三氧化二砷对皮肤成纤维细胞、中性粒细胞MMPs活性,TIMP-1及TGF-β1表达的影响

目的：砒石是化腐生肌的常用中药，其主要成分是三氧化二砷（As₂O₃）。本研究通过观察As₂O₃对基质金属蛋白酶（MMPs）活性、基质金属蛋白酶组织抑制因子-1（TIMP-1）及转化生长因子β₁（TGF-β₁）表达影响，探讨化腐中药能否调节胶原代谢，从而治疗慢性皮肤溃疡。方法:明胶酶谱法检测大鼠中性粒细胞（PMNs）来源的MMP-9活性、人成纤维细胞（hFb）分泌的MMP-1、MMP-2的活性，免疫细胞化学法检测hFb TIMP-1、TGF-β₁的表达。结果:As₂O₃浓度在50 mg/L时可以提高大鼠PMNs来源的MMP-9的活性（P<0.01）；在0.8 mg/L可以提高hFb分泌的MMP-1、MMP-2的活性（分别P<0.01）；同时As₂O₃作用于hFb 6 h、12 h、18 h后，TIMP-1、TGF-β₁表达持续降低（P<0.01）。结论:As₂O₃在一定范围内可提高PMNs来源的MMP-9的活性；也可提高hFb分泌的MMP-1、MMP-2的活性，同时抑制hFbTIMP-1、TGF-β₁的表达。提示砷类制剂可通过提高多种MMPs的活性，降低TIMP-1的表达从而发挥化腐作用。

Effects of arsenic trioxide on activities of MMPs and expression of TIMP-1 and TGF-β1 in skin fibroblasts and polymorphonuclear neutrophils

AIM: To observe the effects of arsenic trioxide (As₂O₃) on activities of matrix metalloproteinases (MMPs), expression of tissue inhibitor of metalloproteinase-1 (TIMP-1) and transforming growth factor beta1 (TGF-β₁) in human fibroblast (hFb), and to discuss weather As₂O₃ promotes the healing of chronic skin ulcer through regulating collagen metabolism. METHODS: Zymography was used for testing activity of MMP-9 deriving from rat polymorphonuclear neutrophils (PMNs) and activities of MMP-1, MMP-2 secreted by hFb. Immunocytochemical method was used to determine the expressions of TIMP-1 and TGF-β₁. RESULTS: At the concentration of 50 mg/L, As₂O₃ elevated the activity of MMP-9 (P<0.01). At the concentration of 0.8 mg/L, As₂O₃ increased the activities of MMP-1 and MMP-2 (P<0.01, respectively). After hFb was cultured with As₂O₃ for 6 h, 12 h and 18 h, the expressions of TIMP-1 and TGF-β₁ decreased continuously (P<0.01). CONCLUSION: As₂O₃ elevates the activities of MMP-1, MMP -2 and MMP-9, also inhibits the expressions of TIMP-1 and TGF-β₁, suggesting that arsenic preparation may exert positive effect on healing chronic skin ulcer through regulating collagen metabolism.