PDGF-BB对UVB诱导的光老化成纤维细胞中ROS的影响
|
摘要
目的:探讨PDGF-BB对紫外线诱导的光老化成纤维细胞中ROS的影响及抑制氧化应激的作用特点。方法:利用CCK-8法检查不同浓度PDGF-BB对成纤维细胞增殖活性的影响,筛选治疗最佳浓度;PDGF-BB预处理成纤维细胞48h后,利用UVB体外重复照射构建成纤维细胞光老化模型;利用β-半乳糖苷酶染色法检测细胞衰老程度;装载活性氧(ROS)检测探针,利用荧光显微镜、流式细胞技术观察和检测各组ROS的表达。结果:PDGF-BB预处理辐照组(UVB+PDGF-BB组)中β-半乳糖苷酶染色细胞的着色比率低于UVB组,差异有统计学意义(P<0.05)。在UVB诱导的应激性老化中,UVB组ROS表达显著高于正常组(P<0.05);PDGF-BB预处理辐照组(UVB+PDGF-BB组)ROS表达显著低于UVB组(P<0.05)。结论:PDGF-BB可降低UVB介导的光老化成纤维细胞中ROS的表达,减少氧化应激反应,可在一定程度上缓解光老化进程。
Objective To explore the effect of PDGF-BB on reactive oxygen species(ROS) in UVB-induced skin fibroblasts and the characteristics of oxidative stress. Methods The effect of PDGF-BB of different dosages on the activity of fibroblasts was detected by CCK-8 assay. After pretreatment of fibroblast 48 h with PDGF-BB, the photoaging model was successfully established with UVB in vitro. The senescence degree of cells was detected by the β-galactosidase staining method. ROS test probe was loaded and oxidative stress of fibroblasts was induced by UVB irradiation in vitro and the expression of ROS was measured by fluorescence microscopy and flow cell technology respectively. Results The coloring percentage of the cells stained by beta galactosidase in the PDGF-BB pretreatment group(UVB+PDGF-BB group) was lower than that in the UVB group, the difference was statistically significant(P<0.05). In UVB induced stress aging, the expression of ROS in UVB group was significantly higher than that in normal group(P<0.05). The expression of ROS in the PDGF-BB pretreatment group(UVB+PDGF-BB group) was significantly lower than that of UVB group(P<0.05). Conclusion PDGF-BB can reduce ROS expression in fibroblasts induced by UVB irradiation and decrease oxidative stress, and it could alleviate photoaging in some extent.
Objective To explore the effect of PDGF-BB on reactive oxygen species(ROS) in UVB-induced skin fibroblasts and the characteristics of oxidative stress. Methods The effect of PDGF-BB of different dosages on the activity of fibroblasts was detected by CCK-8 assay. After pretreatment of fibroblast 48 h with PDGF-BB, the photoaging model was successfully established with UVB in vitro. The senescence degree of cells was detected by the β-galactosidase staining method. ROS test probe was loaded and oxidative stress of fibroblasts was induced by UVB irradiation in vitro and the expression of ROS was measured by fluorescence microscopy and flow cell technology respectively. Results The coloring percentage of the cells stained by beta galactosidase in the PDGF-BB pretreatment group(UVB+PDGF-BB group) was lower than that in the UVB group, the difference was statistically significant(P<0.05). In UVB induced stress aging, the expression of ROS in UVB group was significantly higher than that in normal group(P<0.05). The expression of ROS in the PDGF-BB pretreatment group(UVB+PDGF-BB group) was significantly lower than that of UVB group(P<0.05). Conclusion PDGF-BB can reduce ROS expression in fibroblasts induced by UVB irradiation and decrease oxidative stress, and it could alleviate photoaging in some extent.
引文
[1]俞卓伟,保志军,阮清伟,等.维持抗氧化、抗炎功能稳定与自身稳态和保健延寿[J].老年医学与保健,2015,21(1):1-4.
[2]李春雨,张丽宏,张宁,等.紫外线诱导皮肤光老化的形成机制[J].中国美容医学,2009,18(3):416-419.
[3]Pandel R,Poljsak B,Godic A,et al.Skin photoaging and the role of antioxidants in its prevention[J].ISRN Dermatol,2013,2013:930164.
[4]Zouboulis CC,Makrantonaki E.Clinical aspects and molecular diagnostics of skin aging[J].Clin Dermatol,2011,29(1):3-14.
[5]Fredriksson L,Li H,Eriksson U.The PDGF family:four gene products form five dimeric isoforms[J].Cytokine Growth Factor Rev,2004,15(4):197-204.
[6]Cabezas R,Vega-Vela NE,González-Sanmiguel J,et al.PDGF-BB Preserves Mitochondrial Morphology,Attenuates ROS Production,and Upregulates Neuroglobin in an Astrocytic Model Under Rotenone Insult[J].Mol Neurobiol,2018,55(4):3085-3095.
[7]Zeng JP,Bi B,Chen L,et al.Repeated exposure of mouse dermal fibroblasts at a sub-cytotoxic dose of UVB leads to premature senescence:A robust model of cellular photoaging[J].J Dermatol Sci,2014,73(1):49-56.
[8]陈亮,毕波,曾继平,等.p H对衰老相关β-半乳糖苷酶染色的影响[J].中国美容整形外科杂志,2014,25(12):751-754.
[9]Li L,Tan J,Miao Y,et al.ROS and Autophagy:Interactions and Molecular Regulatory Mechanisms[J].Cell Mol Neurobiol,2015,35(5):615-621.
[10]崔剑,李兆陇,洪啸吟.自由基生物抗氧化与疾病[J].清华大学学报(自然科学版),2000,40(6):9-12.
[11]Pryor WA,Houk KN,Foote CS,et al.Free radical biology and medicine:it's a gas,man![J].Am J Physiol Regul Integr Comp Physiol,2006,291(3):R491-511.
[12]Balaban RS,Nemoto S,Finkel T.Mitochondria,Oxidants,and Aging[J].Cell,2005,120(4):483-495.
[13]Suzuki N,Mittler R.Reactive oxygen species-dependent wound responses in animals and plants[J].Free Radic Biol Med,2012,53(12):2269-2276.
[14]Baxter PS,Hardingham GE.Adaptive regulation of the brain’s antioxidant defences by neurons and astrocytes[J].Free Radic Biol Med,2016,100:147-152.
[15]Tobin DJ.Introduction to skin aging[J].J Tissue Viability,2017,26(1):37-46.
[16]Digiovanni CW,Lin S,Pinzur M.Recombinant human PDGF-BB in foot and ankle fusion[J].Expert Rev Med Devices,2012,9(2):111-122.
[2]李春雨,张丽宏,张宁,等.紫外线诱导皮肤光老化的形成机制[J].中国美容医学,2009,18(3):416-419.
[3]Pandel R,Poljsak B,Godic A,et al.Skin photoaging and the role of antioxidants in its prevention[J].ISRN Dermatol,2013,2013:930164.
[4]Zouboulis CC,Makrantonaki E.Clinical aspects and molecular diagnostics of skin aging[J].Clin Dermatol,2011,29(1):3-14.
[5]Fredriksson L,Li H,Eriksson U.The PDGF family:four gene products form five dimeric isoforms[J].Cytokine Growth Factor Rev,2004,15(4):197-204.
[6]Cabezas R,Vega-Vela NE,González-Sanmiguel J,et al.PDGF-BB Preserves Mitochondrial Morphology,Attenuates ROS Production,and Upregulates Neuroglobin in an Astrocytic Model Under Rotenone Insult[J].Mol Neurobiol,2018,55(4):3085-3095.
[7]Zeng JP,Bi B,Chen L,et al.Repeated exposure of mouse dermal fibroblasts at a sub-cytotoxic dose of UVB leads to premature senescence:A robust model of cellular photoaging[J].J Dermatol Sci,2014,73(1):49-56.
[8]陈亮,毕波,曾继平,等.p H对衰老相关β-半乳糖苷酶染色的影响[J].中国美容整形外科杂志,2014,25(12):751-754.
[9]Li L,Tan J,Miao Y,et al.ROS and Autophagy:Interactions and Molecular Regulatory Mechanisms[J].Cell Mol Neurobiol,2015,35(5):615-621.
[10]崔剑,李兆陇,洪啸吟.自由基生物抗氧化与疾病[J].清华大学学报(自然科学版),2000,40(6):9-12.
[11]Pryor WA,Houk KN,Foote CS,et al.Free radical biology and medicine:it's a gas,man![J].Am J Physiol Regul Integr Comp Physiol,2006,291(3):R491-511.
[12]Balaban RS,Nemoto S,Finkel T.Mitochondria,Oxidants,and Aging[J].Cell,2005,120(4):483-495.
[13]Suzuki N,Mittler R.Reactive oxygen species-dependent wound responses in animals and plants[J].Free Radic Biol Med,2012,53(12):2269-2276.
[14]Baxter PS,Hardingham GE.Adaptive regulation of the brain’s antioxidant defences by neurons and astrocytes[J].Free Radic Biol Med,2016,100:147-152.
[15]Tobin DJ.Introduction to skin aging[J].J Tissue Viability,2017,26(1):37-46.
[16]Digiovanni CW,Lin S,Pinzur M.Recombinant human PDGF-BB in foot and ankle fusion[J].Expert Rev Med Devices,2012,9(2):111-122.