CAJ | 학술논문

目的: 降低创面的高MMPs活性是治疗慢性皮肤溃疡的新途径.本研究主要观察11-羰基-β-乙酰乳香酸(AKBA,中药乳香的一种活性成分)对MMP-1、MMP-2和MMP-9活性的调节作用.方法: 人间质胶原酶(MMP-1)或者明胶酶A(MMP-2)被醋酸氨基苯汞(p-aminophenylmercuric acetate, APMA)激活后,与不同浓度的AKBA共同孵育1h,通过底物裂解法观察其活性的改变.MMP-9在中性粒细胞(polymorphonuclear neutrophils,PMNs)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AKBA共同孵育1 h,通过明胶酶谱法观察其中MMP-9活性的改变.我们建立了3个细胞模型:由TNF-α活化的人皮肤成纤维细胞模型;PMA活化的THP-1细胞模型和成纤维细胞-THP-1共培养细胞模型.AKBA与这3个细胞模型共同孵育24 h后,用ELISA法检测细胞上清中MMP-1、MMP-2和MMP-9的含量,用明胶酶谱法检测细胞上清中 MMP-2、MMP-9的活性.结果: AKBA在0.1-0.8 mmol/L浓度范围内对MMP-1、MMP-2的活性有抑制作用,IC50分别为0.18 mmol/L和0.27 mmol/L;在0.05-0.85mmol/L浓度范围内对MMP-9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.AKBA促进成纤维细胞分泌MMP-2,但是,对THP-1细胞分泌MMP-9表现出抑制作用.在共培养细胞模型中,AKBA对MMP-1、MMP-2和MMP-9的分泌均表现出抑制作用.结论: AKBA作为乳香的一种活性成分,它对MMPs活性的直接抑制作用和对MMPs分泌的抑制作用可能是中药乳香治疗慢性皮肤溃疡的机制之一.
목적: 강저창면적고MMPs활성시치료만성피부궤양적신도경.본연구주요관찰11-탄기-β-을선유향산(AKBA,중약유향적일충활성성분)대MMP-1、MMP-2화MMP-9활성적조절작용.방법: 인간질효원매(MMP-1)혹자명효매A(MMP-2)피작산안기분홍(p-aminophenylmercuric acetate, APMA)격활후,여불동농도적AKBA공동부육1h,통과저물렬해법관찰기활성적개변.MMP-9재중성립세포(polymorphonuclear neutrophils,PMNs)중함량봉부,인차이대서복강PMN작위MMP-9적래원.PMN렬해산물여불동농도적AKBA공동부육1 h,통과명효매보법관찰기중MMP-9활성적개변.아문건립료3개세포모형:유TNF-α활화적인피부성섬유세포모형;PMA활화적THP-1세포모형화성섬유세포-THP-1공배양세포모형.AKBA여저3개세포모형공동부육24 h후,용ELISA법검측세포상청중MMP-1、MMP-2화MMP-9적함량,용명효매보법검측세포상청중 MMP-2、MMP-9적활성.결과: AKBA재0.1-0.8 mmol/L농도범위내대MMP-1、MMP-2적활성유억제작용,IC50분별위0.18 mmol/L화0.27 mmol/L;재0.05-0.85mmol/L농도범위내대MMP-9활성표현출불동정도적억제작용(P<0.01),기억제작용정농도의뢰성.AKBA촉진성섬유세포분비MMP-2,단시,대THP-1세포분비MMP-9표현출억제작용.재공배양세포모형중,AKBA대MMP-1、MMP-2화MMP-9적분비균표현출억제작용.결론: AKBA작위유향적일충활성성분,타대MMPs활성적직접억제작용화대MMPs분비적억제작용가능시중약유향치료만성피부궤양적궤제지일.
AIM: To evaluate the effects of acetyl-11-keto-beta-boswellic acid (AKBA, a main active component from frankincense, one of the traditional Chinese herb for healing wounds) on the activities of matrix metalloproteinase(MMP)-1, MMP-2 and MMP-9.METHODS: Pure human interstitial collagenase (MMP-1) or gelatinase A (MMP-2) was activated by p-aminophenylmercuric acetate (APMA), and was incubated with AKBA for 1 h. The activities of the enzymes were observed by quenched fluorescent substrate. The lysates of rat polymorphonuclear neutrophils [PMNs, rich in gelatinase B (MMP-9)] was incubated with AKBA for 1 h, and activity of MMP-9 was tested by gelatin zymography. Three cell models: activated human dermal fibroblasts by TNF-α, activated THP-1 cells by PMA and fibroblasts-THP-1 co-culture system were established. AKBA was cultured with these cell models for 24 h. The levels of MMP-1, MMP-2 and MMP-9 in the cell culture supernatants were tested by ELISA and activities of MMP-2 and MMP-9 were tested by gelatin zymography assays.RESULTS: AKBA dose-dependently inhibited the activities of human MMP-1 and MMP-2 at the range of 0.1-0.8 mmol/L, with 50% inhibitory concentration (IC50) of 0.18 mmol /L and 0.27 mmol/L, respectively. In the range of 0.05-0.85 mmol/L, AKBA inhibited the MMP-9 activity (P<0.01). Although AKBA promoted fibroblasts to secrete MMP-2, the production of MMP-9 by THP-1 was inhibited. In the cell co-culture system, the inhibitory effects on MMP-1, MMP-2 and MMP-9 productions were also observed.CONCLUSION: AKBA, as a bioactive component of frankincense, has an inhibitory effect on MMPs production and activities, indicating the possible mechanism for healing chronic wounds by frankincense.