Introduction: This study aimed to investigate whether epigallocatechin-3-gallate (EGCG) shows antioxidant activity against angiotensin II (Ang II)-induced human umbilical vein endothelial cell (HUVEC) apoptosis. Materials and Methods: The viability of HUVECs was revealed by MTT and LDH assay. The cell apoptosis was detected by FITC-PI assay. A fluorescent probe assay was used to measure the reactive oxygen species (ROS) generation in HUVECs. Mitochondrial permeability transition pore (MPTP) opening, mitochondrial membrane potential, and caspase-3, -4, -8, -9 activities were also measured. Results: We found that Ang II treatment increased the generation of ROS, enhanced MPTP opening and cytochrome c release, activated caspase-3/9, and consequently induced HUVEC apoptosis. EGCG treatment-suppressed Ang II induces the oxidative stress of HUVECs and mitochondria-related cell apoptosis. We also showed that the antioxidant activity pathway, including cytochrome c release, MPTP opening, and caspase-3/9 activation, is a key endogenous defensive system in HUVECs, provoking Ang II exposure. Our study revealed that increased expression of Nrf2 by EGCG could partially repress Ang II-induced injury effects. Conclusions: All of our findings indicated that EGCG treatment provides a protective effect for Ang II-induced HUVEC apoptosis by decreasing oxidative stress and ameliorating mitochondrial injury.

Ren H, Mu J, Ma J, et al: Selenium inhibits homocysteine-induced endothelial dysfunction and apoptosis via activation of AKT. Cell Physiol Biochem 2016;38:871-882.
Wang F, Pu C, Zhou P, et al: Cinnamaldehyde prevents endothelial dysfunction induced by high glucose by activating Nrf2. Cell Physiol Biochem 2015;36:315-324.
Chen F, Li X, Aquadro E, et al: Inhibition of histone deacetylase reduces transcription of NADPH oxidases and ROS production and ameliorates pulmonary arterial hypertension. Free Radic Biol Med 2016;99:167-178. 2016/10/23.
Kruger R, Mothae M, Smith W: Yia 03-07 reactive oxygen species adversely relates to early vascular changes and arterial stiffness in black normotensive smokers: the African-Predict Study. J Hypertens 2016;34(suppl 1):e205-e206.
Gonzalez GE, Rhaleb NE, D'Ambrosio MA, et al: Cardiac-deleterious role of galectin-3 in chronic angiotensin II-induced hypertension. Am J Physiol Heart Circ Physiol 2016;311:H1287-H1296.
Rosenbaugh EG, Savalia KK, Manickam DS, et al: Antioxidant-based therapies for angiotensin II-associated cardiovascular diseases. Am J Physiol Regul Integr Comp Physiol 2013;304:R917-R928.
Montezano AC, Burger D, Paravicini TM, et al: Nicotinamide adenine dinucleotide phosphate reduced oxidase 5 (Nox5) regulation by angiotensin II and endothelin-1 is mediated via calcium/calmodulin-dependent, rac-1-independent pathways in human endothelial cells. Circ Res 2010;106:1363-1373.
El Bekay R, Alvarez M, Monteseirin J, et al: Oxidative stress is a critical mediator of the angiotensin II signal in human neutrophils: involvement of mitogen-activated protein kinase, calcineurin, and the transcription factor NF-κB. Blood 2003;102:662-671.
Miura S, Saku K: Recent progress in the treatment of cardiovascular disease using olmesartan. Clin Exp Hypertens 2014;36:441-446.
Burrell L: Sy 12-1 renin angiotensin pathway beyond ace and angiotensin II receptors: how it relates to the pathophysiology of hypertension. J Hypertens 2016;34(suppl 1):e367.
Feng W, Zhang K, Liu Y, et al: Apocynin attenuates angiotensin II-induced vascular smooth muscle cells osteogenic switching via suppressing extracellular signal-regulated kinase 1/2. Oncotarget 2016;7:83588-83600.
Shi L, Wu L, Chen Z, et al: MiR-141 activates Nrf2-dependent antioxidant pathway via down-regulating the expression of Keap1 conferring the resistance of hepatocellular carcinoma cells to 5-fluorouracil. Cell Physiol Biochem 2015;35:2333-2348.
Liu W, Wang B, Wang T, et al: Ursodeoxycholic acid attenuates acute aortic dissection formation in angiotensin II-infused apolipoprotein E-deficient mice associated with reduced ROS and increased Nrf2 levels. Cell Physiol Biochem 2016;38:1391-1405.
Gong W, Li J, Chen Z, et al: Polydatin promotes Nrf2-ARE anti-oxidative pathway through activating CKIP-1 to resist HG-induced up-regulation of FN and ICAM-1 in GMCs and diabetic mice kidneys. Free Radic Biol Med 2017;106:393-405.
Yagishita Y, Uruno A, Fukutomi T, et al: Nrf2 improves leptin and insulin resistance provoked by hypothalamic oxidative stress. Cell Rep 2017;18:2030-2044.
Peng S, Hou Y, Yao J, et al: Activation of Nrf2-driven antioxidant enzymes by cardamonin confers neuroprotection of PC12 cells against oxidative damage. Food Funct 2017;8:997-1007.
Rasheed NO, Ahmed LA, Abdallah DM, et al: Nephro-toxic effects of intraperitoneally injected EGCG in diabetic mice: involvement of oxidative stress, inflammation and apoptosis. Sci Rep 2017;7:40617.
Zhang J, Lei Z, Huang Z, et al: Epigallocatechin-3-gallate (EGCG) suppresses melanoma cell growth and metastasis by targeting TRAF6 activity. Oncotarget 2016;7:79557-79571.
Oyama JI, Shiraki A, Nishikido T, et al: EGCG, a green tea catechin, attenuates the progression of heart failure induced by the heart/muscle-specific deletion of MnSOD in mice. J Cardiol 2017;69:417-427.
Li M, Li JJ, Gu QH, et al: EGCG induces lung cancer A549 cell apoptosis by regulating Ku70 acetylation. Oncol Rep 2016;35:2339-2347.
Fang CY, Wu CC, Hsu HY, et al: EGCG inhibits proliferation, invasiveness and tumor growth by up-regulation of adhesion molecules, suppression of gelatinases activity, and induction of apoptosis in nasopharyngeal carcinoma cells. Int J Mol Sci 2015;16:2530-2558.
Adikesavan G, Vinayagam MM, Abdulrahman LA, et al: (-)-Epigallocatechin-gallate (EGCG) stabilize the mitochondrial enzymes and inhibits the apoptosis in cigarette smoke-induced myocardial dysfunction in rats. Mol Biol Rep 2013;40:6533-6545.
Ye R, Yang Q, Kong X, et al: Ginsenoside Rd attenuates early oxidative damage and sequential inflammatory response after transient focal ischemia in rats. Neurochem Int 2011;58:391-398.
Li P, Guo X, Lei P, et al: PI3K/Akt/uncoupling protein 2 signaling pathway may be involved in cell senescence and apoptosis induced by angiotensin II in human vascular endothelial cells. Mol Biol Rep 2014;41:6931-6937.
Hieronimus B, Pfohl J, Busch C, et al: Expression and characterization of membrane-type 4 matrix metalloproteinase (MT4-MMP) and its different forms in melanoma. Cell Physiol Biochem 2017;42:198-210.
Li X, Hong M, Zhu D, et al: OS 29-05 renal denervation attenuates aldosterone expression and associated cardiovascular pathophysiology in angiotensin II-induced hypertension. J Hypertens 2016;34(suppl 1):e254.
Li JM, Shah AM: Mechanism of endothelial cell NADPH oxidase activation by angiotensin II: role of the p47phox subunit. J Biol Chem 2003;278:12094-12100.
Pueyo ME, Gonzalez W, Nicoletti A, et al: Angiotensin II stimulates endothelial vascular cell adhesion molecule-1 via nuclear factor-kappaB activation induced by intracellular oxidative stress. Arterioscler Thromb Vasc Biol 2000;20:645-651.
Piqueras L, Kubes P, Alvarez A, et al: Angiotensin II induces leukocyte-endothelial cell interactions in vivo via AT1 and AT2 receptor-mediated P-selectin upregulation. Circulation 2000;102:2118-2123.
Yi R, Xiao-Ping G, Hui L: Atorvastatin prevents angiotensin II-induced high permeability of human arterial endothelial cell monolayers via ROCK signaling pathway. Biochem Biophys Res Commun 2015;459:94-99.
Maeda R, Noiri E, Isobe H, et al: A water-soluble fullerene vesicle alleviates angiotensin II-induced oxidative stress in human umbilical venous endothelial cells. Hypertens Res 2008;31:141-151.
Hu HJ, Jiang ZS, Qiu J, et al: Protective effects of hydrogen sulfide against angiotensin II-induced endoplasmic reticulum stress in HUVECs. Mol Med Rep 2017;15:2213-2222.
Ahn HY, Kim CH, Ha TS: Epigallocatechin-3-gallate regulates NADPH oxidase expression in human umbilical vein endothelial cells. Korean J Physiol Pharmacol 2010;14:325-329.
Chae YJ, Kim CH, Ha TS, et al: Epigallocatechin-3-O-gallate inhibits the angiotensin II-induced adhesion molecule expression in human umbilical vein endothelial cell via inhibition of MAPK pathways. Cell Physiol Biochem 2007;20:859-866.
Schulz A, Jankowski J, Zidek W, et al: Absolute quantification of endogenous angiotensin II levels in human plasma using ESI-LC-MS/MS. Clin Proteomics 2014;11:37.
Li M, Liu X, He Y, et al: Celastrol attenuates angiotensin II mediated human umbilical vein endothelial cells damage through activation of Nrf2/ERK1/2/Nox2 signal pathway. Eur J Pharmacol 2017;797:124-133.
Griendling KK, FitzGerald GA: Oxidative stress and cardiovascular injury: part I: basic mechanisms and in vivo monitoring of ROS. Circulation 2003;108:1912-1916.
Zhang WJ, Li PX, Guo XH, et al: Role of moesin, Src and ROS in advanced glycation end product-induced vascular endothelial dysfunction. Microcirculation 2017;27:e12358.
Kowald A, Kirkwood TB: Mitochondrial mutations, cellular instability and ageing: modelling the population dynamics of mitochondria. Mutat Res 1993;295:93-103.
Rogers C, Davis B, Neufer PD, et al: A transient increase in lipid peroxidation primes preadipocytes for delayed mitochondrial inner membrane permeabilization and ATP depletion during prolonged exposure to fatty acids. Free Radic Biol Med 2014;67:330-341.
Mateen S, Moin S, Khan AQ, et al: Increased reactive oxygen species formation and oxidative stress in rheumatoid arthritis. PLoS One 2016;11:e0152925.
Zhu Y, Zhang YJ, Liu WW, et al: Salidroside suppresses HUVECs cell injury induced by oxidative stress through activating the Nrf2 signaling pathway. Molecules 2016;21:E1033.
Sedlic F, Muravyeva MY, Sepac A, et al: Targeted modification of mitochondrial ROS production converts high glucose-induced cytotoxicity to cytoprotection: effects on anesthetic preconditioning. J Cell Physiol 2017;232:216-224.
Saha D, Koli S, Reddy KV: Transcriptional regulation of Hb-alpha and Hb-beta through nuclear factor E2-related factor-2 (Nrf2) activation in human vaginal cells: a novel mechanism of cellular adaptability to oxidative stress. Am J Reprod Immunol 2017;77:e12645.
Prasad S, Sajja RK, Kaisar MA, et al: Role of Nrf2 and protective effects of Metformin against tobacco smoke-induced cerebrovascular toxicity. Redox Biol 2017;12:58-69.
Lv H, Liu Q, Zhou J, et al: Daphnetin-mediated Nrf2 antioxidant signaling pathways ameliorate tert-butyl hydroperoxide (t-BHP)-induced mitochondrial dysfunction and cell death. Free Radic Biol Med 2017;106:38-52.
Duan J, Guan Y, Mu F, et al: Protective effect of butin against ischemia/reperfusion-induced myocardial injury in diabetic mice: involvement of the AMPK/GSK-3β/Nrf2 signaling pathway. Sci Rep 2017;7:41491.
Hyung JH, Ahn CB Il, Kim B, et al: Involvement of Nrf2-mediated heme oxygenase-1 expression in anti-inflammatory action of chitosan oligosaccharides through MAPK activation in murine macrophages. Eur J Pharmacol 2016;793:43-48.
Liu J, Zhang FF, Li L, et al: ClC-3 deficiency prevents apoptosis induced by angiotensin II in endothelial progenitor cells via inhibition of NADPH oxidase. Apoptosis 2013;18:1262-1273.
Evangelista AM, Thompson MD, Bolotina VM, et al: Nox4- and Nox2-dependent oxidant production is required for VEGF-induced SERCA cysteine-674 S-glutathiolation and endothelial cell migration. Free Radic Biol Med 2012;53:2327-2334.
Kim YM, Kim SJ, Tatsunami R, et al: ROS-induced ROS release orchestrated by Nox4, Nox2, and mitochondria in VEGF signaling and angiogenesis. Am J Physiol Cell Physiol 2017;312:C749-C764.
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