Compositional differences in flavonoids are varied in the big family of Compositae. By summarizing our previous analytical studies and other scientific evidences, new strategy will be possible to further analyze flavonoids and utilize them for human health. The HPLC analytical method has been established in terms of linearity, sensitivity, accuracy, and precision. Herbs of the family of Compositae have considerable amounts of peroxynitrite (ONOO-)-scavenging effects and their phenolic substances. These effects may contribute to the prevention of disease associated with excess production of ONOO-, depending on the high content of flavonoid substances.
Asteraceae* ; Chromatography, High Pressure Liquid ; Flavonoids ; Humans ; Peroxynitrous Acid ; Phenol

Transient forebrain ischemia results in the delayed neuronal death in the CA1 area of the hippocampus. The present study was performed to determine effects of aminoguanidine, a selective iNOS inhibitor, on the generation of peroxynitrite and delayed neuronal death occurring in the hippocampus following transient forebrain ischemia. Transient forebrain ischemia was produced in the conscious rats by four-vessel occlusion for 10 min. Treatment with aminoguanidine (100 mg/kg or 200 mg/kg, i.p.) or saline (0.4 ml/100 g, i.p.) was started 30 min following ischemia-reperfusion and the animals were then injected twice daily until 12 h before sacrifice. Immunohistochemical method was used to detect 3-nitrotyrosine, a marker of peroxynitrite production. Posttreatment of aminoguanidine (200 mg/kg) significantly attenuated the neuronal death in the hippocampal CA1 area 3 days, but not 7 days, after ischemia-reperfusion. 3-Nitrotyrosine immunoreactivity was enhanced in the hippocampal CA1 area 3 days after reperfusion, which was prevented by the treatment of aminoguanidine (100 mg/kg and 200 mg/kg). Our findings showed that (1) the generation of peroxynitrite in the hippocampal CA1 area 3 days after ischemia-reperfusion was dependent on the iNOS activity; (2) the postischemic delayed neuronal death was attenuated in the early phase through the prevention of peroxynitrite generation by an iNOS inhibitor.
Animals ; Hippocampus* ; Ischemia* ; Neurons ; Peroxynitrous Acid* ; Prosencephalon* ; Rats* ; Reperfusion

Lactuca raddeana (Compositae) is used to treat obesity and complications due to diabetes. The five phenolic compounds including chlorogenic acid, chicoric acid, luteolin 7-O-glucoside, luteolin 7-O-glucuronide, luteolin were qualitatively identified by LC-ESI-MS analysis. The contents were quantitatively determined by HPLC, under the condition of a Capcell Pak C18 column (5 microm, 250 mm x 4.6 mm i.d.) and a gradient elution of 0.05% trifluoroacetic acid (TFA) and 0.05% TFA in MeOH-H2O (60 : 40). The contents of chicoric acid (100.99 mg/g extract) and luteolin 7-O-glucoside (101. 69 mg/g extract) were high, while those of other three phenolic substances were very low. The 3T3-L1 adipocyte cells treated with chicoric acid and luteolin 7-O-glucuronide significantly suppressed the accumulation of fat, suggesting they are effective against obesity. Since high level of peroxynitrite (ONOO) causes cardiovascular disease in obese patients, its scavenging activity was also studied.
Adipocytes* ; Asteraceae ; Cardiovascular Diseases ; Chlorogenic Acid ; Chromatography, High Pressure Liquid ; Humans ; Luteolin ; Obesity ; Peroxynitrous Acid ; Phenol* ; Trifluoroacetic Acid

PURPOSE: To assess the role of nitric oxide in the development of cataract by detection of nitrotyrosine, which is a reaction product of peroxynitrite with tyrosine. METHODS: We have collected aqueous humor and anterior capsule of lens after cataract surgery from 14 eyes. We classified lens opacities into three types such as nuclear sclerosis, cortical and posterior subcapsular opacity. Nitrite, stable oxidative product of nitric oxide, was measured in the aqueous humor and immunohistochemical staining for nitrotyrosine was performed to examine anterior capsules. RESULTS: The concentrations of nitric oxide in aqueous humor were 0.0698+/-0.0036, 0.0652+/-0.0116, and 0.0700+/-0.0022 mM in nuclear sclerosis, cortical and posterior subcapsular opacity, respectively and no significant differences existed among lens opacity types. In lenses with cortical and posterior subcapsular opacity, the expression of nitrotyrosine was observed around epithelial cell and intercellular space. There was no expression of nitrotyrosine in lenses with nuclear sclerosis. CONCLUSIONS: This results suggest that nitration of tyrosine by peroxynitrite is possibly related to the cataractogenesis of lenses with cortical and posterior subcapsular opacity.
Aqueous Humor ; Capsules ; Cataract* ; Epithelial Cells ; Extracellular Space ; Nitric Oxide* ; Peroxynitrous Acid ; Sclerosis ; Tyrosine

Previously, we reported that glucose-deprived astrocytes are more vulnerable to the cytotoxicity of peroxynitrite, the reaction product of nitric oxide and superoxide anion. The augmented vulnerability of glucose-deprived astrocytes to peroxynitrite cytotoxicity was dependent on their proliferation rate. Inhibition of cell cycle progression has been shown to inhibit the apoptotic cell death occurring in cerebral ischemia-reperfusion. In the present study, we demonstrate that the increased death of glucose-deprived astrocytes by peroxynitrte was largely blocked by the cell cycle phase G2/M transition blocker etoposide. However, the cytoprotective effect of etoposide was not associated with its inhibition of cell cycle progression. Instead, etoposide effectively scavenged peroxynitrite. However, etoposide did not scavenge individual nitric oxide and superoxide anion and it did not prevent the hydrogen peroxide-induced cytotoxicity. The present results indicate that etoposide prevents the toxicity of peroxynitrite in astrocytes by directly scavenging peroxynitrite, not by inhibiting cell cycle progression.
Astrocytes ; Cell Cycle ; Cell Death ; Etoposide ; Hydrogen ; Nitric Oxide ; Peroxynitrous Acid ; Superoxides

Many herbs have been used as therapeutics in Korean traditional medicine. In view of their clinical indications, anti-oxidant activity may contribute to their pharmacological effects. However, anti-oxidant information on these plants has not been available. In this study, seventy herbs which have been used in Korean traditional medicine were selected and screened for anti-oxidant activity using their water extracts. The anti-oxidant activity was assessed by their ability to inhibit three oxidation reactions; luminol/Fenton reagent, 2, 7-dichlorodihydrofluorescein (DCHF)/Fenton reagent and DCHF/peroxynitrite. In each assay, 70 herbs were divided into two groups; anti-oxidant group which inhibited the respective oxidation reaction and was majority (about 60 herbs), and pro-oxidant group which enhanced the oxidation reaction but was minority (more or less 10 herbs). When the herbs were listed in the order of their anti-oxidant strength, the orders obtained from each assay were found to be quite similar. The upper top rankers (more or less 10 herbs) in each assay showed strong activity compared to the others. The uppermost rankers in each assay were Rubus coreanus Miquel/ Rubus schizostylus, Schisandra chinensis Baillon/ Schizandra chinensis and Terminalia chebula Retzius/ Terminalia chebula. Of the pro-oxidant herbs, about 4-5 herbs were strongly pro-oxidant, which enhanced the control oxidation reactions to 150-300%. But the meaning of this observation is not known since few of them in one assay were also anti-oxidant in other assays. The results obtained in the present study may serve as information for understanding pharmacological effects of these herbs and developing new drugs from them.
Hydrogen Peroxide ; Iron ; Luminescence ; Medicine, Korean Traditional ; Peroxynitrous Acid ; Schisandra ; Terminalia ; Water

BACKGROUND AND OBJECTIVES: Recent evidence has shown that nitric oxide (NO) levels are increased in allergic rhinitis. However, the role of this molecule in the pathophysiology of allergic rhinitis is still poorly understood. Peroxynitrite (OONO-), the reactive metabolites of NO, causes nitrotyrosine formation by the nitration of tyrosine residues, and promotes deleterious effects on protein function. We attempted in this study to clarify whether NO and nitrotyrosine in the nasal secretion could be increased in the early or the late phase reaction after allergen challenge. MATERIALS AND METHODS: Samples were obtained from thirteen patients with allergic rhinits to Dermatophagoides Pteronyssinus with the filter paper absorption method. The samples were collected right before, 30 minutes after, and 8 hours after the allergen challenge. Then we estimated the concentrations of nitrite (NO2-) and nitrate (NO3-). Nitrotyrosine in nasal secretions was determined by Western blot analysis in three patients. RESULTS: The nitrite/nitrate concentration in nasal secretions did not show significant changes between the baseline, the early, and the late phase (p>0.05). In the Western blot analysis, the concentration of nitrotyrosine was increased in the late phase. CONCLUSIONS: Although the NO was not increased after the allergen challenge, nitrotyrosine, the evidence of the peroxynitrite effect to tyrosine residues of the protein, was increased in the late phase of the reaction rather than the early phase. Because the peroxynitrite is an metabolite of NO, we can estimate that the overall NO effect has an influence on the late phase of the allergic reaction, and it can be presumed that NO has an influence on the long-term deterioration on the nasal mucosa by cytotoxic effect of peroxynitrite, rather than on the immediate reaction of allergic rhinitis.
Absorption ; Blotting, Western ; Dermatophagoides pteronyssinus ; Humans ; Hypersensitivity ; Nasal Mucosa ; Nitric Oxide* ; Peroxynitrous Acid* ; Rhinitis* ; Tyrosine

Peroxynitrite (ONOO-)-scavenging activities of nine Compositae herbs consisting of three Ixeris, two Youngia, two Cirsium and one of each Lactuca and Taraxacum species were evaluated. The contents of their ONOO- scavengers in the extracts were also determined on a HPLC using seven standard compounds, chlorogenic acid (CGA), chicoric acid (CA), luteolin 7-glucoside (luteolin-7-glc), luteolin 7-glucuronide (luteolin-7-glcU), luteolin, linarin and pectolinarin. Five of those compounds exhibited potent ONOO--scavenging activities: IC50, CA (0.76 microM), CGA (1.34 microM), luteolin (0.81 microM), luteolin-7-glc (0.86 microM) and luteolin-7-glcU (3.13 microM). Both CA and luteolin-7-glc were highly contained in I. dentata (19.71 mg/g and 13.58 mg/g, respectively), I. dentata var. albiflora (17.58 mg/g and 23.83 mg/g, respectively) and I. sonchifolia (65.71 mg/g and 6.99 mg/g, respectively). Among the nine herbs, those three Ixeris species had very low IC50 values over the range of 0.48 - 1.74 microg/mL, suggesting that they could be potential therapeutic vegetables, particularly for preventing diabetic complications or obesity, which can be caused by an excess production of ONOO-.
Asteraceae* ; Chlorogenic Acid ; Chromatography, High Pressure Liquid ; Cirsium ; Diabetes Complications ; Inhibitory Concentration 50 ; Luteolin ; Obesity ; Peroxynitrous Acid ; Phenol* ; Taraxacum ; Vegetables

Transient receptor potential cation channel, subfamily V, member 1 (TRPV1, also known as vanilloid receptor 1) is a receptor that detects capsaicin, a pungent component of chili peppers, and noxious heat. Although its function in the primary nociceptor as a pain receptor is well established, whether TRPV1 is expressed in the brain is still under debate. In this study, the responses of primary cortical neurons were investigated. Here, we report that 1) capsaicin induces caspase-3-dependent programmed cell death, which coincides with increased production of nitric oxide and peroxynitrite ; that 2) the prolonged capsaicin treatment induces a steady increase in the degree of capase-3 activation, which is prevented by the removal of capsaicin; 3) and that blocking calcium entry and calcium-mediated signaling prevents capsaicin-induced cell death. These results indicate that cortical neurons express TRPV1 whose prolonged activation causes cell death.
Apoptosis ; Brain ; Calcium ; Capsaicin ; Capsicum ; Caspase 3 ; Cell Death ; Hot Temperature ; Neurons ; Nitric Oxide ; Nociceptors ; Peroxynitrous Acid

The present study was undertaken to find out the role of NO on cataractogenesis in the experimenally-induced uveitis(EIU) model. Nitrite and nitrate, stable oxidative products of nitric oxide(NO), were measured in the aqueous humor and the progression of inflammations and lens opacities were evaluated with slit lamp biomicroscope. Immunoperoxidase staining and immunofluorescent staining for inducible NO synthase(iNOS) and peroxynitrite were performed to confirm the site of production of NO and peroxynitrite. The grades of inflammation were peaked at 24 hours inflammation was gradually decreased after 48 hours and lens opacity after 72 hours. These changes returned to the baseline level by one week after LPS injection. Similarly, NO concentration in aqueous humor was peaked at 24 hours. And it was then decreased after 48 hours and returned to the baseline level by one week. These inflammatory signs and lens opacities were significantly decreased in NG-nitro-L-arginine methyl ester(L-NAME) administrated group. Inflammatory cells in anterior chamber, iris, and ciliary body expressed highly iNOS which was coincide with peroxynitrite immunolocalization. Therefore, these results suggest that cataract formation in EIU is related to the NO production in aqueous humor. Furthermore, lipid peroxidation by peroxynitrite is possibly related with cataractogenesis in EIU. But, we need a further evaluation to seek the relationship between cataractogenesis and increased nitric oxide concentration, combined with studies of other biochemical changes in anterior chamber and lens.
Anterior Chamber ; Aqueous Humor ; Cataract* ; Ciliary Body ; Inflammation ; Iris ; Lipid Peroxidation ; Nitric Oxide ; Nitroarginine ; Peroxynitrous Acid ; Uveitis*