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山东大学学报（医学版） ›› 2015, Vol. 53 ›› Issue (5): 41-45.doi: 10.6040/j.issn.1671-7554.0.2015.152

• 基础医学 • 上一篇    下一篇

Sirt3对人脐静脉内皮细胞衰老的影响

刘辉, 陈桐帅, 李娜, 王舒健, 李静媛, 卜培莉   

  1. 山东大学齐鲁医院心内科, 山东 济南 250012
  • 收稿日期:2015-02-05 修回日期:2015-03-19 出版日期:2015-05-10 发布日期:2015-05-10
  • 通讯作者: 卜培莉。E-mail:peili_bu@163.com E-mail:peili_bu@163.com
  • 基金资助:
    国家自然科学基金(81170135)

Role of Sirt3 on the senescence of human umbilical vein endothelial cells

LIU Hui, CHEN Tongshuai, LI Na, WANG Shujian, LI Jingyuan, BU Peili   

  1. Department of Cardiology, Qilu Hospital of Shandong University, Jinan 250012, Shandong, China
  • Received:2015-02-05 Revised:2015-03-19 Online:2015-05-10 Published:2015-05-10

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摘要： 目的 探讨沉默信息调节因子相关酶3(Sirt3)对人脐静脉内皮细胞(HUVECs)衰老的影响。方法 体外培养HUVECs,合成Sirt3-siRNA序列,Western blotting法筛选出抑制效率最高的序列。将实验分为对照组、Sirt3-siRNA组,Western blotting法检测Sirt3和细胞衰老相关蛋白P16和P21的表达水平,β-半乳糖苷酶染色法检测内皮细胞衰老状态,2',7'-二氯二氢荧光素二乙酯(DCFH-DA)法测定血管内皮细胞活性氧(ROS)的释放。结果 与对照组比较,Sirt3-siRNA组β-gal阳性染色率明显增多,Sirt3的蛋白表达量减少,P16和P21的蛋白表达量升高(P<0.05),ROS释放明显增多。结论 Sirt3抑制HUVECs衰老,其作用机制可能是通过调节ROS的释放而实现的。

关键词: 细胞衰老, 沉默信息调节因子相关酶3, P16蛋白, 活性氧, 人脐静脉内皮细胞, P21蛋白

Abstract: Objective To investigate the role of Sirtuin3(Sirt3) on the senescence of human umbilical vein endothelial cells (HUVECs). Methods HUVECs were cultured in vitro. Sirt3-siRNA was synthesized by reagent company. Sirt3-siRNA with the best inhibition efficiency was screened out with Western blotting assay. HUVECs were divided into negative control group and Sirt3-siRNA group. Western blotting assay was used to evaluate the protein expressions of Sirt3, P16 and P21. Senescence β-gal staining was used to identify the aging status of HUVECs. The intracellular ROS was detected with 2',7'-dichlorodi-hydro-fluorescein diacetate (DCFH-DA). Results Compared with the negative control group, the Sirt3-siRNA group showed decreased protein expression of Sirt3 (P<0.05), increased positive rate of β-gal staining, elevated P16 and P21 expressions (P<0.05), and higher level of intracellular ROS. Conclusion Sirt3 can delay the senescence of HUVECs by regulating ROS generation.

Key words: P21 protein, Sirtuin3, Cell senescence, Human umbilical vein endothelial cells, P16 protein, Reactive oxygen species

中图分类号: 

  • R543.1+2
[1] Paneni F, Costantino S, Cosentino F. Molecular pathways of arterial aging[J]. Clin Sci (Lond), 2015, 128(2): 69-79.
[2] Wang M, Jiang L, Monticone RE, et al. Proinflammation: the key to arterial aging[J]. Trends Endocrinol Metab, 2014, 25(2): 72-79.
[3] Oeseburg H, Iusuf D, van der Harst P, et al. Bradykinin protects against oxidative stress-induced endothelial cell senescence[J]. Hypertension, 2009, 53(2): 417-422.
[4] 时小燕, 杜丽敏. Sirtuin家族成员及其生物学特性[J]. 国际药学研究杂志, 2011,38(5): 349-355. SHI Xiaoyan, DU Limin. Sirtuin family and its biological functions[J]. International Journal of Pharmaceutical Research, 2011, 38(5): 349-355.
[5] Bause AS, Haigis MC. SIRT3 regulation of mitochondrial oxidative stress[J]. Exp Gerontol, 2013, 48(7): 634-639.
[6] Sack MN. The role of SIRT3 in mitochondrial homeostasis and cardiac adaptation to hypertrophy and aging[J]. J Mol Cell Cardiol, 2012, 52(3): 520-525.
[7] Tao R, Vassilopoulos A, Parisiadou L, et al. Regulation of MnSOD enzymatic activity by Sirt3 connects the mitochondrial acetylome signaling networks to aging and carcinogenesis[J]. Antioxid Redox Signal, 2014, 20(10): 1646-1654.
[8] 郝翀, 卫立辛. Sirtuin家族在肝细胞癌中的表达及临床意义[J]. 中华肝胆外科杂志, 2013, 19(10): 789-793. HAO Chong, WEI Lixin. Expression of sirtuin in hepatocellular carcinoma and its clinical significance[J]. Chinese Journal of Hepatobiliary Surgery, 2013, 19(10): 789-793.
[9] Stepanova L, Sorrentino BP. A limited role for p16Ink4a and p19Arf in the loss of hematopoietic stem cells during proliferative stress[J]. Blood, 2005, 106(3): 827-832.
[10] Piotrowska A, Bartnik E. The role of reactive oxygen species and mitochondria in aging[J]. Postepy Biochem, 2014, 60(2): 240-247.
[11] Sundaresan NR, Gupta M, Kim G, et al. Sirt3 blocks the cardiac hypertrophic response by augmenting Foxo3a-dependent antioxidant defense mechanisms in mice[J]. J Clin Invest, 2009, 119(9): 2758-2771.
[12] Chen Y, Zhang J, Lin Y, et al. Tumour suppressor SIRT3 deacetylates and activates manganese superoxide dismutase to scavenge ROS[J]. EMBO Rep, 2011, 12(6): 534-541.
[13] Hirschey MD, Shimazu T, Goetzman E, et al. SIRT3 regulates mitochondrial fatty-acid oxidation by reversible enzyme deacetylation[J]. Nature, 2010, 464(7285): 121-125.
[14] Finley LW, Haas W, Desquiret-Dumas V, et al. Succinate dehydrogenase is a direct target of sirtuin 3 deacetylase activity[J]. PLoS One, 2011, 6(8): e23295. doi: 10.1371/journal.pone.0023295. Epub 2011 Aug 17.
[15] Schlicker C, Gertz M, Papatheodorou P, et al. Substrates and regulation mechanisms for the human mitochondrial sirtuins Sirt3 and Sirt5[J]. J Mol Biol. 2008, 382(3): 790-801.
[16] Someya S, Yu W, Hallows WC, et al. Sirt3 mediates reduction of oxidative damage and prevention of age-related hearing loss under caloric restriction[J]. Cell, 2010, 143(5): 802-812.
[17] Song Z, Liu Y, Hao B, et al. Ginsenoside Rb1 prevents H2O2-induced HUVEC senescence by stimulating sirtuin-1 pathway[J]. PLoS One, 2014, 9(11): e112699. doi: 10.1371/journal.pone.0112699. eCollection 2014.
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