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[Effect of Herba Epimedii Flavone on expression of OPG and RANKL in rat osteoblasts].

OBJECTIVE: To explore the effect of Herba Epimedii Flavone (HEF) on expression of OPG and RANKL in rat osteoblasts. METHODS: Osteoblasts were obtained from new born rat calvaria by digestive enzymes. The different HEF concentrations (0.1, 1, 10, 100 ng/ml)were added into the culture medium. Total mRNA was prepared from osteoblasts after 48 h and 96 h, and mRNA expression of OPG and RANKL were detected by RT-PCR. RESULTS: The mRNA expression of OPG in osteoblasts significantly increased compared with that of the control group, and the V(OPF)/V(RANKL) ratio was deceased markedly in the treatment of 10 ng/ml group after 48 h. After 96 h, the mRNA expression of OPG significantly increased and the V(OPG)/V(RANKL) ratio was also decreased in the treatment group compared with that of the control group. CONCLUSION: HEF suppress the differentiation and maturate of osteoblast. Osteoporosis can be prevented and cured by HEF.

Animals↗

[Studies on the flavones in of Chrozophora sabulosa].

OBJECTIVE: To study the flavone constituents in Chrozophora sabulosa (Xinjiang origin). METHOD: The compounds were extracted with 95% ethyl alcohol, isolated by various column chromatography and identified by spectroscopic methods. RESULT: Seven flavanoids were isolated and identified as quercetin (I), kaempferol (II), apigenin (III), chrysoerid (IV), isoquercitrin (V), chrysoerin-7-O-beta-D-glucoside (VI) and quercetin-3-O-alpha-D-arabinfuranoside (VII). CONCLUSION: All of these seven compounds were obtained from this genus for the first time.

Apigenin↗

[Effect of Herba Epimedii flavone on the osteoblasts metabolism in vitro].

OBJECTIVE: To explore the effect of Herba Epimedii flavone (HEF) on the osteoblast metabolism in vitro. METHOD: Osteoblast were obtained from new born rat calvaria by digestive enzymes. MTF, PNPP and RT-PCR were used to observe the proliferation, activity of ALP and mRNA expression of OPG and RANKL of cultured osteoblasts in vitro. RESULT: It was found that HEF had the effect on stimulating cell proliferation, activity of ALP and the mRNA expression of OPG of cultured osteoblasts (P < 0.01, P < 0.05). CONCLUSION: HEF can promote the proliferation, the differentiation and the expression of OPG mRNA of the osteoblasts cultured in vitro.

Alkaline Phosphatase↗

Inhibition of viral proteases by Zingiberaceae extracts and flavones isolated from Kaempferia parviflora.

In order to identify novel lead compounds with antiviral effect, methanol and aqueous extracts of eight medicinal plants in the Zingiberaceae family were screened for inhibition of proteases from human immunodeficiency virus type 1 (HIV-1), hepatitis C virus (HCV) and human cytomegalovirus (HCMV). In general, the methanol extracts inhibited the enzymes more effectively than the aqueous extracts. HIV-1 protease was strongly inhibited by the methanol extract of Alpinia galanga. This extract also inhibited HCV and HCMV proteases, but to a lower degree. HCV protease was most efficiently inhibited by the extracts from Zingiber officinale, with little difference between the aqueous and the methanol extracts. Many of the methanol extracts inhibited HCMV protease, but the aqueous extracts showed weak inhibition. In a first endeavor to identify the active constituents, eight flavones were isolated from the black rhizomes of Kaempferia parviflora. The most effective inhibitors, 5-hydroxy-7-methoxyflavone and 5,7-dimethoxyflavone, inhibited HIV-1 protease with IC50 values of 19 microM. Moreover, 5-hydroxy-3,7-dimethoxyflavone inhibited HCV protease and HCMV protease with IC50 values of 190 and 250 microM, respectively.

Cytomegalovirus↗

Flavonols and flavones of parsley cell suspension culture change the antioxidative capacity of plasma in rats.

The flavonoid aglycones from an illuminated parsley (Petroselinum crispum (Mill.) Nym.) cell suspension culture were identified and quantified as the flavones apigenin, luteolin and chrysoeriol and the flavonols kaempferol, quercetin and isorhamnetin. Flavonoid extracts from these cultures were purified by solid phase extraction from RP C-18 phase and given by gavage to rats. Only extract from illuminated culture increased the antioxidative capacity (AOC) of blood plasma temporarily with maximum values after 1 h. It is concluded that the course of AOC reflects changes in the plasma content of flavonoids.

Animals↗

Synthesis and antitumour activity of new derivatives of flavone-8-acetic acid (FAA). Part 3: 2-Heteroaryl derivatives.

A range of 14 derivatives of flavone-8-acetic acid (FAA) with a heterocyclic substituent in place of the 2-phenyl group have been prepared and their anti-tumour activity evaluated in vitro against a panel of human and murine tumour cell lines and in vivo against MAC 15A. Some of the compounds, notably 2c,d and s, showed significant in vivo activity and these require further studies in order to evaluate their potential for development.

Animals↗

Synthesis and antitumour activity of new derivatives of flavone-8-acetic acid (FAA). Part 4: Variation of the basic structure.

A range of 11 derivatives of flavone-8-acetic acid (FAA) in which the structure has been substantially altered in different ways have been prepared and their anti-tumour activity evaluated in vitro against a panel of human and murine tumour cell lines and in vivo against MAC 15A. The generally poor activity observed shows that the basic structure cannot be altered much without destroying the activity.

Animals↗

Synthesis and antitumour activity of new derivatives of flavone-8-acetic acid (FAA). Part 1: 6-Methyl derivatives.

A range of 17 derivatives of flavone-8-acetic acid (FAA) with a 6-methyl substituent have been prepared and their anti-tumour activity evaluated in vitro against a panel of human and murine tumour cell lines and in vivo against MAC 15A. While many of the compounds show activity comparable to FAA in vitro, this essentially disappears in vivo, possibly due to degradation before the compounds can reach the tumour site.

Animals↗

Synthesis and antitumour activity of new derivatives of flavone-8-acetic acid (FAA). Part 2: Ring-substituted derivatives.

A range of 18 derivatives of flavone-8-acetic acid (FAA) with substituents on the 2-phenyl group have been prepared and their anti-tumour activity evaluated in vitro against a panel of human and murine tumour cell lines and in vivo against MAC 15A. There was no clear-cut relationship between in vitro and in vivo activity but the activity in each situation was found to be very sensitive to the precise substitution pattern with closely related isomers giving widely different activities. Some of the compounds, notably 10b,c,j, and r, were active in vivo and these require further studies in order to evaluate their potential for development.

Animals↗

Determination of the flavone tricin in human plasma by high-performance liquid chromatography.

Tricin is a flavone constituent of brown rice and rice bran, which interferes potently with the survival of human-derived breast and colon cancer cells in vitro. A specific and simple high-performance liquid chromatographic (HPLC) method was developed for the determination of tricin in human plasma with UV-visible detection. HPLC separation on Hypersil-BDS C(18) (4.6 x 250 mm) was carried out with an isocratic mobile phase of 52% methanol in 0.1 m ammonium acetate, pH 5.10, containing 0.27 mm disodium ethylenediamine tetraacetic acid and detection at 355 nm. The retention times of tricin and quercetin (internal standard) were 14.2 and 7.8 min, respectively. The assay was linear in the range 1-100 microg/mL (r(2 ) > or = 0.995). Tricin in plasma was efficiently extracted with 0.1 m acetic acid in acetone, and the recoveries were in the range 92.6-102.8% (n = 6) with relative standard deviation below 10% for three concentrations of tricin, 5, 10 and 100 microg/mL. The lower limit of quantitation (relative standard deviation <20%) was 1 microg/mL.

Chromatography, High Pressure Liquid↗

A validated HPLC determination of the flavone aglycone diosmetin in human plasma.

Diosmetin, 3',5,7-trihydroxy-4'-methoxy flavone, is the aglycone of the flavonoid glycoside diosmin that occurs naturally in foods of plant origin. Diosmin exhibits antioxidant and anti-inflammatory activities, improves venous tone and it is used for the treatment of chronic venous insufficiency. Diosmin is hydrolyzed by enzymes of intestinal micro flora before absorption of its aglycone diosmetin. A specific, sensitive, precise, accurate and robust HPLC assay for the determination of diosmetin in human plasma was developed and validated. Diosmetin and the internal standard 7-ethoxycoumarin were isolated from plasma by liquid-liquid extraction and separated on a C8 reversed-phase column with methanol-water-acetic acid (55:43:2, v/v/v) as the mobile phase at 43 degrees C. Peaks were monitored at 344 nm. The method was linear in the 10-300 ng/mL concentration range (r > 0.999). Recovery for diosmetin and internal standard was greater than 89.7 and 86.8%, respectively. Intra-day and inter-day precision for diosmetin ranged from 1.6 to 4.6 and from 2.2 to 5.3%, respectively, and accuracy was better than 97.9%.

Chromatography, High Pressure Liquid↗

Determination of the putative cancer chemopreventive flavone tricin in plasma and tissues of mice by HPLC with UV--visible detection.

AHPLC method developed and validated for the determination of tricin in human plasma published previously was cross-validated to allow measurement of the flavone tricin in plasma and tissues of mice. Blank samples of plasma, liver or intestinal mucosa were spiked with tricin at 0.5--4.0, 1.0--8.0 and 5.0--40 microg/mL, respectively. These tricin concentration ranges covered the tricin levels achieved in the mouse tissues in the dose-escalating experiments. Analysis afforded linear calibration curves with regression coefficients of >0.99. Endogenous compounds did not interfere with tricin detection when the detection wavelength was set at 355 nm, the maximum absorbance of tricin. Accuracy and precision were <15% for all concentrations in all matrices except for the precision at the lower limit of quantification (0.5 microg/mL) in mouse plasma, which was 18.4%. Consumption of diet mixed with tricin at 0.05, 0.2 or 0.5% for one week furnished steady-state levels in plasma, liver and small intestine in the 1--3 x 10(-7), 4--22 x 10(-7) and 3--46 x 10(-5) m ranges, respectively.

Animals↗

Selective induction of endothelial cell tissue factor in the presence of a tumour-derived mediator: a potential mechanism of flavone acetic acid action in tumour vasculature.

Flavone acetic acid (FAA) is a potentially useful anti-tumour agent which has been reported to induce changes in tumour vasculature, in particular loss of bloodflow. This led us to examine whether endothelium could be a cellular target of FAA action, with resultant modulation of cell-surface coagulant properties leading to activation of coagulation and blockade of tumour blood flow. Incubation of endothelium with FAA led to the expression of functional tissue factor on the cell surface, in a time-dependent and dose-dependent (half-maximal at 0.6-0.7 mg/ml) manner. Induction of tissue-factor activity resulted from de novo translation of the tissue factor message. To explain the selectivity of FAA's action on tumour vasculature in vivo, we considered its interaction with tumour-derived factors. Starting with serum-free FO-I-melanoma cell-conditioned medium, a co-factor enhancing FAA-mediated induction of endothelial tissue factor (FO-I factor) was partially purified by sequential ion exchange and reverse phase chromatography, followed by preparative SDS-PAGE. The FO-I factor migrates with an apparent Mr of approx. 20 to 25,000 on non-reduced SDS-PAGE, is sensitive to protease K, and augments the effect of FAA on endothelial-cell-tissue factor. This activity is not found in supernatants from non-neoplastically transformed cell lines. These data lead us to hypothesize that FAA exerts its action, at least in part, by promoting activation of coagulation on the endothelial surface, and this effect is selective for the tumour bed by virtue of its interaction with a tumour-derived factor. The interaction of FAA with host factors may be important for optimizing its therapeutic efficacy for a particular tumour.

Blood Coagulation Factors↗

Anti-vascular approaches to solid tumour therapy: evaluation of vinblastine and flavone acetic acid.

Several agents have now been identified which exert their anti-tumour effects in large part via the tumour vasculature; these include TNF alpha and flavone acetic acid (FAA). More recently, Vincristine and Vinblastine have also been shown to cause a prolonged and selective decrease in tumour perfusion. Vinblastine, unlike, FAA, causes no increase in plasma TNF alpha levels in mice bearing the CaNT tumour, suggesting 2 distinct mechanisms of anti-vascular activity for these structurally diverse agents. Since FAA and Vinblastine also show quite different normal tissue toxicities, which are separately dose-limiting, we have examined the strategy of combining these 2 agents. When Vinblastine preceded FAA by 24 hr or less, tumour growth delay was significantly enhanced without a concomitant increase in toxicity. The level of enhancement was not significantly reduced by a 5-fold decrease in Vinblastine dose, though any reduction in the dose of FAA caused a rapid reduction in treatment effectiveness. Investigation of the functional vasculature of treated tumours suggested that increased anti-vascular effects may contribute to the enhanced growth inhibition of the combined treatment. Our results demonstrate the potential benefit of combining 2 different classes of anti-vascular agent, using Vinblastine and FAA (or 5,6-MeXAA) as prototype drugs.

Adenocarcinoma↗

Fragmentation study of rutin, a naturally occurring flavone glycoside cationized with different alkali metal ions, using post-source decay matrix-assisted laser desorption/ionization mass spectrometry.

A post-source decay matrix-assisted laser desorption/ionization mass spectrometric (PSD-MALDI-MS) study of rutin, a naturally occurring flavone glycoside cationized with different alkali metal ions, is reported. The fragmentations of rutin were performed by selecting the [R + Cat]+ peaks for PSD, where R represents a rutin molecule and Cat an alkali metal ion (Li+, Na+, K+). The PSD-MALDI mass spectra showed, depending on Cat, different fragmentation patterns with respect to both the quality and quantity of the fragment ions formed. The intensity of fragmentation decreased in the order Li+ > Na+ > K+. The fragmentation mechanism and an explanation for the observed differences are suggested.

Cations↗

On-line identification of further flavone C- and O-glycosides from sugarcane (Saccharum officinarum L., Gramineae) by HPLC-UV-MS.

HPLC-UV-MS (APCI-MS/MS and CID/MS) was utilised for the identification of eight additional flavone glycosides from sugarcane (Saccharum officinarum L., Gramineae) extracts of bagasse, leaves and juice ('garapa'). Relevant information about substitution patterns was obtained through UV detection using post-column addition of shift reagents, while tandem MS provided structural information confirming the proposed structures of the C-glycosides vitexin, orientin, luteolin-8-C-(rhamnosylglucoside), 4',5'-dimethyl-luteolin-8-C-glycoside and the isomeric pair schaftoside-isoschaftoside, besides the O-glycosides tricin-7-O-neohesperidoside and tricin-7-O-glycoside.

Chromatography, High Pressure Liquid↗

A fragmentation study of a flavone triglycoside, kaempferol-3-O-robinoside-7-O-rhamnoside.

A mass spectrometric method based on the combined use of electrospray ionization, collision-induced dissociation and tandem mass spectrometry has been applied to the structural characterization of the flavone triglycoside, robinin (3,5,7,4'-tetrahydroxyflavone-3-O-robinoside-7-O-rhamnoside). The deprotonated molecule fragments by loss of the rhamnose glycan residue to yield the Y(7) (-) ion (m/z 593) and by scission of the robinose glycan residue to yield the radical anion [Y(3,0)-H](-.) (m/z 430). The Y(7) (-) ion fragments by scission of the robinose glycan residue to yield the radical anion of Y(7)[Y(3,0)-H](-.) (m/z 284). The [Y(3,0)-H](-.) radical anion fragments by loss of the rhamnose glycan residue to yield the radical anion Y(7)[Y(3,0)-H](-.) (m/z 284) and by scission to yield [Y(7)-H][Y(3,0)--H](-) (m/z 283). A fragmentation mechanism has been proposed.

Flavonoids↗