Role of SIRT3 in Angiotensin II-induced human umbilical vein endothelial cells dysfunction

BMC Cardiovascular Disorders

Peili Bu¹  Shujian Wang²  Na Li²  Tongshuai Chen²  Hui Liu² 

[1] Department of Cardiology, Qilu Hospital, Shandong University, No. 107 Wen Hua Xi Road, Jinan 250012, Shandong Province, China;The Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Public Health, Qilu Hospital, Shandong University, Jinan 250012, Shandong Province, China

关键词: HUVECs;    Reactive oxygen species;    AngII;    Endothelial dysfunction;    SIRT3;   

Others  :  1221378
DOI  :  10.1186/s12872-015-0075-4

 received in 2014-12-04, accepted in 2015-07-24,  发布年份 2015

【 摘 要 】

Background

SIRT3, a member of the sirtuin family of NAD ⁺ -dependent deacetylases, resides primarily in the mitochondria and has been shown to deacetylate several metabolic and respiratory enzymes that regulate important mitochondrial functions. Previous researches show an important role of SIRT3 in regulating the production of reactive oxygen species (ROS), and highlight the ability of SIRT3 to protect cells from oxidative damage. A key substance of renin-angiotensin-aldosterone system (RAAS), Angiotensin II (AngII) can induce cells dysfunction by increasing the production of ROS. In this paper, we focus on the role of SIRT3 in AngII-induced human umbilical vein endothelial cells (HUVECs) dysfunction.

Methods

To study the influence of AngII on SIRT3 expression, HUVECs were treated with AngII of 10 ⁻⁷ , 10 ⁻⁶ , 10 ⁻⁵  mol/L for 24 h. SIRT3 expression was detected by wester-blotting analysis and RT-PCR. In addition, to research the role of SIRT3 in AngII-induced HUVECs,we used SIRT3 siRNA to knock down SIRT3 expression in HUVECs. Cells pretreated with negative control siRNA or SIRT3 siRNA were exposed to AngII for 24 h, and endothelial nitric oxide synthase (eNOS) expression, eNOS activity, total level of nitric oxide (NO) and ROS generation of each group were detected.

Results

Here we show that AngII treatment could increase generation of ROS, and decrease eNOS activity and total level of NO, while upregulated eNOS expression as a compensatory mechanism. The stimulation of AngII upregulated the expression of SIRT3 in HUVECs. SIRT3 siRNA worsen the AngII-induced effects above, besides, downregulated eNOS protein expression.

Conclusion

These data suggest that SIRT3 plays a role of protection in AngII-induced HUVECs dysfunction via regulation of ROS generation.

【 授权许可】

   
2015 Liu et al.

【 图 表 】

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【 参考文献 】

• [1]Schachinger V, Britten MB, Zeiher AM: Prognostic impact of coronary vasodilator dysfunction on adverse long-term outcome of coronary heart disease. Circulation 2000, 101(16):1899-906.
• [2]Brunner H, Cockcroft JR, Deanfield J, Donald A, Ferrannini E, Halcox J, Kiowski W, Luscher TF, Mancia G, Natali A, et al.: Endothelial function and dysfunction. Part II: Association with cardiovascular risk factors and diseases. A statement by the Working Group on Endothelins and Endothelial Factors of the European Society of Hypertension. J Hypertens 2005, 23(2):233-46.
• [3]Christensen HM, Schou M, Goetze JP, Faber J, Frystyk J, Flyvbjerg A, Kistorp C: Body mass index in chronic heart failure: association with biomarkers of neurohormonal activation, inflammation and endothelial dysfunction. BMC Cardiovasc Disord 2013, 13:80. BioMed Central Full Text
• [4]John S, Schmieder RE: Impaired endothelial function in arterial hypertension and hypercholesterolemia: potential mechanisms and differences. J Hypertens 2000, 18(4):363-74.
• [5]Rossi R, Chiurlia E, Nuzzo A, Cioni E, Origliani G, Modena MG: Flow-mediated vasodilation and the risk of developing hypertension in healthy postmenopausal women. J Am Coll Cardiol 2004, 44(8):1636-40.
• [6]Cai H, Harrison DG: Endothelial dysfunction in cardiovascular diseases: the role of oxidant stress. Circ Res 2000, 87(10):840-4.
• [7]Fyhrquist F, Metsarinne K, Tikkanen I: Role of angiotensin II in blood pressure regulation and in the pathophysiology of cardiovascular disorders. J Hum Hypertens 1995, 9(Suppl 5):S19-24.
• [8]Touyz RM: Reactive oxygen species and angiotensin II signaling in vascular cells -- implications in cardiovascular disease. Braz J Med Biol Res 2004, 37(8):1263-73.
• [9]Briones AM, Touyz RM: Oxidative stress and hypertension: current concepts. Curr Hypertens Rep 2010, 12(2):135-42.
• [10]Hamanaka RB, Chandel NS: Mitochondrial reactive oxygen species regulate cellular signaling and dictate biological outcomes. Trends Biochem Sci 2010, 35(9):505-13.
• [11]Balaban RS, Nemoto S, Finkel T: Mitochondria, oxidants, and aging. Cell 2005, 120(4):483-95.
• [12]Elnakish MT, Hassanain HH, Janssen PM, Angelos MG, Khan M: Emerging role of oxidative stress in metabolic syndrome and cardiovascular diseases: important role of Rac/NADPH oxidase. J Pathol 2013, 231(3):290-300.
• [13]Bellizzi D, Rose G, Cavalcante P, Covello G, Dato S, De Rango F, Greco V, Maggiolini M, Feraco E, Mari V, et al.: A novel VNTR enhancer within the SIRT3 gene, a human homologue of SIR2, is associated with survival at oldest ages. Genomics 2005, 85(2):258-63.
• [14]Rose G, Dato S, Altomare K, Bellizzi D, Garasto S, Greco V, Passarino G, Feraco E, Mari V, Barbi C, et al.: Variability of the SIRT3 gene, human silent information regulator Sir2 homologue, and survivorship in the elderly. Exp Gerontol 2003, 38(10):1065-70.
• [15]Onyango P, Celic I, McCaffery JM, Boeke JD, Feinberg AP: SIRT3, a human SIR2 homologue, is an NAD-dependent deacetylase localized to mitochondria. Proc Natl Acad Sci U S A 2002, 99(21):13653-8.
• [16]Ahn BH, Kim HS, Song S, Lee IH, Liu J, Vassilopoulos A, Deng CX, Finkel T: A role for the mitochondrial deacetylase Sirt3 in regulating energy homeostasis. Proc Natl Acad Sci U S A 2008, 105(38):14447-52.
• [17]Bell EL, Emerling BM, Ricoult SJ, Guarente L: SirT3 suppresses hypoxia inducible factor 1alpha and tumor growth by inhibiting mitochondrial ROS production. Oncogene 2011, 30(26):2986-96.
• [18]Jacobs KM, Pennington JD, Bisht KS, Aykin-Burns N, Kim HS, Mishra M, Sun L, Nguyen P, Ahn BH, Leclerc J, et al.: SIRT3 interacts with the daf-16 homolog FOXO3a in the mitochondria, as well as increases FOXO3a dependent gene expression. Int J Biol Sci 2008, 4(5):291-9.
• [19]Qiu X, Brown K, Hirschey MD, Verdin E, Chen D: Calorie restriction reduces oxidative stress by SIRT3-mediated SOD2 activation. Cell Metab 2010, 12(6):662-7.
• [20]Sundaresan NR, Gupta M, Kim G, Rajamohan SB, Isbatan A, Gupta MP: Sirt3 blocks the cardiac hypertrophic response by augmenting Foxo3a-dependent antioxidant defense mechanisms in mice. J Clin Invest 2009, 119(9):2758-71.
• [21]Someya S, Yu W, Hallows WC, Xu J, Vann JM, Leeuwenburgh C, Tanokura M, Denu JM, Prolla TA: Sirt3 mediates reduction of oxidative damage and prevention of age-related hearing loss under caloric restriction. Cell 2010, 143(5):802-12.
• [22]Tao R, Coleman MC, Pennington JD, Ozden O, Park SH, Jiang H, Kim HS, Flynn CR, Hill S, Hayes McDonald W, et al.: Sirt3-mediated deacetylation of evolutionarily conserved lysine 122 regulates MnSOD activity in response to stress. Mol Cell 2010, 40(6):893-904.
• [23]Redon J, Oliva MR, Tormos C, Giner V, Chaves J, Iradi A, Saez GT: Antioxidant activities and oxidative stress byproducts in human hypertension. Hypertension 2003, 41(5):1096-101.
• [24]Touyz RM: Reactive oxygen species, vascular oxidative stress, and redox signaling in hypertension: what is the clinical significance? Hypertension 2004, 44(3):248-52.
• [25]Landmesser U, Cai H, Dikalov S, McCann L, Hwang J, Jo H, Holland SM, Harrison DG: Role of p47(phox) in vascular oxidative stress and hypertension caused by angiotensin II. Hypertension 2002, 40(4):511-5.
• [26]Landmesser U, Dikalov S, Price SR, McCann L, Fukai T, Holland SM, Mitch WE, Harrison DG: Oxidation of tetrahydrobiopterin leads to uncoupling of endothelial cell nitric oxide synthase in hypertension. J Clin Invest 2003, 111(8):1201-9.
• [27]Santana HA, Moreira SR, Neto WB, Silva CB, Sales MM, Oliveira VN, Asano RY, Espindola FS, Nobrega OT, Campbell CS, et al.: The higher exercise intensity and the presence of allele I of ACE gene elicit a higher post-exercise blood pressure reduction and nitric oxide release in elderly women: an experimental study. BMC Cardiovasc Disord 2011, 11:71. BioMed Central Full Text
• [28]Brown K, Xie S, Qiu X, Mohrin M, Shin J, Liu Y, Zhang D, Scadden DT, Chen D: SIRT3 reverses aging-associated degeneration. Cell Rep 2013, 3(2):319-27.
• [29]Yang H, Yang T, Baur JA, Perez E, Matsui T, Carmona JJ, Lamming DW, Souza-Pinto NC, Bohr VA, Rosenzweig A, et al.: Nutrient-sensitive mitochondrial NAD+ levels dictate cell survival. Cell 2007, 130(6):1095-107.
• [30]Karamanlidis G, Lee CF, Garcia-Menendez L, Kolwicz SC Jr, Suthammarak W, Gong G, Sedensky MM, Morgan PG, Wang W, Tian R: Mitochondrial complex I deficiency increases protein acetylation and accelerates heart failure. Cell Metab 2013, 18(2):239-50.
• [31]Lin SJ, Ford E, Haigis M, Liszt G, Guarente L: Calorie restriction extends yeast life span by lowering the level of NADH. Genes Dev 2004, 18(1):12-6.
• [32]Weir HJ, Murray TK, Kehoe PG, Love S, Verdin EM, O’Neill MJ, Lane JD, Balthasar N: CNS SIRT3 expression is altered by reactive oxygen species and in Alzheimer’s disease. PLoS One 2012, 7(11):e48225.