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分泌型白细胞蛋白酶抑制剂由人关节软骨软骨细胞和椎间盘纤维软骨细胞产生。

Secretory leucocyte proteinase inhibitor is produced by human articular cartilage chondrocytes and intervertebral disc fibrochondrocytes.

作者信息

Jacoby A S, Melrose J, Robinson B G, Hyland V J, Ghosh P

机构信息

Raymond Purves Bone and Joint Research Laboratories, University of Sydney, Royal North Shore Hospital, Australia.

出版信息

Eur J Biochem. 1993 Dec 15;218(3):951-7. doi: 10.1111/j.1432-1033.1993.tb18452.x.

Abstract

The objective of this study was to examine the expression of the secretory leucocyte proteinase inhibitor (SLPI) gene by human articular cartilage chondrocytes and intervertebral disc fibrochondrocytes. RNA was extracted from human articular cartilage chondrocytes, synovial fibroblasts and fibrochondrocytes of the annulus fibrosus. Expression was analysed by Northern blotting and hybridisation to a SLPI RNA probe. SLPI was isolated from chondrocyte culture supernatant by gel-permeation and cation-exchange chromatography and examined by a solid-phase enzyme-linked immunofiltration assay for SLPI and by SDS/PAGE using Western blotting with polyclonal IgG samples against bronchial mucus leucocyte proteinase inhibitor and SLPI. This identified biochemically and immunologically the major serine proteinase inhibitory protein synthesised by chondrocytes as SLPI. Secretory leucocyte proteinase inhibitor mRNA was detected in RNA preparations from articular cartilage chondrocytes and annulus fibrosus fibrochondrocytes. The results presented here allowed us to deduce that human articular cartilage chondrocytes and intervertebral disc fibrochondrocytes synthesized SLPI whereas human synovial fibroblasts did not.

摘要

本研究的目的是检测人关节软骨软骨细胞和椎间盘纤维软骨细胞中分泌型白细胞蛋白酶抑制剂(SLPI)基因的表达。从人关节软骨软骨细胞、滑膜成纤维细胞和纤维环纤维软骨细胞中提取RNA。通过Northern印迹法和与SLPI RNA探针杂交分析表达情况。通过凝胶渗透和阳离子交换色谱从软骨细胞培养上清液中分离出SLPI,并使用针对支气管黏液白细胞蛋白酶抑制剂和SLPI的多克隆IgG样品通过固相酶联免疫过滤测定法检测SLPI,并通过SDS/PAGE和Western印迹法进行检测。这从生化和免疫方面鉴定出软骨细胞合成的主要丝氨酸蛋白酶抑制蛋白为SLPI。在关节软骨软骨细胞和纤维环纤维软骨细胞的RNA制剂中检测到分泌型白细胞蛋白酶抑制剂mRNA。此处给出的结果使我们推断出人关节软骨软骨细胞和椎间盘纤维软骨细胞合成SLPI,而人滑膜成纤维细胞不合成。

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