In order to explore the application effect of Oldenlandia polysaccharides(OPS)in the production of AA broilers,2007-day-old AA broilers were randomly divided into 4 groups,namely groups Ⅰ,Ⅱ,Ⅲ,and Ⅳ,each including ...In order to explore the application effect of Oldenlandia polysaccharides(OPS)in the production of AA broilers,2007-day-old AA broilers were randomly divided into 4 groups,namely groups Ⅰ,Ⅱ,Ⅲ,and Ⅳ,each including 50 chicks.The control group(group Ⅰ)was fed with basal diet;and the 3 experimental groups were fed based on the basal diet with the addition of OPS at 0.4 g/kg(group Ⅱ),0.8 g/kg(group Ⅲ),and 1.2 g/kg(group Ⅳ)feed for 1 week,respectively,and then fed the basal diet for 4 weeks.At the ages of 14,28,and 42 d,10 chickens were put into death in the control group and the experimental groups,respectively,and the duodenum and jejunum tissue samples were collected for the analysis of the effects of OPS on the structures of chick intestines.The results showed that at the age of 14 d,the villus lengths of duodenum and jejunum in groups Ⅱ,Ⅲ,and Ⅳ were significantly greater than those in group Ⅰ(P<0.05),and the ratios of duodenal or jejunal villus length to crypt depth in groups Ⅱ and Ⅲ were significantly greater than that in group Ⅰ(P<0.05);at the age of 28 d,the villus length of duodenum in group Ⅱ was significantly greater than that in group Ⅰ(P<0.05);and at the age of 42 d,the ratios of jejunual villus length to crypt depth in groups Ⅱ,Ⅲ and Ⅳ were significantly larger than those of group I(P<0.05).It showed that OPS could promote the small intestine development of AA broilers aged 14 to 42 d.展开更多
Objective:To evaluate the apoptosis and cycle arrest effects of Oldenlandia diffusa flavonoids on human gastric cancer cells,determine the action mechanisms in association with the mitochondrial dependent signal trans...Objective:To evaluate the apoptosis and cycle arrest effects of Oldenlandia diffusa flavonoids on human gastric cancer cells,determine the action mechanisms in association with the mitochondrial dependent signal transduction pathway that controls production of reactive oxygen species(ROS),and evaluate the pharmacodynamics of a mouse xenotransplantation model to provide a reference for the use of flavonoids in prevention and treatment of gastric cancer.Methods:Flavonoids were extracted by an enzymatic-ultrasonic assisted method and purified with D-101 resin.Bioactive components were characterized by high-performance liquid chromatography.Cell lines MKN-45,AGS,and GES-1 were treated with different concentrations of flavonoids(64,96,128,and 160μg/mL).The effect of flavonoids on cell viability was evaluated by MTT method,and cell nuclear morphology was observed by Hoechst staining.The apoptosis rate and cell cycle phases were measured by flow cytometry,the production of ROS was detected by laser confocal microscope,the mitochondrial membrane potential(MMP)were observed by fluorescence microscope,and the expression of apoptotic proteins related to activation of mitochondrial pathway were measured by immunoblotting.MKN-45cells were transplanted into BALB/c nude mice to establish a xenograft tumor model.Hematoxylin and eosin staining was used to reveal the subcutaneous tumor tissue.The tumor volume and tumor weight were measured,the expression levels of proliferation markers proliferating cell nuclear antigen(PCNA)and Ki-67were detected by immunohistochemistry,and the expression levels of CA72-4 were measured by enzyme linked immunosorbent assay.Results:Oldenlandia diffusa flavonoids inhibited proliferation of MKN-45 and AGS human gastric cancer cells,arrested the cell cycle in G1/S phase,induced accumulation of ROS in the process of apoptosis,and altered MMP.In addition,flavonoids increased Apaf-1,Cleaved-Caspase-3,and Bax,and decreased Cyclin A,Cdk2,Bcl-2,Pro-Caspase-9,and Mitochondrial Cytochrome C(P<0.05).The MKN-45 cell mouse xenotransplantation model further clarified the growth inhibitory effect of flavonoids towards tumors.The expression levels of PCNA and Ki-67 decreased in each flavonoid dose group,the expression level of CA72-4 decreased(P<0.05).Conclusion:Flavonoids derived from Oldenlandia diffusa can inhibit proliferation and induce apoptosis of human gastric cancer cells by activating the mitochondrial controlled signal transduction pathway.展开更多
Objective:To investigate the anti-invasion efficacy of the ethanol extract of Oldenlandia diffusa Will.(EEOD) on a three-dimensional(3D) human malignant glioma(MG) cell invasion and perfusion model based on microfluid...Objective:To investigate the anti-invasion efficacy of the ethanol extract of Oldenlandia diffusa Will.(EEOD) on a three-dimensional(3D) human malignant glioma(MG) cell invasion and perfusion model based on microfluidic chip culture and the possible mechanism of action of Oldenlandia diffusa Will.(OD).Methods:The comprehensive pharmacodynamic analysis method in this study was based on microfluidic chip 3D cell perfusion culture technology,and the action mechanism of Chinese medicine(CM) on human MG cells was investigated through network pharmacology analysis.First,the components of EEOD were analyzed by ultraperformance liquid chromatography with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF/MS).Then,cell viability and apoptosis were assessed to determine the optimum concentration of EEOD for invasion experiments,and two-dimensional(2D) migration and invasion abilities of U87 and U251 MG cells were evaluated using scratch wound and Transwell assays.The possible mechanism underlying the effects of EEOD on glioma was analyzed through a network pharmacology approach.Results:Thirty-five compounds of EEOD were detected by UPLC-Q-TOF/MS.EEOD suppressed the viability of MG cells,promoted their apoptosis,and inhibited their migratory and invasive potentials(all P<0.05).Network pharmacology analysis showed that OD inhibited the invasion of MG cells by directly regulating MAPK and Wnt pathways through MAPK,EGFR,MYC,GSK3B,and other targets.The anti-invasion effect of OD was also found to be related to the indirect regulation of microtubule cytoskeleton organization.Conclusion:EEOD could inhibit the invasion of human MG cells,and the anti-invasion mechanism of OD might be regulating MAPK and Wnt signaling pathways and microtubule cytoskeleton organization.展开更多
The β-sitosterol imprinted polymer was prepared for selective extraction and analysis of β-sitosterol from Old-enlandia diffusa （0. diffusa） followed by HPLC with UV detection. The imprinted polymers show high aff...The β-sitosterol imprinted polymer was prepared for selective extraction and analysis of β-sitosterol from Old-enlandia diffusa （0. diffusa） followed by HPLC with UV detection. The imprinted polymers show high affinity and selectivity to β-sitosterol. Using this molecularly imprinted polymer （MIP） cartridge as solid-phase extraction （SPE） material, the interferences could be quickly washed out and β-sitosterol was selectively retained and enriched. HPLC analysis method was used to evaluate the characteristics of this MIP material. At the condition of mobile phase composed of MeOH/H20/H3PO4 （99/1/0.1, V/V/V, pH=6.0） and the flow rate of 1.0 mL·min -1, a good linear relationship was demonstrated when the concentrations of β-sitosterol were in the range of 0.50--100.0 μg·mL-1. The recoveries ranged from 75.3% to 86.5% and the inter-day and intra-day relative standard deviations were less than 5%. This developed HPLC method was proved to be acceptable for extraction offl-sitosterol from O. diffusa.展开更多
文摘In order to explore the application effect of Oldenlandia polysaccharides(OPS)in the production of AA broilers,2007-day-old AA broilers were randomly divided into 4 groups,namely groups Ⅰ,Ⅱ,Ⅲ,and Ⅳ,each including 50 chicks.The control group(group Ⅰ)was fed with basal diet;and the 3 experimental groups were fed based on the basal diet with the addition of OPS at 0.4 g/kg(group Ⅱ),0.8 g/kg(group Ⅲ),and 1.2 g/kg(group Ⅳ)feed for 1 week,respectively,and then fed the basal diet for 4 weeks.At the ages of 14,28,and 42 d,10 chickens were put into death in the control group and the experimental groups,respectively,and the duodenum and jejunum tissue samples were collected for the analysis of the effects of OPS on the structures of chick intestines.The results showed that at the age of 14 d,the villus lengths of duodenum and jejunum in groups Ⅱ,Ⅲ,and Ⅳ were significantly greater than those in group Ⅰ(P<0.05),and the ratios of duodenal or jejunal villus length to crypt depth in groups Ⅱ and Ⅲ were significantly greater than that in group Ⅰ(P<0.05);at the age of 28 d,the villus length of duodenum in group Ⅱ was significantly greater than that in group Ⅰ(P<0.05);and at the age of 42 d,the ratios of jejunual villus length to crypt depth in groups Ⅱ,Ⅲ and Ⅳ were significantly larger than those of group I(P<0.05).It showed that OPS could promote the small intestine development of AA broilers aged 14 to 42 d.
基金Supported by the Key Research and Development Program of Gansu Province(No.21YF5FA131)the Innovation Fund of Gansu Universities(No.2021A-084)the Graduate Innovation Fund of Gansu University of Chinese Medicine(No.2021CX49)。
文摘Objective:To evaluate the apoptosis and cycle arrest effects of Oldenlandia diffusa flavonoids on human gastric cancer cells,determine the action mechanisms in association with the mitochondrial dependent signal transduction pathway that controls production of reactive oxygen species(ROS),and evaluate the pharmacodynamics of a mouse xenotransplantation model to provide a reference for the use of flavonoids in prevention and treatment of gastric cancer.Methods:Flavonoids were extracted by an enzymatic-ultrasonic assisted method and purified with D-101 resin.Bioactive components were characterized by high-performance liquid chromatography.Cell lines MKN-45,AGS,and GES-1 were treated with different concentrations of flavonoids(64,96,128,and 160μg/mL).The effect of flavonoids on cell viability was evaluated by MTT method,and cell nuclear morphology was observed by Hoechst staining.The apoptosis rate and cell cycle phases were measured by flow cytometry,the production of ROS was detected by laser confocal microscope,the mitochondrial membrane potential(MMP)were observed by fluorescence microscope,and the expression of apoptotic proteins related to activation of mitochondrial pathway were measured by immunoblotting.MKN-45cells were transplanted into BALB/c nude mice to establish a xenograft tumor model.Hematoxylin and eosin staining was used to reveal the subcutaneous tumor tissue.The tumor volume and tumor weight were measured,the expression levels of proliferation markers proliferating cell nuclear antigen(PCNA)and Ki-67were detected by immunohistochemistry,and the expression levels of CA72-4 were measured by enzyme linked immunosorbent assay.Results:Oldenlandia diffusa flavonoids inhibited proliferation of MKN-45 and AGS human gastric cancer cells,arrested the cell cycle in G1/S phase,induced accumulation of ROS in the process of apoptosis,and altered MMP.In addition,flavonoids increased Apaf-1,Cleaved-Caspase-3,and Bax,and decreased Cyclin A,Cdk2,Bcl-2,Pro-Caspase-9,and Mitochondrial Cytochrome C(P<0.05).The MKN-45 cell mouse xenotransplantation model further clarified the growth inhibitory effect of flavonoids towards tumors.The expression levels of PCNA and Ki-67 decreased in each flavonoid dose group,the expression level of CA72-4 decreased(P<0.05).Conclusion:Flavonoids derived from Oldenlandia diffusa can inhibit proliferation and induce apoptosis of human gastric cancer cells by activating the mitochondrial controlled signal transduction pathway.
基金Supported by the National TCM Characteristic Technology Inheritance Talent Training Program (No. J20194828003).
文摘Objective:To investigate the anti-invasion efficacy of the ethanol extract of Oldenlandia diffusa Will.(EEOD) on a three-dimensional(3D) human malignant glioma(MG) cell invasion and perfusion model based on microfluidic chip culture and the possible mechanism of action of Oldenlandia diffusa Will.(OD).Methods:The comprehensive pharmacodynamic analysis method in this study was based on microfluidic chip 3D cell perfusion culture technology,and the action mechanism of Chinese medicine(CM) on human MG cells was investigated through network pharmacology analysis.First,the components of EEOD were analyzed by ultraperformance liquid chromatography with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF/MS).Then,cell viability and apoptosis were assessed to determine the optimum concentration of EEOD for invasion experiments,and two-dimensional(2D) migration and invasion abilities of U87 and U251 MG cells were evaluated using scratch wound and Transwell assays.The possible mechanism underlying the effects of EEOD on glioma was analyzed through a network pharmacology approach.Results:Thirty-five compounds of EEOD were detected by UPLC-Q-TOF/MS.EEOD suppressed the viability of MG cells,promoted their apoptosis,and inhibited their migratory and invasive potentials(all P<0.05).Network pharmacology analysis showed that OD inhibited the invasion of MG cells by directly regulating MAPK and Wnt pathways through MAPK,EGFR,MYC,GSK3B,and other targets.The anti-invasion effect of OD was also found to be related to the indirect regulation of microtubule cytoskeleton organization.Conclusion:EEOD could inhibit the invasion of human MG cells,and the anti-invasion mechanism of OD might be regulating MAPK and Wnt signaling pathways and microtubule cytoskeleton organization.
文摘The β-sitosterol imprinted polymer was prepared for selective extraction and analysis of β-sitosterol from Old-enlandia diffusa （0. diffusa） followed by HPLC with UV detection. The imprinted polymers show high affinity and selectivity to β-sitosterol. Using this molecularly imprinted polymer （MIP） cartridge as solid-phase extraction （SPE） material, the interferences could be quickly washed out and β-sitosterol was selectively retained and enriched. HPLC analysis method was used to evaluate the characteristics of this MIP material. At the condition of mobile phase composed of MeOH/H20/H3PO4 （99/1/0.1, V/V/V, pH=6.0） and the flow rate of 1.0 mL·min -1, a good linear relationship was demonstrated when the concentrations of β-sitosterol were in the range of 0.50--100.0 μg·mL-1. The recoveries ranged from 75.3% to 86.5% and the inter-day and intra-day relative standard deviations were less than 5%. This developed HPLC method was proved to be acceptable for extraction offl-sitosterol from O. diffusa.
