Resveratrol ameliorates low shear stress‑induced oxidative stress by suppressing ERK/eNOS‑Thr495 in endothelial cells

Wang,Zhimei; Zhang,Junxia; Li,Bing; Gao,Xiaofei; Liu,Yanrong; Mao,Wenxing; Chen,Shao‑Liang

doi:10.3892/mmr.2014.2390

Resveratrol ameliorates low shear stress‑induced oxidative stress by suppressing ERK/eNOS‑Thr495 in endothelial cells

Authors:
- Zhimei Wang
- Junxia Zhang
- Bing Li
- Xiaofei Gao
- Yanrong Liu
- Wenxing Mao
- Shao‑Liang Chen
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Affiliations: Department of Cardiology, Nanjing First Hospital, Nanjing Medical University, Nanjing, Jiangsu 210006, P.R. China
Published online on: July 17, 2014 https://doi.org/10.3892/mmr.2014.2390
Pages: 1964-1972

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Abstract

Fluid shear stress has been revealed to differentially regulate endothelial nitric oxide synthase (eNOS) distribution in vessels. eNOS, a key enzyme in controlling nitric oxide (NO) release, has a crucial role in mediating oxidative stress, and resveratrol (RSV)‑mediated eNOS also attenuates oxidative damage and suppresses endothelial dysfunction. To observe the protective effect of RSV on low shear stress (LSS)‑induced oxidative damage and the potential mechanisms involved, a parallel‑plate flow chamber, which imposed a low level of stress of 2 dynes/cm2 to cells, was employed. Reactive oxygen species (ROS), NO and apoptotic cells were examined in LSS‑treated endothelial cells (ECs) with or without RSV. Western blot analysis was used to examine LSS‑regulated eNOS‑Ser1177, Thr495 and Ser633, which were tightly associated with NO release. To further determine the underlying signaling pathways involved, extracellular signal‑regulated kinase (ERK), a possible upstream target of eNOS‑Thr495, was investigated, followed by examination of eNOS‑Thr495 in ERK‑inhibited cells. Additionally, eNOS mRNA expression levels were analyzed in cells challenged with LSS. The results revealed that RSV markedly decreased LSS‑induced oxidative damage in ECs. Furthermore, eNOS‑Ser1177 and Thr495 as well as phospho‑ERK were time‑dependently activated by LSS. The ERK inhibitor deactivated eNOS‑Thr495, which was accompanied by increased intracellular superoxide dismutase (SOD) levels. Of note, the activation effect of LSS on ERK/eNOS was markedly eliminated by RSV. In conclusion, RSV exerts antioxidant effects by suppressing LSS‑activated ERK/eNOS and may provide a potential therapeutic target for atherosclerosis.

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1	Caro CG, Fitz-Gerald JM and Schroter RC: Arterial wall shear and distribution of early atheroma in man. Nature. 223:1159–1160. 1969. View Article : Google Scholar : PubMed/NCBI
2	VanderLaan PA, Reardon CA and Getz GS: Site specificity of atherosclerosis: site-selective responses to atherosclerotic modulators. Arterioscler Thromb Vasc Biol. 24:12–22. 2004. View Article : Google Scholar : PubMed/NCBI
3	Qin X, Tian J, Zhang P, Fan Y, Chen L, Guan Y, Fu Y, Zhu Y, Chien S and Wang N: Laminar shear stress up-regulates the expression of stearoyl-CoA desaturase-1 in vascular endothelial cells. Cardiovasc Res. 74:506–514. 2007. View Article : Google Scholar : PubMed/NCBI
4	Cheng C, Tempel D, Oostlander A, Helderman F, Gijsen F, Wentzel J, van Haperen R, Haitsma DB, Serruys PW, van der Steen AF, de Crom R and Krams R: Rapamycin modulates the eNOS vs. shear stress relationship. Cardiovasc Res. 78:123–129. 2008. View Article : Google Scholar : PubMed/NCBI
5	Cheng C, van Haperen R, de Waard M, van Damme LC, Tempel D, Hanemaaijer L, van Cappellen GW, Bos J, Slager CJ, Duncker DJ, van der Steen AF, de Crom R and Krams R: Shear stress affects the intracellular distribution of eNOS: direct demonstration by a novel in vivo technique. Blood. 106:3691–3698. 2005. View Article : Google Scholar : PubMed/NCBI
6	Dimmeler S, Fleming I, Fisslthaler B, Hermann C, Busse R and Zeiher AM: Activation of nitric oxide synthase in endothelial cells by Akt-dependent phosphorylation. Nature. 399:601–605. 1999. View Article : Google Scholar
7	Sessa WC: eNOS at a glance. J Cell Sci. 117:2427–2429. 2004. View Article : Google Scholar : PubMed/NCBI
8	Le Brocq M, Leslie SJ, Milliken P and Megson IL: Endothelial dysfunction: from molecular mechanisms to measurement, clinical implications, and therapeutic opportunities. Antioxid Redox Signal. 10:1631–1674. 2008.PubMed/NCBI
9	Atkins GB and Simon DI: Interplay between NF-kappaB and Kruppel-like factors in vascular inflammation and atherosclerosis: location, location, location. J Am Heart Assoc. 2:e0002902013. View Article : Google Scholar : PubMed/NCBI
10	Mount PF, Kemp BE and Power DA: Regulation of endothelial and myocardial NO synthesis by multi-site eNOS phosphorylation. J Mol Cell Cardiol. 42:271–279. 2007. View Article : Google Scholar : PubMed/NCBI
11	Cancel LM and Tarbell JM: The role of mitosis in LDL transport through cultured endothelial cell monolayers. Am J Physiol Heart Circ Physiol. 300:H769–H776. 2011. View Article : Google Scholar : PubMed/NCBI
12	Yu W, Ying H, Tong F, Zhang C, Quan Y and Zhang Y: Protective effect of the silkworm protein 30Kc6 on human vascular endothelial cells damaged by oxidized low density lipoprotein (Ox-LDL). PLoS One. 8:e687462013. View Article : Google Scholar : PubMed/NCBI
13	Guo H, Chen Y, Liao L and Wu W: Resveratrol protects HUVECs from oxidized-LDL induced oxidative damage by autophagy upregulation via the AMPK/SIRT1 pathway. Cardiovasc Drugs Ther. 27:189–198. 2013. View Article : Google Scholar : PubMed/NCBI
14	Malinowska J, Oleszek W, Stochmal A and Olas B: The polyphenol-rich extracts from black chokeberry and grape seeds impair changes in the platelet adhesion and aggregation induced by a model of hyperhomocysteinemia. Eur J Nutr. 52:1049–1057. 2013. View Article : Google Scholar
15	Takizawa Y, Kosuge Y, Awaji H, Tamura E, Takai A, Yanai T, Yamamoto R, Kokame K, Miyata T, Nakata R and Inoue H: Up-regulation of endothelial nitric oxide synthase (eNOS), silent mating type information regulation 2 homologue 1 (SIRT1) and autophagy-related genes by repeated treatments with resveratrol in human umbilical vein endothelial cells. Br J Nutr. 110:2150–2155. 2013. View Article : Google Scholar
16	Quincozes-Santos A, Bobermin LD, Latini A, Wajner M, Souza DO, Goncalves CA and Gottfried C: Resveratrol protects C6 astrocyte cell line against hydrogen peroxide-induced oxidative stress through heme oxygenase 1. PLoS One. 8:e643722013. View Article : Google Scholar : PubMed/NCBI
17	Price NL, Gomes AP, Ling AJ, Duarte FV, Martin-Montalvo A, North BJ, Agarwal B, Ye L, Ramadori G, Teodoro JS, Hubbard BP, Varela AT, Davis JG, Varamini B, Hafner A, Moaddel R, Rolo AP, Coppari R, Palmeira CM, de Cabo R, Baur JA and Sinclair DA: SIRT1 is required for AMPK activation and the beneficial effects of resveratrol on mitochondrial function. Cell Metab. 15:675–690. 2012. View Article : Google Scholar : PubMed/NCBI
18	Rajapakse AG, Yepuri G, Carvas JM, Stein S, Matter CM, Scerri I, Ruffieux J, Montani JP, Ming XF and Yang Z: Hyperactive S6K1 mediates oxidative stress and endothelial dysfunction in aging: inhibition by resveratrol. PLoS One. 6:e192372011. View Article : Google Scholar : PubMed/NCBI
19	Kao CL, Chen LK, Chang YL, Yung MC, Hsu CC, Chen YC, Lo WL, Chen SJ, Ku HH and Hwang SJ: Resveratrol protects human endothelium from H(2)O(2)-induced oxidative stress and senescence via SirT1 activation. J Atheroscler Thromb. 17:970–979. 2010. View Article : Google Scholar : PubMed/NCBI
20	Arunachalam G, Yao H, Sundar IK, Caito S and Rahman I: SIRT1 regulates oxidant- and cigarette smoke-induced eNOS acetylation in endothelial cells: role of resveratrol. Biochem Biophys Res Commun. 393:66–72. 2010. View Article : Google Scholar : PubMed/NCBI
21	Becatti M, Taddei N, Cecchi C, Nassi N, Nassi PA and Fiorillo C: SIRT1 modulates MAPK pathways in ischemic-reperfused cardiomyocytes. Cell Mol Life Sci. 69:2245–2260. 2012. View Article : Google Scholar : PubMed/NCBI
22	Elíes J, Cuíñas A, García-Morales V, Orallo F and Campos-Toimil M: Trans-resveratrol simultaneously increases cytoplasmic Ca(2+) levels and nitric oxide release in human endothelial cells. Mol Nutr Food Res. 8:1237–1248. 2011.
23	Min Z, Kang L, Lin L, Jinghua F, Junna S and Baolin L: Resveratrol restores lysophosphatidylcholine-induced loss of endothelium-dependent relaxation in rat aorta tissue coinciding with inhibition of extracellular-signal-regulated protein kinase activation. Phytother Res. 12:1762–1768. 2010. View Article : Google Scholar
24	Qiu X, Brown K, Hirschey MD, Verdin E and Chen D: Calorie restriction reduces oxidative stress by SIRT3-mediated SOD2 activation. Cell Metab. 12:662–667. 2010. View Article : Google Scholar : PubMed/NCBI
25	Ford RJ and Rush JW: Endothelium-dependent vasorelaxation to the AMPK activator AICAR is enhanced in aorta from hypertensive rats and is NO and EDCF dependent. Am J Physiol Heart Circ Physiol. 300:H64–H75. 2011. View Article : Google Scholar : PubMed/NCBI
26	Conklin BS, Vito RP and Chen C: Effect of low shear stress on permeability and occludin expression in porcine artery endothelial cells. World J Surg. 31:733–743. 2007. View Article : Google Scholar : PubMed/NCBI
27	Furchgott RF and Zawadzki JV: The obligatory role of endothelial cells in the relaxation of arterial smooth muscle by acetylcholine. Nature. 288:373–376. 1980. View Article : Google Scholar : PubMed/NCBI
28	Palmer RM, Ferrige AG and Moncada S: Nitric oxide release accounts for the biological activity of endothelium-derived relaxing factor. Nature. 327:524–526. 1987. View Article : Google Scholar : PubMed/NCBI
29	Greif DM, Kou R and Michel T: Site-specific dephosphorylation of endothelial nitric oxide synthase by protein phosphatase 2A: evidence for crosstalk between phosphorylation sites. Biochemistry. 41:15845–15853. 2002. View Article : Google Scholar
30	Huang A, Yang YM, Yan C, Kaley G, Hintze TH and Sun D: Altered MAPK signaling in progressive deterioration of endothelial function in diabetic mice. Diabetes. 61:3181–3188. 2012. View Article : Google Scholar : PubMed/NCBI
31	Carrizzo A, Puca A, Damato A, Marino M, Franco E, Pompeo F, Traficante A, Civitillo F, Santini L, Trimarco V and Vecchione C: Resveratrol improves vascular function in patients with hypertension and dyslipidemia by modulating NO metabolism. Hypertension. 62:359–366. 2013. View Article : Google Scholar
32	Schmitt CA, Heiss EH and Dirsch VM: Effect of resveratrol on endothelial cell function: molecular mechanisms. Biofactors. 36:342–349. 2010. View Article : Google Scholar : PubMed/NCBI
33	Chan CM, Chang HH, Wang VC, Huang CL and Hung CF: Inhibitory effects of resveratrol on PDGF-BB-induced retinal pigment epithelial cell migration via PDGFRβ, PI3K/Akt and MAPK pathways. PLoS One. 8:e568192013.PubMed/NCBI
34	Skrobuk P, von Kraemer S, Semenova MM, Zitting A and Koistinen HA: Acute exposure to resveratrol inhibits AMPK activity in human skeletal muscle cells. Diabetologia. 55:3051–3060. 2012. View Article : Google Scholar : PubMed/NCBI
35	Boo YC, Hwang J, Sykes M, Michell BJ, Kemp BE, Lum H and Jo H: Shear stress stimulates phosphorylation of eNOS at Ser(635) by a protein kinase A-dependent mechanism. Am J Physiol Heart Circ Physiol. 283:H1819–H1828. 2002.PubMed/NCBI
36	Boo YC, Sorescu G, Boyd N, Shiojima I, Walsh K, Du J and Jo H: Shear stress stimulates phosphorylation of endothelial nitric-oxide synthase at Ser1179 by Akt-independent mechanisms: role of protein kinase A. J Biol Chem. 277:3388–3396. 2002. View Article : Google Scholar : PubMed/NCBI
37	Wang W, Ha CH, Jhun BS, Wong C, Jain MK and Jin ZG: Fluid shear stress stimulates phosphorylation-dependent nuclear export of HDAC5 and mediates expression of KLF2 and eNOS. Blood. 115:2971–2979. 2010. View Article : Google Scholar : PubMed/NCBI