Objective To explore the production and cytotoxicity of the reactive oxygen species(ROS)induced by diallyl trisulfid(DATS)in HL-60 cells.Methods HL-60 cells were either treated with various doses of DATS alone,or DATS...Objective To explore the production and cytotoxicity of the reactive oxygen species(ROS)induced by diallyl trisulfid(DATS)in HL-60 cells.Methods HL-60 cells were either treated with various doses of DATS alone,or DATS combination with Apocynin,a specific NADPH oxidase inhibitor,or with antioxidant N-acetyl-L-cysteine(NAC)for 0,1,3,6,12 and 24 hours,respectively.The intracellular ROS level was measured by flow cytometry.The activity of NADPH oxidase was evaluated by NBT reduction experiment.The content of both malondialdehyde(MDA)and the protein carbonyl was analyzed by spectrophotometer.Results The results from flow cytometry indicated that DATS significantly increased the intracellular ROS level in HL-60 cells(P<0.05),which is a dose-and time-dependent.The fluorescence intensities of ROS reached at maximuam when HL-60 cells were incubated with 150 μmol·L-1 DATS for 3 hours.The NBT reduction experiment showed that DATS activated NADPH oxidase which had highest activity when cell were exposed to 150 μmol·L-1 DATS for 3 hours.Results DATS induced MDA and protein carbonyl production in HL-60 cells.Furrthermore,both MDA and protein carbonyl in the cells exposed to 150 μmol·L-1 DATS for 3 hours reached the highest level.Apocynin and NAC could attenuate the production of MDA and protein carbonyl,which suggested that ROS induced by DATS was involved in the toxicity to cells.Conclusions DATS induce ROS production through activating NADPH oxidase in HL-60 cells.ROS induced by DATS increase the oxidation of the membrane lipid and the protein of HL-60 cell.展开更多
AIM:To investigate the effects of diallyl trisulfide(DATS),a garlic-derived organosulfur compound,in pancreatic cancer cells.METHODS:Human pancreatic cancer cells with wildtype p53 gene(Capan-2)and normal pancreatic e...AIM:To investigate the effects of diallyl trisulfide(DATS),a garlic-derived organosulfur compound,in pancreatic cancer cells.METHODS:Human pancreatic cancer cells with wildtype p53 gene(Capan-2)and normal pancreatic epithelial cells(H6C7)were cultured in RPMI1640.DATS was prepared at a concentration of 100μmol/L.Cell viability was determined via the methyl thiazolyl tetrazolium assay.Apoptotic cells were detected by TUNEL assay.Cell cycle analysis was performed using flow cytometry.Protein expression was determined by Western blot.Bax and Bcl-2 expression was detected by immunofluorescence.Apoptosis genes and cell cycle were assessed by quantitative real-time polymerase chain reaction.RESULTS:DATS suppressed the viability of cultured human pancreatic cancer cells(Capan-2)by increasing the proportion of cells in the G2/M phase and induced apoptotic cell death.Western blot analysis indicated that DATS enhanced the expression of Fas,p21,p53and cyclin B1,but downregulated the expression of Akt,cyclin D1,MDM2 and Bcl-2.DATS induced cell cycle inhibition which was correlated with elevated levels of cyclin B1 and p21,and reduced levels of cyclin D1 in Capan-2 cells and H6C7 cells.DATS-induced apoptosis was markedly elevated in Capan-2 cells compared with H6C7 cells,and this was correlated with elevated levels of cyclin B1 and p53,and reduced levels of Bcl-2.DATS-induced apoptosis was correlated with downregulation of Bcl-2,Akt and cyclin D1 protein levels,and up-regulation of Bax,Fas,p53 and cyclin B protein levels in Capan-2 cells.CONCLUSION:DATS induces apoptosis of pancreatic cancer cells(Capan-2)and non-tumorigenic pancreatic ductal epithelial cells(H6C7).展开更多
Breast cancer is one of the leading causes of cancer-related deaths in women worldwide.It is a cancer that originates from the mammary ducts and involves mutations in multiple genes.Recently,the treatment of breast ca...Breast cancer is one of the leading causes of cancer-related deaths in women worldwide.It is a cancer that originates from the mammary ducts and involves mutations in multiple genes.Recently,the treatment of breast cancer has become increasingly challenging owing to the increase in tumor heterogeneity and aggressiveness,which gives rise to therapeutic resistance.Epidemiological,populationbased,and hospital-based case-control studies have demonstrated an association between high intake of certain Allium vegetables and a reduced risk in the development of breast cancer.Diallyl disulfide(DADS)and diallyl trisulfide(DATS)are the main allyl sulfur compounds present in garlic,and are known to exhibit anticancer activity as they interfere with breast cancer cell proliferation,tumor metastasis,and angiogenesis.The present review highlights multidrug resistance mechanisms and their signaling pathways in breast cancer.This review discusses the potential anticancer activities of DADS and DATS,with emphasis on drug resistance in triple-negative breast cancer(TNBC).Understanding the anticancer activities of DADS and DATS provides insights into their potential in targeting drug resistance mechanisms of TNBC,especially in clinical studies.展开更多
At high concentration (50 μg/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 μg/ml). DATS augmente...At high concentration (50 μg/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 μg/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 μg/ml) , DATS can inhibit the production of NO by macrophas,es (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 DAIS 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.展开更多
The protective effects of diallyl trisulfide on liver were examined in rats with sepsis. Sepsis was reproduced in rats by cecum ligation and puncture (CLP). Fifty-six male Wistar rats were randomly divided into sham-o...The protective effects of diallyl trisulfide on liver were examined in rats with sepsis. Sepsis was reproduced in rats by cecum ligation and puncture (CLP). Fifty-six male Wistar rats were randomly divided into sham-operated group (group S, n=8), sepsis model group (group C, n=24), diallyl trisulfide (DATS)-treated group (group D, n=24). Animals in groups C and D were further divided into three subgroups according to different observation time points, with 8 rats in each sub-group.Rats in group D and C were intravenously injected with normal saline or DATS respectively at a dose of 20 mg/kg after the establishment of sepsis model. Eight rats in groups C and D were sacrificed at 3, 6 and 24 h post-CLP and their livers were harvested for detection of interleukin (IL)-1 receptor associated kinase-4 (IRAK-4), nuclear factor-κB (NF-κB), c-fos, c-jun, malondialdehydethhe (MDA) and superoxide dismutase (SOD), tumor necrosis factor alpha (TNF-α) and for pathological examination. The results showed that the levels of serum IRAK-4, NF-κB and TNF-α in hepatic tissues were higher in group C than group S (control group) (P<0.05). After DATS treatment, the levels of IRAK-4 and NF-κB in the hepatic tissues and serum TNF-α in group D were lower than those in group C (P<0.05). The levels of c-fos and c-jun and MDA in the hepatic tissues were higher in group C than in group S (P<0.05). After DATS treatment, the levels of c-fos and c-jun and MDA in the hepatic tissues were significantly lower in group D than in group C (P<0.05). When compared with group S group, concentration of SOD in the hepatic tissues in group C was significantly lower (P<0.05). After DATS treatment, the concentration of SOD in the hepatic tissues was higher in group D than in group C (P<0.05). These findings suggested that treatment with DATS could ameliorate sepsis-induced liver injury in rats. The protective effect might be related to its ability to inhibit the signal pathway of IRAK-4 and NF-κB, thereby decreasing the production of oxygen free radicals and down-regulating the expression of c-fos and c-jun.展开更多
Objective To explore the effect and mechanism of diallyl trisulfide on the activity of NADPH oxidase in Hl-60 cells.Methods HL-60 cells were treated with DATS at a indicated concentration for 0,1,3,6,12 hours,respecti...Objective To explore the effect and mechanism of diallyl trisulfide on the activity of NADPH oxidase in Hl-60 cells.Methods HL-60 cells were treated with DATS at a indicated concentration for 0,1,3,6,12 hours,respectively.The activity of NADPH oxidase was measured by the reduction of the yellow dye nitroblue tetrazolium(NBT).The mRNA expression of NADPH oxidase subunits,including gp91phox,p47 phox,p22 phox,Rac2 and Rac1,was detected by RT-PCR.The protein expression of p67 phox,gp91 phox and Rac2 was analyzed by Western blot.The cell membrane fractions were prepared according to the instruction of Mem-PER kit from Pierce Corp.Results The results showed that reduction ability of HL-60 cells for NBT markedly increased in a concentration-dependent manner following DATS incubation for 3 and 6 hours(P<0.05).HL-60 cells treated by DATS at a concentration of 150 μM for 3 hours have a maximal reduction effect for NBT.The results from RT-PCR indicated that mRNA expression of NADPH oxidase subunits,including p47phox,gp91 phox,p22 phox,Rac2 and Rac1,significantly increased in a concentration-dependent manner in HL-60 cells treated by DATS.The results from western blot showed that HL-60 cells following DATS incubation have a higher level expression of Rac2 and gp91phox,compared with untreated-HL-60 cells.Our results also indicated that a maximal expression level of p47phox,gp91 phox,p22 phox,Rac2 and Rac1in HL-60 cells is present at 3 hours following DATS incubation.We found that levels of both Rac2 and p67phox was reduced in the cytosolic fraction and meanwhile increased in the membrane fraction following HL-60 exposed to DATS,Which is dependent on the concentration and time of DATS treatment.Furthermore,the level of both Rac2 and p67phox located to the plasma membrane translocation was maximized following 150 μM of DATS incubation for 3 hours.Conclusions DATS induce the activation of NADPH oxidase by both up-regulating the expression of NADPH oxidase subunit and translocating the cytosolic Rac2 and p67 phox subunit to the plasma membrane in HL-60 cells.展开更多
Mosl of anticancer drugs are mutagenic. A possible exeeption is diallyl .trisulfide(DAT ), a component of garlic. It is an antimutagenic anticunccr chemical although it ismainly uscd as antibiotic. Its modifying eff...Mosl of anticancer drugs are mutagenic. A possible exeeption is diallyl .trisulfide(DAT ), a component of garlic. It is an antimutagenic anticunccr chemical although it ismainly uscd as antibiotic. Its modifying effeci on induction of UDS by mutagenicmitomycin C (MMC), cyclophosphamide (CP) and cis-diamine dichloroplatin (DDP) was invcstigiltcd with the UDS assay in the primary cultures of Wistar rat hepatocytes (hpc)using the autoradiographic technique. Resultsshowed that 1.0-4.0 nmol/ml of DAT didnot inducc UDS and that MMC, CP and DDP resulted in a significant induction ofdosc-dependent UDS. DAT enhanced induction of UDS by these drugs. A dose-effectrclationship was observed betwecn dose of DAT and enhancement of induction of UDS.Howcvcr, thc mcchanism of the enhancement is not clear.展开更多
文摘Objective To explore the production and cytotoxicity of the reactive oxygen species(ROS)induced by diallyl trisulfid(DATS)in HL-60 cells.Methods HL-60 cells were either treated with various doses of DATS alone,or DATS combination with Apocynin,a specific NADPH oxidase inhibitor,or with antioxidant N-acetyl-L-cysteine(NAC)for 0,1,3,6,12 and 24 hours,respectively.The intracellular ROS level was measured by flow cytometry.The activity of NADPH oxidase was evaluated by NBT reduction experiment.The content of both malondialdehyde(MDA)and the protein carbonyl was analyzed by spectrophotometer.Results The results from flow cytometry indicated that DATS significantly increased the intracellular ROS level in HL-60 cells(P<0.05),which is a dose-and time-dependent.The fluorescence intensities of ROS reached at maximuam when HL-60 cells were incubated with 150 μmol·L-1 DATS for 3 hours.The NBT reduction experiment showed that DATS activated NADPH oxidase which had highest activity when cell were exposed to 150 μmol·L-1 DATS for 3 hours.Results DATS induced MDA and protein carbonyl production in HL-60 cells.Furrthermore,both MDA and protein carbonyl in the cells exposed to 150 μmol·L-1 DATS for 3 hours reached the highest level.Apocynin and NAC could attenuate the production of MDA and protein carbonyl,which suggested that ROS induced by DATS was involved in the toxicity to cells.Conclusions DATS induce ROS production through activating NADPH oxidase in HL-60 cells.ROS induced by DATS increase the oxidation of the membrane lipid and the protein of HL-60 cell.
基金Supported by Fund for Science and Technology Program of Shaanxi Province,China,No.2010K01-141
文摘AIM:To investigate the effects of diallyl trisulfide(DATS),a garlic-derived organosulfur compound,in pancreatic cancer cells.METHODS:Human pancreatic cancer cells with wildtype p53 gene(Capan-2)and normal pancreatic epithelial cells(H6C7)were cultured in RPMI1640.DATS was prepared at a concentration of 100μmol/L.Cell viability was determined via the methyl thiazolyl tetrazolium assay.Apoptotic cells were detected by TUNEL assay.Cell cycle analysis was performed using flow cytometry.Protein expression was determined by Western blot.Bax and Bcl-2 expression was detected by immunofluorescence.Apoptosis genes and cell cycle were assessed by quantitative real-time polymerase chain reaction.RESULTS:DATS suppressed the viability of cultured human pancreatic cancer cells(Capan-2)by increasing the proportion of cells in the G2/M phase and induced apoptotic cell death.Western blot analysis indicated that DATS enhanced the expression of Fas,p21,p53and cyclin B1,but downregulated the expression of Akt,cyclin D1,MDM2 and Bcl-2.DATS induced cell cycle inhibition which was correlated with elevated levels of cyclin B1 and p21,and reduced levels of cyclin D1 in Capan-2 cells and H6C7 cells.DATS-induced apoptosis was markedly elevated in Capan-2 cells compared with H6C7 cells,and this was correlated with elevated levels of cyclin B1 and p53,and reduced levels of Bcl-2.DATS-induced apoptosis was correlated with downregulation of Bcl-2,Akt and cyclin D1 protein levels,and up-regulation of Bax,Fas,p53 and cyclin B protein levels in Capan-2 cells.CONCLUSION:DATS induces apoptosis of pancreatic cancer cells(Capan-2)and non-tumorigenic pancreatic ductal epithelial cells(H6C7).
基金supported by UGC-DAE-CSR,Kolkata(Grant No.:KC/CRS/19/RB-04/1047).
文摘Breast cancer is one of the leading causes of cancer-related deaths in women worldwide.It is a cancer that originates from the mammary ducts and involves mutations in multiple genes.Recently,the treatment of breast cancer has become increasingly challenging owing to the increase in tumor heterogeneity and aggressiveness,which gives rise to therapeutic resistance.Epidemiological,populationbased,and hospital-based case-control studies have demonstrated an association between high intake of certain Allium vegetables and a reduced risk in the development of breast cancer.Diallyl disulfide(DADS)and diallyl trisulfide(DATS)are the main allyl sulfur compounds present in garlic,and are known to exhibit anticancer activity as they interfere with breast cancer cell proliferation,tumor metastasis,and angiogenesis.The present review highlights multidrug resistance mechanisms and their signaling pathways in breast cancer.This review discusses the potential anticancer activities of DADS and DATS,with emphasis on drug resistance in triple-negative breast cancer(TNBC).Understanding the anticancer activities of DADS and DATS provides insights into their potential in targeting drug resistance mechanisms of TNBC,especially in clinical studies.
文摘At high concentration (50 μg/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 μg/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 μg/ml) , DATS can inhibit the production of NO by macrophas,es (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 DAIS 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.
基金supported by grants from the Natural Science Foundation of Hubei Province (No. 2011CDB196)
文摘The protective effects of diallyl trisulfide on liver were examined in rats with sepsis. Sepsis was reproduced in rats by cecum ligation and puncture (CLP). Fifty-six male Wistar rats were randomly divided into sham-operated group (group S, n=8), sepsis model group (group C, n=24), diallyl trisulfide (DATS)-treated group (group D, n=24). Animals in groups C and D were further divided into three subgroups according to different observation time points, with 8 rats in each sub-group.Rats in group D and C were intravenously injected with normal saline or DATS respectively at a dose of 20 mg/kg after the establishment of sepsis model. Eight rats in groups C and D were sacrificed at 3, 6 and 24 h post-CLP and their livers were harvested for detection of interleukin (IL)-1 receptor associated kinase-4 (IRAK-4), nuclear factor-κB (NF-κB), c-fos, c-jun, malondialdehydethhe (MDA) and superoxide dismutase (SOD), tumor necrosis factor alpha (TNF-α) and for pathological examination. The results showed that the levels of serum IRAK-4, NF-κB and TNF-α in hepatic tissues were higher in group C than group S (control group) (P<0.05). After DATS treatment, the levels of IRAK-4 and NF-κB in the hepatic tissues and serum TNF-α in group D were lower than those in group C (P<0.05). The levels of c-fos and c-jun and MDA in the hepatic tissues were higher in group C than in group S (P<0.05). After DATS treatment, the levels of c-fos and c-jun and MDA in the hepatic tissues were significantly lower in group D than in group C (P<0.05). When compared with group S group, concentration of SOD in the hepatic tissues in group C was significantly lower (P<0.05). After DATS treatment, the concentration of SOD in the hepatic tissues was higher in group D than in group C (P<0.05). These findings suggested that treatment with DATS could ameliorate sepsis-induced liver injury in rats. The protective effect might be related to its ability to inhibit the signal pathway of IRAK-4 and NF-κB, thereby decreasing the production of oxygen free radicals and down-regulating the expression of c-fos and c-jun.
文摘Objective To explore the effect and mechanism of diallyl trisulfide on the activity of NADPH oxidase in Hl-60 cells.Methods HL-60 cells were treated with DATS at a indicated concentration for 0,1,3,6,12 hours,respectively.The activity of NADPH oxidase was measured by the reduction of the yellow dye nitroblue tetrazolium(NBT).The mRNA expression of NADPH oxidase subunits,including gp91phox,p47 phox,p22 phox,Rac2 and Rac1,was detected by RT-PCR.The protein expression of p67 phox,gp91 phox and Rac2 was analyzed by Western blot.The cell membrane fractions were prepared according to the instruction of Mem-PER kit from Pierce Corp.Results The results showed that reduction ability of HL-60 cells for NBT markedly increased in a concentration-dependent manner following DATS incubation for 3 and 6 hours(P<0.05).HL-60 cells treated by DATS at a concentration of 150 μM for 3 hours have a maximal reduction effect for NBT.The results from RT-PCR indicated that mRNA expression of NADPH oxidase subunits,including p47phox,gp91 phox,p22 phox,Rac2 and Rac1,significantly increased in a concentration-dependent manner in HL-60 cells treated by DATS.The results from western blot showed that HL-60 cells following DATS incubation have a higher level expression of Rac2 and gp91phox,compared with untreated-HL-60 cells.Our results also indicated that a maximal expression level of p47phox,gp91 phox,p22 phox,Rac2 and Rac1in HL-60 cells is present at 3 hours following DATS incubation.We found that levels of both Rac2 and p67phox was reduced in the cytosolic fraction and meanwhile increased in the membrane fraction following HL-60 exposed to DATS,Which is dependent on the concentration and time of DATS treatment.Furthermore,the level of both Rac2 and p67phox located to the plasma membrane translocation was maximized following 150 μM of DATS incubation for 3 hours.Conclusions DATS induce the activation of NADPH oxidase by both up-regulating the expression of NADPH oxidase subunit and translocating the cytosolic Rac2 and p67 phox subunit to the plasma membrane in HL-60 cells.
文摘Mosl of anticancer drugs are mutagenic. A possible exeeption is diallyl .trisulfide(DAT ), a component of garlic. It is an antimutagenic anticunccr chemical although it ismainly uscd as antibiotic. Its modifying effeci on induction of UDS by mutagenicmitomycin C (MMC), cyclophosphamide (CP) and cis-diamine dichloroplatin (DDP) was invcstigiltcd with the UDS assay in the primary cultures of Wistar rat hepatocytes (hpc)using the autoradiographic technique. Resultsshowed that 1.0-4.0 nmol/ml of DAT didnot inducc UDS and that MMC, CP and DDP resulted in a significant induction ofdosc-dependent UDS. DAT enhanced induction of UDS by these drugs. A dose-effectrclationship was observed betwecn dose of DAT and enhancement of induction of UDS.Howcvcr, thc mcchanism of the enhancement is not clear.
