| The dried heartwood of Caesalpinia Sappan L.which is called Sappan Lignum belongs to the family of Leguminosae.Sappan L.is a traditional Chinese medicine in my country.The Chinese Pharmacopoeia records that it has the effects of removing blood stasis and detumescence,etc.It is mainly used for the treatment of fracture and injury of tendons and bruises.In recent years,there have been a large number of reports about Sappan L.extract inhibiting a variety of tumor cells,especially in the treatment of bladder cancer after treatment,and it has been proved that its active ingredients are mainly brazilin and protosappanin B.However,there are few reports on the research of breast cancer,the world’s number one cancer incidence among women.Therefore,in this study,breast cancer cells were used as the research object,and the active components of Sappan L.in anti-breast cancer were screened and structurally identified;and its anti-cancer effect was verified in vitro and in vivo,in order to provide scientific basis for the clinical development and application of Sappan L.extract and its active components.The specific research contents and results are as follows:(1)Screening,structure identification and effect verification of anti-breast cancer active ingredients from Sappan L.First,the extracts from Sappan L.(Sample No.1-16)were prepared by using different extraction methods,and the main components in the extracts were separated and their relative content were determined by HPLC.Secondly,the inhibitory rates of different extracts from Sappan L.(Sample No.1-16)on cells MDA-MB-231 for 24 h and 48 h were determined by MTT method.Then,the correlation study was carried out on the inhibitory rate of each component concentration of the extract to determine that the component whose concentration is positively correlated with the inhibitory rate is the anti-breast cancer active component.Finally,the structure identification and effect verification of the active ingredients were carried out.The results showed that the inhibitory rates of different extracts from Sappan L.on MDA-MB-231 cells for 24 h and 48 h were significantly positively related to the content of the component with a retention time of 14 min in HPLC,and the correlation coefficients were 0.845 and 0.740,respectively(P<0.01),and its inhibition rate increased with the prolongation of the action time,and the active component has been structurally identified as brazilin.(2)Effect of brazilin on the proliferation,movement,migration and invasion ability of different breast cancer cellsHuman breast cancer MDA-MB-231 cells and mouse breast cancer 4T1 cells were selected as the research objects,the cell proliferation experiment was used to detect the effect of brazilin on cell proliferation at different concentrations and different time.The wound healing experiment was used to detect the effect of brazilin on the motility of breast cancer cells,and the transwell migration test and invasion test were used to detect the effect of brazilin on the migration and invasion of breast cancer cells.The results showed that brazilin inhibited the proliferation of different breast cancer cells in a dose-and time-dependent manner compared with the control group.In addition,brazilin can significantly inhibit the motility,migration and invasion capabilities of different breast cancer cells(P<0.05)in a dose-dependent manner.In short,brazilin has a significant inhibitory effect on the proliferation and metastasis of different breast cancer cells which further confirm that brazilin is the active component of Sappan L.against breast cancer.(3)Study on the anti-metastasis effect of Sappan L.extract in breast tumorThe effect of Sappan L.extract on breast tumor metastasis was explored.The mice were divided into blank group,model group,5-Fu group,high,medium,and low dose groups of Sappan L.extract.Except for the blank control group,the remaining mice were injected with 4T1 cell suspension through the tail vein to establish a mouse breast tumor lung metastasis model.The blank control group and the model group were intraperitoneally injected with normal saline,0.2 m L/mouse,once a day;the 5-Fu group(125 mg/kg)was intraperitoneally injected with 0.1 m L/10 g mouse body weight,once a week;The high,medium and low dose groups of Sappan L.extract were given 200 mg/kg,100 mg/kg and 50 mg/kg respectively,and 0.1 m L/10 g mouse body weight was injected intraperitoneally once a day for a total of 21 days.The changes in body mass,survival time and survival rate,organ index and total number of metastatic nodules on the surface of lung were detected.The results showed that the extract of Sappan L.had no significant effect on the body weight of mice.Compared with the control group,the average survival days and spleen index of the model group were significantly decreased,while the lung index and the number of pulmonary metastatic nodules were significantly increased(P<0.05);Compared with the model group,the average survival days and spleen index of each dose of Sappan L.extract group were significantly increased(P<0.05),and the lung index and the number of pulmonary metastatic nodules were significantly decreased(P<0.05).In conclusion,Sappan L.extract can significantly inhibit the metastasis of mouse breast tumor and prolong the survival time of mice. |
PDF Full Text Request