| In recent years,with the research at home and abroad on the anti-tumor,anti-depressant,liver protection and kidney protection,hypoglycemic regulation and other pharmacological effects of Saponins of Rhizoma Polygonati,the annual consumption of sealwort is also gradually increasing,and the extraction of Saponins of Rhizoma Polygonati by ethanol is the main method to obtain Saponins of Rhizoma Polygonati,but the extraction rate of Saponins of Rhizoma Polygonati is low.In this paper,the extraction process of astrasin was extracted by ultrasound-assisted extraction,the optimal extraction process was optimized by the response surface method,the extraction process of xanthine saponin was optimized from four factors:ultrasonic temperature,ultrasonic time,ethanol concentration and liquid-to-material ratio,and the best extraction process was obtained by analyzing and comparing the results,which provided theoretical basis and technical support for the comprehensive utilization of xanthin,the macroporous resin was used to separate and purify the Saponins of Rhizoma Polygonati,the purity of the obtained components was analyzed using high performance liquid phase and were identified by nuclear magnetic hydrogen spectroscopy and carbon spectroscopy.MTT method,scratch test,flow cytometry,Western Blot and other experiments were used to further explore the effect of dioscin on human breast cancer MCF-7 cells and how to exert anti-tumor effects.The specific results of the study are as follows:1、The best extraction process for ultrasonic extraction of Saponins of Rhizoma Polygonati:extraction time,extraction temperature,ethanol concentration and liquid-tomaterial ratio were 1.6h,60℃,81%,respectively,and the yield was 3.18%.2、The Saponins of Rhizoma Polygonati were separated and purified by column chromatography,the purity was determined to be 94.17%by HPLC and identified as dioscin by nuclear magnetic hydrogen spectrometry and nuclear magnetic carbon spectrometry.3、MTT results showed that dioscin inhibited cell proliferation after acting on human breast cancer MCF-7 cells,and was concentration-dependent,and half of its inhibitory concentration(IC50)was 1.524μg/ml.4、The results of scratch experiments showed that a certain dose of Dioscin within 48 hours showed that it inhibited the migration of MCF-7 cells in breast cancer.5、Flow cytometry showed that dioscin could effectively inhibit the cell cycle of MCF-7 cells in G2/M phase to promote apoptosis.6、The results of Western Blot showed that dioscin could significantly upregulate the expression levels of P53,caspase-3,caspase-9 and Bax proteins in MCF-7 cells,downregulate the expression levels of Bcl-2 proteins,and upregulate the Bax/Bcl-2 ratio,which promoted apoptosis in breast cancer MCF-7 cells.In this paper,the ultrasound-assisted extraction of Saponins of Rhizoma Polygonati was optimized by the response surface method,and the results showed that the extraction rate of Saponins of Rhizoma Polygonati reached 3.18%after optimization.The isolated product was identified as dioscin.Dioscin can inhibit the proliferation and migration of human breast cancer MCF-7 cells,inhibit the MCF-7 cell cycle in the G2/M phase and promote apoptosis,and the expression level of some proteins in MCF-7 cells will increase,such as P53,caspase3,Bax and other proteins,and the expression level of Bcl-2 protein will be reduced,so as to promote the apoptosis of human breast cancer cells MCF-7.Finally,it was found that dioscin may be a potential active ingredient in anti-breast cancer in sealwort,which provides a certain theoretical basis for the development of anti-tumor drugs using sealwort saponin. |
PDF Full Text Request