CAJ | 학술논문

目的:基质金属蛋白酶家族(matrix metalloproteinases, MMPs)是一类活性依赖于金属锌离子的蛋白酶,它可以降解绝大多数细胞外基质成分.MMPs水解蛋白质的特性有利于创伤愈合过程中坏死组织的清除,以及细胞的迁移.降低慢性创面升高的MMPs活性可能是治疗慢性皮肤溃疡的新途径.丹参外用可治疗慢性皮肤溃疡,丹酚酸B(salvianolic acid B, Sal B)是丹参中含量丰富且具有较高生物活性的水溶性成分.本研究主要观察Sal B在体外对MMPs的活性是否有直接影响.方法:人MMP-1和MMP-2被醋酸氨基苯汞(p-aminophenylmercuric acetate, APMA)激活后,与不同浓度的Sal B共同孵育1 h,通过底物裂解法观察其活性的改变.MMP-9在中性粒细胞(polymorphonuclear neutrophil, PMN)中含量丰富,因此以大鼠腹腔PMN作为MMP-9的来源.PMN裂解产物与不同浓度的Sal B共同孵育1 h,通过明胶酶谱法,观察其中MMP-9活性的改变.结果:Sal B在0.002 4～0.3 g/L浓度范围内对MMP-1和MMP-2的活性有抑制作用,半抑制浓度(50% inhibiting concentration, IC50)分别为(0.090±0.015)g/L和(0.080±0.005)g/L;在0.003～0.3 g/L浓度范围内对MMP-9的活性表现出不同程度的抑制作用(P<0.01),其抑制作用呈浓度依赖性.结论:Sal B可抑制MMPs的活性,改善慢性创面高酶活性是丹参治疗慢性皮肤溃疡的作用机制之一.
목적:기질금속단백매가족(matrix metalloproteinases, MMPs)시일류활성의뢰우금속자리자적단백매,타가이강해절대다수세포외기질성분.MMPs수해단백질적특성유리우창상유합과정중배사조직적청제,이급세포적천이.강저만성창면승고적MMPs활성가능시치료만성피부궤양적신도경.단삼외용가치료만성피부궤양,단분산B(salvianolic acid B, Sal B)시단삼중함량봉부차구유교고생물활성적수용성성분.본연구주요관찰Sal B재체외대MMPs적활성시부유직접영향.방법:인MMP-1화MMP-2피작산안기분홍(p-aminophenylmercuric acetate, APMA)격활후,여불동농도적Sal B공동부육1 h,통과저물렬해법관찰기활성적개변.MMP-9재중성립세포(polymorphonuclear neutrophil, PMN)중함량봉부,인차이대서복강PMN작위MMP-9적래원.PMN렬해산물여불동농도적Sal B공동부육1 h,통과명효매보법,관찰기중MMP-9활성적개변.결과:Sal B재0.002 4～0.3 g/L농도범위내대MMP-1화MMP-2적활성유억제작용,반억제농도(50% inhibiting concentration, IC50)분별위(0.090±0.015)g/L화(0.080±0.005)g/L;재0.003～0.3 g/L농도범위내대MMP-9적활성표현출불동정도적억제작용(P<0.01),기억제작용정농도의뢰성.결론:Sal B가억제MMPs적활성,개선만성창면고매활성시단삼치료만성피부궤양적작용궤제지일.
Objective: Matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases, which as a group can degrade essentially all extracellular matrix components. The proteolytic property of the MMPs is important during wound healing to remove debris and facilitate cell migration. Targeting towards the decreased MMPs activities is a new treatment strategy for healing chronic wounds. Salvia miltiorrhiza is a popular Chinese herb that could promote chronic ulcers healing for topical use. Salvianolic acid B (Sal B) is the most abundant bioactive component in Salvia miltiorrhiza. The research was designed to explore the inhibitory effects of Sal B on MMP-1, MMP-2 and MMP-9 activities.Methods: Pure human interstitial collagenase (MMP-1) or gelatinase A (MMP-2) was activated by p-aminophenylmercuric acetate (APMA), and was incubated with Sal B for 1 h. The activities were observed by quenched fluorescent substrate. Gelatinase B (MMP-9) is rich in polymorphonuclear neutrophils (PMN), so the rat PMN was used as a source of MMP-9 for MMPs activity assays. In vitro MMP-9 from rats' PMN lysate was incubated with Sal B for 1 h, and its activity was tested by gelatin zymography.Results: Sal B dose-dependently inhibited the human MMP-1 and MMP-2 activities in the range of 0.002 4 to 0.3 g/L, with 50% inhibiting concentration (IC50) of (0.090±0.015) g/L and (0.080±0.005) g/L respectively. In the range of 0.003 to 0.3 g/L, Sal B could inhibit the MMP-9 activity (P<0.01).Conclusion: The broad-spectrum inhibitory effects of Sal B on MMPs may reveal one of the mechanisms for the effects of Salvia miltiorrhiza on chronic wounds.