Study On Isolation And Antitumor Activities Of Components From The Roots Of Arctiumlappa

Objective:To explore primarily methods of extraction and isolation of the roots of Arctiumlappa and the research of antitumor activities.Methods:Different components of the roots of Arctiumlappa chloro form extract were separated by silica gel column elution.MTT assay was applied to determine the antitumor active components by S180 cells,K562 cells,NCI-H1975 cells and MCF-7 cells.Flow cytometry was used to detect the effects of antitumor active components on cell cycle and apoptosis of K562 cells,and Western blotting assay to determine the expression of apoptosis related gene.And lymphocyte transformation explore the effect of active components on immune function in Mice.MTT assay observe the effect of reversing drug resistance.Cell scratch method was used to detect the ability of migration of NCI-H1975 cells in each group.Transwell assay to determine the cell invasion potential in vitro Results:Four components including Steroidal components,Triperpenoids components,L-3 and L-4 were separated by column chromatography of the roots of Arctiumlappa chloroform extract.Four components have strong inhibitory effect on cell proliferation of S180 cells,the IC50 value are below 10 μg/mL.The IC50 of the four components for NCI-H1975 cells were 137.95,260.95,245.03 and>500 μg/mL,respectively;The IC50 of the four components for MCF-7 cells were 282.76,140.32,>500,and>500 μg/mL,respectively.Steroidal components and Triperpenoids components were identified to have stronger inhibition effect on K562 cell by MTT assay,the IC50 value are 204.50μg/mL and 117.56μg/mL.Triperpenoids components significantly increased the proportion of G0/G1 phase cells from 45.52%to 53.82%and decreased the proportion of S phase cells from 45.68%to 37.11%in K562 cells(P<0.05).Steroidal components and Triperpenoids components could significantly increase the apoptotic rate of K562 cells from 5.1%to 21.8%and 38.0%respectively.Western blot show that Bax expression was enhanced by Steroidal components or Triperpenoids components treatment,while caspase-3,Bcl-2 expression was suppressed.Triperpenoids components promote lymphocyte proliferation of mice.Steroidal componentsc an reverse the drug resistance of K562/O2 cell,and the resistance index decreased from 1165 to 468(P<0.01).Compared with control group,the wound healing rates in different components were significantly decreased(P<0.05),and the effect of Steroidal components and Triperpenoids components was more significant than the other components.The wound healing rates was 30.32±1.10%in the control group,23.58±1.01%in the L-4 group,23.06±2.07%in the L-3 group,16.08±2.86%in the L-1 group and 15.99±2.09%in the L-2 group.The results of cell invasion experiment showed that the number of transmembrane cells decreased significantly in a certain concentration of four components compared with the control group(P<0.05).The number of transmembrane cells was 52.00 ± 7.24 in the control group,36.500±5.40 in the L-4 group,27.00±4.90 in the L-1 group,25.00±9.66 in the L-3 group and 18.80± 3.54 in the L-2 group(P<0.05).Conclusion:Steroidal components and Triperpenoids components of chloroform extracts from the roots of Arctium lappa exert antitumor effect by blocking cycle arrest,inducing apoptosis,regulating immunity,reversing multidrug resistance and decreasing the invasive ability.