Composition, stability, and bioavailability of garlic products used in a clinical trial

In support of a new clinical trial designed to compare the effects of crushed fresh garlic and two types of garlic supplement tablets (enteric-coated dried fresh garlic and dried aged garlic extract) on serum lipids, the three garlic products have been characterized for (a) composition (14 sulfur and 2 non-sulfur compounds), (b) stability of suspected active compounds, and (c) availability of allyl thiosulfinates (mainly allicin) under both simulated gastrointestinal (tablet dissolution) conditions and in vivo. The allyl thiosulfinates of blended fresh garlic were stable for at least 2 years when stored at -80 degrees C. The dissolution release of thiosulfinates from the enteric-coated garlic tablets was found to be >95%. The bioavailability of allyl thiosulfinates from these tablets, measured as breath allyl methyl sulfide, was found to be complete and equivalent to that of crushed fresh garlic. S-Allylcysteine was stable for 12 months at ambient temperature. The stability of the suspected active compounds under the conditions of the study and the bioavailability of allyl thiosulfinates from the dried garlic supplement have validated the use of these preparations for comparison in a clinical trial.     

Low allicin release from garlic supplements: a major problem due to the sensitivities of alliinase activity

Most garlic supplements are standardized on allicin potential and are enteric-coated to prevent gastric acid inactivation of the allicin-producing enzyme, alliinase. To determine whether these products release the claimed amount of allicin under simulated gastrointestinal conditions, USP dissolution method 724A for drug release was applied to all 24 known brands of enteric-coated tablets. It was found that nearly all brands employed effective coatings and that they met their claims for allicin potential when crushed and suspended in water. However, all brands except one gave low dissolution allicin release, with 83% of the brands releasing less than 15% of their potential. The low allicin release was found to be due to both impaired alliinase activity, mostly caused by tablet excipients, and to slow tablet disintegration, which also impairs alliinase activity. Only when tablets had high alliinase activity and disintegrated rapidly did they show high allicin release. The ability of USP 724A to estimate allicin release in vivo was validated by monitoring breath levels of the allicin metabolite, allyl methyl sulfide. In conclusion, garlic powder supplements should no longer be standardized on allicin potential, but rather on dissolution allicin release.     

Biological and chemical stability of garlic-derived allicin

This study verifies the instability of garlic ( Allium sativum L.)-derived allyl 2-propenylthiosulfinate (allicin) in various aqueous and ethanolic solutions as well as in vegetable oil through chemical and biological analyses performed simultaneously. Crushed fresh garlic cloves generated antibacterial activity and chemically detectable allicin, a major antibacterial principle, and both declined on a daily basis in aqueous and ethanolic solutions at room temperature, showing biological and chemical half-lives of about 6 and 11 days, respectively. Allicin was more stable in 20% alcohol than in water, but surprisingly unstable in vegetable oil, with an activity half-life 0.8 h, as estimated from its antibacterial activity toward Escherichia coli, and a chemical half-life of 3.1 h, based on chromatographic quantification. In alcoholic and aqueous extracts, the biological half-life of allicin tended to be longer than the chemical one, suggesting the occurrence of bioactive compounds other than allicin in the extracts.     

Quantitative determination of allicin in garlic: supercritical fluid extraction and standard addition of alliin

A quantitative method is described for the determination of allicin (2-propene-1-sulfinothioic acid S-2-propenyl ester) in garlic, using standard additions of alliin (l-(+)-S-allylcysteine sulfoxide) in conjunction with supercritical fluid extraction (SFE) and high performance liquid chromatography analysis with UV-vis absorbance detection. Optimum CO(2)-SFE conditions provided 96% recovery for allicin with precision of 3% (RSD) for repeat samples. The incorporation of an internal standard (allyl phenyl sulfone) in the SFE step resulted in a modest improvement in recovery (99%) and precision (2% RSD). Standard additions of alliin were converted to allicin in situ by endogenous alliinase (l-(+)-S-alk(en)ylcysteine sulfoxide lyase, EC 4.4.1.4). Complete conversion of the spiked alliin to allicin was achieved by making additions after homogenization-induced conversion of the naturally occurring cysteine sulfoxides to thiosulfinates had taken place, thus eliminating the likelihood of competing reactions. Concentration values for allicin determined in samples of fresh garlic (Allium sativum L. and Allium ampeloprasum) and commercially available garlic powders (Allium sativum L.) by standard addition of alliin were found in all cases to be in statistical agreement (95% confidence interval) with values determined using a secondary allicin standard (concentration determined using published extinction coefficients). This method provides a convenient alternative for assessing the amount of allicin present in fresh and powdered garlic, as alliin is a far more stable and commercially prevalent compound than allicin and is thus more amenable for use as a standard for routine analysis.     

Effect of cooking on garlic (Allium sativum L.) antiplatelet activity and thiosulfinates content

The raw form of garlic and some of its preparations are widely recognized as antiplatelet agents that may contribute to the prevention of cardiovascular disease. Herein, we examined the in-vitro antiaggregatory activity (IVAA) of human blood platelets induced by extracts of garlic samples that were previously heated (in the form of crushed versus uncrushed cloves) using different cooking methods and intensities. The concentrations of allicin and pyruvate, two predictors of antiplatelet strength, were also monitored. Oven-heating at 200 degrees C or immersing in boiling water for 3 min or less did not affect the ability of garlic to inhibit platelet aggregation (as compared to raw garlic), whereas heating for 6 min completely suppressed IVAA in uncrushed, but not in previously crushed, samples. The latter samples had reduced, yet significant, antiplatelet activity. Prolonged incubation (more than 10 min) at these temperatures completely suppressed IVAA. Microwaved garlic had no effect on platelet aggregation. However, increasing the concentration of garlic juice in the aggregation reaction had a positive IVAA dose response in crushed, but not in uncrushed, microwaved samples. The addition of raw garlic juice to microwaved uncrushed garlic restored a full complement of antiplatelet activity that was completely lost without the garlic addition. Garlic-induced IVAA was always associated with allicin and pyruvate levels. Our results suggest that (1) allicin and thiosulfinates are responsible for the IVAA response, (2) crushing garlic before moderate cooking can reduce the loss of activity, and (3) the partial loss of antithrombotic effect in crushed-cooked garlic may be compensated by increasing the amount consumed.     

Modulation of cytokine secretion by garlic oil derivatives is associated with suppressed nitric oxide production in stimulated macrophages

We previously described that garlic oil derivatives differentially suppress the production of nitric oxide (NO) and prostaglandin E(2) (PGE(2)) in activated macrophages. In the present study, we investigated the effects of the garlic derivatives, diallyl sulfide (DAS), diallyl disulfide (DADS), and allyl methyl sulfide (AMS), on cytokine production in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells, and the association between modulation of cytokines and inhibition of NO production was also assessed. The results indicated that these garlic compounds had different effects on the secretion of activated cytokines, including proinflammatory tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1beta, and IL-6, as well as the antiinflammatory, IL-10. DAS inhibited the production of all stimulated cytokines in a concentration-dependent manner, and the inhibition was closely associated with the suppression of NO and PGE(2) production. DADS repressed the production of stimulated TNF-alpha and IL-10 and increased the production of activated IL-1beta and, to a lesser extent, IL-6; but only the decreased IL-10 production was associated with DADS-induced NO inhibition. Yet, the DAS- and DADS-suppressed NO production was independent of TNF-alpha. AMS, on the other hand, slightly suppressed the stimulated TNF-alpha but enhanced IL-10 production, and such modulation was closely associated with the decrease in NO production.     

Identification of two novel pigment precursors and a reddish-purple pigment involved in the blue-green discoloration of onion and garlic

By using a model reaction system representing blue-green discoloration that occurs when purees of onion (Allium cepa L.) and garlic (Allium sativum L.) are mixed, we isolated two pigment precursors (PPs) and a reddish-purple pigment (PUR-1) and determined their chemical structures. PPs were isolated from a heat-treated solution containing color developer (CD) and either l-valine or l-alanine, and their structures were determined as 2-(3,4-dimethylpyrrolyl)-3-methylbutanoic acid (PP-Val), and 2-(3,4-dimethyl-1H-pyrrolyl) propanoic acid (PP-Ala), respectively. Next, PUR-1 was isolated from a heat-treated solution containing PP-Val and allicin, and its structure was determined as (1E)-1-(1-((1S)-1-carboxy-2-methylpropyl)-3,4-dimethyl-1H-pyrrol-2-yl)-prop-1-enylene-3-(1-((1S)-1-carboxy-2-methylpropyl)-3,4-dimethyl-1H-pyrrol-2-ylidenium). The structure of PUR-1 suggested that PP molecules containing a 3,4-dimethyl pyrrole ring had been cross-linked by an allyl group of allicin to form conjugated pigments. While PUR-1 is a dipyrrole compound exhibiting a reddish-purple color, a color shift toward blue to green can be expected as the cross-linking reaction continues to form, for example, tri- or tetrapyrrole compounds.     

Comparison of the bioactive compounds and antioxidant potentials of fresh and cooked Polish, Ukrainian, and Israeli garlic

Garlic (Allium sativum L.) is an essential part of Polish, Ukrainian, and Israeli cuisine. The aim of this investigation was to compare the changes in bioactive compounds, proteins, and antioxidant potentials in fresh Polish, Ukrainian, and Israeli garlic samples after subjection to cooking temperature. Dietary fiber and essential trace elements were comparable. The antioxidant potentials were determined by four scavenging methods using beta-carotene, 1,1-diphenyl-2-picrylhydrazyl (DPPH), nitric oxide (NO), and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS(*)(+)) radical cation with K(2)S(2)O(8) or MnO(2) assays. Polyphenols, tocopherols, proteins, and antioxidant potentials were higher in Polish garlic, but not significantly (P > 0.05). The SDS- and native-PAGE electrophoretic patterns of all three fresh garlic samples were without significant differences. Most of the proteins were in the molecular mass range of 24-97 kDa, and the more intensive major bands were concentrated at 50 and 12 kDa. The 50 kDa protein nearly disappears and the intensity of the 12 kDa lectin bands slightly decreases during cooking. It was observed that the bioactive compounds, antioxidant potential, and proteins in garlic decrease significantly after 20 min of cooking at 100 degrees C (P < 0.05). In conclusion, (a) the bioactive compounds, electrophoretic patterns, and antioxidant potential of fresh Polish, Ukrainian, and Israeli garlic samples are comparable; (b) garlic samples subjected to 100 degrees C during 20 min preserve their bioactive compounds, antioxidant potential, and protein profile and are comparable with fresh garlic; and (c) fresh garlic should be added to dishes cooked at 100 degrees C in the last 20 min of the cooking process.     

Deodorization with ku-ding-cha containing a large amount of caffeoyl quinic acid derivatives

Caffeoyl quinic acid (CQA) derivatives in ku-ding-cha, mate, coffee, and related plants were determined by HPLC. One ku-ding-cha contained a large amount of 3,5-dicaffeoylquinic acid (3,5-diCQA, 10.6% in dry weight) as well as 3-CQA (1.7%), 4-CQA (1.1%), 5-CQA (6.3%), 3,4-diCQA (1.8%), and 4,5-diCQA (4.3%). In this ku-ding-cha, the total caffeic acid moiety was 90.3 mmol/100 g of dry weight. The leaves of Ilex latifolia, which is one original species of ku-ding-cha, and another plant of the same genus, I. rotunda, also contained 3,5-diCQA (9.5 and 14.6%), 3-CQA (4.3 and 1.9%), and 5-CQA (4.8 and 3.8%), respectively, whereas raw coffee bean contained 5.5% 5-CQA and other low CQA derivatives. 3,5-DiCQA and 5-CQA with an apple acetone powder (AP) containing polyphenol oxidase showed high capturing activities toward thiols, and two addition compounds between 3,5-diCQA and methane thiol were also identified. Ku-ding-cha indicated extremely strong capturing activities toward methanethiol, propanethiol, and 2-propenethiol in the presence of apple AP. Furthermore, drinking ku-ding-cha reduced the amount of allyl methyl sulfide gas, well-known to persist as malodorous breath long after the ingestion of garlic.     

紫色土增施单质硫对大蒜生长发育和硫素营养的影响

对紫色土施用单质硫条件下大蒜生长发育及硫素营养吸收和代谢进行了研究。结果表明,大蒜株高、叶面积、经济产量(蒜头)和经济系数以中等供硫水平最高,而茎粗和生物产量则以高硫水平最大。大蒜能耐高浓度的硫素供应,在0～120kghm-2供硫情况下,其全硫(TS)、水溶性硫(SS)和无机硫(Io-S)含量均随供硫量的增加而上升,小分子水溶性含硫氨基酸含量(Ws-S)以低硫水平最高,而大分子蛋白质硫含量(Wis-S)以中等供硫水平时最高,与大蒜素含量的变化一致。大蒜素含量与不同硫组分比率的关系分析发现,大蒜素含量与Ws-S/TS呈极显著的正相关(r=0.752 ),与Io-S/SS呈显著的正相关(r=0.702 )。全氮含量与全硫含量变化趋势一致,全磷含量以低硫水平时最高,全钾含量以中等供硫水平时最高。     

Differential inhibition of human platelet aggregation by selected Allium thiosulfinates

Thiosulfinates (TSs) have been implicated as a principle source of the antiplatelet property of raw onion and garlic juice. The in vitro responses of human platelets to dosages of four TSs were measured using whole blood aggregometry and compared by regression analysis. Of the compounds evaluated, methyl methane-TS (MMTS), propyl propane-TS (PPTS), and 2-propenyl 2-propene-TS (allicin) are present in freshly cut Allium vegetables, whereas ethyl ethane-TS (EETS) has not been detected. All TSs were synthesized using a model reaction system. PPTS and allicin had the strongest antiplatelet activity at 0.4 mM, inhibiting aggregation by 90 and 89%, respectively. At the same concentration, EETS and MMTS were significantly weaker, inhibiting 74 and 26%, respectively. Combinations of TSs were not additive in their inhibition of aggregation, indicating that the antiplatelet potential of Allium extracts cannot be easily predicted by quantifying organosulfur components. EETS, PPTS, and allicin were significantly more potent platelet inhibitors than aspirin at nearly equivalent concentrations.     

Isolation and identification of organosulfur compounds oxidizing canine erythrocytes from garlic (Allium sativum)

Five compounds oxidizing canine erythrocytes were isolated from an aqueous ethanol garlic extract by silica gel column chromatography and preparative thin-layer chromatography. On the basis of nuclear magnetic resonance, infrared spectroscopy, and mass spectrometry, they were identified as three known compounds: bis-2-propenyl trisulfide (1), bis-2-propenyl tetrasulfide (2), and bis-2-propenyl pentasulfide (3) as well as two novel compounds, bis-2-propenyl thiosulfonate (4) and trans-sulfuric acid allyl ester 3-allylsulfanyl-allyl ester (5). A mixture of compounds 1-3 and compounds 4 and 5 induced methemoglobin formation in canine erythrocyte suspension in vitro resulting in the oxidation of canine erythrocytes. These groups of characteristic organosulfur compounds contained in garlic probably contribute to oxidations in blood. The constituents of garlic have the potential to oxidize erythrocytes and hemoglobin, suggesting that foods containing quantities of garlic should be avoided for feeding dogs.     

Differential effects of garlic oil and its three major organosulfur components on the hepatic detoxification system in rats.

The objective of this study was to compare the modulatory effect of garlic oil and its three organosulfur compounds, diallyl sulfide (DAS), diallyl disulfide (DADS), and diallyl trisulfide (DATS), on rat hepatic detoxification enzyme activity, and protein and mRNA expression. Rats were orally administered garlic oil (80 or 200 mg/kg bw), DAS (20 or 80 mg/kg bw), DADS (80 mg/kg bw), or DATS (70 mg/kg bw) three times a week for 6 weeks. Control rats received corn oil. According to the results, garlic oil and DAS in dosages of 200 and 80 mg/kg bw, respectively, significantly increased pentoxyresorufin O-dealkylase (PROD) activity as compared with the that of the control rats (P < 0.05). In contrast, N-nitrosodimethylamine demethylase activity in rats that received DADS and DATS was significantly lower than that in the control rats (P < 0.05). Ethoxyresorufin O-deethylase and erythromycin demethylase activities were not influenced by garlic oil, DAS, DADS, or DATS. To the phase II enzyme, garlic oil, DADS, and DATS significantly increased the glutathione S-transferase (GST) activity toward ethacrynic aicd (P < 0.05). Immunoblot assay showed that the protein contents of cytochrome P450 1A1, 2B1, and 3A1 were increased by garlic oil and each of three allyl sulfides, and the change among the allyl sulfides was in the order of DAS > DADS > DATS. The placental form of GST (PGST) level was also increased by garlic oil and the three allyl sulfides, but the increase among the allyl sulfides was DATS congruent with DADS > DAS. P450 2E1, however, was suppressed by each garlic component. Northern blot results indicated that the changes in P450 1A1, 2B1, 3A1, and PGST mRNA levels by garlic components were similar to those noted in the protein levels. These results indicate that the modulatory effect of garlic oil on hepatic drug-metabolizing enzymes can be attributed to its three major allyl sulfide components DAS, DADS, and DATS. These three allyl sulfides vary in modulatory activity, and this variation is related to the number of sulfur atoms in the molecule.     

生物质炭浸提液对大蒜生长品质及土壤的影响

[目的]证实生物质炭中的水溶性有机碳对作物生长和品质的作用,为生物质炭的应用开辟新的途径。[方法]在实验室制取400,450,500℃小麦秸秆生物质炭,以KOH溶液浸提水溶性有机碳获得生物质炭浸提液W_(400),W_(450),W_(500),并应用于盆栽大蒜试验,研究其对大蒜生长、品质及土壤的影响。[结果](1)W_(400),W_(450),W_(500)对大蒜的生长都没有显著影响;(2)W_(400)处理使大蒜可溶性糖的含量显著提高了27.53%,土壤有机质和速效磷含量分别提高37%,26%;(3)W_(450)处理下大蒜维生素C、大蒜素含量分别显著提高34.9%和8.2%。[结论]生物质炭浸提液对大蒜生长没有影响,但可以提高大蒜品质。     

硒对大蒜生理特性、含硒量及品质的影响

以金蒜3号为试材,采用盆栽试验,通过叶面喷施不同浓度、不同次数的亚硒酸钠研究了硒对大蒜生理特性、含硒量和品质的影响。结果表明:在生长期4月5日、15日和25日累计喷施1次、2次、3次3个浓度（5、10、15 mg/L）的亚硒酸钠溶液,均不同程度影响大蒜生理特性、含硒量和品质。与对照相比,叶面喷施2次10 mg/L的亚硒酸钠溶液,可显著提高叶片中光合色素含量和光合性能;能显著改善蒜薹和鳞茎品质,单薹鲜质量和单头鲜质量分别提高68.25%和29.00%,可溶性糖、可溶性蛋白、维生素C含量分别提高73.05%、104.66%、18.95%和82.01%、51.27%、69.82%;并可有效提高鳞茎中游离氨基酸含量（38.91%）,降低蒜薹中游离氨基酸含量（36.95%）;同时可降低蒜薹和鳞茎中大蒜素含量,但差异不显著。蒜薹和鳞茎硒含量随硒浓度和喷施次数的增加而显著提高,最高可达19.81和23.96 mg/kg,为对照的3.85和4.41倍。综合各因素指标,大蒜以叶面喷施2次10 mg/L的亚硒酸钠为宜。     

硫对青蒜生长及品质的影响

以金乡白皮蒜为材料,采用Hoagland营养液水培方法,探究硫对青蒜生长发育、品质及主要矿质元素吸收的影响。结果表明:硫可显著提高青蒜植株的叶片数、叶长和叶面积;当硫处理浓度为1.50 mmol/L时,叶片中大蒜素含量最高,较不施硫处理增加97.96%;假茎中大蒜素含量以硫浓度为2.25 mmol/L时最高;当营养液中硫处理浓度为2.25 mmol/L时,青蒜叶片和假茎内可溶性糖和维生素C含量最大,可溶性糖比不施硫处理分别增加2.64和0.53个百分点,维生素C含量分别增加14.98%和24.57%;青蒜植株（叶片和假茎）中全氮和全硫含量随着营养液硫浓度的升高而增加,呈显著正相关;全磷和全钾含量以硫浓度为2.25 mmol/L时最高。总之,硫能显著促进青蒜叶片生长,提高青蒜的营养品质,增加矿质元素的积累,营养液硫处理浓度为2.25 mmol/L时各项指标表现最好。     

适宜的氮硫配施提高基质栽培大蒜鳞茎品质

试验采用蛭石-珍珠岩盆栽方式,研究了不同浓度氮(5、10、20 mmol/L,记作N1、N2、N3)、硫(2、4、8 mmol/L,记作S1、S2、S3)配施条件对大蒜鳞茎品质的影响,并采用隶属函数法对大蒜品质进行综合评价,分析鳞茎品质与氮硫互作水平的响应关系。试验结果表明,氮素、硫素对大蒜主要营养成分含量影响不尽相同,氮硫配施能够不同程度提高大蒜鳞茎中可溶性蛋白、VC、大蒜多糖、游离氨基酸的含量,且氮、硫单一因素对大蒜鳞茎品质影响远低于元素间交互作用。但在氮素适宜水平下(10~20 mmol/L),硫素水平对大蒜鳞茎中大蒜辣素含量有显著影响,提高硫浓度能够显著增加大蒜辣素含量,而在硫素浓度过高时(8 mmol/L)则明显抑制该物质合成。隶属函数法综合评价以N3S2处理组大蒜鳞茎品质最优,隶属函数值为0.81,对照组(CK)最差。综合分析可见,无机基质栽培条件下氮硫配施显著影响大蒜鳞茎品质,氮素浓度20 mmol/L(N3)、硫素浓度4 mmol/L(S2)时最利于大蒜鳞茎品质的提升,为大蒜无机基质栽培最佳氮、硫配施水平,且两元素配施存在明显的互作效应。该结论为大蒜栽培氮、硫元素合理配施提供试验指导。     

Allium discoloration: precursors involved in onion pinking and garlic greening

Precursors involved in the formation of pink and green-blue pigments generated during onion and garlic processing, respectively, have been studied. It has been confirmed that the formations of both pigments are of very similar natures, with (E)-S-(1-propenyl)cysteine sulfoxide (isoalliin) serving as the primary precursor. Upon disruption of the tissue, isoalliin and other S-alk(en)ylcysteine sulfoxides are enzymatically cleaved, yielding 1-propenyl-containing thiosulfinates [CH3CH=CHS(O)SR; R = methyl, allyl, propyl, 1-propenyl] among others. The latter compounds have been shown to subsequently react with amino acids to produce the pigments. Whereas the propyl, 1-propenyl, and methyl derivatives form pink, pink-red, and magenta compounds, those containing the allyl group give rise to blue products after reacting with glycine at pH 5.0. The role of other thiosulfinates [RS(O)SR'] (R, R' = methyl, allyl, propyl) and (Z)-thiopropanal S-oxide (the onion lachrymatory principle) in the formation of the pigments is also discussed.     

钾素营养对大蒜生长、光合特性及品质的影响

采用溶液培养试验探讨了钾对大蒜苗期生长、生理生化指标及品质的影响。供试营养液K+浓度分别为0、3.0、6.0、9.0和12.0 mmol/L。结果表明,K+浓度在0～6.0 mmol/L范围内,大蒜苗期生长量、色素含量、净光合速率、可溶性蛋白质及大蒜素含量随K+浓度升高而增加;当达到9.0 mmol/L时,上述指标则呈下降趋势。营养液中K+浓度为6.0 mmol/L时,大蒜叶片及假茎生长量、光合色素含量、光合速率及蒸腾速率均最大,假茎及叶片大蒜素含量分别比0 mmol/L时高出63.57%和62.04%,叶片可溶性蛋白质含量明显高于其他处理;假茎及叶片可溶性糖含量在K+浓度为9.0 mmol/L时最高。在施钾各处理间游离氨基酸含量差异不显著,但均显著低于不施钾处理。总之,钾能明显促进大蒜苗期假茎和叶片生长,增加叶片色素含量,极大改善叶片光合特性,提高大蒜幼苗营养品质。营养液K+浓度以6.0 mmol/L时表现最好。     

Diallyl disulfide and diallyl trisulfide up-regulate the expression of the pi class of glutathione S-transferase via an AP-1-dependent pathway

Garlic organosulfur compounds are recognized as potential chemopreventive compounds. This protection is related to the induction of phase II detoxification enzymes. We previously reported that diallyl disulfide (DADS) and diallyl trisulfide (DATS) up-regulate the gene expression of the pi class of glutathione S-transferase (GSTP) and that an enhancer element named GPE I is required for this induction. In the present study, we further investigated the signal pathway involved in DADS and DATS up-regulation of this detoxification enzyme in Clone 9 cells. Cells were cultured with 25-200 micromol/L of DADS or DATS for 24 h. Western and Northern blots showed that both garlic allyl sulfides concentration dependently induced GSTP protein and mRNA expression, respectively. Changes in GST activity toward ethacrynic acid were consistent with the increase in GSTP expression (P < 0.05). Electromobility gel shift assay showed that the DNA binding activity of nuclear activator protein-1 (AP-1) is concentration-dependently increased in the presence of DADS and DATS as compared with that of the control cells. The phosphorylation of c-Jun NH2-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK), but not of p38, was stimulated in the presence of both garlic allyl sulfides. Pretreatment with SP600125 and PD98059, which are JNK and ERK inhibitors, respectively, abolished the increase in AP-1-DNA binding activity and also the induction of GSTP protein by either allyl sulfide. Our results indicate that the effectiveness of DADS and DATS on GSTP expression is likely related to the JNK-AP-1 and ERK-AP-1 signaling pathways and, thus, that DADS and DATS enhance the binding of AP-1 to GPE I.