Study On Extraction And Separation Of Active Components From Solanum Nigrum And Its Anti-Tumor Effect

Objective:The best extraction and isolation method of Solanum nigrum was screened by CCK-8 experiment,and the anti-tumor mechanism of Solanum nigrum extract on intestinal cancer DLD-1 in vivo and in vitro was studied.Method:The cell inhibition rate obtained by CCK-8 method was used as a reference to select the best extracting solvent and method of Solanum nigrum;Selection of the best separation method of Solanum nigrum by CCK-8 experiment;the effects of Solanum nigrum extract on the proliferation,migration and apoptosis of intestinal cancer DLD-1were studied by CCK8 experiment,cell migration experiment,AO-EB double staining experiment and flow cytometry experiment.The effect of Solanum nigrum extract on nude mice tumors was studied by Immunohistochemical method.Results:1.When Solanum nigrum was heated and refluxed with 80%ethanol solution,its extract inhibited the proliferation of DLD-1 cells.Within the range of administration,the highest inhibitory rate was 85.5%,and its IC₅₀0 value was 0.83 mg/mL.Therefore,80%ethanol reflux was the best extraction method.When 80%alcohol extract of Solanum nigrum was eluted by LSA-10 macroporous resin,the IC₅₀0 value of 50%alcohol eluent was 1.056 mg/mL.When LSA-10 macroporous resin was eluted,50%eluent was the best separation component.2.Study on the anti-tumor effect of Solanum nigrum extract on colon cancer in vitro:According to the IC50 value obtained from CCK-8 experiment,when the inhibition rate of cell proliferation was 50%,the concentration of Pentafluorouracil was 0.46 mg/mL,that of Solanum nigrum extract was 0.94 mg/mL,and that of solanine was 0.86 mg/mL.Cell scratch test confirmed that the drug group had the ability to inhibit cell migration.AO-EB double-staining fluorescence staining was used to detect the apoptotic morphology of cells under administration.The cells of blank control group showed normal cell state.The cells of Solanum nigrum extract group showed full morphology and decreased number.Cells in solanine group showed apoptotic phenomena,and the number of cells decreased.The apoptotic cells of pentafluorouracil group were more serious and the number of cells decreased,the volume decreased significantly.When the apoptotic rate was detected by flow cytometry,it was found that apoptotic phenomena were induced in all groups.3.Study on the anti-tumor effect of Solanum nigrum extract on colon cancer in vivo:Solanum nigrum extract,solanine,combined administration and pentafluorouracil can inhibit the mass and volume of nude mice tumors to a certain extent.After immunohistochemical staining of nude mice tumor tissue sections,a large number of brown spots or plaques appeared in the tissue sections under the optical microscope,which is a phenomenon of positive cell reaction.The positive expression rate of the drug group was lower than that of the blank control group.The results showed that all the drug groups had inhibitory effects on the tumors of nude mice.Conclusion:Combined with CCK-8 experiment,the best extraction and separation method of anti-tumor active ingredients was screened.The anti-tumor mechanism of extract in vitro showed that Solanum nigrum extract could inhibit the proliferation and migration of cells to a certain extent,affect the morphology and quantity of cells,and induce cell apoptosis.The anti-tumor mechanism of Solanum nigrum extract in vivo showed that Solanum nigrum extract could inhibit the weight and volume of nude mice tumors,and the positive rate of cells was low.