PDGF—BB对UVB诱导的光老化成纤维细胞中ROS的影响(1)
[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法检查不同濃度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。
[关键词]光老化；PDGF-BB；成纤维细胞；ROS；UVB
, http://www.100md.com
[中图分类号]R758.1 [文献标志码]A [文章编号]1008-6455(2018)05-0109-03
Effect of PDGF-BB on Reactive Oxygen Species in UVB-induced Skin Fibroblasts
REN Run-jian1，FANG Lei1，JIA Chuan-long2，ZHAO Hu1，CHEN Liang2，QIN Deng-ke2，ZHOU Hui-min1，BI Bo1
(1.Department of Clinical Laboratory Medicine；2.Department of Plastic and Aesthetic Surgery，Huadong Hospital Affiliated to Fudan University，Shanghai 200040，China)
, 百拇医药
Abstract： 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 48h 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.
, http://www.100md.com
Key words： photoaging； platelet derived growth factor BB(PDGF-BB)； fibroblasts； reactive oxygen species(ROS)； ultraviolet radiation B(UVB)
皮肤光老化作为皮肤老化的重要组成部分，严重影响人们的外观和身体健康[1-2]。在其发生发展进程中，紫外线，尤其是波长范围290～320nm的中波紫外线(Ultraviolet radiation B， UVB)成为主要诱因[3]。UVB引起的皮肤光老化主要表现为弹性缺失，皱纹加深，色素沉着，甚至诱发黑色素瘤等[4]。在这一复杂病理生理过程中，成纤维细胞(Fibroblasts， FBs)受到损伤，出现一系列衰老表型，如细胞肥大铺展，β-半乳糖苷酶染色阳性，细胞周期阻滞等。有研究认为UVB诱导活性氧(Reactive oxygen species， ROS)的异常生成，光子能量转化为化学能量所造成的氧化应激是导致光老化的主要机制。血小板衍生因子(Platelet derived growth factor， PDGF)是由两条多肽链组成的二聚体，相对分子质量为28～35KD，主要由成熟的血小板分泌，存在于多种组织中如血小板、骨骼肌细胞、内皮细胞、成骨细胞，是一种促有丝分裂刺激源[5]。同时，PDGF有很强的趋化作用，对骨骼肌细胞、未分化间充质细胞、单核细胞等都有很强的趋化功能。近来有研究发现在星形胶质细胞中，PDGF-BB可抑制自由基活性氧(ROS)产生，保护线粒体超微结构对抗氧化应激，从而发挥其保护作用[6]。然而，PDGF-BB能否改善UVB造成的氧化应激，从而延缓光老化进程尚不清楚。本研究应用此前在体外建立的成纤维细胞光老化模型(Stress-induced premature senescence， SIPS)，予以外源性PDGF-BB处理，观察其对SIPS-FBs的生物学作用。, 百拇医药(任润健 方磊 贾传龙 赵虎 陈亮 秦登科 周慧敏 毕波)
[关键词]光老化；PDGF-BB；成纤维细胞；ROS；UVB
, http://www.100md.com
[中图分类号]R758.1 [文献标志码]A [文章编号]1008-6455(2018)05-0109-03
Effect of PDGF-BB on Reactive Oxygen Species in UVB-induced Skin Fibroblasts
REN Run-jian1，FANG Lei1，JIA Chuan-long2，ZHAO Hu1，CHEN Liang2，QIN Deng-ke2，ZHOU Hui-min1，BI Bo1
(1.Department of Clinical Laboratory Medicine；2.Department of Plastic and Aesthetic Surgery，Huadong Hospital Affiliated to Fudan University，Shanghai 200040，China)
, 百拇医药
Abstract： 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 48h 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.
, http://www.100md.com
Key words： photoaging； platelet derived growth factor BB(PDGF-BB)； fibroblasts； reactive oxygen species(ROS)； ultraviolet radiation B(UVB)
皮肤光老化作为皮肤老化的重要组成部分，严重影响人们的外观和身体健康[1-2]。在其发生发展进程中，紫外线，尤其是波长范围290～320nm的中波紫外线(Ultraviolet radiation B， UVB)成为主要诱因[3]。UVB引起的皮肤光老化主要表现为弹性缺失，皱纹加深，色素沉着，甚至诱发黑色素瘤等[4]。在这一复杂病理生理过程中，成纤维细胞(Fibroblasts， FBs)受到损伤，出现一系列衰老表型，如细胞肥大铺展，β-半乳糖苷酶染色阳性，细胞周期阻滞等。有研究认为UVB诱导活性氧(Reactive oxygen species， ROS)的异常生成，光子能量转化为化学能量所造成的氧化应激是导致光老化的主要机制。血小板衍生因子(Platelet derived growth factor， PDGF)是由两条多肽链组成的二聚体，相对分子质量为28～35KD，主要由成熟的血小板分泌，存在于多种组织中如血小板、骨骼肌细胞、内皮细胞、成骨细胞，是一种促有丝分裂刺激源[5]。同时，PDGF有很强的趋化作用，对骨骼肌细胞、未分化间充质细胞、单核细胞等都有很强的趋化功能。近来有研究发现在星形胶质细胞中，PDGF-BB可抑制自由基活性氧(ROS)产生，保护线粒体超微结构对抗氧化应激，从而发挥其保护作用[6]。然而，PDGF-BB能否改善UVB造成的氧化应激，从而延缓光老化进程尚不清楚。本研究应用此前在体外建立的成纤维细胞光老化模型(Stress-induced premature senescence， SIPS)，予以外源性PDGF-BB处理，观察其对SIPS-FBs的生物学作用。, 百拇医药(任润健 方磊 贾传龙 赵虎 陈亮 秦登科 周慧敏 毕波)