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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.“,”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.