关键词: 基质金属蛋白酶, 肿瘤坏死因子转换酶, 肿瘤坏死因子, 大肠杆菌内毒素, 热毒清

Abstract: BACKGROUND ＆ AIM: To study the function and role of two subclasses in MPs family, MMPs and ADAMs, for processing of precursor TNF-α, and explore the anti-inflammatory mechanisms of Chinese herbal preparation Reduqing (RDQ). MATERIAL AND METHODS: The in vitro study was carried out on HL-60、U937 cells and macrophages of murine abdominal cavity treated by LPS and RDQ. Using MTT colorimetry,in situ hybridization and Gelatin-PAGE to detect transcriptional levels of MMPs,ADAM17, TNF-α mRNA and the changes of enzymatic activity. RESULTS: ①After stimulation by LPS, the expression level of MMPs and electrophoresis zymogram on gelatin-PAGE decreased with increased stimulation time; ②ADAM17 (TACE) played a more important role in precursor processing of proTNF-α of HL-60 cells compared with MMPs; ③RDQ had obvious inhibitory effects on enhancing secretion of TNF-α induced by LPS stimulation (P<0.01) at the transcriptional level of ADAM17 mRNA in HL-60 cells. CONCLUSION: ①ADAM17 of MPs family had a main role in TNF-α secretion induced by LPS. It is a key step to choose and design the correct probe or primers aiming specifically at the disintegrin domain for ADAM17, in order to eliminate the interference of MMPs and reflect the true expression level of ADAM17 gene; ②The mechanism of RDQ against inflammation may be its inhibitory effect on ADAM17 mRNA expression activated by LPS. ADAM17 is able to represent the novel target for studing the mechanism and the therapeutic pathway for inflammation.

Key words: matrix metalloproteinases, TNF-α, Lipopolysacharides, Reduging