角膜基质细胞基质金属蛋白酶1,2活性与组织因子途径抑制物2的效应(英文)

Effect of tissue factor pathway inhibitor-2 on the expressions of matrix metalloproteinase 1 and 2 in keratocytes

背景:有研究表明,基质金属蛋白酶所参与的细胞外基质降解在角膜新生血管形成过程中起关键作用,组织因子途径抑制物2是新近发现的一种新型的丝氨酸蛋白酶抑制物,能有效抑制基质金属蛋白酶的活性。目的:观察组织因子途径抑制物2对体外角膜基质细胞表达基质金属蛋白酶活性的关系。方法:在体外对兔角膜基质细胞进行原代及传代培养,用脂质体介导的人类组织因子途径抑制物2真核表达载体转染兔角膜基质细胞,G418筛选阳性细胞。结果与结论:RT-PCR,Western blot及明胶酶谱法分析结果显示,转染后角膜基质细胞组织因子途径抑制物2 mRNA和蛋白质的表达均上调(P<0.05),而基质金属蛋白酶1,2的活性下降(P<0.05)。结果提示,组织因子途径抑制物2可明显抑制角膜基质细胞中基质金属蛋白酶1,2的活性。

BACKGROUND： The degradation of extracellular matrix, which is mediated by matrix metalloproteinases, plays a crucial role in the corneal neovascularization. Tissue factor pathway inhibitor 2, a new type serine proteinase inhibitor, can effectively inhibit the activity of matrix metalloproteinases. OBJECTIVE： To elucidate the effect of tissue factor pathway inhibitor 2 on the expressions of matrix metalloproteinases in keratocytes in vitro. METHODS： Rabbit keratocytes were primarily cultured and subcultured in vitro. Plasmid vector pBos-Cite-neo/TFPI-2 was transfected into keratocytes with Lipofectamine 2000. The positive cells were selected using G418. RESULTS AND CONCLUSION： The results of reverse transcription-polymerase chain reaction, western blot analysis and gelatinase zymography analysis showed that, expression of mRNA and protein of tissue factor pathway inhibitor 2 was upregulated in the transfected keratocytes （P 〈 0.05）, while activity of matrix metalloproteinase 1 and 2 was significantly decreased （P 〈 0.05）. Experimental findings indicate that that tissue factor pathway inhibitor 2 strongly inhibits the activity of matrix metalloproteinase 1 and 2 in keratocytes.