雌激素对人成纤维细胞基质金属蛋白酶-12、赖氨酸氧化酶基因表达的影响

Effects of estrogen on the expression of matrix metalloproteinase-12、lysyl oxidase gene in human dermal fibroblasts in vitro

目的:通过研究雌激素对体外培养的人皮肤成纤维细胞基质金属蛋白酶-12(MMP-12)及赖氨酸氧化酶(LOX)的mRNA转录水平的影响,探讨雌激素减缓皮肤老化的机制。方法:采用酶消化法进行体外人皮肤成纤维细胞原代培养,然后将不同浓度的雌激素(0 mol/L,1×10-5 mol/L,1×10-4 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比值来判断人成纤维细胞中MMP-12及LOX mRNA的表达水平。结果:对体外培养的人成纤维细胞进行雌激素干预,经β-act i n内参校正后,RT-PCR示对照组、低剂量组、高剂量组MMP-12表达水平三组间差异无统计学意义(F=3.711,P=0.056),LOX mRNA表达水平三组间差异具有统计学意义(F=4.337,P=0.038),SNK-q检验显示,低剂量组与对照组、低剂量组与高剂量组相比,差异无统计学意义(P>0.05)、高剂量组与对照组相比,差异具有统计学意义(P<0.05)。结论:雌激素可以上调LOX基因的转录水平,提示雌激素可能具有促进皮肤成纤维细胞弹力纤维的合成,逆转或缓解皮肤老化的作用。

Objective To expore the mechanism of estrogen on delaying skin aging by observing MMP-12 and LOX mRNA expression of in human dermal fibroblasts in vitro. Methods Human skin fibroblast were cultured in vitro.these fibroblast cells were then divided into 3 groups, lower dimensional estrogen was added in the medium in the second group, higher dimensional estrogen added in the medium in the third group. A few days later,we collected the fibroblast which was index number growth phase form different groups,extracted RNA, then amplified by RT-PCR. The PCR product was determined by agorose electrophoresis, to analysis the level of MMP-12,LOX mRNA expression. Results RT-PCR showed that there was no significant difference of the expression MMP-12 mRNA among control group,lowdose group and high-dose group （F=3.711, P=0.056）. The difference of the expression LOX mRNA among these three groups was statistically significant （F=4.337, P=0.038）.SNK-q test showed that no significant difference （P〉0.05） was found in LOX mRNA expression between the low-dose group and control group as well as between the low-dose group and the high-dose group; LOX mRNA expression was significantly higher （P〈0.05）in the high-dose group than the control group. Conclusion Estrogen can enhance the level of LOX mRNA expression.