吡咯喹啉醌对UVA诱导人皮肤成纤维细胞衰老的保护作用及机制研究
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南京医科大学科技发展基金重点项目(2011NJMU242)


Protective effect and mechanism of pyrroloquinoline quinine(PQQ) against UVA induced human skin fibroblasts aging
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    摘要:

    目的:研究吡咯喹啉醌(pyrroloquinoline quinine,PQQ)对UVA诱导人皮肤成纤维细胞衰老的保护作用及其机制。方法:体外使用6孔板培养人皮肤成纤维细胞(human skin fibroblasts,HSF),予以9 J/cm2照射,照射前后实验组加入80 ng/ml的8-甲氧基补骨脂(8-methoxypsoralan,8-MOP)和浓度为50-100-200 ng/ml的PQQ。UVA照射72 h后,对细胞进行衰老β-半乳糖苷酶染色-JC-1染色荧光显微镜检测线粒体膜电位变化-JC-1染色流式细胞仪检测线粒体膜去极化状况-real-time PCR检测衰老相关基因mmp1-mmp3的表达。结果:低剂量PQQ(50 ng/ml)实验组β-半乳糖苷酶染色呈阴性,JC-1染色后低剂量PQQ(50 ng/ml)实验组细胞线粒体膜电位改变的趋势有回转,JC-1染色后流式细胞仪检测PQQ对UVA诱导的衰老细胞线粒体去极化有保护作用,而且低剂量PQQ(50 ng/ml)实验组衰老相关基因mmp1-mmp3的表达也有明显降低。结论:吡咯喹啉醌对由UVA诱导的人皮肤成纤维细胞衰老有保护作用,而且这一作用可能是通过线粒体途径发挥作用的。

    Abstract:

    Objective:To study the protective effect and mechanism of pyrroloquinoline quinine(PQQ) against UVA induced human skin fibroblasts. Methods:Human skin fibroblasts were cultured in 6-well plate in vitro with an equal number of cells(104~106 per well). HSF exposed to UV irradiation at a dose of 9 J/cm2. Pyrroloquinoline quinine(50,100,200 ng/ml) and 8-methoxypsoralan (80 ng/ml) were added into the cell culture medium before and after the irradiation. 8-MOP was used to enhance UVA absorption. After 72 hours of irradiation, SA-β-Gal staining was performed to evaluate the senescence state,change of mitochondria membrane potential was examined by JC-1 staining using fluorescence microscope,JC-1 staining was also used to test mitochondria membrane depolarization by flow cytometry(FCM),and real-time PCR was used to determine mRNA expression of senescence-associated gene of mmp1,mmp3. Results:Cells cultured with lower dosage PQQ (50 ng/ml) exhibited negative SA-β-Gal staining. JC-1 staining showed the change of mitochondria membrane potential was reversed by lower dosage PQQ (50 ng/ml). Mitochondria membrane depolarization was relieved lower dosage PQQ(50 ng/ml). Meanwhile,the gene expression of senescence-associated signals of mmp1,mmp3 decreased in PQQ group compared with control group. Conclusion:Pyrroloquinoline quinine can protect human skin fibroblasts from aging induced by UVA irradiation via mitochondria pathway.

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张春丽,邹 阳,林金德,温传俊,沈 干.吡咯喹啉醌对UVA诱导人皮肤成纤维细胞衰老的保护作用及机制研究[J].南京医科大学学报(自然科学版),2013,(3):337-341

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  • 收稿日期:2012-11-13
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  • 在线发布日期: 2013-06-04
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