Diallyl trisufide (DATS) suppresses high glucose-induced cardiomyocyte apoptosis by inhibiting JNK/NFκB signaling via attenuating ROS generation☆
Introduction
Hyperglycemia, a major feature of type I and type II diabetes mellitus, can cause oxidative stress, which is associated with a variety of diabetic organ damage, including cardiovascular diseases. Cardiac cells are susceptible to oxidative stress, which can induce cardiomyocyte apoptosis and decrease cardiac function under high glucose conditions, leading to diabetic cardiomyopathy [1].
Studies have demonstrated that NADPH oxidase-related reactive oxygen species (ROS) are involved in the detrimental effect of high glucose-induced cell damage [2]. In endothelial cells and cardiomyocytes, NADPH oxidase isoforms generate reactive oxygen species as modulators of redox-sensitive signaling pathways [3], [4]. Over activation of NADPH oxidase, however, can result in loss of the redox balance, thereby leading to abnormal ROS production. Structurally, NADPH oxidase comprises multiple subunits. Cytochrome b558, the membrane-bound component, consists of the subunits p22phox and gp91phox, which are responsible for enzyme activity. The enzyme complex also includes at least four cytosolic subunits, p47phox, p67phox, p40phox, and the small guanosine triphosphatase Rac1, which translocate to the cell membrane and associate with the membrane-bound components forming an active complex [5]. NADPH oxidase inhibitors, such as apocynin, inhibit the formation of the oxidase complex and its subsequent activation by preventing the association of p47phox with the membrane-bound components [6].
Garlic oil has been shown be a potent antioxidant and to have cardioprotective effects [7]. Garlic oil comprises three organosulfur compounds, namely diallyl sulfide (DAS), diallyl disulfide (DADS), and diallyl trisulfide (DATS). Liu et al. [8] reported that the antioxidant potential of these organosulfur compounds is related to the number of sulfur atoms and is in the order DATS > DADS > DAS. In addition to the hypolipidemic effects by decreasing fatty acid synthesis and lipid accumulation [9], DATS has been shown to attenuate oxidative stress-induced liver injury in mice [12] and suppress oxidized LDL-induced adhesion molecule expression in vascular endothelial cells [10]. Studies have also provided evidence that DATS can inhibit LPS-induced activation of NF-κB in the lung [11] and in macrophages [8]. Stent coated with DATS improved the damage of endothelial function in the coronary by increasing the eNOS expression to raise the NO level [12]. Furthermore, using myocardial ischemia–reperfusion injury (MI-R) animal model, DATS treatment could reduce infarct size, increased nitric oxide bioavailability, reversed myocardial contractile function [13]. In our previous study we demonstrated that garlic oil dose-dependently ameliorated hyperglycemia-induced cardiac dysfunction and apoptosis [14]. However, the mechanisms underlying the protective effects of DATS against hyperglycemia-induced cardiac damage are not known.
Excessive production of ROS induced by high glucose has been documented to be an important mechanism in the pathogenesis of diabetic cardiomyopathy [15]. In a previous study using apocynin, and JNK and NF-κB small interfering RNAs, we found that ROS-induced apoptosis was mediated via the JNK-dependent activation of NF-κB in cardiomyocytes exposed to high glucose [16]. In the present study, we investigated the mechanisms governing the anti-apoptotic effect of DATS in cardiac cells exposed to high glucose in vitro and in vivo.
Section snippets
Cell culture and treatments
H9c2 cell lines purchased from American Type Culture Collection (ATCC), with passages between 8 and 10 were used in this study. Cells were cultured in Dulbecco's modified essential medium (DMEM) with 10% fetal bovine serum (FBS), 2 mM glutamine, 100 units/ml penicillin, 100 μg/ml streptomycin, and 1 mM pyruvate in humidified air (5% CO2) at 37 °C. During the experiment period, H9c2 cells were cultured in minimal essential medium with glucose at normal concentrations (5.5 mM) for 12 h, then followed by
DATS inhibited the protein expression of p22phox and gp91phox as well as the production of ROS in HG-treated H9c2 cells
The subunits that comprise the membrane-bound component of NADPH oxidase, namely p22phox and gp91phox, play a critical role in the activity and stability of the enzyme complex. We found that exposure to HG resulted in a marked increase in the protein expression of p22phox and gp91phox and that treatment with DATS (1 to 10 μM) attenuated the HG-induced increase in the expression of those proteins (Fig. 1A). In addition, HG also resulted in an increase in the productions of DCF-sensitive ROS and
Discussion
Numerous lines of evidence suggest that over production of ROS is a key early event in the long-term development of diabetic complications [18]. In our previous study we demonstrated that high concentrations of glucose led to a marked increase in levels of NADPH oxidase-generated ROS and that this increase in ROS induced apoptosis in cardiomyocytes by activating the JNK/NF-κB signaling pathway [16]. The results of the in vitro and in vivo experiments in the present study show that DATS at low
Acknowledgment
We thank Dr Cheng-Wen Lin for kindly providing his plasmid for luciferase assay. We also thank Chien-Chung Wang for his final revision data polish. This study is supported by the National Science Council of Republic of China (grant NSC99-2320-B-039-033-MY3), China Medical University (grants CMU99-COL-28-1, CMU99-COL-28-2, and CMU98-S-07) and the Taiwan Department of Health Clinical Trial and Research Center of Excellence (DOH101-TD-B-111-004) . The authors of this manuscript have certified that
References (46)
Selective targeting of NADPH oxidase for cardiovascular protection
Curr Opin Pharmacol
(2009)- et al.
The vascular NAD(P)H oxidases as therapeutic targets in cardiovascular diseases
Trends Pharmacol Sci
(2003) - et al.
Diallyl trisulfide suppresses the adipogenesis of 3T3–L1 preadipocytes through ERK activation
Food Chem Toxicol
(2012) - et al.
Diallyl disulfide and diallyl trisulfide suppress oxidized LDL-induced vascular cell adhesion molecule and E-selectin expression through protein kinase A- and B-dependent signaling pathways
J Nutr
(2008) - et al.
High glucose-induced apoptosis in human vascular endothelial cells is mediated through NF-kappaB and c-Jun NH2-terminal kinase pathway and prevented by PI3K/Akt/eNOS pathway
Cell Signal
(2006) - et al.
Roles of p38 and JNK mitogen-activated protein kinase pathways during cantharidin-induced apoptosis in U937 cells
Biochem Pharmacol
(2004) - et al.
Activation of c-Jun N-terminal kinase is essential for oxidative stress-induced Jurkat cell apoptosis by monochloramine
Leuk Res
(2009) - et al.
Modulation of the JNK pathway in liver affects insulin resistance status
J Biol Chem
(2004) - et al.
The c-Jun NH(2)-terminal kinase promotes insulin resistance during association with insulin receptor substrate-1 and phosphorylation of Ser(307)
J Biol Chem
(2000) - et al.
Regulating the regulator: NF-kappaB signaling in heart
J Mol Cell Cardiol
(2006)
Diallyl trisulfide protects rats from carbon tetrachloride-induced liver injury
J Nutr
Diallyl sulfide enhances antioxidants and inhibits inflammation through the activation of Nrf2 against gentamicin-induced nephrotoxicity in Wistar rats
Eur J Pharmacol
Differential effects of diallyl disulfide on neuronal cells depend on its concentration
Toxicology
Protective effect of diallyl disulfide on oxidative stress-injured neuronally differentiated PC12 cells
Brain Res Mol Brain Res
Antidiabetic effect of garlic oil but not diallyl disulfide in rats with streptozotocin-induced diabetes
Food Chem Toxicol
Diabetic cardiomyopathy revisited
Circulation
Vascular NAD(P)H oxidase activation in diabetes: a double-edged sword in redox signalling
Cardiovasc Res
Localizing NADPH oxidase-derived ROS
Sci STKE
Compartmentalization of redox signaling through NADPH oxidase-derived ROS
Antioxid Redox Signal
Use of alternative medicines in diabetes mellitus
Diabet Med
DATS reduces LPS-induced iNOS expression, NO production, oxidative stress, and NF-kappaB activation in RAW 264.7 macrophages
J Agric Food Chem
Effect of diallyl trisulfide on tumor necrosis factor-alpha expression and nuclear factor-kappaB activity in mice with acute lung injury induced by lipopolysaccharide
Zhongguo Wei Zhong Bing Ji Jiu Yi Xue
Effect of stent coated with diallyl trisulfide on endothelial structure and function after coronary injury: experiment with dogs
Zhonghua Yi Xue Za Zhi
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This work was supported by the National Science Council, Taiwan (NSC99-2320-B-039-033-MY3) and China Medical University (DOH101-TD-B-111-004 and CMU98-S-07).
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These authors contributed equally to this work.