Objective： To observe the effect of extracts from Radix Ginseng, Radix Notoginseng and Rhizoma Chuanxiong （EXT） on delaying vascular smooth muscle cells （VSMCs） aging in aged rats. Methods： VSMCs were obtained b...Objective： To observe the effect of extracts from Radix Ginseng, Radix Notoginseng and Rhizoma Chuanxiong （EXT） on delaying vascular smooth muscle cells （VSMCs） aging in aged rats. Methods： VSMCs were obtained by the modified tissue explants technique and were shown to be positive for smooth muscle α-actin （SM-α-actin） by immunohistochemistry staining. VSMCs obtained from the young rats were served as the young control group; VSMCs obtained from the old rats were treated with no drug （the old group）, with low dose extracts （20 mg/L, the EXT low-concentration group） and high dose extracts （40 mg/L, the EXT highconcentration group）, and with Probucal （106 mol/L, the Probucal group） as a positive control. All groups were cultured for 24 h in the medium with 10% serum for 24 h followed by another 24 h in the serum-free medium. At the end of the 48-h culture, the following analyses were performed including determination of senescenceassociated β-galactosidase （SA β-Gal） activity, flow cytometry analysis of cell cycle, real-time quantitative reverse transcription polymerase chain reaction （RT-PCR） analyses of p16, Cyclin D1, cyclin-dependent kinase 4 （CDK4） and retinoblastoma （Rb） mRNA expression, and Western blotting analyses of p16, cyclin D1, CDK4 and phosphoretinoblastoma （pRb） protein expressions. Results： （1） In comparison to the younger rats, VSMCs from aged rats had significantly more SA β-Gal positive cells （P〈0.01） and more cells in S phase （P〈0.05）. VSMCs from the all treated groups showed a significant decrease in both SA β-Gal positive cells （P〈0.05） and S phase （P〈0.05） compared to the old rats. （2） Compared with the young group, VSMCs in the old group had a significant decrease in p16 and Rb mRNA expression and a significant increase in Cyclin D1 and CDK4 mRNA expression. Compared with the old group, VSMCs in the treated groups had a significant increase in p16 and Rb mRNA expression and a significant decrease in Cyclin D1 and CDK4 mRNA expression （P〈0.05）. （3） Compared with the young group, VSMCs in the old group had a significant decrease in p16 protein expression and a significant increase in Cyclin D1, CDK4 and pRb protein expressions （P〈0.05）. Compared with the old group, VSMCs in the treated groups had a significant increase in p16 protein expression and a significant decrease in cyclinD1, CDK4 and pRb protein expressions （P〈0.05）. Conclusions： VSMCs obtained from old rats showed typical signs of cellular senescence and vascular aging. EXT had an effect on delaying senescence of VSMCs in vitro by altering the p16-cyclinD/CDK-Rb pathway.展开更多
Microwave-assisted extraction was optimized with response surface methodology for HPLC-fluorescence determination of puerarin and daidzein in Radix Puerariae thomsonii.The optimized extraction procedure was achieved b...Microwave-assisted extraction was optimized with response surface methodology for HPLC-fluorescence determination of puerarin and daidzein in Radix Puerariae thomsonii.The optimized extraction procedure was achieved by soaking the sample with 70% methanol（1∶15,v/v）for 30 min,and then microwave irradiation for 11 min at a power of 600 W.Coupling the extraction process with HPLC-fluorescence presented good recovery,satisfactory precision,and good linear relation.Compared with a method from the Chinese Pharmacopoeia,the proposed method enables higher extraction efficiency and more accurate analytical results.It can be of potential value in quality assessment of Radix Puerariae thomsonii medicinal materials.展开更多
The feasibility of removal of the organochlorine pesticides residues of hexachlorocyclohexane(BHC) from radix ginseng with supercritical CO2 was explored. Some factors, such as extraction pressure, extraction temperat...The feasibility of removal of the organochlorine pesticides residues of hexachlorocyclohexane(BHC) from radix ginseng with supercritical CO2 was explored. Some factors, such as extraction pressure, extraction temperature, and kinds of co-solvents were investigated. The experimental results indicate that it is possible to reduce BHC residues in radix ginseng to the level of 0.1 × 10^-6 with supercritical CO2 in the presence of suitable amount of co-solvent, such as water.展开更多
基金Supported by the National Basic Research Program of China (Program 937,No.2007CB507400)the National Natural Science Foundation of China(No.30973828)the Independent Topic Program of China Academy of Chinese Medical Sciences (No.Z02151)
文摘Objective： To observe the effect of extracts from Radix Ginseng, Radix Notoginseng and Rhizoma Chuanxiong （EXT） on delaying vascular smooth muscle cells （VSMCs） aging in aged rats. Methods： VSMCs were obtained by the modified tissue explants technique and were shown to be positive for smooth muscle α-actin （SM-α-actin） by immunohistochemistry staining. VSMCs obtained from the young rats were served as the young control group; VSMCs obtained from the old rats were treated with no drug （the old group）, with low dose extracts （20 mg/L, the EXT low-concentration group） and high dose extracts （40 mg/L, the EXT highconcentration group）, and with Probucal （106 mol/L, the Probucal group） as a positive control. All groups were cultured for 24 h in the medium with 10% serum for 24 h followed by another 24 h in the serum-free medium. At the end of the 48-h culture, the following analyses were performed including determination of senescenceassociated β-galactosidase （SA β-Gal） activity, flow cytometry analysis of cell cycle, real-time quantitative reverse transcription polymerase chain reaction （RT-PCR） analyses of p16, Cyclin D1, cyclin-dependent kinase 4 （CDK4） and retinoblastoma （Rb） mRNA expression, and Western blotting analyses of p16, cyclin D1, CDK4 and phosphoretinoblastoma （pRb） protein expressions. Results： （1） In comparison to the younger rats, VSMCs from aged rats had significantly more SA β-Gal positive cells （P〈0.01） and more cells in S phase （P〈0.05）. VSMCs from the all treated groups showed a significant decrease in both SA β-Gal positive cells （P〈0.05） and S phase （P〈0.05） compared to the old rats. （2） Compared with the young group, VSMCs in the old group had a significant decrease in p16 and Rb mRNA expression and a significant increase in Cyclin D1 and CDK4 mRNA expression. Compared with the old group, VSMCs in the treated groups had a significant increase in p16 and Rb mRNA expression and a significant decrease in Cyclin D1 and CDK4 mRNA expression （P〈0.05）. （3） Compared with the young group, VSMCs in the old group had a significant decrease in p16 protein expression and a significant increase in Cyclin D1, CDK4 and pRb protein expressions （P〈0.05）. Compared with the old group, VSMCs in the treated groups had a significant increase in p16 protein expression and a significant decrease in cyclinD1, CDK4 and pRb protein expressions （P〈0.05）. Conclusions： VSMCs obtained from old rats showed typical signs of cellular senescence and vascular aging. EXT had an effect on delaying senescence of VSMCs in vitro by altering the p16-cyclinD/CDK-Rb pathway.
基金the National Natural Science foundationof China(No.20875060) for financial support
文摘Microwave-assisted extraction was optimized with response surface methodology for HPLC-fluorescence determination of puerarin and daidzein in Radix Puerariae thomsonii.The optimized extraction procedure was achieved by soaking the sample with 70% methanol（1∶15,v/v）for 30 min,and then microwave irradiation for 11 min at a power of 600 W.Coupling the extraction process with HPLC-fluorescence presented good recovery,satisfactory precision,and good linear relation.Compared with a method from the Chinese Pharmacopoeia,the proposed method enables higher extraction efficiency and more accurate analytical results.It can be of potential value in quality assessment of Radix Puerariae thomsonii medicinal materials.
文摘The feasibility of removal of the organochlorine pesticides residues of hexachlorocyclohexane(BHC) from radix ginseng with supercritical CO2 was explored. Some factors, such as extraction pressure, extraction temperature, and kinds of co-solvents were investigated. The experimental results indicate that it is possible to reduce BHC residues in radix ginseng to the level of 0.1 × 10^-6 with supercritical CO2 in the presence of suitable amount of co-solvent, such as water.
