The Effect Of Valeriana Jatamansi Jones Extract In Human Colon Cancer Cell SW480 In Vitro Research

Objective:To investigate the effects of inhibit proliferation and anti-metastasis of human colon cancer cell SW480 by Valeriana jatamansi Jones extract. Method:The optimum technological conditions of the extraction was determined by orthogonal test according to the content of total flavonoids,then purified by macroporous adsorptive resins to obtain extract of Valeriana jatamansi Jones.To determine the content of total flavonoids of Valeriana jatamansi Jones and purified extract by ultraviolet spectrophotometry.To determine the content of hesperidin,quercetin and apigenin by HPLC.To observe apoptosis induction and proliferation inhibition effects on human colon cancer cells SW480 by MTT assay,cell detachment test,light microscope and flow cytometry test.And to observe anti-metastasis effect on colon cancer cells SW480 by scratch test and cell adhesion test.Results:According to the content of total flavonoids,the optimum technological conditions were as follows:ethanol concentration 65%,ratio of solid to liquid 1:20,extracting duration 20 minutes,extraction times was 3.The content of total flavonoids of Valeriana jatamansi Jones determined by ultraviolet spectrophotometry was 2.27%,and the purified extract was 3.47%.The content of hesperidin and apigenin of Valeriana jatamansi Jones were 0.34% and 0.0009%,the content of quercetin was unattained.The content of hesperidin, quercetin and apigenin of the purified extract were 2.99%,0.04% and 0.05%.To observe Apoptosis induction effect of the purified extract,hesperidin, apigenin and 5-FU on colon cancer cells SW480 by MTT assay.24 hours after the administration,all drug groups could effectively inhibit proliferation of colon cancer cells SW480 according to the experiment dose,there was no significant difference of purified extract,hesperidin and apigenin compared with 5-FU.Compared with blank control group,all drug groups could increase the separation rates and inhibit proliferation in cell detachment test, there was the most significant difference(P<0.01).In flow cytometry test:24 hours after the administration, all drug groups could effectively elevated the inhibiting rate compared with blank control group(P<0.01) Compared with blank control group,the purified extract and hesperidin could decrease cell number of phase G0/G1 and increase phases of S and G2/M. the morphological changes of SW480 cells were observed by light microscopy,the cells of all groups showed characteristic of Tumor cell,such as irregular shape,large nucleus,large proportion of nucleus and plasma,multi nucleoli and so on. apoptotic cells were occasionally seen in the blank control group,but the other groups were seen typical features of apoptotic cells,such as volumetric reduction, cells turned round,intact cell membrane, karyopyknosis, chromatin agglutination and anachromasis and so on.Scratch test showed that moving ability of colon cancer cells SW480 could be effectively decreased (P<0.05) by purified extract and apigenin,apigenin had strongest function.There were no obvious differences of adhesive ratios between different groups.Conclusions:According to orthogonal test,the optimum technological conditions were as follows: ethanol concentration 65%,ratio of solid to liquid 1:20,extracting duration 20 minutes,extraction times was 3.The extract of Valeriana jatamansi Jones purified by macroporous adsorptive resins.The content of total flavonoids, hesperidin, quercetin and apigenin in the purified extract were 3.47%,2.99%,0.04% and 0.05% respectively.The purified extract,hesperidin, apigenin could effectively inhibit proliferation of human colon cancer cells SW480,induce apoptosis,arrest the cells at S and G2/M phases, effectively decrease moving ability. The results showed that purified extract and its active components could effectively inhibit proliferation and anti-metastasis of human colon cancer cells SW480.