hucMSC-exo对UVB诱导的光老化成纤维细胞的保护作用

Protective Effect of hucMSC-exo on UVB-Induced Photoaging in Human Dermal Fibroblasts

目的探讨人脐带间充质干细胞来源外泌体(exosomes derived from human umbilical cord mesenchymal stem cells,hucMSC-exo)对中波紫外线(UVB)诱导的光老化人真皮成纤维细胞(human dermal fibroblasts,HDF)的影响。方法使用超高速离心法从hucMSC条件培养基中分离hucMSC-exo。体外培养HDF,用UVB照射建立细胞光老化模型。实验分为正常对照组、模型组和实验组。使用hucMSC-exo于UVB干预前24 h对HDF进行预处理。培养48 h后,使用流式细胞仪检测细胞周期,使用CCK-8法检测细胞增殖活力,使用微丝绿色荧光探针标记细胞骨架;培养72 h后,使用β-半乳糖苷酶染色检测细胞衰老水平,分别使用实时荧光PCR和Western blot检测基质金属蛋白酶-1(matrix metalloproteinases,MMP-1)、Ⅰ型胶原蛋白(typeⅠcollagen,COL-Ⅰ)和白介素1(interleukin-1,IL-1)的相对表达水平。结果hucMSC-exo可以提高光老化HDF增殖活力,调控细胞周期,缓解细胞衰老的改变。hucMSC-exo可以下调MMP-1 mRNA和蛋白的表达并上调COL-Ⅰ的mRNA和蛋白的表达,实验组同模型组及对照组相比,差异均具有统计学意义(P<0.05)。结论hucMSC-exo预处理对皮肤光老化HDF具有保护作用。这可能与hucMSC-exo参与调控MMPs与COL-Ⅰ的表达相关。

Objective To investigate the effect of exosomes derived from human umbilical cord mesenchymal stem cells(hucMSC-exo)on medium-wave ultraviolet(UVB)-induced photoaging in human dermal fibroblasts(HDF).Methods The hucMSC-exo was isolated from hucMSC conditioned medium by ultra-high-speed centrifugation.HDF were cultured in vitro and UVB irradiation was used to establish a cellular photoaging model.The experiment was divided into normal control group,model group and experimental group.HDF were pretreated with hucMSC-exo at 24 h before UVB intervention.At 48 h after culture,the cell cycle was detected by flow cytometry,the cell proliferation was detected by CCK-8 assay,and the cytoskeleton was labeled with microfilament green fluorescent probe.At 72 h after culture,the cell senescence level was detected byβ-galactosidase staining,and the relative expression levels of matrix metalloproteinases-1(MMP-1),typeⅠcollagen(COL-Ⅰ)and interleukin-1(IL-1)were detected by real-time PCR and Western blot,respectively.Results HucMSC-exo could improve the proliferation activity of photoaging HDF,regulate cell cycle and alleviate the changes of cell senescence.HucMSC-exo could down-regulate the expression of MMP-1 mRNA and protein and up-regulate the expression of COL-ⅠmRNA and protein.Compared with the model group and the control group,the differences in the experimental group were significant(P<0.05).Conclusion The hucMSC-exo pretreatment can protect skin photoaging HDF.This may be related to the involvement of hucMSC-exo in regulating the expression of MMPs and COL-Ⅰ.