miRNA-17对UVA诱导的皮肤成纤维细胞光老化的调节作用

Regulation effect of miR-17 on ultraviolet A-induced photoageing of human dermal fibroblasts

目的检测miR-17在长波紫外线(UVA)诱导的光老化中的调节作用。方法分离人皮肤成纤维细胞(HDFs),用0 J/cm^2(对照组),2.5,5.0,7.5 J/cm^2剂量的UVA辐射诱导光老化模型,用实时定量PCR和Western blot分别检测miR-17和基质金属蛋白酶3(MMP3)的表达水平。对HDFs细胞分别转染miR-17的功能模拟物、miR-17阴性对照物、miR-17的功能抑制物和miR对照的功能抑制物,转染24 h后UVA辐射诱导光老化模型,检测MMP3蛋白表达水平。结果经不同剂量UVA辐射后,HDFs中MMP3表达与对照组相比显著升高(P<0.01),且miR-17表达与对照组相比显著降低(P<0.01),其中2.5 J/cm^2组的MMP3蛋白表达水平和miR-17表达显著低于5.0 J/cm^2组和7.5 J/cm^2组(P<0.05)。miR-17转染HDFs后再用UVA辐射,miR-17转染组的MMP3的蛋白水平显著低于miR对照组(P<0.05),而miR-17抑制物转染组MMP3的蛋白水平与miR对照组相比无显著的改变。结论人皮肤成纤维细胞光老化模型中UVA可导致miR-17表达的下降,从而负性调节MMP3的表达。

Objective To investigate the effects of miR-17 on ultraviolet A（ UVA）-induced skin photoageing. Methods The primary human dermal fibroblasts（ HDFs） from human skin were isolated and cultured. After exposed to different doses of UVA（ 0,2. 5,5. 0,7. 5 J / cm^2）,the mRNA and protein levels of metal matrix proteinase 3（ MMP3） in HDFs were detected by real-time PCR and Western blot respectively,and the level of miR-17 was detected by real-time PCR. MiR-17 mimic,miR mimic control,anti-miR-17 and anti-miR mimic control were respectively transfected into HDFs before UVA exposure,and then the protein level of MMP3 was detected. Results After exposed to UVA,the expression of MMP3 was significantly increased（ P 〈0. 01）,while the level of miR-17 was significantly decreased（ P 〈0. 01）. The expression of MMP3 and the level of miR-17 in 2. 5 J / cm^2 group were significantly lower than in 5. 0 J / cm^2 group and 7. 5 J / cm^2 group（ P〈0. 05）. When HDFs were exposed to UVA after transfected with miR-17 mimic,the protein level of MMP3 in miR-17 mimic group was significantly lower than in miR-control group（ P〈0. 05）,but there was no difference between anti-miR-17 group and its control group. Conclusion UVA could down-regulate the levels of miR-17 and miR-17 thus down-regulating the expression of MMP3 in HDFs photoageing model.