Chinese Medicine The Daji Chemical Ingredients, Its Anti-proliferation Of Lung Cancer A549 Cells

Circium japonicum DC., a traditional Chinese medicine, is a perennial plant. The whole plant or root in Circium japonicum DC. can be used as medicine. It's chemical components are complex, which mainly contain flavones, flavonoid glycoside, volatile oils, triter pene, sterol. In the well-established folk recipes, Circium japonicum DC. is usually used in the treatment of cancers. For example, the root of Circium japonicum DC. and rhizome of Chinese lizard tail are combined to treat liver cancer; the ground fresh leaves of Circium japonicum DC. and the egg white are together applied to affected part to treat breast cancer. Since the chemical constituents of Circium japonicum DC. are complicated, many antitumor active compounds have not been found yet, while the relationship between chemical compounds and clinical uses should be further elucidated.The objectives of this study were to isolate constituents and evaluate their antitumor activities from Circium japonicum. Ethanol extract was suspended with water and fractionated with same volume of methylene trichloride, ethyl acetate, petroleum ether, and water. Nine compounds were isolated from F. suspensa using silica gel column chromatography, Sephadex LH-20chromatography and semi-preparative HPLC methods. Using UV, ESIMS,'H-NMR and13C-NMR spectroscopy, they were identified as inulolide; taraxast-20-ene-3β,30-diol;6-o-(2-methylpropen -oyl)-3β-hydroxy-4(15),10(14),11(13)-guaiatrien-12,8-olide; taraxaster--ol3-o-acetate;(±)evofolin B;3β-hydroxy-taraxaster-20-en-30-aldehyde; atractyligenin;21a-hydroxy-taraxasterol。 From them, inulolide, taraxast--erol3-O-acetate, atractyligenin have not been reported from this plant yet.The antitumor activity for different fractions and compound1-9were assayed by MTT method using human lung carcinoma A549cells, accompanied with the morphological observation of the cells under light microscope. The result indicated that the petroleum fraction, EtoAc fraction, MeoH fraction, Chloroform fraction and water fraction all have antitumor activities in high concentration group. Besides, petroleum fraction and EtoAc fraction showed anti-proliferation activity compared with the DMEM medium-treated group in a concentration-dependent manner. The inhibition ratio of them were93.57%,.94.8%at a concentration of2mg/mL, with the IC50value of322.55μg/mL,222.29μg/mL, respectively. In addition, among the compounds obtained,1,3,5,8,9showed the antitumor effect on the A549cells to a different extent. Among them,5showed the highest activity on the proliferation of A549cells with the IC5O value of70.5μg/mL; Compound3,8,9moderately inhibited the proliferation of A549cells with the inhibition ratio of45.5%,47.6%, and47.4%at a concentration of2mg/mL respectively; while the others had weak effect towards A549cells. Compound6,7had no obvious anti-proliferation activity, without a concentration-dependent manner. Petroleum fraction, EtoAc fraction, and compound3,5,8,9were remarkable compared with the control group (p<0.01). However, the MeoH fraction, water fraction and compound1,4,6,7did not show anti-proliferation activity, but they all could affect the growth of A549cells in different extent, and high concentration group of them were also remarkable compared with control group(p<0.01or p<0.05).