溶菌酶对体外培养的人成纤维细胞基质金属蛋白酶-1和12及赖氨酸氧化酶基因表达的影响  

Influences of lysozyme on the gene expressions of matrix metaHoproteinase （MMP）-1, -12 and lysyl oxidase in cultured human dermal fibroblasts in vitro

作　　者：赵利媛[1] 刘原君[1] 范丽云[1] 张俊艳[1] 亓玉清[1] 刘全忠[1] 

机构地区：[1]天津医科大学总医院皮肤科、天津性传播疾病研究所,300052

出　　处：《中华皮肤科杂志》2010年第1期36-39,共4页Chinese Journal of Dermatology

基　　金：天津市自然科学基金（003610211）

摘　　要：目的探讨溶菌酶对体外培养的人成纤维细胞基质金属蛋白酶-1和12及赖氨酸氧化酶基因表达的影响。方法采用酶消化法进行体外人皮肤成纤维细胞原代培养，然后将不同浓度的溶菌酶（0，1×10^-8，1×10^-7mol／L）]加入体外培养的人皮肤成纤维细胞中，待药物作用后，提取总RNA，通过逆转录反应获得cDNA并进行体外扩增，对扩增产物进行琼脂糖凝胶电泳，根据条带的平均光密度A值的比值来判断人成纤维细胞中基质金属蛋白酶（MMP）-1、MMP-12及赖氨酸氧化酶（LOX）mRNA的表达水平。结果对体外培养的人成纤维细胞进行溶菌酶干预，经B肌动蛋白内参校正后，RT-PCR示对照组、低剂量组、高剂量组MMP-1、MMP-12 mRNA表达水平三组间差异具有统计学意义（F值分别为6．98和4．44，P值均〈0．05）。SNK—q检验显示，低剂量组与对照组、高剂量组比较，差异均无统计学意义（P〉0．05），高剂量组与对照组相比，差异有统计学意义（P〈0．05）。LOX mRNA表达水平三组间差异具有统计学意义（F=5．24，P〈0．05），SNK—q检验显示，低剂量组、高剂量组与对照组相比，差异具有统计学意义（P〈0．05），低剂量组与高剂量组相比，差异无统计学意义（P〉0．05）。结论溶菌酶可以下调MMP-1和MMP-12及上调LOX基因的转录水平。Objective To investigate the influences of lysozyme on the mRNA expressions of MMP-1, -12 and lysyl oxidase （LOX） in cultured fibroblasts in vitro. Methods Primarily cultured fibroblasts isolated from human skin were treated with three concentrations （0, 1 × 10^-8, 1 × 10^-7 mol/L） of lysozyme followed hy another 24-hour culture. Subsequently, total RNA was extracted from the fibroblasts and subjected to RT-PCR for the detection of MMP-1, -12 and LOX mRNA. Results There was a significant difference in the mRNA expressions of MMP-1, -12 and LOX among the fibroblasts treated with the three concentrations of lysozyme （F = 6.98, 4.44, 5.24, respectively, all P 〈 0.05）. SNK-q test showed that untreated fibroblasts differed significantly from those treated with lysozyme of 1 × 10^-7 mol/L in the mRNA expression of MMP-1 and MMP-12 （P 〈 0.05）, and from those treated with lysozyme of 1 ×10^-7 mol/L and 1 × 10^-8 mol/L in the mRNA expression of LOX （both P 〈 0.05）, whereas no significant difference was observed between fibroblasts treated with lysozyme of 1 × 10^-8 mol/L and untreated fibroblasts or those with lysozyme of 1 × 10^-7 mol/L in the mRNA expression of MMP-I and MMP-12, or between fibroblasts treated with lysozyme of 1 × 10^-8 mol/L and those with that of 1 ×10^-7 mol/L in the expression of LOX （all P 〉 0.05）. Conclusions Lysozyme upregulates the mRNA expression of MMP-1 and MMP-12 but downregulates the mRNA expression of LOX in cultured fibroblasts in vitro.

关 键 词：成纤维细胞 胞壁质酶 基质金属蛋白酶1 基质金属蛋白酶12 蛋白赖氨酸6-氧化酶 

分 类 号：R751[医药卫生—皮肤病学与性病学]