irisolidone and kakkalide

This study was designed to determine whether irisolidone and its glycoside kakkalide, which are the major constituents of the flower of Pueraria lobata (Kudzu) can attenuate ethanol-induced gastritic injury in mice.. Irisolidone and kakkalide inhibited IL-8 secretion and NF-κB activation in lipopolysaccharide-stimulated KATO III cells. Therefore, we investigated their protective effects against ethanol-induced gastric injury in mice. Pretreatment with kakkalide or irisolidone decreased the area of hemorrhagic ulcerative lesions caused by ethanol and suppressed stomach myeloperoxidase activity, CXCL4 secretion, and NF-κB activation. The ameliorating effect of irisolidone was more potent than that of kakkalide.. Irisolidone may attenuate ethanol-induced gastritis by inhibiting the infiltration of immune cells, particularly neutrophils, through the regulation of CXCL-4 or IL-8 secretion.

Topics: Animals; Cell Line; Ethanol; Flavonoids; Gastritis; Glycosides; Interleukin-8; Isoflavones; Lipopolysaccharides; Male; Mice, Inbred ICR; Neutrophils; NF-kappa B; Peroxidase; Platelet Factor 4; Protective Agents; Tumor Necrosis Factor-alpha

Kakkalide and irisolidone, the main isoflavones of Flos Puerariae, exhibit a wide spectrum of bioactivities. Intestinal bacteria biotransformation plays an important role in the metabolic pathways of flavones, and is directly related to the bioactivities of the prodrugs after oral administration. To the best of our knowledge, the metabolic pathways of kakkalide and irisolidone in vitro have not been comprehensively studied yet. This paper describes the strategy using ultra-high performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UHPLC/Q-TOF MS) for the rapid analysis of the metabolic profiles of kakkalide and irisolidone after incubated with human and rat intestinal bacteria. Bacteria incubated samples were prepared and analyzed after incubated under anaerobic conditions for 48 h. A total of 17 metabolites, including parent compounds, were detected in human and rat intestinal bacteria incubated samples. The results obtained indicate that hydrolysis, dehydroxylation, demethoxylation, demethylation, hydroxylation, decarbonylation, and reduction were the detected metabolic pathways of kakkalide and irisolidone in vitro. The conversion rate of irisolidone in human and rat bacteria was 8.57% and 6.51%, respectively. Biochanin A was the relatively main metabolite of irisolidone, and the content of biochanin A in human and rat bacteria was 3.68% and 4.25%, respectively. The conversion rate of kakkalide in human and rat bacteria was 99.92% and 98.58%, respectively. Irisolidone was the main metabolite of kakkalide, and the content of irisolidone in human and rat bacteria was 89.58% and 89.38%, respectively. This work not only provides the evidence of kakkalide and irisolidone metabolites in vivo, but also demonstrates a simple, fast, sensitive, and inexpensive method for identification of metabolites of other compounds transformed by intestinal bacteria.

Topics: Adult; Animals; Bacteria; Biotransformation; Chromatography, High Pressure Liquid; Feces; Flavonoids; Flowers; Glycosides; Humans; Intestinal Mucosa; Intestines; Isoflavones; Male; Mass Spectrometry; Molecular Structure; Plant Extracts; Pueraria; Rats; Rats, Sprague-Dawley

This study investigated the metabolic fate of kakkalide (irisolidone 7-xylosylglucoside), a major isoflavone found in extracts of Pueraria lobata flowers, and in rat urine, bile, and feces. Using HPLC/UV or LC/MS/MS methods, seven metabolites, tectorigenin-7-O-glucuronide, tectorigenin-7-O-sulfate, tectorigenin-4'-O-sulfate, 6-OH biochanin A-glucuronide, irisolidone-7-O-glucuronide, tectorigenin, and irisolidone were identified in rat urine after oral administration of kakkalide. Furthermore, irisolidone-7-O-glucuronide was found in bile, and irisolidone and kakkalide were found in feces. An HPLC/UV method for simultaneous quantification of all the metabolites and kakkalide in urine, bile, and feces was developed using daidzein or apigenin as the internal standard. Over a 72-h period, 13.2 ± 2.8 % of the kakkalide was excreted as seven metabolites in urine. Over the same time period, irisolidone-7-O-glucuronide excretion in bile accounted for 3.8 ± 1.1 % of the dose, while kakkalide and irisolidone excretion in feces accounted for 2.1 ± 0.7 % and 0.7 ± 0.1 % of the dose, respectively. The results indicate that urine is the primary route of kakkalide elimination in vivo and that extensive metabolism may be one of the reasons for the low bioavailability of kakkalide.

Topics: Administration, Oral; Animals; Bile; Chromatography, High Pressure Liquid; Chromatography, Liquid; Feces; Flavonoids; Glycosides; Isoflavones; Male; Pueraria; Rats; Rats, Wistar; Tandem Mass Spectrometry

The anti-inflammatory activities of kakkalide, a major constituent of the flower of Pueraria thunbergiana, and irisolidone, a metabolite of kakkalide produced by intestinal microflora, against carrageenan-induced inflammation in air pouches on the backs of mice and in lipopolysaccharide (LPS)-stimulated peritoneal macrophages were investigated. Kakkalide and irisolidone down-regulated the gene expression of cytokines [tumor necrosis factor alpha (TNF-α) and interleukin-1 beta (IL-1β)] and cyclooxygenase-2 (COX-2) and the production of pro-inflammatory cytokines, TNF-α and IL-1β, and inflammatory mediators, NO and prostaglandin E(2) (PGE(2)), in LPS-stimulated peritoneal macrophages. These agents also inhibited the phosphorylation of IκB-α and the nuclear translocation of nuclear factor-kappa B (NF-κB). Orally administered kakkalide and irisolidone significantly reduced carrageenan-induced inflammatory markers, leukocyte number, and protein amount in the exudates of the air pouch. These constituents also inhibited PGE(2) production and COX-2 inducible nitric oxide synthase, IL-1β, and TNF-α expression. These agents also inhibited NF-κB activation. The anti-inflammatory effects of irisolidone were more potent than those of kakkalide. Based on these findings, kakkalide and irisolidone may inhibit inflammatory reactions via NF-κB pathway, and irisolidone, a metabolite of kakkalide, may more potently inhibit these inflammatory reactions.

Topics: Animals; Anti-Inflammatory Agents; Carrageenan; Cells, Cultured; Cyclooxygenase 2; Dinoprostone; Flavonoids; Glycosides; Inflammation; Inflammation Mediators; Interleukin-1beta; Isoflavones; Lipopolysaccharides; Macrophages, Peritoneal; Male; Mice; Mice, Inbred C57BL; NF-kappa B; Nitric Oxide Synthase Type II; Signal Transduction; Tumor Necrosis Factor-alpha

The protective properties of irisolidone (a metabolite of kakkalide by intestinal bacteria) against hydrogen peroxide (H(2)O(2)) induced cell damage were investigated. Irisolidone was found to scavenge 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, and the intracellular reactive oxygen species (ROS), thereby preventing lipid peroxidation and DNA damage. Irisolidone inhibited apoptosis in Chinese hamster lung fibroblast (V79-4) cells induced by H(2)O(2) via radical scavenging activity. This was achieved by the activation of the extracellular signal regulated kinase (ERK) and DNA binding activity of activator protein-1 (AP-1) (a downstream transcription factor of ERK) by irisolidone.

Topics: Animals; Antioxidants; Apoptosis; Cell Line; Cricetinae; Cytoprotection; DNA Damage; Enzyme Activation; Extracellular Signal-Regulated MAP Kinases; Flavonoids; Free Radicals; Glycosides; Humans; Hydrogen Peroxide; Intestinal Mucosa; Intestines; Isoflavones; Lipid Peroxidation; Molecular Structure; Transcription Factor AP-1

As part of our search for anti-arteriosclerosis agents from traditional Chinese medicines, the 3-hydroxy-3-methylglutaryl (HMG)-CoA reductase (HCR)-inhibitory constituent, kakkalide, was isolated from the flower of Pueraria thunbergiana (PT, family Leguminosae). The antihyperlipidemic effects of kakkalide and its metabolite, irisolidone, which may be a bioactive form in vivo and potently inhibit the HCR activity, were investigated in vivo. Both the oral and interperitoneal administrations of kakkalide and irisolidone, with the exception of intraperitoneally treated kakkalide, potently lowered the serum levels of total cholesterol (TC) and triglyceride (TG) in Trition WR1339-induced hyperlipidemic mice. The oral administrations of kakkalide and irisolidone in hyperlipidemic mice induced, by the long-term feeding of a high fat diet, also potently reduced the serum levels of TC and TG and epididymal fat pad weight. These findings suggest that PT can improve hyperlipidemia, and the hypolipidemic effect may be due to HMG-CoA reductase.

Topics: Animals; Cholesterol; Cholesterol, HDL; Flavonoids; Flowers; Glycosides; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Hyperlipidemias; Isoflavones; Male; Mice; Mice, Inbred ICR; Plant Extracts; Polyethylene Glycols; Pueraria; Rats; Rats, Sprague-Dawley; Spectrometry, Mass, Fast Atom Bombardment; Surface-Active Agents; Triglycerides

To understand the relationship between the metabolism and estrogenic activity of kakkalide and tectoridin, main isoflavones in the flower of Pueraria thunbergiana (family Leguminosae), these isoflavones and their metabolites by human intestinal microflora as well as their estrogenic effects were investigated. All human fecal specimens metabolized kakkalide and tectoridin. All isolated kakkalide-hydrolyzing intestinal bacteria also hydrolyzed kakkalide and tectoridin to irisolidone and tectorigenin, respectively. When the estrogenic effects of kakkalide and tectoridin were compared with those of their metabolites irisolidone and tectorigenin, the metabolites more potently increased proliferation of MCF-7 cells than kakkalide and tectoridin. These metabolites also potently induced estrogen-response c-fos and pS2 mRNA expression. These results suggest that kakkalide and tectoridin may be metabolized mainly to irisolidone and tectorigenin, respectively, by intestinal microflora in the intestines, and which may be subsequently absorbed into the blood where they can express their estrogenic effect.

Topics: Adult; Bifidobacterium; Cell Line, Tumor; Cell Proliferation; Feces; Flavonoids; Flowers; Glycosides; Humans; Intestines; Isoflavones; Male; Phytoestrogens; Pueraria; Substrate Specificity; Xylosidases; Young Adult