中华皮肤科杂志 ›› 2010, Vol. 43 ›› Issue (1): 36-39.

• 论著 • 上一篇    下一篇

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

赵利媛1,刘原君2,范丽云3,张俊艳3,亓玉清4,刘全忠2   

  1. 1. 天津医科大学总医院
    2. 天津医科大学总医院皮肤性病科
    3. 天津医科大学总医院皮肤科
    4. 天津医科大学总医院临床检验科
  • 收稿日期:2009-02-27 修回日期:2009-03-19 出版日期:2010-01-15 发布日期:2010-01-05
  • 通讯作者: 赵利媛 E-mail:zlyuan2007@126.com
  • 基金资助:

    天津市自然科学基金（003610211）

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

  • Received:2009-02-27 Revised:2009-03-19 Online:2010-01-15 Published:2010-01-05

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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 × 10-7 mol/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β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。

关键词: 溶菌酶, 基质金属蛋白酶-1, 基质金属蛋白酶-12, 赖氨酸氧化酶, 成纤维细胞

Abstract:

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 by 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-1 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 fibro-blasts in vitro.