摘要
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 neu-trophils,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1h,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24h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0.1-0.8mmol/L浓度范围内对MMP-1、MMP-2的活性有抑制作用,IC50分别为0.18mmol/L和0.27mmol/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。
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 fi- broblasts - 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 tbe 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.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2009年第10期2004-2011,共8页
Chinese Journal of Pathophysiology
基金
Beijng Natural Science Foundation(No.7072014)