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Mutants of phycomyces with decreased gallic acid content.

Most plants and some fungi accumulate phenols. Two hydroxybenzoic acids, gallic and protocatechuic acids, are abundant in the giant sporangiophores of the zygomycete Phycomyces blakesleeanus, much more so than in the basal mycelium or the culture medium. The actual concentrations vary with illumination, age of the culture, and composition of the medium. We devised a simple screening procedure to isolate hba mutants whose sporangiophores contained less gallic acid than the wild type. The most useful mutant had very low concentrations of hydroxybenzoic acids in the sporangiophores, but about the same as the wild type in the basal mycelium and the medium. The mutant was only slightly different from the wild type in growth and morphology. Mutant and wild-type sporangiophores grew away from ultraviolet C sources (260 nm) equally well. Contrary to previous conjectures, ultraviolet tropism does not depend on the ultraviolet absorption of gallic acid or other free hydroxybenzoic acids in the sporangiophore. Against expectations, phenols did not impair DNA extraction: sporangiophores produced better DNA preparations than basal mycelia and the hba mutant only slightly better than the wild type.

DNA, Fungal↗

Antitumor-promoting activities of tannic acid, ellagic acid, and several gallic acid derivatives in mouse skin.

Naturally occurring plant phenols with antimutagenic and anticarcinogenic activities were tested for their abilities to inhibit the biochemical and biological effects of the potent tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate (TPA) in mouse epidermis in vivo. When applied topically to mouse skin, tannic acid (TA), ellagic acid, and several gallic acid derivatives all inhibit TPA-induced ornithine decarboxylase activity, hydroperoxide production, and DNA synthesis, three biochemical markers of skin tumor promotion. Moreover, in the two-step initiation-promotion protocol, the same phenolic compounds also inhibit the incidence and yield of skin tumors promoted by TPA. TA is the most effective of these treatments. Since they are already known to inhibit tumor initiation, the plant phenols protecting against skin tumor promotion by TPA may be universal inhibitors of multistage carcinogenesis. TA and other polyphenols, therefore, might be valuable in cancer therapy and/or prevention.

Animals↗

Structure-activity relationship analysis of antioxidant ability and neuroprotective effect of gallic acid derivatives.

Gallic acid and its derivatives are a group of naturally occurring polyphenol antioxidants which have recently been shown to have potential healthy effects. In order to understand the relationship between the structures of gallic acid derivatives, their antioxidant activities, and neuroprotective effects, we examined their free radical scavenging effects in liposome and anti-apoptotic activities in human SH-SY5Y cell induced by 6-hydrodopamine autooxidation. It was found that these polyphenol antioxidants exhibited different hydrophobicity and could cross through the liposome membrane to react with 1,1-diphenyl-2-picryl-hydrazyl (DPPH) free radical in a time and dose-dependent manner. At the same time, the structure-antioxidant activity relationship of gallic acid derivatives on scavenging DPPH free radical in the liposome was also analyzed based on theoretical investigations. Analysis of cell apoptosis, intracellular GSH levels, production of ROS and the influx of Ca(2+) indicated that the protective effects of gallic acid derivatives in cell systems under oxidative stress depend on both their antioxidant capacities and hydrophobicity. However, the neuroprotective effects of gallic acid derivatives seem to depend more on their molecular polarities rather than antioxidant activities in the human SH-SY5Y cell line. In conclusion, these results reveal that compounds with high antioxidant activity and appropriate hydrophobicity are generally more effective in preventing the injury of oxidative stress in neurodegenerative diseases.

Antioxidants↗

Ca2+-Dependent caspase activation by gallic acid derivatives.

Gallic acid (GA) derivatives, 3,4-methylenedioxyphenyl 3,4,5-trihydroxybenzoate (GD-1) and S-(3,4-methylenedioxyphenyl)3,4,5-trihydroxythiobenzoate (GD-3), were previously reported to induce apoptosis in tumor cells with IC50s of 14.5 microm and 3.9 microm, respectively. To elucidate the mechanism by which these gallic acid derivatives (GDs) induce apoptosis, we studied whether GD-1 and GD-3 can activate caspases. When promyelocytic leukemia HL-60RG cells were treated with GD-1 and GD-3, poly(ADP-ribose)polymerase (PARP), a substrate of caspase-3, was cleaved into 85 kDa of degradative product with increasing incubation time. GA also activated PARP cleavage, which was inhibited by catalase, N-acetyl-L-cysteine (NAC), and intracellular Ca2+ chelator 1,2-bis(2-aminophenoxyethane)-N,N,N,N'-tetraacetic acid tetrakis (acetoxymethyl ester) (BAPTA-AM), in addition to a caspase inhibitor, Z-VAD-FMK. Its inhibitory pattern was identical with that of hypoxanthine/xanthine oxidase. On the other hand, GD-1- and GD3-induced PARP cleavage was not suppressed by catalase or NAC, but by BAPTA-AM. This suggested that the GD-elicited signaling pathway is different from GA's. Taken together, GDs activated caspase-3 following intracellular Ca2+ elevation independent of reactive oxygen species. Thus, it became evident that the signaling pathway leading to apoptosis was regulated by GDs in a different manner from GA.

Acetylcysteine↗

2-pyrone-4,6-dicarboxylic acid, a catabolite of gallic acids in Pseudomonas species.

2-Pyrone-4,6-dicarboxylate hydrolase was purified from 4-hydroxybenzoate-grown Pseudomonas testosteroni. Gel filtration and electrophoretic measurements indicated that the preparation was homogeneous and gave a molecular weight of 37,200 for the single subunit of the enzyme. Hydrolytic activity was dependent upon a functioning sulfhydryl group(s) and was freely reversible; the equilibrium position was dependent upon pH, with equimolar amounts of pyrone and open-chain form present at pH 7.9. Since the hydrolase was strongly induced when the nonfluorescent organisms P. testosteroni and P. acidovorans grew with 4-hydroxybenzoate, it is suggested that 2-pyrone-4,6-dicarboxylate is a normal intermediate in the meta fission degradative pathway of protocatechuate. Laboratory strains of fluorescent pseudomonads did not metabolize 2-pyrone-4,6-dicarboxylate, but a strain of P. putida was isolated from soil that utilized this compound for growth; the hydrolase was then induced, but it was absent from extracts of 4-hydroxybenzoate-grown cells that readily catabolized protocatechuate by ortho fission reactions. 2-Pyrone-4,6-dicarboxylic acid was the major product formed when gallic acid was oxidized by purified protocatechuate 3,4-dioxygenase. Protocatechuate 4,5-dioxygenase gave only the open-chain ring fission product when gallic acid was oxidized, but the enzyme attacked 3-O-methylgallic acid, giving 2-pyrone-4,6-dicarboxylic acid as the major product. Cell suspensions of 4-hydroxybenzoate-grown P. testosteroni readily oxidized 3-O-methylgallate with accumulation of methanol.

Carboxylic Ester Hydrolases↗

Proton dissociation is important to understanding structure-activity relationships of gallic acid antioxidants.

Gallic acid derivatives (GADs) can efficiently scavenge free radicals, which is partially responsible for their neuroprotective effects. As GADs tend to deprotonate to give birth to GAD anions, which has big influence on the radical-scavenging behaviors of GADs, to understand the structure-activity relationships (SARs) of GAD antioxidants, the anions should be taken into consideration. In this paper, a combined density functional theory method, labeled as (RO)B3LYP/6-311+G(2d,2p)//AM1/AM1, was employed to calculate homolytic O-H bond dissociation enthalpies and adiabatic ionization potentials for GADs and derived anions in solvent (ethanol), by which the experimentally observed SARs of GADs were better elucidated.

Antioxidants↗

Assessment of the no-observed-adverse-effect level (NOAEL) of gallic acid in mice.

Gallic acid is a naturally occurring plant phenol obtained by the hydrolysis of tannins and is known to show some pharmacological activities. The purpose of this paper is to establish the safety of gallic acid in mice. In this study, acute administration of gallic acid even at a dose as high as 5 g/kg body weight did not produce any signs of toxicity or mortality. In the subacute study, gallic acid at a dose of 1000 mg/kg body weight did not significantly alter the hematological parameters. Further, no appreciable change was noted in the various biochemical parameters such as SGOT and SGPT, as well as many serum constituents such as protein, cholesterol, urea and bilirubin. Therefore, from this study, it may be concluded that gallic acid is non-toxic up to a level of 5000 mg/kg body weight, when given orally. In addition, the subacute study indicated the absence of cumulative toxicity, as reflected by the non-significant alterations in the parameters investigated. The NOAEL was 5000 mg/kg body weight, the highest dose tested.

Administration, Oral↗

Modes of action of gallic acid in suppressing food intake of rats.

Gallic acid (3,4,5,trihydroxybenzoic acid) is a naturally occurring polyphenol comprising the major hydrolytic product of tannic acid. Gallic acid and tannic acid were previously shown to suppress food intake of animals to a similar extent. However, the mechanism by which this effect takes place has not been previously reported. Reported here is that the effect of gallic acid on food intake is not mediated entirely through taste aversion or through other gastrointestinal factors, since a continuous daily infusion of a gallic acid solution (18 ml; 2%) resulted in a significant reduction of food intake. The catechol moiety of gallic acid plays an important role in its suppression of food intake since administration of tis 4-0 methyl derivative was significantly less effective is suppressing food intake (P less than 0.01). The effectiveness of gallic acid in suppressing food intake diminishes with time, indicating adaptation to the consumption of this polyphenol. Propyl gallate is a more potent suppressor of food intake than gallic acid. Adaptation to the consumption of this polyphenol, it it exists at all, is much slower than with gallic.

Animals↗

Modification of apoptosis-inducing activity of gallic acid by saliva.

Gallic acid, a structural unit of tannin, induced apoptotic cell death, characterized by nuclear condensation and caspase activation in human oral tumor cell lines (HSC-2, HSG). Agarose gel electrophoresis demonstrated that gallic acid produced large DNA fragments in these cells, as well as in T-cell leukemia (MOLT-4) and erythroleukemia (K-562) cells, whereas it induced internucleosomal DNA fragmentation in human myelogenous leukemic cell lines (HL-60, ML-1, U-937, THP-1). This indicates that induction of internucleosomal DNA fragmentation or production of large DNA fragments depends on both target cells and inducers. Addition of total saliva dose-dependently reduced the cytotoxicity induction by gallic acid. These data suggested that the biological action of gallic acid might be modified by physiological fluids.

Antioxidants↗

Gene expression profiles of inducible nitric oxide synthase and cytokines in Leishmania major-infected macrophage-like RAW 264.7 cells treated with gallic acid.

The effects of gallic acid on the gene expressions of inducible nitric oxide synthase (iNOS) and the cytokines interleukin (IL)-1, IL-10, IL-12, IL-18, TNF-alpha, and interferon (IFN)-gamma were investigated by reverse-transcription polymerase chain reaction (RT-PCR). The experiments were performed in parallel in non-infected and in L. major-infected RAW 264.7 cells and the expression profiles were compared with those mediated by IFN-gamma plus lipopolysaccharide (LPS). The infection per se induced the expression first of IL-1 and TNF-alpha mRNA, later that of IL-10 mRNA. Gallic acid induced low and transient levels of TNF-alpha and IL-10 in non-infected cells, and it clearly enhanced and prolonged iNOS and cytokine mRNA expressions in Leishmania-parasitised cells. Interestingly, and in contrast to activation by IFN-gamma/LPS, gallic acid also stimulated Leishmania-infected cells to produce IFN-gamma mRNA. For IFN-alpha, a sandwich immunoassay was performed to determine its amount present in the supernatant of gallic acid-stimulated RAW 264.7 cells. In showing predominant stimulation of infected cells and the induction especially of IFN-gamma, a cytokine that plays a central role in antimicrobial macrophage and T cell regulation, these data provide the basis for an immunological concept of gallic acid and possibly other plant polyphenols for their beneficial effects in various infectious conditions.

Animals↗

Use of methanolysis for the determination of total ellagic and gallic acid contents of wood and food products.

Anhydrous methanolic HCl has been found to be an excellent reagent for releasing ellagic acid and gallic acid (as methyl gallate) from biomass substrates. Optimization of both the reaction conditions and the gradient HPLC analysis has led to the development of a new protocol. The method provides ellagic acid yields significantly higher than those obtained previously, indicating total ellagic acid contents of several substrates have previously been underestimated.

Alcoholic Beverages↗

Trypanocidal effects of gallic acid and related compounds.

Gallic acid (3,4,5-trihydroxybenzoic acid) is a naturally abundant plant phenolic compound and it is well known as a component of hydrolyzable tannins. We report here that gallic acid and related compounds have trypanocidal activity against Trypanosoma brucei brucei (GUTat 3.1) in both the long slender bloodstream forms and the procyclic forms, in vitro. LD50 values of gallic acid are 46.96 +/- 1.28 microM for bloodstream forms and 30.02 +/- 3.49 for procyclic forms, respectively. A study of structurally related compounds suggested that the pyrogallol moiety could be responsible for this activity.

Animals↗

Inhibition of CYP3A-mediated oxidation in human hepatic microsomes by the dietary derived complex phenol, gallic acid.

Plant polyphenols, such as gallic acid, have been reported to have a range of biological activities including antimutagenic effects. Previously, we reported that gallic acid (3,4,5-trihydroxybenzoic acid), an agent found in wine and tea, inhibits androstenedione 6beta-hydroxylase activity (Ki 70 microm), a cytochrome P450 (CYP3A) marker in human liver microsomes. The pre-incubation of gallic acid (100 microM) with human liver microsomes in the absence of NADPH, as compared with the presence of NADPH, before assay of androstenedione 6beta-hydroxylase activity significantly increased the inhibitory effects of the gallic acid (0.03 +/- 0.03 nmol (mg microsomal protein)(-1) min(-1) compared with 0.20 +/- 0.06 nmol (mg microsomal protein) (-1) min(-1) (P < 0.05)). The antioxidant ascorbic acid and the radical scavenger glutathione prevented this observed increase in inhibition. Removal of gallic acid-derived products from the incubation completely restored CYP3A activity. In contrast, the activities of CYP1A and CYP2E, and non-CYP mediated reductive microsomal 17beta-hydroxysteroid dehydrogenase activity were refractory to inhibition by gallic acid.

17-Hydroxysteroid Dehydrogenases↗

Phenol biosynthesis in higher plants. Gallic acid.

The biosynthesis of gallic acid in a number of higher plants was investigated by using l-[U-(14)C]phenylalanine, (-)-[G-(14)C]shikimic acid, d-[1-(14)C]glucose and d-[6-(14)C]glucose as tracers. The results are compared with those obtained similarly for caffeic acid and are interpreted in terms of the dehydrogenation of 5-dehydroshikimic acid as a normal route of metabolism for gallic acid.

Cinnamates↗

Metabolic fate of gallic acid orally administered to rats.

The metabolic behavior of orally administered gallic acid was investigated by HPLC and 4-O-methyl gallic acid was found to be the main metabolite in rat peripheral blood and urine. After oral administration of gallic acid, maximum concentration in portal vein and inferior vena cava occurred at 15 and 30 min, respectively. In portal vein, gallic acid was preferentially detected relative to 4-O-methyl gallic acid, whereas gallic acid and 4-0-methyl gallic acid were equally detected in inferior vena cava. On the other hand, 4-O-methyl gallic acid but not gallic acid was found in liver. The contents of gallic acid and 4-O-methyl gallic acid in urine were nearly 100 times higher than those in blood. The ratio of 4-O-methyl gallic acid to total gallic acid metabolites in urine was from 0.55 to 0.76, indicating that a considerable amount of gallic acid was excreted without being metabolized. In this study we found that gallic acid administered orally existed in the blood for 6 h at most, and more than half was metabolized to 4-O-methyl gallic acid, followed by excretion into urine.

Administration, Oral↗

Potentiation of vasoconstrictor response and inhibition of endothelium-dependent vasorelaxation by gallic acid in rat aorta.

In the isolated rat thoracic aorta, gallic acid potentiated the vasoconstrictor response to phenylephrine. The potentiation produced by gallic acid was absent in endothelium-denuded arteries. The potentiation was abolished by N(G)-nitro-L-arginine methyl ester, an inhibitor of nitric oxide synthesis, and slightly attenuated by an addition of L-arginine, while indomethacin or BQ610 had no effect. The potentiation of response to phenylephrine was not found for structural modifications of gallic acid, except for caffeic acid. Gallic acid also inhibited vasorelaxation induced by acetylcholine, sodium nitroprusside or prostacyclin, especially that by acetylcholine. The effect on vasorelaxation induced by acetylcholine was decreased by esterification of the carboxy group of gallic acid, and in the absence or by the methylation of the o-dihydroxy group. Caffeic acid inhibited the vasorelaxation, though the effect was smaller than that of gallic acid. These findings indicate that gallic acid produces a potentiation of contractile response and inhibition of vasorelaxant responses, probably through inactivation of nitric oxide (NO), in which endothelially produced NO is principally involved, and that the modification of functional groups of the gallic acid molecule abolishes the potentiation of contractile response and attenuates the inhibition of vasorelaxant responses.

Acetylcholine↗

Radical intensity and cytotoxic activity of curcumin and gallic acid.

Natural phenolic compounds, curcumin and gallic acid, were compared for their cytotoxic activity in relation to their radical modulating activity. These two compounds induced apoptotic cell death in human promyelocytic leukemic HL-60 cells and human oral squamous carcinoma HSC-4 cells. Curcumin was more cytotoxic than gallic acid. Catalase reduced significantly the cytotoxic activity of gallic acid, but not that of curcumin. ESR spectroscopy demonstrated that curcumin produced radicals under alkaline conditions, scavenged the superoxide anion radical, and enhanced the radical intensity of sodium ascorbate at higher concentrations. As compared with curcumin, gallic acid produced higher amounts of radicals and more efficiently scavenged the superoxide anion radical. Gallic acid reduced the radical intensity of sodium ascorbate, suggesting a possible interaction between these two compounds. These data suggest that curcumin and gallic acid induce apoptosis by different mechanisms.

Antineoplastic Agents↗