝胶电泳显示每个组织均有凋亡特征性的DNA梯带。原位末端标记(ISEL)用于识别冰冻切片中与凋亡一致的DNA链断裂。通过ISEL研究了12个RA和4个骨关节炎ST,所有样本均呈阳性,但4个正常组织中只有2个呈阳性。凋亡细胞的主要位置是滑膜衬里。一些滑膜下层细胞也呈阳性,但明显没有淋巴样聚集染色。免疫组织化学与ISEL相结合显示,有DNA链断裂的衬里细胞主要是巨噬细胞,尽管一些成纤维样细胞也有标记。有DNA片段化的滑膜下层细胞包括巨噬细胞和成纤维细胞,但表达大量bcl-2的淋巴样聚集中的T细胞未受累。使用ISEL评估培养的成纤维样滑膜细胞中的DNA链断裂。放线菌素D、抗fas抗体、IL-1和TNF-α可诱导凋亡,但IFN-γ不能。使用流式细胞术也在成纤维样滑膜细胞上检测到了Fas表达。因此,关节炎患者的滑膜中会发生DNA链断裂。细胞因子调节这一过程,RA中的细胞因子谱(高IL-1/TNF;低IFN-γ)以及局部氧化损伤可能有利于诱导凋亡。