骨髓间充质干细胞来源的外泌体miR-29a-3p减轻UV照射诱导的皮肤成纤维细胞光老化  

Bone Marrow Mesenchymal Stem Cell-derived Exosome miR-29a-3p Alleviates UV Irradiation-induced Photoaging in Skin Fibroblast

作　　者：严婷婷 颜韵灵 陶旌晶 黄莉宁 钟益萍 王晓华 YAN Tingting;YAN Yunling;TAO Jingjing;HUANG Lining;ZHONG Yiping;WANG Xiaohua(Department of Medical Cosmetology,Dermatology Hospital of Southern Medical University,Guangzhou 510091,China)

机构地区：[1]南方医科大学皮肤病医院医学科,广东广州510091

出　　处：《皮肤科学通报》2023年第5期564-573,共10页Dermatology Bulletin

摘　　要：目的间充质干细胞衍生的外泌体(msc-exo)被发现可以减少光老化。本研究的目的是探讨来自骨髓msc-exo(BMSCs-exo)的microRN A(miR)-29a-3p在光老化中的潜在作用。方法从骨髓间质干细胞中分离外泌体并进行western blot验证。UVB照射人真皮成纤维细胞(HDFs)构建体外光老化细胞模型。采用实时荧光定量PCR(RT-qPCR)检测miR-29a-3p、Ⅰ型胶原蛋白和基质金属蛋白酶(MMPs)mRNA水平。CCK-8、Transwe ll和流式细胞术检测细胞活力、迁移和凋亡。用于测量氧化应激指标水平的商用试剂盒。最后,采用双荧光素酶报告基因检测来验证miR-29a-3p的靶标。结果提取的外泌体HSP70和CD9阳性。HDFs的存活率以外泌体浓度依赖的方式增加。与对照组相比,UVB照射抑制miR-29a-3p水平,但BMSCs-exo治疗恢复miR-29a-3p水平(P<0.05)。此外,BMSCs-exo-miR-29a-3p逆转了UVB照射对HDFs迁移和氧化应激的抑制,以及促进细胞凋亡。然而,这种逆转通过抑制miR-29a-3p而减弱(P<0.05)。此外,BMSCs-exo-miR-29a-3p也抑制了Ⅰ型胶原的降解和光老化过程中MMPs的产生,并且它们也被减少的miR-29a-3p所消除。最后,MMP-2是miR-29a-3p的靶基因。结论我们的研究提出了BMSCs-exo-miR-29a-3p在改善皮肤光老化功能中的新作用,这些发现可能为皮肤光老化的靶向治疗提供新的见解。Objective Mesenchymal stem cells-derived exosome(MSCs-exo)was identified to reduce photoaging.The purpose of this study was to investigate the potential role of microRNA(miR)-29a-3p derived from bone marrow MSCs-exo(BMSCs-exo)in photoaging.Methods Exosomes were isolated from BMSCs and verified by western blot.Construction of an in vitro photoaging cell model by UVB irradiation of human dermal fibroblasts(HDFs).Quantitative real-time PCR(RT-qPCR)was performed to detect the mRNA levels of miR-29a-3p,collagen type I,and matrix metalloproteinases(MMPs).CCK-8,Transwell,and flow cytometry were applicated to examine cell viability,migration,and apoptosis.Commercial kits to measure levels of oxidative stress indicators.Finally,a dual-luciferase reporter assay was applied to validate the target of miR-29a-3p.Results Extracted exosomes were positive for HSP70 and CD9.Survival of HDFs increased in an exosome concentration-dependent manner.UVB irradiation inhibited miR-29a-3p levels compared to controls,but BMSCs-exo treatment restored miR-29a-3p levels(P<0.05).Additionally,BMSCs-exo-miR-29a-3p reversed the inhibition of HDFs migration and oxidative stress by UVB irradiation,as well as the promotion of apoptosis.However,this reversal was attenuated by the suppression of miR-29a-3p(P<0.05).Furthermore,BMSCs-exo-miR-29a-3p also inhibited the degradation of collagen type I and the production of MMPs in photoaging,and they were also eliminated by the reduced miR-29a-3p.Finally,MMP-2 was the target gene of miR-29a-3p.Conclusion Our study presented a novel role for BMSCs-exo-miR-29a-3p in improving skin photoaging function,and these findings may provide new insights into the targeted treatment of skin photoaging.

关 键 词：光老化 BMSC-exo miR-29a-3p UVB照射 

分 类 号：R751[医药卫生—皮肤病学与性病学]