白藜芦醇对D-半乳糖诱导的WI-38衰老细胞的保护作用
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摘要
目的探讨白藜芦醇对D-半乳糖诱导的WI-38衰老细胞的保护作用以及对MORF4基因表达的影响。方法D-半乳糖诱导WI-38细胞构建衰老模型;不同浓度白藜芦醇(25μmol/L,50μmol/L,100μmol/L,200μmol/L)对衰老模型作用24h,并与对照组比较:MTT法检测细胞增殖活力,SA-β-半乳糖苷酶染色法检测细胞衰老程度,比色法检测SOD活性;反转录半定量PCR检测MORF4基因的表达。结果与衰老模型组相比,白藜芦醇组(25μmol/L、50μmol/L、100μmol/L、200μmol/L)细胞存活率升高(F=81.95,P<0.05),SA-β-半乳糖苷酶染色率下降(P<0.05),白藜芦醇各组总SOD活力均高于衰老模型组(P<0.05)及对照组(P<0.05),以100μmol/L白藜芦醇浓度最明显。100μmol/L白藜芦醇能明显上调MORF4基因的表达(P<0.05)。结论白藜芦醇对衰老WI-38细胞有保护作用,以100μmol/L白藜芦醇的保护作用最强,可以明显上调衰老细胞MORF4基因的表达。
Objective To explore the Resveratrol protective effect on WI-38 senescence cells induced by D-galactose and MORF4 gene express. Methods D-galactose was used to induce WI-38 cells senescence model,the model cell was treated with different Resveratrol concentrations( 25μmol/L,50μmol/L,100μmol/L,200μmol/L) for 24 h to observe the protective effect compared with the control group. The viability of WI-38 cell was detected by MTT. Senescence-Associated-β-galactosidase staining was used to show the degree of cell ageing and the activity of superoxide dismutase( SOD) in cells was measured by colourimetric method. Reverse transcription semi-quantitative PCR was used to detect the mRNA expression of MORF4 gene. Data for each experimental group were analyzed with SPSS16. 0 by ANOVA. Results Compared with the ageing model group,the survival rate of Resveratrol group( 25μmol/L,50μmol/L,100μmol/L,200μmol/L) increased( F = 81. 95,P< 0. 05),the number of SA-β-galactosidase staining was decreased( P < 0. 05). The level of SOD activity in Resveratrol protection group was significantly increased,compared with the ageing group( P < 0. 05) and normal group( P < 0. 05),especially the 100μmol/L concentration,respectively. Compared with the ageing group,the expression of MORF4 was significantly enhanced in Resveratrol 100μmol/L protection group( P < 0. 05). Conclusion Resveratrol protects senescent WI-38 cells with the strongest protective effect at 100 μmol/L,which can significantly up-regulate the expression of MORF4 gene in senescent cells.
Objective To explore the Resveratrol protective effect on WI-38 senescence cells induced by D-galactose and MORF4 gene express. Methods D-galactose was used to induce WI-38 cells senescence model,the model cell was treated with different Resveratrol concentrations( 25μmol/L,50μmol/L,100μmol/L,200μmol/L) for 24 h to observe the protective effect compared with the control group. The viability of WI-38 cell was detected by MTT. Senescence-Associated-β-galactosidase staining was used to show the degree of cell ageing and the activity of superoxide dismutase( SOD) in cells was measured by colourimetric method. Reverse transcription semi-quantitative PCR was used to detect the mRNA expression of MORF4 gene. Data for each experimental group were analyzed with SPSS16. 0 by ANOVA. Results Compared with the ageing model group,the survival rate of Resveratrol group( 25μmol/L,50μmol/L,100μmol/L,200μmol/L) increased( F = 81. 95,P< 0. 05),the number of SA-β-galactosidase staining was decreased( P < 0. 05). The level of SOD activity in Resveratrol protection group was significantly increased,compared with the ageing group( P < 0. 05) and normal group( P < 0. 05),especially the 100μmol/L concentration,respectively. Compared with the ageing group,the expression of MORF4 was significantly enhanced in Resveratrol 100μmol/L protection group( P < 0. 05). Conclusion Resveratrol protects senescent WI-38 cells with the strongest protective effect at 100 μmol/L,which can significantly up-regulate the expression of MORF4 gene in senescent cells.
引文
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[12]Chen M,Tominaga K,Pereira-Smith OM.Emerging role of the MORF/MRG gene family in various biological processes,including aging[J].Annals of the New York Academy of Sciences,2010,1197(11):134-141.
[13]Zou C,Li J,Xiong S,et al.Mortality factor 4 like 1 protein mediates epithelial cell death in a mouse model of pneumonia[J].Science Translational Medicine,2015,7(311):171-183.
[14]Garcia SN,Pereira-Smith O.MRGing chromatin dynamics and cellular senescence[J].Cell Biochemistry and Biophysics,2008,50(3):133-141.
[15]Tominaga K,Tominaga E,Ausserlechner MJ,et al.The cell senescence inducing gene product MORF4 is regulated by degradation via the ubiquitin/proteasome pathway[J].Experimental Cell Research,2010,316(1):92-102.
[2]Kavas GO,Ayral PA,Elhan AH.The effects of resveratrol on oxidant/antioxidant systems and their cofactors in rats[J].Advances in clinical and experimental medicine:official organ Wroclaw Medical University,2013,22(2):151-155.
[3]Kasiotis KM,Pratsinis H,Kletsas D,et al.Resveratrol and related stilbenes:their anti-aging and anti-angiogenic properties[J].Food and chemical toxicology:an international journal published for the British Industrial Biological Research Association,2013,61(3):112-120.
[4]肖林霞,卢其能,李润根.白藜芦醇的研究进展[J].现代农业科技,2015(24):264-265.
[5]李博,宁光,张志国.白藜芦醇在抗衰老领域的研究进展[J].国际内分泌代谢杂志,2013,33(3):189-191,198.
[6]Pena AN,Pereira-Smith OM.The role of the MORF/MRG family of genes in cell growth,differentiation,DNA repair,and thereby aging[J].Annals of the New York Academy of Sciences,2007,1100(8):299-305.
[7]葛亮,王桦,曾尔亢,等.衰老相关基因研究进展[J].中国老年学杂志,2014(22):6529-6532.
[8]Kilic Eren M,Kilincli A,Eren?.Resveratrol induced premature senescence is associated with DNA damage mediated SIRT1 and SIRT2 Down-Regulation[J].PLOS One,2015,10(4):e0124837.
[9]蒲秀瑛,于双,樊文博,等.归芪多糖对早衰WI-38细胞端粒酶活性的影响[J].中国中医药信息杂志,2015,22(11):50-53.
[10]Sohal RS,Mockett RJ,Orr WC.Mechanisms of aging:an appraisal of the oxidative stress hypothesis[J].Free radical biology&medicine,2002,33(5):575-586.
[11]张翠利,付丽娜,杨小云,等.活性氧自由基与细胞衰老关系的研究进展[J].广州化工,2015(19):5-7.
[12]Chen M,Tominaga K,Pereira-Smith OM.Emerging role of the MORF/MRG gene family in various biological processes,including aging[J].Annals of the New York Academy of Sciences,2010,1197(11):134-141.
[13]Zou C,Li J,Xiong S,et al.Mortality factor 4 like 1 protein mediates epithelial cell death in a mouse model of pneumonia[J].Science Translational Medicine,2015,7(311):171-183.
[14]Garcia SN,Pereira-Smith O.MRGing chromatin dynamics and cellular senescence[J].Cell Biochemistry and Biophysics,2008,50(3):133-141.
[15]Tominaga K,Tominaga E,Ausserlechner MJ,et al.The cell senescence inducing gene product MORF4 is regulated by degradation via the ubiquitin/proteasome pathway[J].Experimental Cell Research,2010,316(1):92-102.