医学分子生物学杂志2007，Vol.4Issue(3)：189-194,6.
两类金属蛋白酶在炎性细胞模型中的变化及中药RDQ的天然抑制效应
Change in the Activity of MMPs and ADAMs (TACE) in Inflammation Cell Model and Natural Suppressive Effect of Chinese Herb RDQ on These Two Subclasses of Metalloproteinase Family
摘要
Abstract
Objective This study was aimed to investigate the altered activity of two subclasses of metalloproteinases (MPs) family,matrix metalloproteinases (MMPs) and a disintegrin and metalloproteinases( ADAMs),during the transformation of precursor TNF-α (mTNF-α) to mature TNF-α (sTNF-α),and to explore the innate inhibitory effect of the Chinese herbal Re Du Qing (RDQ) on inflammation.Methods HL-60 cells were treated in vitro by lipopolvsaccharide (LPS) and RDQ.MPs activity was measured on SDS-PAGE substrate gel embedded with gelatin (zymography, Gelatin-PAGE).The mRNA expression of MMPs (MMP2,MMP7 and MMP9),ADAM17 (also called TNF-α converting enzyme,TACE),and sTNF-α, at various time points after LPS stimulation, was examined by Dot blot hybridization and RT-PCR,respectively.The amount of released sTNF-α was detected by MTT colorimetric assay.RT-PCR was used to assess the inhibitory effect of RDQ on TACE at mRNA level.Result Zymographic analysis revealed that the activity of MMPs in HL-60 cells gradually decreased with the time of LPS stimulation, whereas the alteration of MMPs at mRNA expression level was not significant, as detected by RT-PCR.Dot-blot results indicated that the expression of TACE and TNF-α at mRNA level increased simultaneously.The Chinese herb RDQ displayed obvious inhibitry effect on the cytotoxicity of sTNF-α induced by LPS stimulation at the transcriptional level of TACE gene.Conclusion TACE subclass of MPs family plays a crucial role in controlling the secretion of TNF-α induced by LPS.The Chinese herb RDQ can regulate TACE mRNA expression and inhibit sTNF-α secretion.TACE may serve as a novel target for inflammatory therapy.关键词
肿瘤坏死因子/脂多糖/肿瘤坏死因子转换酶/金属蛋白酶/热毒清Key words
tumor necrosis factor-α/inflammation/TNF-α converting enzyme/matrix metalloproteinases:Re Du Qing分类
生物科学引用本文复制引用
李小鸥,闫媛,王震,王开富,杨渝珍,匡宏伟..两类金属蛋白酶在炎性细胞模型中的变化及中药RDQ的天然抑制效应[J].医学分子生物学杂志,2007,4(3):189-194,6.基金项目
国家自然科学基金(No.30371309)This work was supported by a grant from the National Natural Sciences Foundation of China(No.30371309). （No.30371309）
