紫檀芪抑制血管平滑肌細胞遷移和基質金屬蛋白酶的機制探討

血管平滑肌細胞的遷移和基質金屬蛋白酶(matrix metalloproteinase ; MMP) 的活化，在動脈粥狀硬化的發展中扮演一個重要的角色。紫檀芪 (Pterostilbene) 是存在葡萄與藍莓中的典型黃酮類化合物，目前許多研究發現，紫檀芪具抗發炎、抗氧化與抗癌的功效。因此，本研究欲探討紫檀芪抑制血管平滑肌細胞遷移與基質金屬蛋白酶的相關性。本研究利用MTT assay來評估紫檀芪對平滑肌細胞株(A7r5)存活率的影響；接著利用migration/invasion assay、wound healing assay 及zymography 來評估紫檀芪對平滑肌細胞株(A7r5)，其細胞移動及基質金屬蛋白酶活性的影響；最後以西方墨點法評估紫檀芪抑制細胞外基質(ECM) 分解酵素的機制分析，如影響侵襲能力的基質金屬蛋白水解酶活性有關的訊息傳遞MAPK family ERK 1/2、p38及JNK 1/2、phosphatidylinositol 3-kinase (PI3K)/Akt pathway。Gelatin zymography與西方墨點法皆顯示紫檀芪會降低MMP-2的分泌與蛋白質表現，而migration/invasion assay與wound healing assay的遷移試驗顯示紫檀芪可有效抑制平滑肌細胞的遷移。另外，在訊息傳遞路徑的影響，可從西方墨點法的結果證實紫檀芪可誘發ERK磷酸化，故再利用ERK抑制劑去觀察其對細胞遷移與MMP-2的影響，結果也顯示紫檀芪是透過ERK的活化來抑制細胞遷移與MMP-2的表現。綜合以上結果可知，紫檀芪可有效抑制血管平滑肌的增生與遷移，以延緩動脈粥狀硬化的病程發展，因此，在臨床上可建議心血管相關疾病之患者可適量補充莓果(藍莓與葡萄)，將有助於預防動脈粥狀硬化。

關鍵字

動脈粥狀硬化；平滑肌細胞遷移；基質金屬蛋白酶紫檀芪

並列摘要

Smooth muscle cells (SMCs) migration and matrix metalloproteinase-2 (MMP-2) activation are main roles in atherosclerosis. Pterostilbene (trans-3, 5-dimethoxy-4-hydroxystilbene), a flavonoid extensively found in blueberries and grapes, confers potent antioxidant, anti-inflammatory and anticarcinogenic properties. The present study aimed to investigate the anti-atheroscleroic property of pterostilbene in the rat smooth muscle cell (SMC) A7r9 cell lines and the underlying mechanisms. The cytotoxic effect of pterostilbene on smooth muscle cell line (A7r5) was investigated using 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyl tetrazolium bromide (MTT) assay. The mobility of vascular smooth muscle cells through the extracellular matrix was determined by migration assay and wound healing assay. Evaluate of matrix metalloproteinase (MMP) activation and protein expression by gelatin zymography and western blot. In this study, pterostilbene treatment significantly inhibited migration/invasion capacities of in A7r9 cell. Pterostilbene was also found to significantly decreased MMP-2 activity and expression by gelatin zymography and western blot assay in SMC. In the MAPK signaling pathway, western blot assay also indicated that pterostilbene up-regulated the phosphorylation of extracellular-signal- regulated kinase (Erk) 1/2. Moreover, inhibition of Erk1/2 by specific inhibitors significantly abolished the pterostilbene-decreased expression of MMP-2 and migration/invasion capacities. These findings suggest that pterostilbene inhibited SMC migration and that MMP-2 activation could be mediated via Erk1/2 phosphorylation. It is further possible that pterostilbene could play a novel role in the treatment of atherosclerosis.

並列關鍵字

Pterostilbene ； smooth muscle cells ； migration ； matrix metalloproteinase-2

參考文獻

Riche, D. M., McEwen, C. L., Riche, K. D., Sherman, J. J., Wofford, M. R., Deschamp, D., & Griswold, M. (2013). Analysis of safety from a human clinical trial with pterostilbene. J Toxicol, 2013, 463595. doi: 10.1155/2013/463595

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紫檀芪抑制血管平滑肌細胞遷移和基質金屬蛋白酶的機制探討

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