Hydrogen-rich medium suppresses the generation of reactive oxygen species, elevates the Bcl-2/Bax ratio and inhibits advanced glycation end product-induced apoptosis

Jiang,Hong; Yu,Pan; Qian,De-Hui; Qin,Zhe-Xue; Sun,Xue-Jun; Yu,Jie; Huang,Lan

doi:10.3892/ijmm.2013.1334

Hydrogen-rich medium suppresses the generation of reactive oxygen species, elevates the Bcl-2/Bax ratio and inhibits advanced glycation end product-induced apoptosis

Authors:
- Hong Jiang
- Pan Yu
- De-Hui Qian
- Zhe-Xue Qin
- Xue-Jun Sun
- Jie Yu
- Lan Huang
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Affiliations: Institute of Cardiovascular Diseases of PLA, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, P.R. China, Department of Burns and Plastic Surgery, Jinling Hospital, School of Medicine, Nanjing University, Nanjing, Jiangsu 210002, P.R. China, Department of Diving Medicine, Faculty of Naval Medicine, Second Military Medical University, Shanghai 200433, P.R. China
Published online on: April 5, 2013 https://doi.org/10.3892/ijmm.2013.1334
Pages: 1381-1387

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Abstract

The purpose of the present study was to determine whether using hydrogen-rich medium (HRM) to increase hydrogen levels in endothelial cells (ECs) protects ECs from apoptosis induced by advanced glycation end products (AGEs). The thoracic aorta was removed from 2-3-year-old Sprague-Dawley rats, and ECs were isolated and cultured. After culturing ECs in the presence of AGEs and/or with HRM for 24 h, Annexin V/7-AAD and TUNEL staining were carried out to detect apoptosis. Intracellular ROS were detected by fluorescent probe and quantified by flow cytometry. The expression of antioxidative enzymes (superoxide dismutase, glutathione peroxidase) was determined by real-time PCR analysis and enzymatic assay. The relative expression levels of Bcl-2 and Bax were analyzed by western blotting. The addition of AGEs increased the apoptosis of ECs in a concentration-dependent manner and HRM reduced the AGE (400 µg/ml)-induced apoptosis from 21.61±2.52 to 11.32±1.75%. HRM also significantly attenuated the AGE-induced intracellular ROS induction and decrease in the expression of antioxidative enzymes. In conclusion, hydrogen exhibits significant protective effects against AGE-induced EC injury possibly through reducing ROS generation, intracellular antioxidant enzyme system protection and elevation of the Bcl-2/Bax ratio.

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1	Faria AM, Papadimitriou A, Silva KC, Lopes de Faria JM and Lopes de Faria JB: Uncoupling endothelial nitric oxide synthase is ameliorated by green tea in experimental diabetes by re-establishing tetrahydrobiopterin levels. Diabetes. 61:1838–1847. 2012. View Article : Google Scholar
2	Caporali A, Pani E, Horrevoets AJ, et al: Neurotrophin p75 receptor (p75NTR) promotes endothelial cell apoptosis and inhibits angiogenesis: implications for diabetes-induced impaired neovascularization in ischemic limb muscles. Circ Res. 103:e15–e26. 2008. View Article : Google Scholar
3	Csiszar A and Ungvari Z: Endothelial dysfunction and vascular inflammation in type 2 diabetes: interaction of AGE/RAGE and TNF-alpha signaling. Am J Physiol Heart Circ Physiol. 295:H475–H476. 2008. View Article : Google Scholar : PubMed/NCBI
4	Goldin A, Beckman JA, Schmidt AM and Creager MA: Advanced glycation end products: sparking the development of diabetic vascular injury. Circulation. 114:597–605. 2006. View Article : Google Scholar : PubMed/NCBI
5	Xiang M, Yang M, Zhou C, Liu J, Li W and Qian Z: Crocetin prevents AGEs-induced vascular endothelial cell apoptosis. Pharmacol Res. 54:268–274. 2006. View Article : Google Scholar : PubMed/NCBI
6	Chen J, Huang L, Song M, Yu S, Gao P and Jing J: C-reactive protein upregulates receptor for advanced glycation end products expression and alters antioxidant defenses in rat endothelial progenitor cells. J Cardiovasc Pharmacol. 53:359–367. 2009. View Article : Google Scholar
7	Tan AL, Forbes JM and Cooper ME: AGE, RAGE, and ROS in diabetic nephropathy. Semin Nephrol. 27:130–143. 2007. View Article : Google Scholar : PubMed/NCBI
8	Ohsawa I, Ishikawa M, Takahashi K, et al: Hydrogen acts as a therapeutic antioxidant by selectively reducing cytotoxic oxygen radicals. Nat Med. 13:688–694. 2007. View Article : Google Scholar
9	Xie K, Yu Y, Huang Y, et al: Molecular hydrogen ameliorates lipopolysaccharide-induced acute lung injury in mice through reducing inflammation and apoptosis. Shock. 37:548–555. 2012.PubMed/NCBI
10	Cai J, Kang Z, Liu K, et al: Neuroprotective effects of hydrogen saline in neonatal hypoxia-ischemia rat model. Brain Res. 1256:129–137. 2009. View Article : Google Scholar : PubMed/NCBI
11	Chen H, Sun YP, Li Y, et al: Hydrogen-rich saline ameliorates the severity of l-arginine-induced acute pancreatitis in rats. Biochem Biophys Res Commun. 393:308–313. 2010. View Article : Google Scholar : PubMed/NCBI
12	Liu Q, Shen WF, Sun HY, et al: Hydrogen-rich saline protects against liver injury in rats with obstructive jaundice. Liver Int. 30:958–968. 2010. View Article : Google Scholar : PubMed/NCBI
13	Sun Q, Kang Z, Cai J, et al: Hydrogen-rich saline protects myocardium against ischemia/reperfusion injury in rats. Exp Biol Med. 234:1212–1219. 2009. View Article : Google Scholar : PubMed/NCBI
14	Li J, Wang C, Zhang JH, Cai JM, Cao YP and Sun XJ: Hydrogen-rich saline improves memory function in a rat model of amyloid-beta-induced Alzheimer’s disease by reduction of oxidative stress. Brain Res. 1328:152–161. 2010.PubMed/NCBI
15	Sun Q, Cai J, Zhou J, Tao H, Zhang JH, Zhang W and Sun XJ: Hydrogen-rich saline reduces delayed neurologic sequelae in experimental carbon monoxide toxicity. Crit Care Med. 39:765–769. 2011. View Article : Google Scholar : PubMed/NCBI
16	Sun H, Chen L, Zhou W, et al: The protective role of hydrogen-rich saline in experimental liver injury in mice. J Hepatol. 54:471–480. 2011. View Article : Google Scholar : PubMed/NCBI
17	Terasaki Y, Ohsawa I, Terasaki M, et al: Hydrogen therapy attenuates irradiation-induced lung damage by reducing oxidative stress. Am J Physiol Lung Cell Mol Physiol. 301:L415–L426. 2011. View Article : Google Scholar : PubMed/NCBI
18	Ohsawa I, Nishimaki K, Yamagata K, Ishikawa M and Ohta S: Consumption of hydrogen water prevents atherosclerosis in apolipoprotein E knockout mice. Biochem Biophys Res Commun. 377:1195–1198. 2008. View Article : Google Scholar : PubMed/NCBI
19	Qin ZX, Yu P, Qian DH, et al: Hydrogen-rich saline prevents neointima formation after carotid balloon injury by suppressing ROS and the TNF-α/NF-κB pathway. Atherosclerosis. 220:343–350. 2012.PubMed/NCBI
20	Wu X, Zhou Q, Huang L, Sun A, Wang K, Zou Y and Ge J: Ageing-exaggerated proliferation of vascular smooth muscle cells is related to attenuation of Jagged1 expression in endothelial cells. Cardiovasc Res. 77:800–808. 2008. View Article : Google Scholar : PubMed/NCBI
21	Tang X, Guo Y, Nakamura K, et al: Nitroalkenes induce rat aortic smooth muscle cell apoptosis via activation of caspase-dependent pathways. Biochem Biophys Res Commun. 397:239–244. 2010. View Article : Google Scholar : PubMed/NCBI
22	Chen J, Song M, Yu S, Gao P, Yu Y, Wang H and Huang L: Advanced glycation endproducts alter functions and promote apoptosis in endothelial progenitor cells through receptor for advanced glycation endproducts mediate overpression of cell oxidant stress. Mol Cell Biochem. 335:137–146. 2010. View Article : Google Scholar
23	Marsh SA, Laursen PB, Pat BK, Gobe GC and Coombes JS: Bcl-2 in endothelial cells is increased by vitamin E and alpha-lipoic acid supplementation but not exercise training. J Mol Cell Cardiol. 38:445–451. 2005. View Article : Google Scholar : PubMed/NCBI
24	Oltvai ZN, Milliman CL and Korsmeyer SJ: Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell. 74:609–619. 1993. View Article : Google Scholar : PubMed/NCBI
25	Jandeleit-Dahm K, Watson A and Soro-Paavonen A: The AGE/RAGE axis in diabetes-accelerated atherosclerosis. Clin Exp Pharmacol Physiol. 35:329–334. 2008. View Article : Google Scholar
26	Kajiyama S, Hasegawa G, Asano M, et al: Supplementation of hydrogen-rich water improves lipid and glucose metabolism in patients with type 2 diabetes or impaired glucose tolerance. Nutr Res. 28:137–143. 2008. View Article : Google Scholar : PubMed/NCBI
27	Qian L, Li B, Cao F, Huang Y, Liu S, Cai J and Gao F: Hydrogen-rich PBS protects cultured human cells from ionizing radiation-induced cellular damage. Nucl Technol Radiat Prot. 25:23–29. 2010. View Article : Google Scholar
28	Yu P, Wang Z, Sun X, Chen X, Zeng S, Chen L and Li S: Hydrogen-rich medium protects human skin fibroblasts from high glucose or mannitol induced oxidative damage. Biochem Biophys Res Commun. 409:350–355. 2011. View Article : Google Scholar : PubMed/NCBI
29	Yu Ys and Zheng H: Chronic hydrogen-rich saline treatment reduces oxidative stress and attenuates left ventricular hypertrophy in spontaneous hypertensive rats. Mol Cell Biochem. 365:233–242. 2012. View Article : Google Scholar : PubMed/NCBI
30	Zhang Y, Sun Q, He B, Xiao J, Wang Z and Sun X: Anti-inflammatory effect of hydrogen-rich saline in a rat model of regional myocardial ischemia and reperfusion. Int J Cardiol. 148:91–95. 2011. View Article : Google Scholar : PubMed/NCBI
31	Zong C, Song G, Yao S, et al: Administration of hydrogen-saturated saline decreases plasma low-density lipoprotein cholesterol levels and improves high-density lipoprotein function in high-fat diet-fed hamsters. Metabolism. 61:794–800. 2012. View Article : Google Scholar
32	Schoenfeld MP, Ansari RR, Zakrajsek JF, et al: Hydrogen therapy may reduce the risks related to radiation-induced oxidative stress in space flight. Med Hypotheses. 76:117–118. 2011. View Article : Google Scholar : PubMed/NCBI
33	Hess DA and Hegele RA: Linking diabetes with oxidative stress, adipokines, and impaired endothelial precursor cell dunction. Can J Cardiol. 28:629–630. 2012. View Article : Google Scholar : PubMed/NCBI
34	Sharpe PC, Liu WH, Yue KK, et al: Glucose-induced oxidative stress in vascular contractile cells: comparison of aortic smooth muscle cells and retinal pericytes. Diabetes. 47:801–809. 1998. View Article : Google Scholar : PubMed/NCBI
35	Li BY, Li XL, Cai Q, et al: Induction of lactadherin mediates the apoptosis of endothelial cells in response to advanced glycation end products and protective effects of grape seed procyanidin B2 and resveratrol. Apoptosis. 16:732–745. 2011. View Article : Google Scholar : PubMed/NCBI
36	Deng X, Gao F and May WS Jr: Bcl2 retards G1/S cell cycle transition by regulating intracellular ROS. Blood. 102:3179–3185. 2003. View Article : Google Scholar : PubMed/NCBI