ABSTRACT
To investigate the effect of Orostachys (O.) japonicus, a perennial herbaceous plant of the Family Crassulaceae, on biofilm formed by methicillin-resistant Staphylococcus aureus (MRSA). Methods: Powdered O. japonicus was extracted by 95% methanol, concentrated, and then, systematically fractionated with n-heane, dichloromethane (DCM), ethyl acetate (EtOAc), n-butanol, and H2O according to polarity. Among them, the flavonoid-rich EtOAc fraction demonstrated the highest antibacterial activity and was used in this study. Using the biofilm inhibition assay, cell-surface attachment assay, confocal laser scanning microscopy, latex agglutination assay, and real time qRT-PCR, we examined whether the EtOAc fraction inhibited the formation of MRSA biofilm. Results: The EtOAc fraction exhibited distinct activity against biofilm formation and cell-surface attachment of MRSA up to 1 mg/mL through down-regulating the expression of mecA gene and the production and agglutination of penicillin-binding protein 2a as solidly observed in biofilm inhibition assay, cell-suface attachment assay, confocal laser scanning microscopy, latex agglutination assay, and real time qRT-PCR analysis. Conclusions: These results suggest that O. japonicus could be utilized as a potential resource for the development of new antibiofilm formation of MRSA and antibacterial agents in the future.
ABSTRACT
Objective: To determine the bioactive compounds of Orostachys japonicus (O. japonicus) (Maxim.) A. Berger extraction by different solvents (70％ ethanol or water) and to evaluate the in vitro antioxidant and anti-inflammatory activities.Methods: Total polyphenol, flavonoid, and anthocyanin contents in O. japonicus extract were measured. Antioxidant activities were determined by DPPH, ABTS, and superoxide radical-scavenging ability assays. Anti-inflammatory activities were evaluated by nitric oxide production, tumor necrosis factor-α, and interleukin-6 expression techniques. Results: Extraction with 70％ ethanol yielded the highest total polyphenol (60.03 mg/g dry weight) and flavonoid (55.50 mg/g dry weight) contents. The total anthocyanin contents in 70％ ethanol and water extracts were 57.25 and 91.71 mg/g dry weight, respectively. The 70％ ethanol extract also showed stronger antioxidant activity than the water extract. Antioxidant activity and reducing power increased with the increasing concentration of O. japonicus extract. O. japonicus extract at 0-400 μg/mL did not affect the growth of RAW 264.7 cells, whereas dose-dependent inhibition of nitric oxide, tumor necrosis factor-α, and interleukin-6 production was observed. Conclusions: O. japonicus inhibits oxidative and inflammatory reactions with potentially positive health-related effects.
ABSTRACT
To investigate the effect of Orostachys (O.) japonicus, a perennial herbaceous plant of the Family Crassulaceae, on biofilm formed by methicillin-resistant Staphylococcus aureus (MRSA). Methods: Powdered O. japonicus was extracted by 95% methanol, concentrated, and then, systematically fractionated with n-heane, dichloromethane (DCM), ethyl acetate (EtOAc), n-butanol, and H2O according to polarity. Among them, the flavonoid-rich EtOAc fraction demonstrated the highest antibacterial activity and was used in this study. Using the biofilm inhibition assay, cell-surface attachment assay, confocal laser scanning microscopy, latex agglutination assay, and real time qRT-PCR, we examined whether the EtOAc fraction inhibited the formation of MRSA biofilm. Results: The EtOAc fraction exhibited distinct activity against biofilm formation and cell-surface attachment of MRSA up to 1 mg/mL through down-regulating the expression of mecA gene and the production and agglutination of penicillin-binding protein 2a as solidly observed in biofilm inhibition assay, cell-suface attachment assay, confocal laser scanning microscopy, latex agglutination assay, and real time qRT-PCR analysis. Conclusions: These results suggest that O. japonicus could be utilized as a potential resource for the development of new antibiofilm formation of MRSA and antibacterial agents in the future.
ABSTRACT
To determine the bioactive compounds of Orostachys japonicus (O. japonicus) (Maxim.) A. Berger extraction by different solvents (70% ethanol or water) and to evaluate the in vitro antioxidant and anti-inflammatory activities. Methods: Total polyphenol, flavonoid, and anthocyanin contents in O. japonicus extract were measured. Antioxidant activities were etermined by DPPH, ABTS, and superoxide radical-scavenging ability assays. Anti-inflammatory activities were evaluated by nitric oxide production, tumor necrosis factor-a, and interleukin-6 expression techniques. Results: Extraction with 70% ethanol yielded the highest total polyphenol (60.03 mg/g dry weight) and flavonoid (55.50 mg/g dry weight) contents. The total anthocyanin contents in 70% ethanol and water extracts were 57.25 and 91.71 mg/g dry weight, respectively. The 70% ethanol extract also showed stronger antioxidant activity than the water extract. Antioxidant activity and reducing power increased with the increasing concentration of O. japonicus extract. O. japonicus extract at 0-400 ug/mL did not affect the growth of RAW 264.7 cells, whereas dose-dependent inhibition of nitric oxide, tumor necrosis factor-a, and interleukin-6 production was observed. Conclusions: O. japonicus inhibits oxidative and inflammatory reactions with potentially positive health-related effects.
ABSTRACT
Objective: To investigate the anti-colon cancer effects of ethylacetate fraction from Orostachys japonicus (O. japonicus) on HT-29 cancer cells. Methods: The viability of HT-29 cells was assayed by the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) method. Apoptosis induction and cell cycle inhibition were confirmed by fluorescein isothiocyanate and propidium iodide staining using flow cytometry. Morphological changes in the nucleus were observed, using a fluorescence microscope with 4′,6-diamidino-2-phenylindole (DAPI) nuclear staining. The expression levels of the upstream and downstream proteins involved in the anti-cancer mechanism were confirmed by Western blotting. Results: After treating HT-29 cells with different concentrations of ethylacetate fraction from O. japonicus, the viability of cells decreased in a concentration-dependent manner, while apoptosis induction and apoptotic body formation increased. Cell cycle analysis showed that the arrest occurred at the sub-G
ABSTRACT
Objective:To investigate the anti-colon cancer effects of ethylacetate fraction from Orostachys japonicus (O. japonicus) on HT-29 cancer cells.Methods:The viability of HT-29 cells was assayed by the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) method. Apoptosis induction and cell cycle inhibition were confirmed by fluorescein isothiocyanate and propidium iodide staining using flow cytometry. Morphological changes in the nucleus were observed, using a fluorescence microscope with 4′,6-diamidino-2-phenylindole (DAPI) nuclear staining. The expression levels of the upstream and downstream proteins involved in the anti-cancer mechanism were confirmed by Western blotting.Results:After treating HT-29 cells with different concentrations of ethylacetate fraction from O. japonicus, the viability of cells decreased in a concentration-dependent manner, while apoptosis induction and apoptotic body formation increased. Cell cycle analysis showed that the arrest occurred at the sub-GConclusions:Combining the above results, it is thought that the survival of HT-29 cells is suppressed by ethylacetate fraction from O. japonicus through mitochondrial regulation-induced caspase cascade activation, induction of apoptosis and cell cycle arrest.
ABSTRACT
The hypolipidemic and hypoglycemic effects of two dietary dosages (0.1% and 0.5%) of water and 80% ethanol extracts from hot-air dried Orostachys japonicus A. Berger were evaluated in the serum and organ tissues of streptozotocin-induced diabetic rats. The STZ-induced diabetic groups supplemented with the O. japonicus extracts showed significantly higher body weight compared to a diabetic control group at the end of experiment. The extracts exhibited substantial hypoglycemic effects by significant reductions of fasting blood glucose levels at all time points tested compared to the initial stage before treatment of the extracts. Declines of serum and hepatic triglyceride levels were greater than declines of total cholesterol in the groups treated with the 0.5% O. japonicus extract (DBW2 and DBE2) when compared to the DBC group. Hepatic glycogen content was higher in the groups treated with O. japonicus extract, while lipid peroxide content was decreased in these treated groups compared to the DBC group. Hepatic antioxidant activity was significantly increased in the groups supplemented with the O. japonicus ethanol extract. The hypolipidemic and hypoglycemic effects of the O. japonicus ethanol extract were significantly greater than the effects of the water extract. Based on this study, it seems that O. japonicus ethanol extract, due to its higher phenolic and flavonoid components than the water extract, may control blood glucose and alleviate hyperlipidemia in diabetes.