Effects Of Diallyl Trisulfide Research Articles

Effects of Diallyl Trisulfide on TNF‐α‐stimulated Genetically different Triple‐Negative Breast Cancer Cells

Many studies have demonstrated the potential anticancer properties of garlic and their respective constituents through in vitro and in vivo studies. Diallyl trisulfide (DATS), an organosulfur compound of garlic, has been shown to prevent and inhibit breast carcinogenesis through various signaling pathways. The triple‐negative breast cancer (TNBC) subtype, which accounts for approximately 15% of all breast cancers, usually behaves more aggressively than other types of breast cancer and is more challenging to treat. Cancer progression is associated with the overexpression of the monocyte chemotactic protein‐1 (MCP‐1. MCP‐1, also known as C‐C motif chemokine ligand‐2 (CCL2), plays a significant role in breast cancer cell signaling, leading to increased cell proliferation, immune suppression, epithelial‐to‐mesenchymal transition (EMT), and angiogenesis. This study explored DATS effects on the genetically different triple‐negative breast cancer cells, MDA‐MB‐231 and MDA‐MB‐468. DATS effects on TNF‐α induced TNBC cells were examined via trypan blue exclusion test, wound‐healing assay, human cytokine arrays, ELISA, and RT‐PCR. The results showed that DATS induced cytotoxicity in a dose and time‐response way in MDA‐MB‐231 and MDA‐MB‐468 cell lines. Wound healing assays demonstrated that DATS significantly inhibited cell migration after a 12 h exposure in both cell lines. Protein expression of CCL2/MCP‐1, IL‐6, PDGF‐BB, NT‐3, and GM‐CSF was increased in the TNF‐α‐treated cells. However, the cotreatment with DATS and TNF‐α showed that the compound significantly decreased the expression of CCL2/MCP‐1 in MDA‐MB‐231 but not in MDA‐MB‐468 cells; data confirmed with ELISA. Moreover, DATS significantly down‐regulated mRNA expression of IKBKE and MAPK8 in both cell lines, indicating a possible effect in genes involved in the NF‐κB and MAPK signaling. The data show that genetically different cells may respond in a different way to DATS treatment. In conclusion, DATS may be a potential candidate for breast cancer therapy to slow TNBC progression.

Effect of Diallyl Trisulfide on TNF-α-induced CCL2/MCP-1 Release in Genetically Different Triple-negative Breast Cancer Cells

Diallyl trisulfide (DATS) has been shown to prevent and inhibit breast carcinogenesis. CCL2/MCP-1 has been shown to play a significant role in breast cancer. This study explored DATS efficacy on triple-negative breast cancer (TNBC) cells. DATS efficacy on TNF-α induced TNBC cells were examined via trypan blue exclusion test, wound-healing assay, human cytokine arrays, ELISA, and RT-PCR. DATS significantly induced cell death and inhibited cell migration. Expression of CCL2/MCP-1, IL-6, PDGF-BB, NT-3, and GM-CSF in TNF-α-treated cells increased. However, DATS significantly decreased the expression of CCL2/MCP-1 in TNF-α-treated MDA-MB-231 but not in MDA-MB-468 cells. DATS significantly down-regulated mRNA expression of IKBKE and MAPK8 in both cell lines, indicating a possible effect in genes involved in the NF-κB and MAPK signaling. DATS may have a role in TNBC therapy and prevention by targeting CCL2.

Diallyl trisulfide inhibited tobacco smoke-mediated bladder EMT and cancer stem cell marker expression via the NF-κB pathway in vivo.

ObjectiveThis study examined the effect of the NF-κB pathway on tobacco smoke-elicited bladder epithelial–mesenchymal transition (EMT) and cancer stem cell (CSC) marker expression in vivo. The effect of diallyl trisulfide (DATS) treatment was also examined.MethodsBALB/c mice were exposed to tobacco smoke and treated with an NF-κB inhibitor and DATS. Western blotting, quantitative real-time PCR, and immunohistochemical staining were used to detect the changes of relevant indices.ResultsPhosphorylated inhibitor of kappa-B kinase alpha/beta expression and p65 and p50 nuclear transcription were increased by tobacco smoke exposure, whereas inhibitor of kappa-B expression was decreased. In addition, tobacco smoke reduced the expression of epithelial markers but increased that of mesenchymal and CSC markers. Our study further demonstrated that tobacco smoke-mediated EMT and CSC marker expression were attenuated by inhibition of the NF-κB pathway. Moreover, DATS reversed tobacco smoke-induced NF-κB pathway activation, EMT, and the acquisition of CSC properties in bladder tissues.ConclusionsThese data suggested that the NF-κB pathway regulated tobacco smoke-induced bladder EMT, CSC marker expression, and the protective effects of DATS.

Effects of diallyl trisulfide, an active substance from garlic essential oil, on structural chemistry of chitin in Sitotroga cerealella (Lepidoptera: Gelechiidae)

The environmental pollution, evolution of resistance, and risks to human and aquatic animal health associated with pesticide application have attracted much attention globally. Herein, we tested the capacity of diallyl trisulfide (DAT) from garlic essential oil to control the destructive stored-product pest, Sitotroga cerealella. The effects of DAT on the total content of cuticular chitin and structure of adults S. cerealella were evaluated. This study was the first to investigate changes in chitin structure in adults due to exposure to DAT through Fourier-transform infrared spectroscopy, thermogravimetric analysis, X-ray diffraction, and differential scanning calorimetry. The results of these analyses revealed that the cuticular chitin content of pests decreased after DAT treatment. DAT treatment also reduced thermal stability and crystallinity of chitin. These findings indicate that DAT is a potent biopesticide that is active against the moth, and establishes the basis for its use as an IPM and alternative to chitin synthesis inhibitors.

Effect of Diallyl Trisulfide on the Reproductive Behavior of the Grain Moth, Sitotroga cerealella (Lepidoptera: Gelechiidae).

The Angoumois grain moth, Sitotroga cerealella (Olivier, 1789) (Lepidoptera: Gelechiidae), is primarily a pest of stored products, that feeds inside the grain as larvae inducing significant economic loss in various stored commodities. Our previous studies proved that garlic essential oil and its active substances inhibit oviposition in moths. To further explore the effect on reproductive behavior and accurately interpret the results in terms of effective control of the moth population, moths were treated with diallyl trisulfide (DATS), an active substance of garlic essential oil, at a dose of 0.015 µL/L in air (LC20, sub-lethal concentration). The results showed that fecundity and the proportion of viable eggs significantly decreased when the moths were treated with LC20 DATS. Furthermore, female circadian mating rhythms and calling periodicity changed significantly after treatment. Compared with controls, the peak in mating occurred approximately 1 h earlier on the first day after DATS treatment, while it was significantly later on days 2 and 3. Moreover, mating frequency declined in presence of DATS compared with the controls. The percentage of females engaging in calling behavior decreased significantly with time, to less than 50%, 2 days after treatment, while a high calling percentage (>80%) was recorded for control moths on all 4 days. In addition, DATS had an inhibitory effect on the mating duration of S. cerealella. Moreover, a significant reduction was observed in the amount of sex pheromones extracted 8 h and 9 h after treatment. Our findings suggested that DATS has the potential to manipulate the moth population at LC20 and would be an efficient alternative to synthetic insecticides for the control of pests having low toxicity to non-target organisms and ecosystems.

Diallyl Trisulfide Enhances Benzo[a]pyrene-induced CYP1A1 Expression and Metabolic Activation in Hepatic HepG2 Cells.

Benzo[a]pyrene (BaP), an environmental pollutant produced by combustion processes, induces expression of cytochrome P450 (CYP) 1A1 via the activation of aryl hydrocarbon receptor (AHR). Induced CYP1A1 is involved in BaP metabolism, resulting in either detoxification or metabolic activation in a context-dependent manner. The effect of diallyl trisulfide (DATS), a garlic-derived organosulfur compound, on BaP metabolism has not been investigated. The combined effect of DATS and BaP on BaP metabolism in hepatocyte-derived HepG2 cells was examined. DATS enhanced BaP-induced CYP1A1 and CYP1B1 mRNA expression, BaP hydroxylation and BaP-DNA adduct formation. Combined treatment of BaP and DATS also increased reactive oxygen species levels. DATS enhanced BaP-induced AHR recruitment and histone H3 acetylation on the CYP1A1 promoter. DATS combined treatment enhances BaP metabolic activation through an AHR-modulating mechanism.

Protective effect of diallyl trisulfide on acute lung injury in rats

Objective Observing the protective effect of diallyl trisulfide (DATS) on acute lung injury induced by cecal ligation and puncture (CLP). Methods The animal model of acute lung injury was induced by CLP. The 30 male SD rats were fasted 16 h before surgery, then was anesthetized by intraperitoneal injection of 10% chloral hydrate (4 ml/kg). Then all rats were randomly divided into 3 groups (n=10), sham group (group Ⅰ), CLP+normal saline (NS) group (groupⅡ) and CLP+DATS group (group Ⅲ). Rats in groupⅠ and group Ⅱ were injected with saline 10 ml/kg through caudal vein, but group Ⅲ were injected with DATS 10 mg/kg. The arterial cannulation was used in carotid artery of the rat to measure the blood gas after the surgery at 0 h (T0), 6 h (T6) and 12 h (T12). All male SD rats were subcutaneously injected with normal saline 0.05 ml/g. Subsequently, the male SD rats were sacrificed at 12 h after operation by the blooding and collected samples of lung tissue. The lung tissue wet/dry weight ratio (W/D), myeloperoxidase (MPO) activity, malondialdehyde (MDA), superoxide dismutase (SOD) and nitric oxide (NO) levels, expression of nuclear factor-κB(NF-κB) P65 and IκB-α in the cytoplasm of lung tissue, expression of NF-κB P65 in the nucleus of lung tissue were measured. Pathological changes of lung tissue was observed under light microscope. Another 30 male SD rats were grouped randomly and received the same experiment methods. The survival rate at 24 h was observed. Results Compared with group Ⅰ, the arterial partial pressure of oxygen (PaO2) and activity of SOD were significantly decreased (P<0.05), but the arterial partial pressure of carbon dioxide (PaCO2), lung tissue W/D, activity of MPO, concentration of MDA and NO in the group Ⅱ and group Ⅲ at T6 and T12 were significantly increased (P<0.05). Inflammatory reaction was significantly increased under light microscope in the group Ⅱ. The expression of cytoplasmic NF-κB P65 and IκB-α was decreased (P<0.05), but the expression of nucleus NF-κB P65 was increased in the group Ⅱ(P<0.05). The survival rate of the group Ⅱ in 24 h was lower than the rate in group Ⅰ (P<0.05). Compared with group Ⅱ, the arterial PaO2 and activity of SOD was increased (P<0.05), but the arterial PaCO2, lung tissue W/D, activity of MPO, concentration of MDA and NO were decreased in the group Ⅲ at T6 and T12(P<0.05). Inflammatory reaction was significantly decreased under light microscope in group Ⅲ. The expression of cytoplasmic NF-κB P65 and IκB-α were increased (P<0.05), but the expression of nucleus NF-κB P65 were decreased in group Ⅲ (P<0.05). The survival rate of the group Ⅲ in 24 h was improved (P<0.05). Conclusions DATS has a protective effect on acute lung injury in SD rats. Key words: Diallyl trisulfide; Cecum; Perforation of intestine; Acute lung injury

Effect of Diallyl Trisulfide on Ischemic Tissue Injury and Revascularization in a Diabetic Mouse Model.

Background and Objective:Allitridin [diallyl trisulfide (DATS)] is an extract from garlic (Allium sativum) that putatively improves endothelial function and is protective against cardiovascular diseases. Endothelial dysfunction after tissue ischemia in diabetic patients is partially due to poor angiogenic response. This study investigated whether DATS may improve angiogenesis in a diabetic mouse model with hind limb ischemia.Methods:Streptozotocin was administered by intraperitoneal injection to establish the model of diabetes in male C57BL/6 mice. After 14 days, nondiabetic and diabetic mice (n = 24, each) underwent unilateral hind limb ischemia by femoral artery ligation. The mice were apportioned to 4 groups: nondiabetic treated (or not) with DATS and diabetic treated (or not) with DATS. DATS treatment consisted of a single daily intraperitoneal injection of 500 μg·kg−1·d−1 for 14 days, beginning on the day of induced ischemia. Ischemia was scored by standard criteria. Blood perfusion was determined using thermal infrared imaging. Tissue capillary density and oxidative stress levels were measured by immunohistochemistry and immunofluorescence, respectively. Serum lipids were measured by enzymatic colorimetric assay. Fasting serum insulin was detected using an insulin enzyme-linked immunosorbent assay kit. Nitric oxide (NO) metabolites and protein carbonyls in tissues were determined by enzyme-linked immunosorbent assay. Targeted protein concentrations were measured by western blotting.Results:At 14 days after ligation, the ischemic skeletal muscle of the streptozotocin-induced diabetic mice had lower levels of endothelial NO synthase, phosphorylated endothelial NO synthase, and vascular endothelial growth factor compared with nondiabetic group. In addition, the hind limb blood perfusion, capillary density, and NO bioactivity were lower in the diabetic group, whereas oxidative stress and protein carbonyl levels were higher. These changes were ameliorated by DATS treatment.Conclusions:DATS treatment of diabetic mice promoted revascularization in ischemic tissue.

Effects of diallyl trisulfide on induction of apoptotic death in murine leukemia WEHI‐3 cells in vitro and alterations of the immune responses in normal and leukemic mice in vivo

Diallyl trisulfide (DATS), a chemopreventive dietary constituent and extracted from garlic, has been shown to against cultured many types of human cancer cell liens but the fate of apoptosis in murine leukemia cells in vitro and immune responses in leukemic mice remain elusive. Herein, we clarified the actions of DATS on growth inhibition of murine leukemia WEHI-3 cells in vitro and used WEHI-3 cells to generate leukemic mice in vivo, following to investigate the effects of DATS in animal model. In in vitro study, DATS induced apoptosis of WEHI-3 cells through the G0/G1 phase arrest and induction of caspase-3 activation. In in vivo study DATS decreased the weight of spleen of leukemia mice but did not affect the spleen weight of normal mice. DATS promoted the immune responses such as promotions of the macrophage phagocytosis and NK cell activities in WEHI-3 leukemic and normal mice. However, DATS only promotes NK cell activities in normal mice. DATS increases the surface markers of CD11b and Mac-3 in leukemia mice but only promoted CD3 in normal mice. In conclusion, the present study indicates that DATS induces cell death through induction of apoptosis in mice leukemia WHEI-3 cells. DATS also promotes immune responses in leukemia and normal mice in vivo.

Abstract 3670: The role of reactive oxygen species in anticancer effects of diallyl trisulfide, a bioactive component of garlic.

Abstract Diallyl trisulfide (DATS) is a bioactive component of processed garlic with in vivo activity against chemically-induced as well as oncogene-driven cancer in rodent models. However, the mechanisms underlying anticancer effects of DATS (eg, growth inhibition, apoptosis induction, and suppression of migration in cancer cells) are not fully understood. The present study reveals a critical role for reactive oxygen species (ROS) in cellular anticancer responses to DATS. Exposure of human breast cancer cells (MDA-MB-231 and MCF-7) as well as a cell line derived from spontaneously developing mammary tumor of a MMTV-neu transgenic mouse (BRI-JM04) to DATS resulted in dose-dependent inhibition of cell viability and apoptosis induction. Interestingly, a non-tumorigenic normal human mammary cell line (MCF-10A) was resistant to growth inhibition and apoptosis induction by DATS. The DATS-induced apoptosis in MDA-MB-231, MCF-7, and BRI-JM04 cells was associated with ROS production as evidenced by fluorescence microscopy and flow cytometry using a chemical probe (MitoSOX Red). Ectopic expression of Cu,Zn-superoxide dismutase (Cu,Zn-SOD) as well as Mn-SOD conferred significant protection against DATS-induced ROS production and apoptotic cell death. The DATS treatment caused activation of Bax and Bak, and activation of Bak was nearly fully abolished by overexpression of Mn-SOD. Likewise, inhibition of MDA-MB-231 and MCF-7 cell migration resulting from DATS treatment was partially but statistically significantly attenuated by stable overexpression of Cu,Zn-SOD and Mn-SOD. This protection against inhibition of cell migration was attributable to abrogation of DATS-mediated induction of E-cadherin and suppression of vimentin. These results indicate that ROS play an important role in anticancer effects of DATS at least in human breast cancer cells. This study was supported by grant CA113363-07 awarded by the National Cancer Institute. Citation Format: Julie A. Arlotti, Kumar Chandra-Kuntal, Shivendra V. Singh. The role of reactive oxygen species in anticancer effects of diallyl trisulfide, a bioactive component of garlic. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3670. doi:10.1158/1538-7445.AM2013-3670

The DNA injury-repair of esophageal cancer EC109 cells to diallyl trisulfide (DATS) combining radiation

Objective The aim of our study was to investigate the effect of diallyl trisulfide (DATS) combining radiation on DNA injury-repair of Esophageal cancer EC109 cells.

Effect of diallyl trisulfide on the pharmacokinetics of dipyridamole in rats

This study was aimed to evaluate the effect of diallyl trisulfide (DATS), a major component derived from garlic used to inhibit platelet thromboxane formation, on the pharmacokinetics of dipyridamole. Pharmacokinetic parameters of dipyridamole were determined in rats following intragastric (80 mg/kg suspension or 40 mg/kg solution) or intravenous (3 mg/kg) administration of dipyridamole with coadministration (20 mg/kg) and long-term pretreatment of DATS (10 or 20 mg/kg/day for 15 consecutive days). In addition, everted gut sac models were used to assess transepithelial transport of dipyridamole and the effect of DATS on the intestinal absorption of dipyridamole. After coadministration and long-term pretreatment of DATS, significantly lower Cmax and AUC(0-24 h) were observed for intragastric administration of dipyridamole, whereas little change was noted after intravenous dipyridamole administration. After adding DATS (10 and 50 μg/mL) in the everted gut sacs, absorption of dipyridamole was remarkably decreased in the ileum and jejunum (p < 0.01). In conclusion, DATS reduced the oral exposure of dipyridamole in rats likely by the modification of the dissolution rate and intestinal absorption of dipyridamole, indicating that combined use of DATS or DATS-containing supplements with dipyridamole may require caution as low plasma concentrations of dipyridamole may lead to a subtherapeutic effect of this agent.

Sodium 2-propenyl thiosulfate derived from garlic induces phase II detoxification enzymes in rat hepatoma H4IIE cells

There is evidence that onions and garlic protect against cancer in humans. It has been suggested that this effect is partly due to the organosulfur compounds in Allium vegetables and that these substances act through induction of phase II detoxification enzymes. Here, we hypothesized that alk(en)yl thiosulfates, sodium n-propyl thiosulfate (NPTS), and sodium 2-propenyl thiosulfate (2PTS), which were identified in onions and garlic, respectively, may induce phase II enzymes. Therefore, rat hepatoma cells (H4IIE) were cultured with 1 to 100 micromol/L of NPTS or 2PTS for 48 hours at 37 degrees C; and the activities and messenger RNA (mRNA) expression levels of phase II enzymes in H4IIE cells were investigated. The effects of diallyl trisulfide and tert-butylhydroquinone, known as phase II inducers, were also examined as positive controls and compared with the responses of NPTS and 2PTS. Quinone reductase (QR) activity and mRNA expression levels of QR and epoxide hydrolase 1 were significantly increased by 2PTS (P < .05-.005). In particular, QR activity was increased at a relatively low concentration of 2PTS (10 micromol/L). However, glutathione S-transferase activity and mRNA expression levels of glutathione S-transferase A5 and uridine diphosphate glucuronosyl transferase 1A1 were not changed by 2PTS. In contrast, NPTS did not affect the activities and mRNA expression levels of these phase II enzymes. These results show that 2PTS can induce phase II enzymes, and its inductive effect is comparable or superior to that of diallyl trisulfide and tert-butylhydroquinone.

Inhibitory effects of diallyl trisulfide on the growth of hepatocellular carcinoma in rats after orthotopic liver transplantation

Objective To study the effect of diallyl trisulfide (DATS) on the growth of hepato-cellular carcinoma(HCC)after orthotopie liver transplantation(OLTx)in rats. Methods Twenty-four rats bearing subcutaneous xenograft HCC after OLTx were randomly divided into 3 groups: the control group,the DATS group Ⅰ(DATS 1 mg/kg every day) and DATS group Ⅱ(DATS 5 mg/kg every day). The effects of DATS on the growth of the tumor were evaluated after 2 weeks,including the tumor size and weight. Fluorescent quantitation ploymerase chain reaction was used to detect the expression of vascular en-dothelial growth factor (VEGF) mRNA and matrix metalloproteinase 2(MMP-2)mRNA. The effects of DATS on the liver and kidney in rats were assayed by biochemical methods. Results DATS inhibited the growth of HCC in rats after OLTx. Tumor size : the control group(0. 78±0.25)cm3, DATS group Ⅰ (0.42±0.17) cm3 ,and DATS group Ⅱ (0.38±0.21)cm3 (P 0.05 ). Conclusion DATS can inhibit the growth of HCC in rats af-ter OLTx without side effects in vivo by suppressing the expression of VEGF mRNA and MMP-2 mRNA inHCC. Key words: Diallyl trisulfide; Carcinoma,hepatocellular; Vascular endothelial growth factor; Matrix metalloproteinase

Effect of diallyl trisulfide on the activation of T cell and macrophage-mediated cytotoxicity.

At high concentration (50 micrograms/ml), diallyl trisulfide (DATS) had an inhibitory effect on T cell activation (compared with control group, P < 0.05). But at appropriate concentrations (3.125-12.5 micrograms/ml), DATS augmented the activation of T lymphocytes by Con A (compared with control group, P < 0.01). The augmentation of T cell activation by DATS was related to its inhibitory effect on the production of nitric oxide (NO) by macrophages. In a wide range of concentrations (1-100 micrograms/ml), DATS can inhibit the production of NO by macrophages (P < 0.05, P < 0.01). In addition, DATS can antagonize the inhibition of tumor-derived immunosuppressive factors produced by S180 cells and Ehrlich ascitic cancer cells on the activation of T cells, and reduce the inhibitory rate significantly (P < 0.01). DATS, despite its inhibition of the production of NO by macrophages, can significantly enhance the production of hydrogen peroxide (H2O2) by macrophages. When macrophages were pretreated with DATS for 24 h, the cytotoxicity % of macrophages to three tumor cell lines was significantly higher than that in corresponding control group (P < 0.05, P < 0.01). In the presence of both DATS and LPS, the cytotoxicity of macrophages was further enhanced so that the cytotoxicity % of macrophages to tumor cells was significantly higher than either that in the presence of DATS alone or that in the presence of LPS alone (P < 0.05, P < 0.01). These results indicate that DATS can augment the activation of T cells and enhance the anti-tumor function of macrophage, suggesting that DATS may be potentially useful in tumor therapy.