Allicin prevents H2O2-induced apoptosis of HUVECs by inhibiting an oxidative stress pathway

BMC Complementary and Alternative Medicine

Hui Chai³  Like Wo¹  Lin Zhang³  Ruyi Chen³  Ying Qian³  Yuye Tang²  Sisi Chen³ 

[1] The First Affiliated Hospital of Zhejiang Chinese Medicine University, Hangzhou 310006, China;Hangzhou pharmavaxin co., LTD, Hangzhou 310052, China;College of Life Science, Zhejiang Chinese Medical University, Hangzhou 310053, China

关键词: H₂O₂;    Anti-apoptosis;    Human umbilical vein endothelial cell (HUVEC);    Allicin;   

Others  :  1086839
DOI  :  10.1186/1472-6882-14-321

 received in 2014-07-17, accepted in 2014-08-27,  发布年份 2014

【 摘 要 】

Background

Allicin, a primary ingredient of garlic, has been proposed to possess cardioprotective properties, which are commonly mediated by improved endothelial function.

Methods

To investigate the effect and mechanism of allicin on the apoptosis of human umbilical vein endothelial cells (HUVECs), we used Propidium iodide (PI) staining and Annexin V/ PI staining assays to establish a model of oxidative stress apoptosis induced by H₂O₂. MTT, RT-PCR and western-blot assays were used to detect the effects and mechanism of allicin on the model.

Results

PI staining, Annexin V/ PI staining assays and morphological assessment suggest that the cell death induced by 0.5 mM H₂O₂ is primarily apoptotic. Conversely, allicin reverses the effect of H₂O₂ on cell death, suggesting a role in protecting HUVECs from apoptosis. We demonstrated that H₂O₂ activates PARP cleavage, reduces pro-Caspase-3 levels and activates Bax expression; however, allicin inhibits each of these apoptotic signaling indicators. Allicin also reduces the levels of malondialdehyde and increases the levels of superoxide dismutase, nitric oxide release and endothelial nitric oxide synthase mRNA, but has no significant effect on inducible nitric oxide synthase mRNA levels.

Conclusion

These results demonstrate that allicin has powerful effects in protecting HUVECs from apoptosis and suggest that protection occurs via a mechanism involving the protection from H₂O₂-mediated oxidative stress.

【 授权许可】

   
2014 Chen et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Chan JY, Yuen AC, Chan RY, Chan SW: A review of the cardiovascular benefits and antioxidant properties of allicin. Phytother Res 2013, 27(5):637-646.
• [2]Lusis AJ: Atherosclerosis. Nature 2000, 407(6801):233-241.
• [3]Libby P: Inflammation in atherosclerosis. Nature 2002, 420(6917):868-874.
• [4]Herman AG, Moncada S: Therapeutic potential of nitric oxide donors in the prevention and treatment of atherosclerosis. Eur Heart J 2005, 26(19):1945-1955.
• [5]Osterud B, Bjorklid E: Role of monocytes in atherogenesis. Physiol Rev 2003, 83(4):1069-1112.
• [6]Wang YK, Hong YJ, Wei M, Wu Y, Huang ZQ, Chen RZ, Chen HZ: Curculigoside attenuates human umbilical vein endothelial cell injury induced by H2O2. J Ethnopharmacol 2010, 132(1):233-239.
• [7]Roberto D, Micucci P, Sebastian T, Graciela F, Anesini C: Antioxidant activity of limonene on normal murine lymphocytes: relation to H2O2 modulation and cell proliferation. Basic Clin Pharmacol Toxicol 2010, 106(1):38-44.
• [8]Arzanlou M, Bohlooli S, Jannati E, Mirzanejad-Asl H: Allicin from garlic neutralizes the hemolytic activity of intra- and extra-cellular pneumolysin O in vitro. Toxicon 2011, 57(4):540-545.
• [9]Eilat S, Oestraicher Y, Rabinkov A, Ohad D, Mirelman D, Battler A, Eldar M, Vered Z: Alteration of lipid profile in hyperlipidemic rabbits by allicin, an active constituent of garlic. Coron Artery Dis 1995, 6(12):985-990.
• [10]Abramovitz D, Gavri S, Harats D, Levkovitz H, Mirelman D, Miron T, Eilat-Adar S, Rabinkov A, Wilchek M, Eldar M, Vered Z: Allicin-induced decrease in formation of fatty streaks (atherosclerosis) in mice fed a cholesterol-rich diet. Coron Artery Dis 1999, 10(7):515-519.
• [11]Oliver FJ, de la Rubia G, Rolli V, Ruiz-Ruiz MC, De Murcia G, Murcia JM: Importance of poly (ADP-ribose) polymerase and its cleavage in apoptosis. Lesson from an uncleavable mutant. J Biol Chem 1998, 273(50):33533-33539.
• [12]Boulares AH, Yakovlev AG, Ivanova V, Stoica BA, Wang G, Iyer S, Smulson M: Role of poly (ADP-ribose) polymerase (PARP) cleavage in apoptosis. Caspase 3-resistant PARP mutant increases rates of apoptosis in transfected cells. J Biol Chem 1999, 274(33):22932-22940.
• [13]Hoetelmans R, Van Slooten HJ, Keijzer R, Erkeland S, van de Velde CJ, Dierendonck JH: Bcl-2 and Bax proteins are present in interphase nuclei of mammalian cells. Cell Death Differ 2000, 7(4):384-392.
• [14]Liu YZ TB, Zheng YL, Ma KJ, Xu SZ, Qiu FZ: Screening methods for waterlogging tolerance at maize (Zea mays L. ) seedling stage. Agric Sci 2010, 9(3):362-369.
• [15]Du XMYW, Zhang H: The progress of super oxide dismutase. J China Biotechnol 2002, 23(1):77-79.
• [16]Liao JK, Shin WS, Lee WY, Clark SL: Oxidized low-density lipoprotein decreases the expression of endothelial nitric oxide synthase. J Biol Chem 1995, 270(1):319-324.
• [17]Chen CA, Wang TY, Varadharaj S, Reyes LA, Hemann C, Talukder MA, Chen YR, Druhan LJ, Zweier JL: S-glutathionylation uncouples eNOS and regulates its cellular and vascular function. Nature 2010, 468(7327):1115-1118.
• [18]Knowles RG, Moncada S: Nitric oxide synthases in mammals. Biochem J 1994, 298(Pt 2):249-258.
• [19]Lockshin RA, Zakeri Z: Programmed cell death and apoptosis: origins of the theory. Nat Rev Mol Cell Biol 2001, 2(7):545-550.
• [20]Chu YL, Ho CT, Chung JG, Rajasekaran R, Sheen LY: Allicin induces p53-mediated autophagy in Hep G2 human liver cancer cells. J Agric Food Chem 2012, 60(34):8363-8371.
• [21]Al-Qattan KK, Alnaqeeb MA, Ali M: The antihypertensive effect of garlic (Allium sativum) in the rat two-kidney–one-clip Goldblatt model. J Ethnopharmacol 1999, 66(2):217-222.
• [22]Yin MCCW: Antioxidant activity of several Allium members. J Agric Food Chem 1998, 46(10):4097-4101.
• [23]Gardner CD, Chatterjee LM, Carlson JJ: The effect of a garlic preparation on plasma lipid levels in moderately hypercholesterolemic adults. Atherosclerosis 2001, 154(1):213-220.
• [24]Vimal V, Devaki T: Hepatoprotective effect of allicin on tissue defense system in galactosamine/endotoxin challenged rats. J Ethnopharmacol 2004, 90(1):151-154.
• [25]Prasad K, Laxdal VA, Yu M, Raney BL: Evaluation of hydroxyl radical-scavenging property of garlic. Mol Cell Biochem 1996, 154(1):55-63.
• [26]Rabinkov A, Miron T, Mirelman D, Wilchek M, Glozman S, Yavin E, Weiner L: S-Allylmercaptoglutathione: the reaction product of allicin with glutathione possesses SH-modifying and antioxidant properties. Biochim Biophys Acta 2000, 1499(1–2):144-153.
• [27]Agarwal KC: Therapeutic actions of garlic constituents. Med Res Rev 1996, 16(1):111-124.
• [28]Wang J, Lenardo MJ: Roles of caspases in apoptosis, development, and cytokine maturation revealed by homozygous gene deficiencies. J Cell Sci 2000, 113(Pt 5):753-757.
• [29]Spears JR, Prcevski P, Xu R, Li L, Brereton G, DiCarli M, Spanta A, Crilly R, Lavine S, Vander Heide R: Aqueous oxygen attenuation of reperfusion microvascular ischemia in a canine model of myocardial infarction. ASAIO J 2003, 49(6):716-720.