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

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。

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

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 cuhure.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 signifi-cantly 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 iysozyme of 1×10~(-7) mol/L.and 1×10~(-8)mol/L in the mRNA expres-sion of LOX(both P<0.05),whereas no significant difference was ohserved between fibroblasts treated with lysozyme of 1×10~(-8) mol/L and untreated fibrohlasts or those with lysozyme of 1 x 10~(-7)mol/L in the mRNA expression of MMP-1 and MMP-12.or between fibroblasts treated with lysozyme of 1 x 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.