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  

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

作　　者：张春丽[1] 邹阳[2] 林金德[1] 温传俊[2] 沈干[3] 

机构地区：[1]南京医科大学附属友谊整形外科医院,江苏南京210029 [2]南京师范大学生命科学学院细胞所,江苏南京210029 [3]南京医科大学第二附属医院整形科,江苏南京210003

出　　处：《南京医科大学学报（自然科学版）》2013年第3期337-341,共5页Journal of Nanjing Medical University(Natural Sciences)

基　　金：南京医科大学科技发展基金重点项目(2011NJMU242)

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。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-[3-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 mmpl ,romp3. 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 mmpl, 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.

关 键 词：吡咯喹啉醌 UVA照射 细胞衰老 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学]