Objective: To investigate the anti-colon cancer effects of ethylacetate fraction from Orostachys japonicus(0. japonicus) on HT-29 cancer cells. Methods: The viability of HT-29 cells was assayed by the 3-(4,5-dimethylt...Objective: To investigate the anti-colon cancer effects of ethylacetate fraction from Orostachys japonicus(0. 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)-2 H-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 with4',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_1 and S phase. Among the upstream and downstream proteins involved in anti-cancer activity, the level of B cell lymphoma-2 decreased, and the bcl-2-associated x protein increased. The level of pro-caspase-3, pro-caspase-8, and pro-caspase-9 decreased, while the level of cleaved-caspase-3, cleaved-caspase-8, and cleaved-caspase-9 increased. Moreover, the phosphorylation, that is, activation of extracellular signal regulated kinase 1/2, Jun-N-terminal kinase, and p38 increased. Conclusions: Combining the above results, it is thought that the survival of HT-29 cells is suppressed by ethylacetate fraction from0. japonicus through mitochondrial regulation-induced caspase cascade activation, induction of apoptosis and cell cycle arrest.展开更多
This study aimed to investigate the tissue culture and rapid propagation techniques of Orostachys fimbriata, a medicinal and ornamental herbaceous perennial herb belonging to the family Crassulaceae. The leaves of O. ...This study aimed to investigate the tissue culture and rapid propagation techniques of Orostachys fimbriata, a medicinal and ornamental herbaceous perennial herb belonging to the family Crassulaceae. The leaves of O. fimbriata were used as explants to investigate the effects of plant growth regulators such as thidiazuron (TDZ), 6-benzylamino purine (6-BA) and 1-naphthaleneacetic acid (NAA), on the induction of callus, differentiation of adventitious buds and rooting of shoots. Our results showed that optimum callus induction medium was MS medium supplemented with 0.5 mg·L<sup>-1</sup> TDZ and 0.2 mg·L<sup>-1</sup> NAA. 1.5 mg·L<sup>-1</sup> 6-BA and 0.2 mg·L<sup>-1</sup> NAA was the optimum hormone combination for differentiation of adventitious buds. And 0.1 mg·L<sup>-1</sup> NAA could efficiently promote rooting. The survival rate of transplants reached about 90%. In this study, using leaves of O. fimbriata as explants, high efficient tissue culture and regeneration system of O. fimbriata were established, and the period from leave to transplant plantlet was about 3 months. The presently developed protocol could be used for large scale clonal propagation and germplasm conservation of O. fimbriata. The efficient tissue culture system of O. fimbriata would provide technical support for its utilization.展开更多
Objective: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.japonicu...Objective: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-hexane,dichloromethane(DCM),ethyl acetate(EtOAc),n-butanol,and H2 O 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/m L through down-regulating the expression of mec A gene and the production and agglutination of penicillin-binding protein 2 a 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.展开更多
基金supported by the 2016 Inje University research grant
文摘Objective: To investigate the anti-colon cancer effects of ethylacetate fraction from Orostachys japonicus(0. 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)-2 H-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 with4',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_1 and S phase. Among the upstream and downstream proteins involved in anti-cancer activity, the level of B cell lymphoma-2 decreased, and the bcl-2-associated x protein increased. The level of pro-caspase-3, pro-caspase-8, and pro-caspase-9 decreased, while the level of cleaved-caspase-3, cleaved-caspase-8, and cleaved-caspase-9 increased. Moreover, the phosphorylation, that is, activation of extracellular signal regulated kinase 1/2, Jun-N-terminal kinase, and p38 increased. Conclusions: Combining the above results, it is thought that the survival of HT-29 cells is suppressed by ethylacetate fraction from0. japonicus through mitochondrial regulation-induced caspase cascade activation, induction of apoptosis and cell cycle arrest.
文摘This study aimed to investigate the tissue culture and rapid propagation techniques of Orostachys fimbriata, a medicinal and ornamental herbaceous perennial herb belonging to the family Crassulaceae. The leaves of O. fimbriata were used as explants to investigate the effects of plant growth regulators such as thidiazuron (TDZ), 6-benzylamino purine (6-BA) and 1-naphthaleneacetic acid (NAA), on the induction of callus, differentiation of adventitious buds and rooting of shoots. Our results showed that optimum callus induction medium was MS medium supplemented with 0.5 mg·L<sup>-1</sup> TDZ and 0.2 mg·L<sup>-1</sup> NAA. 1.5 mg·L<sup>-1</sup> 6-BA and 0.2 mg·L<sup>-1</sup> NAA was the optimum hormone combination for differentiation of adventitious buds. And 0.1 mg·L<sup>-1</sup> NAA could efficiently promote rooting. The survival rate of transplants reached about 90%. In this study, using leaves of O. fimbriata as explants, high efficient tissue culture and regeneration system of O. fimbriata were established, and the period from leave to transplant plantlet was about 3 months. The presently developed protocol could be used for large scale clonal propagation and germplasm conservation of O. fimbriata. The efficient tissue culture system of O. fimbriata would provide technical support for its utilization.
基金supported by the 2016 creative research program of Inje University.
文摘Objective: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-hexane,dichloromethane(DCM),ethyl acetate(EtOAc),n-butanol,and H2 O 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/m L through down-regulating the expression of mec A gene and the production and agglutination of penicillin-binding protein 2 a 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.
