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目的探讨金属蛋白酶(MPs)家族的MMPs和ADAMs两亚类成分在前体TNF-α(mTNF-α)向成熟型TNF-α(sTNF-α)转化过程所起的作用,MMPs在炎症过程中活性的变化和中药RDQ抗炎保护功效的分子机制,为人工干预炎症过程提供依据和手段。方法以LPS刺激HL-60细胞为天然模型,用细胞学和分子生物学技术在蛋白质和mRNA水平上研究MMPs、ADAM17、TNF-α在炎症过程中的表达和活性变化。结果①LPS刺激后,HL60细胞上清中MMPs的活性随刺激时间延长逐渐减弱,胞浆裂解物中的MMPs则随刺激时间延长活性增加但LPS刺激对MMPs mRNA的表达影响不大;②LPS刺激能使HL-60细胞中ADAM17mRNA和TNF-αmRNA表达在6h同时达高峰;③RDQ能明显降低LPS诱导的ADAM17 mR- NA的增加,并能抑制sTNF-α的分泌。结论①ADAM17对sTNF-α的产生起主导作用;②纯中药注射液热毒清(RDQ)能调节TACE的表达,最终抑制TNF-α的分泌。
Objective To investigate the role of the two subclasses of MMPs and ADAMs in the metalloproteinases (MPs) family in the process of transforming precursor TNF-α (mTNF-α) into mature TNF-α (sTNF-α). MMPs are involved in inflammation. The changes in activity and the molecular mechanisms of the anti-inflammatory protective efficacy of traditional Chinese medicine RDQ provide a basis and means for human intervention in the inflammatory process. Methods LPS-stimulated HL-60 cells were used as a natural model. Cytogenetic and molecular biological techniques were used to study the expression and activity of MMPs, ADAM17 and TNF-α in the process of inflammation at the protein and mRNA levels. Results 1 After LPS stimulation, the activity of MMPs in the supernatant of HL60 cells gradually weakened with the prolongation of the stimulation time. The MMPs in cytoplasmic lysates increased with the stimulation time but the activity of LPS stimulation had little effect on the expression of MMPs mRNA; 2LPS stimulation could make The expression of ADAM17 mRNA and TNF-α mRNA in HL-60 cells reached the peak at the same time at 6h. 3RDQ could significantly reduce the increase of ADAM17 mR-NA induced by LPS and inhibit the secretion of sTNF-α. Conclusion 1 ADAM17 plays a leading role in the production of sTNF-α.2 Pure Chinese herbal injection RDQ can regulate the expression of TACE and ultimately inhibit the secretion of TNF-α.