| The perennial herb Ornithogalum thyrsoides belong to the branch of Liliaceae under Ornithogalum,which is native to Southern Africa,and now is widely distributed in North China,the saponins are the main constituents of it and also the main therapeutic material basis of antitumor.O.thyrsoides,the total saponins have significant effects of antitumor in the treatment of liver cancer,breast cancer,leukemia and other research through multi-target and multi-channel.The adverse reactions of total saponins are limited.The development and use of its medicinal value can be improved effectively along with the expansion of the anti-cancer spectrum research.Male patients' morbidity and mortality rates of the lung cancer is first among the multiple cancer,although the treatment of Ornithogalum in cancer has been widely investigated,but the research of its effects on lung cancer is rare.Studies had been found that the IC50 of OSW-1 in vitro to non-small cell lung cancer A549 cells was low,which indicates that it has a good effect of anti-cancer,however,studies were not carried further due to the synthesis of OSW-1 cannot be achieved in large scale.The effect of Ornithogalum total saponins on lung cancer treatment has not been reported.The O.thyrsoides grown in Angola of Southern Africa was used in this study for the first time in China to test its antitumor activity on non-small cell lung cancer(A549 cell line)in vitro and in vivo and investigate the anticancer mechanisms of the extract of the total saponins and evaluate its safety.1.Fresh stem of O.thyrsoides was dried and crushed into powder,then reflux extraction method with 80%ethanol was used to extract saponins.The total saponins was separated using D-101 macroporous adsorption column from the aqueous phase,final product rate was 1.02%.The Vanillin-perchloric acid colorimetric method and spectrophotometric method were used to determine the total contents of saponins.The final product rate was higher than that using similar technology(1.02%VS 37.0%)and it showed the total soap content of the O.thyrsoides species originating from the Angola was higher.2.MTT assay method was used to screen the anti-tumor cells.It was found that the total saponins had strong inhibitory effects on the non-small cell lung cancer cell line(A549),human gastric cancer cell line(BGC-823),colorectal cancer cell line(HT-29)and human hepatic carcinoma cell line(HepG-2),and that the strongest inhibitory effects was found on A549 and there were a dose-dependent manner,the highest inhibition rate was up to 91.6%,the optimal time was 48h,and IC5O=22.28?g/mL.3.The non-small cell lung cancer cell line(A549)was used to investigate the in vitro antitumor activity of the total saponins.The results showed that the decrease of the vital cells as well as cytoplasmic shrinkage,the increase of refractive index and cell volume shrinking were observed using inverted fluorescence microscopy with the increase the dose of the total saponins(15.6?g/mL to 125.0?g/mL).Hoechst fluorescence staining showed the cell morphology changed as chromatin highly pyknosis agglutination,marginalization,which indicated a typical morphological characteristics of apoptosis cells.As the total saponins concentration increasing from 15.6?g/mL to 125.0?g/mL,the DNA content decreased in the G0/G1 period and G2/M phase,while increased significantly and arrested in S phase.The results suggested that total saponins can inhibit the proliferation of A549 cell line and induce cell apoptosis.4.Non-small cell lung cancer A549 tumor strains were transplanted into mice to establish the animal model to investigate the in vivo anti-tumor activity of the saponins.The inhibition rate of each dose group was>40%,even up to 51.61%.Significant difference was observed in viscera index and immune blood cells between the dose groups,including thymus and spleen index,the count and percentage of LYM,MON and neutral cells.Hb,WBC and PLT counts elevated in blood cell parameters.The results showed that the total saponins can directly inhibit A549 in vivo,without obvious inhibition on immune organs,and the normal adverse effects of general antitumor drugs wrer not observed.5.Preliminary safety evaluation experiments were performed by injecting mice intraperitoneally with the total saponins.Bliss method was used to determine the median lethal dose LD50 values,the LD50 was 23.808mg/kg and the most animal death time within 8h?48h due to the organ congestion,the organs of animal had scattered hemorrhage.The O.thyrsoides grown in Angola of Southern Africa was used in this study for the first time to expand the anti-tumor spectrum.The results showed that the total saponins of O.thyrsoides.had inhibitory proliferation and pro-apoptosis effects on non-small cell lung cancer A549 cell line both in vivo and in vitro.Safety evaluation study suggested that the drug had high safety and insignificant adverse reactions effects without obvious inhibition on immune organs,and the normal adverse effects of general antitumor drugs were not observed.Evaluation of anti-tumor effects and safety also provided theoretical and experimental basis for anticancer drug development and reasonable application.The study also provided a potential treatment of lung cancer using traditional Chinese medicine. |
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