Gle most important contributor to this growing burden of cardiovascular disease [3]. Endothelial dysfunction is considered an early indicator of atherosclerosis, preceding angiographic or ultrasonic evidence of atherosclerotic plaques [4]. In addition to managing anabolism and exchange of blood and tissue fluids, endothelial cells also act as an* Correspondence: [email protected]; [email protected]; [email protected] Equal contributors 1 College of Life Science, Zhejiang Chinese Hexanoyl-Tyr-Ile-Ahx-NH2 side effects Medical University, Hangzhou 310053, China 3 The First Affiliated Hospital of Zhejiang Chinese Medicine University, Hangzhou 310006, China Full list of author information is available at the end of the articleendocrine gland. Endothelial cells produce and secrete multiple biologically active substances that help to maintain normal angiostasis and balance of blood. Considerable evidence indicates that oxidized low-density lipoprotein PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/26024392 (ox-LDL) can cause the apoptosis of vascular endothelial cells through multiple pathways [5]. However the production process of ox-LDL is complicated, and it is stable for only 1 month at 4 . Conversely, H2O2 is economical, simple and practical, so it is commonly used in injury models to replicate the effects of ox-LDL [6,7]. Natural antioxidants are important for the prevention and treatment of atherosclerosis. Garlic has been studied extensively for its cardioprotective properties with very promising results [8]. Its primary active ingredient, 2propene-1-sulfinothioic acid S-2-propenyl ester (also known as allicin), has been shown to alter the composition of fatty acids in mice or rats fed a high fatty acid diet [9,10]. The aim of present study was to use H2O2 instead?2014 Chen et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons FCCP web Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.Chen et al. BMC Complementary and Alternative Medicine 2014, 14:321 http://www.biomedcentral.com/1472-6882/14/Page 2 ofof ox-LDL to establish a model of oxidative stress and apoptosis in which to observe the intervention effect of allicin on endothelial cell apoptosis. The characterization of a new antioxidant drug may be beneficial as a novel strategy for the treatment of atherosclerosis.flow cytometry was performed using a FACSCalibur (Beckmancoulter, USA). Data were analyzed using CellQuest software (Becton ickinson). The amount of apoptosis was determined as the percentage of annexin V-positive cells/PI-negative cells.MTT assayMethodsCell cultureHUVECs were obtained from Cambrex (Shanghai Biological Technology Co., Ltd., China) and were grown in Dulbecco’s modified eagle medium (DMEM) supplemented with 10 heat-inactivated FBS (Hangzhou Sijiqing biological engineering materials Co., Ltd., China) at 37 in a humidified atmosphere of 5 CO2. Cells were used at passage 4? for all experiments.Propidium iodide (PI) stainingHUVECs were cultured in 6 well plates (BD Falcon, USA) at a density of 2.0 ?105 cells/well in DMEM supplemented with 10 FBS. One day after plating, the cells were washed and incubated in serum-free medium for 12 hours. The cells were then washe.Gle most important contributor to this growing burden of cardiovascular disease [3]. Endothelial dysfunction is considered an early indicator of atherosclerosis, preceding angiographic or ultrasonic evidence of atherosclerotic plaques [4]. In addition to managing anabolism and exchange of blood and tissue fluids, endothelial cells also act as an* Correspondence: [email protected]; [email protected]; [email protected] Equal contributors 1 College of Life Science, Zhejiang Chinese Medical University, Hangzhou 310053, China 3 The First Affiliated Hospital of Zhejiang Chinese Medicine University, Hangzhou 310006, China Full list of author information is available at the end of the articleendocrine gland. Endothelial cells produce and secrete multiple biologically active substances that help to maintain normal angiostasis and balance of blood. Considerable evidence indicates that oxidized low-density lipoprotein PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/26024392 (ox-LDL) can cause the apoptosis of vascular endothelial cells through multiple pathways [5]. However the production process of ox-LDL is complicated, and it is stable for only 1 month at 4 . Conversely, H2O2 is economical, simple and practical, so it is commonly used in injury models to replicate the effects of ox-LDL [6,7]. Natural antioxidants are important for the prevention and treatment of atherosclerosis. Garlic has been studied extensively for its cardioprotective properties with very promising results [8]. Its primary active ingredient, 2propene-1-sulfinothioic acid S-2-propenyl ester (also known as allicin), has been shown to alter the composition of fatty acids in mice or rats fed a high fatty acid diet [9,10]. The aim of present study was to use H2O2 instead?2014 Chen et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.Chen et al. BMC Complementary and Alternative Medicine 2014, 14:321 http://www.biomedcentral.com/1472-6882/14/Page 2 ofof ox-LDL to establish a model of oxidative stress and apoptosis in which to observe the intervention effect of allicin on endothelial cell apoptosis. The characterization of a new antioxidant drug may be beneficial as a novel strategy for the treatment of atherosclerosis.flow cytometry was performed using a FACSCalibur (Beckmancoulter, USA). Data were analyzed using CellQuest software (Becton ickinson). The amount of apoptosis was determined as the percentage of annexin V-positive cells/PI-negative cells.MTT assayMethodsCell cultureHUVECs were obtained from Cambrex (Shanghai Biological Technology Co., Ltd., China) and were grown in Dulbecco’s modified eagle medium (DMEM) supplemented with 10 heat-inactivated FBS (Hangzhou Sijiqing biological engineering materials Co., Ltd., China) at 37 in a humidified atmosphere of 5 CO2. Cells were used at passage 4? for all experiments.Propidium iodide (PI) stainingHUVECs were cultured in 6 well plates (BD Falcon, USA) at a density of 2.0 ?105 cells/well in DMEM supplemented with 10 FBS. One day after plating, the cells were washed and incubated in serum-free medium for 12 hours. The cells were then washe.