[1]田黎明,彭　圆,胡荣毅,等.衰老标记蛋白30对过氧化氢所致人皮肤成纤维细胞衰老表型的影响[J].新乡医学院学报,2016,33(6):452-456.[doi:10.7683/xxyxyxb.2016.06.002]
　TIAN Li-ming,PENG Yuan,HU Rong-yi,et al.Effect of senescence marker protein 30 on senescent phenotypes in hydrogen peroxide-induced senescence human skin fibroblasts[J].Journal of Xinxiang Medical University,2016,33(6):452-456.[doi:10.7683/xxyxyxb.2016.06.002]
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衰老标记蛋白30对过氧化氢所致人皮肤成纤维细胞衰老表型的影响
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
33
期数:
2016年6
页码:
452-456
栏目:
基础研究
出版日期:
2016-06-05

文章信息/Info

Title:
Effect of senescence marker protein 30 on senescent phenotypes in hydrogen peroxide-induced senescence human skin fibroblasts
作者:
田黎明1; 彭　圆2; 胡荣毅1; 张　平1; 姜红浩1; 许　斌1; 田庆均1; 张　翀2; 王　平2
（1.湖北中医药大学中西医结合医院　武汉市中西医结合医院皮肤科，湖北　武汉　430022；2.湖北中医药大学基础医学院，湖北　武汉　430065）
Author(s):
TIAN Li-ming1; PENG Yuan2; HU Rong-yi1; ZHANG Ping1; JIANG Hong-hao1; XU Bin1; TIAN Qing-jun1; ZHANG Zhong2; WANG Ping2
（1.Department of Dermatology，Integrative Medicine Hospital of Wuhan City,Integrative Medicine Hospital of Hubei University of Chinese Medicine，Wuhan 430022，Hubei Province，China；2.College of Basic Medicine，Hubei University of Chinese Medicine，Wuhan 430065，Hubei Province，China）
关键词:
衰老标记蛋白30; 过氧化氢; 成纤维细胞; 氧化应激; 细胞衰老
Keywords:
senescence marker protein 30; hydrogen peroxide; fibroblasts; oxidative stress; cell aging
分类号:
R334.5
DOI:
10.7683/xxyxyxb.2016.06.002
文献标志码:
A
摘要:
目的　观察高表达的衰老标记蛋白30（SMP30）对过氧化氢（H2O2）所致的正常人皮肤成纤维细胞（NHSF）衰老表型的影响。方法　实验分为转染H2O2处理组（NHSF+SMP30+H2O2组）、转空载体H2O2处理组（NHSF+空载体+H2O2组）和转空载体组（NHSF+空载体组）。NHSF细胞分别转染人SMP30 cDNA 或空载体 pcDNA3.1 （分别命名为NHSF+SMP30、NHSF+空载体），然后 150 μmol·L-1 H2O2处理2 h。应用反转录-聚合酶链反应（RT-PCR）和蛋白印迹法分析 SMP30 的表达，显微镜观察细胞形态，并检测与衰老相关的 β-半乳糖苷酶 （SA-β-gal）、细胞活性氧 （ROS） 及超氧化物歧化酶 （SOD） 的活性。结果　人SMP30 cDNA 明显上调了NHSF中 SMP30 的表达。NHSF+空载体+H2O2组 SMP30 mRNA和蛋白表达水平明显低于NHSF+空载体组（P<0.05）；NHSF+SMP30+H2O2组SMP30 mRNA和蛋白表达水平高于NHSF+空载体+H2O2组（P<0.05）。NHSF+空载体+H2O2组较NHSF+空载体组SA-β-gal染色率明显增高（P<0.05）；NHSF+SMP30+H2O2组较NHSF+空载体+H2O2组SA-β-gal染色率明显下降（P<0.05）。NHSF+空载体+H2O2组较NHSF+空载体组ROS活性明显增高（P<0.05），而SOD水平明显降低（P<0.05）；NHSF+SMP30+H2O2组较NHSF+空载体+H2O2组ROS活性明显下降，而SOD水平明显上升（P<0.05）。结论　高表达的SMP30能够抑制细胞SA-β-gal和ROS的表达，同时上调SOD的活性。
Abstract:
Objective　To observe the effect of high expression of senescence marker protein 30 （SMP30） on senescent phenotypes in normal human skin fibroblasts （NHSF） induced by hydrogen peroxide （H2O2）.Methods　Cultured NHSF were classified into three groups.NHSF+SMP30+H2O2 group was transfected with a recombinant plasmid human SMP30 cDNA and treated with H2O2 of 150 μmol·L-1 for two hours;NHSF+empty vector+H2O2 group was transfected with the empty vector pcDNA3.1 and treated by H2O2 of 150 μmol·L-1 for two hours;and NHSF+empty vector group was transfected with the empty vector pcDNA3.1 and receiving no treatment.Reverse transcription-reverse transcription-polymerase chain reaction （RT-PCR） and Western blot were performed to quantify the mRNA and protein expressions of SMP30 in these cells，microscopy was used to observe the morphological changes of cells.The activity of senescence-associated β-galactosidase （SA-β-gal） and superoxide dismutase （SOD） as well as the level of reactive oxygen species （ROS） were detected by using commercial kits.Results　The expression of SMP30 was significantly upregulated in NHSF transfected with the recombinant plasmid human SMP30 cDNA.Both the mRNA and protein expression levels of SMP30 in NHSF+empty vector+H2O2 group were significantly lower in the NHSF+empty vector group（P<0.05），while the mRNA and protein expression levels of SMP30 in NHSF+SMP30+H2O2 group were higher than that in NHSF+empty vector+H2O2 group（P<0.05）.The percentage of SA-β-gal-positive cells in NHSF+empty vector+H2O2 group was higher than that in NHSF+empty vector group（P<0.05），while the percentage of SA-β-gal-positive cells in NHSF+SMP30+H2O2 group was lower than that in NHSF+empty vector+H2O2 group（P<0.05）.The ROS activity significantly increased，but SOD level significantly decreased in the NHSF+empty vector+H2O2 group compared with the NHSF group （P<0.05），while the ROS activity significantly decreased，but SOD level significantly increased in the NHSF+SMP30+H2O2 group compared with the NHSF+empty vector+H2O2 group （P<0.05）.Conclusion　The overexpression of SMP30 can downregulate the activity of SA-β-gal and ROS levels，but enhance the activity of SOD.

参考文献/References:

［1］　ARUN P，ALETI V，PARIKH K，et al.Senescence marker protein 30 （SMP30） expression in eukaryotic cells：existence of multiple species and membrane localization［J］.PLoS One，2011，6（2）：e16545.
［2］　MARUYAMA N，ISHIGAMI A，KONDO Y.Molecular abnormality in aging：its contribution to clinical pathology［J］.Rinsho Byori，2005，53（8）：728-734.
［3］　TIAN L M，XIE H F，XIAO X，et al.Study on the roles of β-catenin in hydrogen peroxide-induced senescence in human skin fibroblasts［J］.Exp Dermatol，2011，20（10）：836-838.
［4］　田黎明，谢红付，李吉，等.氧化应激所致的人皮肤成纤维细胞衰老中β-连环蛋白的表达研究［J］.中华皮肤科杂志，2011，44（4）：36-40.
［5］　田黎明，谢红付，李吉，等.β联蛋白对体外过氧化氢所致人皮肤成纤维细胞衰老表型影响［J］.中华皮肤科杂志，2013，46（7）：485-488.
［6］　TOBISAWA M，TSURUSAKI Y，YAMAGUCHI M.Decrease in regucalcin level and enhancement of protein tyrosine phosphatase activity in rat brain microsomes with increasing age［J］.Int J Mol Med，2003，12（4）：577-580.
［7］　KAGAMI Y，KONDO Y，HANDA S，et al.Senescence marker protein-30/gluconolactonase expression in the mouse ovary during gestation［J］.Biol Pharm Bull，2013，36（12）：2005-2008.
［8］　SATO Y，AMANO A，KISHIMOTO Y，et al.Ascorbic acid prevents protein oxidation in livers of senescence marker protein-30/gluconolactonase knockout mice［J］.Geriatr Gerontol Int，2014，14（4）：989-995.
［9］　MIZUKAMI H，SAITOH S，MACHII H，et al.Senescence marker protein-30 （SMP30） deficiency impairs myocardium-induced dilation of coronary arterioles associated with reactive oxygen species［J］.Int J Mol Sci，2013，14（5）：9408-9423.
［10］　LEE J H，YOO J H，OH S H，et al.Knockdown of moesin expression accelerates cellular senescence of human dermal microvascular endothelial cells［J］.Yonsei Med J，2010，51（3）：438-447.
［11］　TREIBER N，MAITY P，SINGH K，et al.The role of manganese superoxide dismutase in skin aging［J］.Dermatoendocrinol，2012，4（3）：232-235.
［12］　BOTTAI G，MANCINA R，MURATORI M，et al.17β-estradiol protects human skin fibroblasts and keratinocytes against oxidative damage［J］.J Eur Acad Dermatol Venereol，2013，27（10）：1236-1243.
［13］　POON H F，CALABRESE V，SCAPAGNINI G，et al.Free radicals and brain aging［J］.Clin Geriatr Med，2004，20（2）：329-359.
［14］　任翔，傅廷亮，马明明，等.黄芩苷对肠上皮细胞缺氧复氧损伤后屏障功能的影响［J］.中华实用儿科临床杂志，2015，30（7）：494-497.
［15］　MULLER F L，LUSTGARTEN M S，JANG Y，et al.Trends in oxidative aging theories［J］.Free Radic Biol Med，2007，43（4）：477-503.
［16］　WONDRAK G T，JACOBSON M K，JACOBSON E L.Endogenous UVA-photosensitizers：mediators of skin photodamage and novel targets for skin photoprotection［J］.Photochem Photobiol Sci，2006，5（2）：215-237.

更新日期/Last Update: 2016-06-05