NAD(P)H oxidases were detected in suspension cultured cells of ginseng (Panax ginseng C. A. Meyer). The activities of these enzymes were induced by an elicitor (Cle) extracted from cell walls of Col-letotrichum lagera...NAD(P)H oxidases were detected in suspension cultured cells of ginseng (Panax ginseng C. A. Meyer). The activities of these enzymes were induced by an elicitor (Cle) extracted from cell walls of Col-letotrichum lagerarium. In addition, Cle induced an oxidative burst and enhanced the synthesis of saponin, activity of phenylalanine ammonialyase (PAL) , accumulation of chalcone synthase (CHS) and the transcription of a hydroxyproline-rich glycoprotein gene ( hrgp ) . Pre-treatments with DPI and quinacrine (two inhibitors of mammalian neutrophil plasma membrane NADPH oxidase) for 30 min prior to Cle addition blocked the NAD(P)H oxidase activity induced by Cle. These inhibitors also inhibited the release of H2C2, the synthesis of saponin, PAL activity and CHS accumulation. Our data revealed homology between plasma membrane NAD(P)H oxidases of mammalian neutrophil cells and ginseng suspension cells. They also indicated that deactivated NAD(P)H oxidases catalysed the release of H2O2 and that H2O2 was functioning as a second messenger stimulating PAL activity, saponin synthesis and hrgp transcription. Elevations of Ca2 + and protein phos-phorylation/dephosphorylation were required for this defense process. We propose that NAD(P)H oxidases mediate the processes of Cle-induced defense responses in ginseng suspensions, and postulate the existence of a signalling cascade including extracellular Cle stimulation, activation of plasma membrane NAD(P)H oxidases, release of H2O2, and the intracellular responses of metabolism and gene transcription in ginseng suspension cells.展开更多
Endogenous elicitor, termed cellulase-degraded cell wall (CDW), was prepared from the cell wall of suspension-cultured ginseng (Panax ginseng C.A. Meyer) cells via cellulase degradation. CDW activated the NADPH oxidas...Endogenous elicitor, termed cellulase-degraded cell wall (CDW), was prepared from the cell wall of suspension-cultured ginseng (Panax ginseng C.A. Meyer) cells via cellulase degradation. CDW activated the NADPH oxidase activity of isolated plasma membranes and stimulated in vivo H2O2 generation in ginseng cell suspensions. CDW also increased the activity of phenylalanine ammonia lyase (PAL), expression of a P. ginseng squalene epoxidase (sqe) gene and saponin synthesis. NADPH oxidase inhibitors inhibited both in vitro NADPH oxidase activity and in vivo H2O2 generation. Induction of PAL activity, saponin synthesis and sqe gene expression were all inhibited by such inhibitor treatments and reduced by incubation with catalase and HA scavengers. These data indicate that activation of NADPH oxidase and generation of H2O2 are essential signalling events mediating defence responses induced by the endogenous elicitor(s) present in CDW.展开更多
The removal of lead from ginseng ethanol extracts by a fixed-bed column filled with an adsorbent bearing amine and carboxyl groups was investigated. The Pb2+ content was determined by inductively coupled plasma mass s...The removal of lead from ginseng ethanol extracts by a fixed-bed column filled with an adsorbent bearing amine and carboxyl groups was investigated. The Pb2+ content was determined by inductively coupled plasma mass spectrometry. When the flowrate increased from 0.12 to 0.34 ml·min-1 , the column exhibited a marked increase in percentage of lead removal from 54.9% to 92.3%. Further increase in the flowrate did not bring evident changes to the lead removal, whereas an increase in the temperature could reinforce adsorption further, suggesting that the adsorption process was controlled by external film diffusion below the flowrate of 0.34 ml·min-1 , and by the intraparticle pore diffusion of lead ions when the flowrate exceeded it. A low remaining lead amount in extracts such as 0.11 mg·kg-1 (extracts powder) was achieved. The adsorbents also adsorbed effective constituents to some extent. But 88% of constituents adsorbed were taken off using a 70% ethanol aqueous solution.展开更多
Objective：The aim of this study was to investigate the af ecting of Rg3 to secreted VEGF of human laryngeal carcinoma Hep-2 cells and its mechanism of inhibition to tumor angiogenesis. Methods：Cultured human larynge...Objective：The aim of this study was to investigate the af ecting of Rg3 to secreted VEGF of human laryngeal carcinoma Hep-2 cells and its mechanism of inhibition to tumor angiogenesis. Methods：Cultured human laryngeal cancer cellline Hep-2 and human vascular endothelial cells in vitro, cells got into the period of exponential phase of growth, was diviced into 3 groups：group I （control group）, group II （DDP group）, group III （Rg3 group）. Added to the Hep-2 cells Rg3 and DDP, made Rg3 final concentration was 300μg/mL, and DDP was 3μg/mL. 48 h later, specimens from sample to be done immunocytochemistry, and the protein of VEGF in Hep-2 cells to be detected. Col ecting Hep-2 cells supernatant, some was used to measure the protein level of VEGF in Hep-2 cells supernatant by ELISA. Some was used to culture HVEC. 24 h later, cellgrowth inhibition rate of human vascular endothelial was determined by MTT. Results：The protein level of VEGF was evi-dently higher in group I compared to group II and group III, it was not only in Hep-2 cells, but also in supernatant of Hep-2 cells. There was no significantly dif erent between group II and group III. MTT results showed that, the human vascular endothelial cellgrowth inhibition rate of group I was significantly lower than that of group II and group III （P〈0.05）. At the same time the HVEC growth inhibition rate of group II was significantly lower than that of group III （P〈0.05）. Conclusion：The inhibition to tumor angiogenesis of Rg3 is stronger than traditional chemotherapy drug cisplatin. It worke by reducing the biological ef ects of secreted VEGF, But the ef ecting worke by reducing the activity of secreted VEGF itself or af ecting endothelial function of VEGF receptor or some other ways to be further studied.展开更多
This study reports an increase in power generation of a MFC （microbial fuel cell） by the addition of Korean ginseng （Panax ginseng）. It was noted that the use of ginseng enhances the microbial anaerobic degradatio...This study reports an increase in power generation of a MFC （microbial fuel cell） by the addition of Korean ginseng （Panax ginseng）. It was noted that the use of ginseng enhances the microbial anaerobic degradation of cellobiose, a disaccharide that was used as a substrate in the anode chamber of the MFC. The power output of the MFC where ginseng was added showed noticeable enhancement compared to the control MFC. The increase slowly ramped at the initial days and became appreciably higher after the 11th day of incubation in an experiment set up for 16 days duration. It is attributed that the ginseng increases the CO2 production by accelerating the fermentation process. Decrease in CH4/CO2 ratio was observed also due to decrease in methane production per digested cellobiose, the proton donor in the current study. Four ring steroid-like structural moiety Ginsenoside of Panax ginseng seemed to play a beneficial role in the electron transfer from ceilobiose to the anode, perhaps by rendering easier electron transfer due to favorable energy level alignments.展开更多
The spectroscopy characteristics and the fluorescence lifetime for the chloroplasts isolated from the pseudo ginseng, water hyacinth and spinach plant leaves have been studied by absorption spectra, low temperature st...The spectroscopy characteristics and the fluorescence lifetime for the chloroplasts isolated from the pseudo ginseng, water hyacinth and spinach plant leaves have been studied by absorption spectra, low temperature steady-state fluorescence spectroscopy and single photon counting measurement under the same conditions and by the same methods. The similarity of the absorption spectra for the chloroplasts at room temperature suggests that different plants can efficiently absorb light of the same wavelength. The fluorescence decays in PS II measured at the natural QA state for the chloroplasts have been fitted by a three-exponential kinetic model. The three fluorescence lifetimes are 30, 274 and 805 ps for the pseudo ginseng chloroplast; 138, 521 and 1494 ps for the water hyacinth chloroplast; 197, 465 and 1459 ps for the spinach chloroplast, respectively. The slow lifetime fluorescence component is assigned to a collection of associated light harvesting Chl a/b proteins, the fast lifetime component to the reaction center of PS II and the middle lifetime component to the delay fluorescence of recombination of P+ 680 and Pheo-. The excitation energy conversion efficiency(η) in PS II RC is defined and calculated on the basis of the 20 ps electron transfer time constant model, 60%, 87% and 91% for the pseudo ginseng, water hyacinth and spinach chloroplasts, respectively. This interesting result is in unconformity with what is assumed to be 100% efficiency in PS II RC. Our result in this work stands in line with the 20 ps electron transfer time constant in PS II rather sound and the water hyacinth plant grows slower than the spinach plant does as envisaged on the efficiency. But, our results predict that those plants can perform highly efficient transfer of photo-excitation energy from the light-harvesting pigment system to the reaction center (closely to 100%). The conclusion contained in this paper reveals the plant growth characteristics expressed in the primary processes of photosynthesis and a relationship between a plant growing rate and its spectroscopy characteristics and fluorescence lifetimes, namely, the slower a plant grows, the less excitation energy conversation efficiency used might be anticipated.展开更多
OBJECTIVE: To study the mechanism of the inhibitory effect of ginsenoside Rg3 on colon cancer cell migration.METHODS: Transwell migration assays were performed to investigate the inhibitory effect of ginsenoside Rg3 o...OBJECTIVE: To study the mechanism of the inhibitory effect of ginsenoside Rg3 on colon cancer cell migration.METHODS: Transwell migration assays were performed to investigate the inhibitory effect of ginsenoside Rg3 on SW480 cell migration. Electrophoretic mobility shift assays(EMSAs) and dual luciferase reporter assays were used to study the suppression capability of Rg3 on nuclear factor kappa B(NF-κB) activity. Western blotting was adopted to determine protein levels.RESULTS: Two-hundred micromolar ginsenoside Rg3 significantly inhibited SW480 cell migration(P < 0.05). EMSA showed that Rg3 suppressed the DNA binding ability of NF-κB. Dual luciferase reporter assay showed that Rg3 decreased NF-κB-regulated gene transcription(P < 0.01). Western blots indicated that Rg3 down-regulated expression of the NF-κB-regulated matrix metalloproteinase 9,cyclooxygenase-2 and C-Myc. An NF-κB inhibitor,pyrrolidine dithiocarbamate,enhanced the inhibitory effect of Rg3 on SW480 cell migration.CONCLUSION: Ginsenoside Rg3 has a strong antitumor migration capability by suppressing NF-κB activity and expression of NF-κB-regulated gene products. It could be a good adjuvant for colon cancer patients during the course of chemotherapy.展开更多
文摘NAD(P)H oxidases were detected in suspension cultured cells of ginseng (Panax ginseng C. A. Meyer). The activities of these enzymes were induced by an elicitor (Cle) extracted from cell walls of Col-letotrichum lagerarium. In addition, Cle induced an oxidative burst and enhanced the synthesis of saponin, activity of phenylalanine ammonialyase (PAL) , accumulation of chalcone synthase (CHS) and the transcription of a hydroxyproline-rich glycoprotein gene ( hrgp ) . Pre-treatments with DPI and quinacrine (two inhibitors of mammalian neutrophil plasma membrane NADPH oxidase) for 30 min prior to Cle addition blocked the NAD(P)H oxidase activity induced by Cle. These inhibitors also inhibited the release of H2C2, the synthesis of saponin, PAL activity and CHS accumulation. Our data revealed homology between plasma membrane NAD(P)H oxidases of mammalian neutrophil cells and ginseng suspension cells. They also indicated that deactivated NAD(P)H oxidases catalysed the release of H2O2 and that H2O2 was functioning as a second messenger stimulating PAL activity, saponin synthesis and hrgp transcription. Elevations of Ca2 + and protein phos-phorylation/dephosphorylation were required for this defense process. We propose that NAD(P)H oxidases mediate the processes of Cle-induced defense responses in ginseng suspensions, and postulate the existence of a signalling cascade including extracellular Cle stimulation, activation of plasma membrane NAD(P)H oxidases, release of H2O2, and the intracellular responses of metabolism and gene transcription in ginseng suspension cells.
文摘Endogenous elicitor, termed cellulase-degraded cell wall (CDW), was prepared from the cell wall of suspension-cultured ginseng (Panax ginseng C.A. Meyer) cells via cellulase degradation. CDW activated the NADPH oxidase activity of isolated plasma membranes and stimulated in vivo H2O2 generation in ginseng cell suspensions. CDW also increased the activity of phenylalanine ammonia lyase (PAL), expression of a P. ginseng squalene epoxidase (sqe) gene and saponin synthesis. NADPH oxidase inhibitors inhibited both in vitro NADPH oxidase activity and in vivo H2O2 generation. Induction of PAL activity, saponin synthesis and sqe gene expression were all inhibited by such inhibitor treatments and reduced by incubation with catalase and HA scavengers. These data indicate that activation of NADPH oxidase and generation of H2O2 are essential signalling events mediating defence responses induced by the endogenous elicitor(s) present in CDW.
基金Supported by the National＇Natural Science Foundation of China （20976012, 20806009）, and the Specialized Research Fund for the Doctoral Program of Higher Education （20070007055, 20091101110035）.
文摘The removal of lead from ginseng ethanol extracts by a fixed-bed column filled with an adsorbent bearing amine and carboxyl groups was investigated. The Pb2+ content was determined by inductively coupled plasma mass spectrometry. When the flowrate increased from 0.12 to 0.34 ml·min-1 , the column exhibited a marked increase in percentage of lead removal from 54.9% to 92.3%. Further increase in the flowrate did not bring evident changes to the lead removal, whereas an increase in the temperature could reinforce adsorption further, suggesting that the adsorption process was controlled by external film diffusion below the flowrate of 0.34 ml·min-1 , and by the intraparticle pore diffusion of lead ions when the flowrate exceeded it. A low remaining lead amount in extracts such as 0.11 mg·kg-1 (extracts powder) was achieved. The adsorbents also adsorbed effective constituents to some extent. But 88% of constituents adsorbed were taken off using a 70% ethanol aqueous solution.
文摘Objective：The aim of this study was to investigate the af ecting of Rg3 to secreted VEGF of human laryngeal carcinoma Hep-2 cells and its mechanism of inhibition to tumor angiogenesis. Methods：Cultured human laryngeal cancer cellline Hep-2 and human vascular endothelial cells in vitro, cells got into the period of exponential phase of growth, was diviced into 3 groups：group I （control group）, group II （DDP group）, group III （Rg3 group）. Added to the Hep-2 cells Rg3 and DDP, made Rg3 final concentration was 300μg/mL, and DDP was 3μg/mL. 48 h later, specimens from sample to be done immunocytochemistry, and the protein of VEGF in Hep-2 cells to be detected. Col ecting Hep-2 cells supernatant, some was used to measure the protein level of VEGF in Hep-2 cells supernatant by ELISA. Some was used to culture HVEC. 24 h later, cellgrowth inhibition rate of human vascular endothelial was determined by MTT. Results：The protein level of VEGF was evi-dently higher in group I compared to group II and group III, it was not only in Hep-2 cells, but also in supernatant of Hep-2 cells. There was no significantly dif erent between group II and group III. MTT results showed that, the human vascular endothelial cellgrowth inhibition rate of group I was significantly lower than that of group II and group III （P〈0.05）. At the same time the HVEC growth inhibition rate of group II was significantly lower than that of group III （P〈0.05）. Conclusion：The inhibition to tumor angiogenesis of Rg3 is stronger than traditional chemotherapy drug cisplatin. It worke by reducing the biological ef ects of secreted VEGF, But the ef ecting worke by reducing the activity of secreted VEGF itself or af ecting endothelial function of VEGF receptor or some other ways to be further studied.
文摘This study reports an increase in power generation of a MFC （microbial fuel cell） by the addition of Korean ginseng （Panax ginseng）. It was noted that the use of ginseng enhances the microbial anaerobic degradation of cellobiose, a disaccharide that was used as a substrate in the anode chamber of the MFC. The power output of the MFC where ginseng was added showed noticeable enhancement compared to the control MFC. The increase slowly ramped at the initial days and became appreciably higher after the 11th day of incubation in an experiment set up for 16 days duration. It is attributed that the ginseng increases the CO2 production by accelerating the fermentation process. Decrease in CH4/CO2 ratio was observed also due to decrease in methane production per digested cellobiose, the proton donor in the current study. Four ring steroid-like structural moiety Ginsenoside of Panax ginseng seemed to play a beneficial role in the electron transfer from ceilobiose to the anode, perhaps by rendering easier electron transfer due to favorable energy level alignments.
基金Acknowledgements The authors thank Prof. Xia Zongju in Peking University and Prof. Peng Hangcheng in the Institute of Biophysics of Chinese Academy of Sciences for their help in carrying out the single photon counting experiment, and Dr. Lin Su in Photos
文摘The spectroscopy characteristics and the fluorescence lifetime for the chloroplasts isolated from the pseudo ginseng, water hyacinth and spinach plant leaves have been studied by absorption spectra, low temperature steady-state fluorescence spectroscopy and single photon counting measurement under the same conditions and by the same methods. The similarity of the absorption spectra for the chloroplasts at room temperature suggests that different plants can efficiently absorb light of the same wavelength. The fluorescence decays in PS II measured at the natural QA state for the chloroplasts have been fitted by a three-exponential kinetic model. The three fluorescence lifetimes are 30, 274 and 805 ps for the pseudo ginseng chloroplast; 138, 521 and 1494 ps for the water hyacinth chloroplast; 197, 465 and 1459 ps for the spinach chloroplast, respectively. The slow lifetime fluorescence component is assigned to a collection of associated light harvesting Chl a/b proteins, the fast lifetime component to the reaction center of PS II and the middle lifetime component to the delay fluorescence of recombination of P+ 680 and Pheo-. The excitation energy conversion efficiency(η) in PS II RC is defined and calculated on the basis of the 20 ps electron transfer time constant model, 60%, 87% and 91% for the pseudo ginseng, water hyacinth and spinach chloroplasts, respectively. This interesting result is in unconformity with what is assumed to be 100% efficiency in PS II RC. Our result in this work stands in line with the 20 ps electron transfer time constant in PS II rather sound and the water hyacinth plant grows slower than the spinach plant does as envisaged on the efficiency. But, our results predict that those plants can perform highly efficient transfer of photo-excitation energy from the light-harvesting pigment system to the reaction center (closely to 100%). The conclusion contained in this paper reveals the plant growth characteristics expressed in the primary processes of photosynthesis and a relationship between a plant growing rate and its spectroscopy characteristics and fluorescence lifetimes, namely, the slower a plant grows, the less excitation energy conversation efficiency used might be anticipated.
基金Supported by the Talents Training Joint Program of the National Natural Science Foundation of China(NSFC)the Science and Technology Agency of Henan Province(Role and Mechanism of mi R-31 in Neoangiogenesis of Colorectal Cancers,No.U1204818)
文摘OBJECTIVE: To study the mechanism of the inhibitory effect of ginsenoside Rg3 on colon cancer cell migration.METHODS: Transwell migration assays were performed to investigate the inhibitory effect of ginsenoside Rg3 on SW480 cell migration. Electrophoretic mobility shift assays(EMSAs) and dual luciferase reporter assays were used to study the suppression capability of Rg3 on nuclear factor kappa B(NF-κB) activity. Western blotting was adopted to determine protein levels.RESULTS: Two-hundred micromolar ginsenoside Rg3 significantly inhibited SW480 cell migration(P < 0.05). EMSA showed that Rg3 suppressed the DNA binding ability of NF-κB. Dual luciferase reporter assay showed that Rg3 decreased NF-κB-regulated gene transcription(P < 0.01). Western blots indicated that Rg3 down-regulated expression of the NF-κB-regulated matrix metalloproteinase 9,cyclooxygenase-2 and C-Myc. An NF-κB inhibitor,pyrrolidine dithiocarbamate,enhanced the inhibitory effect of Rg3 on SW480 cell migration.CONCLUSION: Ginsenoside Rg3 has a strong antitumor migration capability by suppressing NF-κB activity and expression of NF-κB-regulated gene products. It could be a good adjuvant for colon cancer patients during the course of chemotherapy.
