Screening of Anti-tumor Effective Extracts from Sedum bulbiferum Makino and HPLC Determination of Quercetin and Kaempferol in This Herbal Medicine
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摘要
[Objectives] To investigate the anti-tumor activity of Sedum bulbiferum Makino in vitro,and establish a HPLC method for determination of quercetin and kaempferol in S. bulbiferum. [Methods] The inhibitory activities of different extracts and total flavonoids of S. bulbiferum on proliferation of three kinds of cancer cells( Hep G2,EC109,SW480) were tested by MTT assay. HPLC method for determination of quercetin and kaempferol in S. bulbiferum was established. [Results]The growth and proliferation of the three kinds of cancer cells were all significantly inhibited by ethyl acetate fraction and total flavonoids isolated from S. bulbiferum. With each extraction concentration increasing,stronger anti-tumor activity was found. The linear ranges of quercetin and kaempferol were 0. 03-0. 36 μg( R = 0. 999 9) and0. 08-0. 96 μg( R = 0. 999 9),and the average recoveries were 98. 90%( RSD = 1. 15%) and 98. 27%( RSD = 1. 70%),respectively.[Conclusions]S. bulbiferum has significant anti-tumor activity in vitro. The HPLC method established was accurate,reproducible,and could be used for quality control of this crude drug.
[Objectives] To investigate the anti-tumor activity of Sedum bulbiferum Makino in vitro,and establish a HPLC method for determination of quercetin and kaempferol in S. bulbiferum. [Methods] The inhibitory activities of different extracts and total flavonoids of S. bulbiferum on proliferation of three kinds of cancer cells( Hep G2,EC109,SW480) were tested by MTT assay. HPLC method for determination of quercetin and kaempferol in S. bulbiferum was established. [Results]The growth and proliferation of the three kinds of cancer cells were all significantly inhibited by ethyl acetate fraction and total flavonoids isolated from S. bulbiferum. With each extraction concentration increasing,stronger anti-tumor activity was found. The linear ranges of quercetin and kaempferol were 0. 03-0. 36 μg( R = 0. 999 9) and0. 08-0. 96 μg( R = 0. 999 9),and the average recoveries were 98. 90%( RSD = 1. 15%) and 98. 27%( RSD = 1. 70%),respectively.[Conclusions]S. bulbiferum has significant anti-tumor activity in vitro. The HPLC method established was accurate,reproducible,and could be used for quality control of this crude drug.
[Objectives] To investigate the anti-tumor activity of Sedum bulbiferum Makino in vitro,and establish a HPLC method for determination of quercetin and kaempferol in S. bulbiferum. [Methods] The inhibitory activities of different extracts and total flavonoids of S. bulbiferum on proliferation of three kinds of cancer cells( Hep G2,EC109,SW480) were tested by MTT assay. HPLC method for determination of quercetin and kaempferol in S. bulbiferum was established. [Results]The growth and proliferation of the three kinds of cancer cells were all significantly inhibited by ethyl acetate fraction and total flavonoids isolated from S. bulbiferum. With each extraction concentration increasing,stronger anti-tumor activity was found. The linear ranges of quercetin and kaempferol were 0. 03-0. 36 μg( R = 0. 999 9) and0. 08-0. 96 μg( R = 0. 999 9),and the average recoveries were 98. 90%( RSD = 1. 15%) and 98. 27%( RSD = 1. 70%),respectively.[Conclusions]S. bulbiferum has significant anti-tumor activity in vitro. The HPLC method established was accurate,reproducible,and could be used for quality control of this crude drug.
引文
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[5]WEI HL,YANG Q,WEI HY.Determination of quercetin in hawthorn leaves by HPLC[J].Chinese Journal of Experimental Traditional Medicine Formulae,2006,2006,12(8):10-11.(in Chinese).
[6]FU HZ,WAN KH,LUO YH.RP-HPLC determination of kaempferide in Fuyang tablets[J].Chinese Journal of Experimental Traditional Medicine Formulae,2008,14(1):8-9.(in Chinese).
[7]SHU Y,TAN T,ZHANG SY,et al.Advances in pharmacology of quercetin[J].West China Journal of Pharmaceutical Sciences,2008,23(6):689-691.(in Chinese).
[8]NAIR HK,RAO KV,AALINKEEL R,et al.Inhibition of prostate cancer cell colony formation by the flavonoid quercetin correlates with modulation of specific regulatory genes[J].Clinical and Diagnostic Laboratory Immunology,2004,11(1):63-69.
[9]GRANADO-SERRANO AB,MARTIN MA,BRAVO L,et al.Quercetin induces apoptosis via caspase activation,regulation of Bcl-2,and inhibition of PI-3-kinase/Akt and ERK pathways in a human hepatoma cell line(Hep G2)[J].Journal of Nutrition,2006,136(11):2715-2721.
[10]ZHANG YM,ZHENG ZW.Advances in molecular mechanisms of antitumor effects of flavonoids[J].Chinese Journal of Drug Application and Monitoring,2006,3(6):51.(in Chinese).
[11]VIJAYABABU MR,ARUNKUMAR A,KANAGARA P.Quercetin downregulates matrix metalloproteinases 2 and 9 proteins expression in prostate cancer cells(PC-3)[J].Molecular and Cellular Biochemistry,2006,287(1-2):109-116.
[12]JI YB.Pharmacologically active substances and application of traditional Chinese medicine[M].Harbin:Heilongjiang Science and Technology Publishing House,1995.(in Chinese).
[13]CHEN YH,ZHOU KY,YUAN HY.Review of kaempferol:pharmacodynamics[J].Guangdong Medical Journal,2010,31(8):1064-1066.(in Chinese).
[2]CHEN YJ,LIN QX,WAN DR,et al.Study on the anti-tumor activities of the different extract fractions and total flavones of the three sedum plant drugs[J].Journal of the Central University for Nationalities(Natural Sciences Edition),2011,20(2):88-92.(in Chinese).
[3]YANG B,LI ZY,YOU QD.The development in the research of antitumor mechanism of flavonoids[J].Progress in Pharmaceutical Sciences,2008,32(9):391-397.(in Chinese).
[4]TANG Y,HE W,WANG YZ,et al.Studies on the anti-tumor activity of the extract of Spatholobus suberctus Dunn[J].Chinese Journal of Experimental Traditional Medicine Formulae,2007,13(2):51-54.(in Chinese).
[5]WEI HL,YANG Q,WEI HY.Determination of quercetin in hawthorn leaves by HPLC[J].Chinese Journal of Experimental Traditional Medicine Formulae,2006,2006,12(8):10-11.(in Chinese).
[6]FU HZ,WAN KH,LUO YH.RP-HPLC determination of kaempferide in Fuyang tablets[J].Chinese Journal of Experimental Traditional Medicine Formulae,2008,14(1):8-9.(in Chinese).
[7]SHU Y,TAN T,ZHANG SY,et al.Advances in pharmacology of quercetin[J].West China Journal of Pharmaceutical Sciences,2008,23(6):689-691.(in Chinese).
[8]NAIR HK,RAO KV,AALINKEEL R,et al.Inhibition of prostate cancer cell colony formation by the flavonoid quercetin correlates with modulation of specific regulatory genes[J].Clinical and Diagnostic Laboratory Immunology,2004,11(1):63-69.
[9]GRANADO-SERRANO AB,MARTIN MA,BRAVO L,et al.Quercetin induces apoptosis via caspase activation,regulation of Bcl-2,and inhibition of PI-3-kinase/Akt and ERK pathways in a human hepatoma cell line(Hep G2)[J].Journal of Nutrition,2006,136(11):2715-2721.
[10]ZHANG YM,ZHENG ZW.Advances in molecular mechanisms of antitumor effects of flavonoids[J].Chinese Journal of Drug Application and Monitoring,2006,3(6):51.(in Chinese).
[11]VIJAYABABU MR,ARUNKUMAR A,KANAGARA P.Quercetin downregulates matrix metalloproteinases 2 and 9 proteins expression in prostate cancer cells(PC-3)[J].Molecular and Cellular Biochemistry,2006,287(1-2):109-116.
[12]JI YB.Pharmacologically active substances and application of traditional Chinese medicine[M].Harbin:Heilongjiang Science and Technology Publishing House,1995.(in Chinese).
[13]CHEN YH,ZHOU KY,YUAN HY.Review of kaempferol:pharmacodynamics[J].Guangdong Medical Journal,2010,31(8):1064-1066.(in Chinese).