Efficacy of curcumin therapy against Angiostrongylus cantonensis-induced eosinophilic meningitis.

Angiostrongylus cantonensis can invade the central nervous system, leading to human eosinophilic meningitis or eosinophilic meningoencephalitis. Curcumin is a natural product which has the effects of anti-inflammation, anti-oxidation and anti-carcinogensis, while the administration of curcumin has been reported to possibly relieve the symptoms of meningitis. The present study tested the potential efficacy of curcumin in A. cantonensis-induced eosinophilic meningitis of BALB/c mice. Assay indicators for the therapeutic effect included the larvicidal effect, eosinophil counts and matrix metalloproteinase-9 (MMP-9) activity in angiostrongyliasis. Eosinophils were mildly reduced in treatment groups compared with infected-untreated mice. However, there were no significant differences in larvicidal effects or MMP-9 activity. This study suggests that anti-inflammatory treatment with curcumin alone has low efficacy, but the treatment does not interfere with MMP-9 expression and is not useful for larvicidal effects. The possible reasons include low curcumin across the blood-brain barrier and also those larvae that survive stimulate MMP-9 production, which promotes blood-brain barrier damage, with leukocytes then crossing the blood-brain barrier to cause meningitis. Further studies will be required to test these possibilities.

Suppressive effect of penta-acetyl geniposide on the development of gamma-glutamyl transpeptidase foci-induced by aflatoxin B(1) in rats.

The suppressive effects of penta-acetyl geniposide, (Ac)(5)-GP, on the hepatotoxic lesions-induced by aflatoxin B(1) (AFB(1)) were investigated in male Wistar rats. Rats were divided into six groups: groups I and II served as normal and solvent control, respectively; group III was given AFB(1) (2 mg/kg body weight) alone; group IV was given (Ac)(5)-GP (2 mg/kg) alone; and groups V and VI received both AFB(1) (2 mg/kg body weight) and (Ac)(5)-GP (1 mg and 2 mg/kg body weight, respectively). Rats received treatments for 8 weeks, then were maintained on basal diet for 32 weeks. At the end of the experiment (week 40), the liver lesions (e.g. fatty change, eosinophilic and bile duct dilation) and preneoplastic changes in rats of groups V and VI were reduced when they were compared with group III. There were no liver lesions and preneoplastic changes in rats treated with (Ac)(5)-GP alone. Although no differences in the total number of gamma-glutamyl transpeptidase (GGT)-positive foci was observed between the groups treated with AFB(1) along with or without (Ac)(5)-GP, the treatment of (Ac)(5)-GP significantly reduced the number of AFB(1)-induced GGT positive foci (with diameter larger than 0.3 mm). These results indicated that the protective effect of (Ac)(5)-GP on early hepatocarcinogenesis-induced by AFB(1) was associated with the inhibition of GGT foci development.

Development of brain injury in mice by Angiostrongylus cantonensis infection is associated with the induction of transcription factor NF-kappaB, nuclear protooncogenes, and protein tyrosine phosphorylation.

Eosinophilic meningitis or meningoencephalitis caused by Angiostrongylus cantonensis is endemic to the Pacific area of Asia, especially Taiwan, Thailand, and Japan. Although eosinophilia is an important clinical manifestation of A. cantonensis infection, the role of eosinophils in the progress of the infection remains to be elucidated. In this experiment, we showed that A. cantonensis-caused eosinoplia and inflammation might lead to the induction of NF-kappaB and protooncogene expression via activation of the tyrosine phosphorylation signal pathway. After mice were infected daily with 30 third-stage larvae of A. cantonensis by oral adminstration for 6 weeks, no significant differences PKC-alpha, MEK-1, ERK-2, JNK, and p38 protein expression were found between the control and infected mice. However, the protein tyrosine phosphorylation levels, NF-kappaB, and iNOS protein products were significantly increased by 3.5-, 3.3-, and 6.3-fold, respectively, after 3 weeks of A. cantonensis infection. The same pattern was found for c-Myc, c-Jun, and c-Fos proteins, which were elevated by 3.2-, 2.3-, and 3.4-fold, respectively, compared to control animals after 3 weeks. The expression potency of these proteins started increasing in week 1, reaching maximal induction in week 3, and then declining in week 5 after A. cantonensis infection. Another consistent result was noted in the pathological observations, including eosinophilia, leukocyte infiltration, granulomatous reactions, and time responses in brain tissues of infected mice. These data suggest that the development of brain injury by eosinophlia of A. cantonensis infection is associated with NF-kappaB and/or nuclear protooncogenes expression, which is activated by the tyrosine phosphorylation pathway.

Suppression of the TPA-induced expression of nuclear-protooncogenes in mouse epidermis by crocetin via antioxidant activity.

Crocetin, a major component of the fruit of Gardenia jasminoides Ellis, was investigated for its antitumor promoting effect on 12-O-tetradecanoylphorbol-13-acetate-promoted mouse skin carcinogenesis. Topical application of 5 nmol TPA to CD-1 mice once daily for 5 days caused epidermal hyperplasia, and increases in the levels of c-Fos, c-Jun and c-Myc in the suprabasal layer of epidermis and the muscle layer of dermis. Immunocytolochemical examination showed that pretreatment of 1 mumol crocetin repressed the TPA-induced epidermal hyperplasia and the expressions of c-Jun, c-Fos and c-Myc to the extent of 47, 44 and 45% respectively. Crocetin of 3.0 mumol exhibited stronger inhibition on the induced hyperplasia and the oncoproteins levels (by 60, 53 and 55% respectively). Western blotting analysis confirmed this inhibitory effect of crocetin. Pretreatment of crocetin also repressed the TPA-induced H2O2 production and myeloperoxidase activity. These data indicate that crocetin suppresses the TPA-induced skin carcinogenesis maybe via its antioxidant property which, in turn, leads to a reduction in the TPA-induced expressions of c-Jun, c-Fos and c-Myc in mouse epidermis.

Inhibitory effect of crocetin on benzo(a)pyrene genotoxicity and neoplastic transformation in C3H10T1/2 cells.

Crocetin is a major component in the fruit of Gardenia jasminoides Ellis, a Chinese herbal medicine. In the work, we investigate the protective action and mechanism against benzo(a)pyrene [B(a)P]-induced genotoxicity and neoplastic transformation with a non-toxic dose of crocetin (0.01-0.10 mM) for 1 hour prior to the administration of 0.1 mM B(a)P. B(a)P genotoxicity was inhibited significantly by crocetin in a dose responsive manner. Pretreating C3H10T1/2 cells with crocetin (0.1 mM) also caused a decrease in the covalent binding of B(a)P-diol-epoxide to DNA, to about half that of cells without crocetin treatment. Crocetin also inhibited B(a)P-induced transformations. When the culture was treated with crocetin (0.01, 0.05 and 0.10 mM) for 7 days, the transformation frequencies were lower than that of the culture without crocetin treatment. Furthermore, pretreating cells with crocetin (0.01-0.10 mM) also caused an increase in the activity of GSH S-transferase (GST) to 18-71% that of the cells without crocetin treatment. These results suggest that the inhibition by crocetin of B(a)P-induced genotoxicity and neoplastic transformation in C3H10T1/2 cells is due to a mechanism that increases the activity of GST and decreases the formation of a B(a)P-DNA adduct.

Crocetin protects against oxidative damage in rat primary hepatocytes.

Crocetin is a major component in the fruit of Gardenia jaminoides Ellis, a Chinese herbal medicine. Its protective action and mechanism against oxidative damage were investigated and mechanism against oxidative damage were investigated. Reactive oxygen species (ROS) were generated enzymatically in the xanthine-xanthine oxidase (X/XO 5 microM/0.01 u/ml) system and non-enzymatically in the paraquat (PQ 5 mM) system. Both systems increased leakage of lactate dehydrogenase (LDH) and alanine transaminase (ALT) in rat primary hepatocytes, but the hepatotoxicity was significantly suppressed on pretreatment with crocetin (10, 20 microM). Crocetin decreased formation of malondialdehyde (MDA) as an index of lipid peroxidation induced by ROS. The oxyradical generation by X/XO or PQ caused DNA damage evaluated with unscheduled DNA synthesis (UDS) in rat primary hepatocytes. The addition of crocetin decreased genotoxicity evaluated with UDS in both systems. The data showed that crocetin also inhibited the formation of superoxide anion in the X/XO system and bleached the free radical 1, 1-diphenyl-2- picrylhydrazyl (DPPH). The protective action of crocetin operated via quenching of the superoxide anion and/or free radical.

N-Nitroso-N-(3-keto-1,2-butanediol)-3'-nitrotyramine. A new genotoxic agent derived from the reaction of tyrosine and glucose in the presence of sodium nitrite.

N-Nitroso-N-(3-keto-1,2-butanediol)-3'-nitrotyramine (NO-NTA) is a product of a model browning system in the presence of sodium nitrite. In this study, the chemical structure is confirmed by spectral studies, including UV, mass spectrometry, nuclear magnetic resonance and infrared spectroscopy. NO-NTA is strongly genotoxic to the rat hepatocyte and is moderately cytotoxic to mouse C3H10T1/2 cells. Results obtained in this study indicate that NO-NTA inflicted DNA damage through the formation of a DNA adduct. In addition, C3H10T1/2 cells were treated with NO-NTA and, following addition of 12-O-tetradecanoylphorbol-13-acetate (TPA) as promotor, the increase of transformed foci indicated that NO-NTA could possibly be an initiator [corrected] of TPA tumor promotion. A transformed cell line from NO-NTA initiated and TPA promoted foci increased saturation density and growth ability in soft agar reactive to the control line. These results suggest that the formation of a genotoxic agent of nitroso-derivatives may take place in a nitrite-containing food system during processing and cooking.

Effects of crocetin on the hepatotoxicity and hepatic DNA binding of aflatoxin B1 in rats.

The effects of crocetin pretreatment on both hepatic aflatoxin B1 (AFB1)-DNA binding and AFB1 hepatotoxicity in rats has been examined. For these studies, male Wistar rats were treated with AFB1 (2 mg/kg) by i.p. administration, and the different degrees of hepatic damage were revealed by the elevations of levels of serum marker enzymes such as aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase and gamma-glutamyltranspeptidase. After pretreatment of the animals with crocetin (2 or 6 mg/kg) daily for three consecutive days, the enzyme elevations were significantly suppressed. This suggested that the crocetin possessed chemopreventive effects on the early acute hepatic damage induced by AFB1. Under these experimental conditions, consistent elevations of hepatic glutathiones (GSH) and activities of glutathione S-transferase (GST) and glutathione peroxidase (GSH-Px) were observed. Crocetin treatment also decreased AFB1-DNA adduct formation in AFB1-treated animals. From these results, we suggest that the protective effect of crocetin on AFB1 hepatotoxicity in rats might be due to the hepatic tissues' defense mechanisms that elevated the cytosol GSH and the activities of GST and GSH-Px.

Effects of san-huang-hsieh-hsin-tang on central monoaminergic and GABAergic systems in rats.

In this paper, the effects of San-Huang-Hsieh-Hsin-Tang on central monoaminergic and GABAergic systems were studied in rats. It was found that San-Huang-Hsieh-Hsin-Tang significantly (1) prolonged the period from the onset of clonic to tonic convulsions induced by picrotoxin, (2) prolonged the sleep duration induced by hexobarbital, (3) inhibited central catecholaminergic activity, (4) promoted central GABAergic activity, and (5) decreased the turnovers of central norepinephrine and dopamine.