Antitumor Effect Of Polysaccharides From Goutou Jujube And Its Effect On The Absorption And Transport Of Genistein

Jujube(Ziziphus jujuba Mill.),as the traditional Chinese medicine named jujuba,is the fruit of jujube in the genus Rhamnaceae.Jujube is a widely-used food in daily life and a common Chinese medicine.It has the functions of strengthening the spleen and stomach,nourishing blood,calming nerves,and relieving medicinal properties.It is often used to treat various diseases such as weakness of the spleen and stomach,lack of food,loose stools,deficiency of blood,and ease the effects of the medicine.Polysaccharides are the highest content of biologically active components in jujube.In recent years,studies have reported that natural polysaccharides have various biological activities such as antioxidant,antitumor,anti-inflammatory,and immune regulation.However,there are few reports on the antitumor activity of jujube polysaccharides,and antitumor mechanism of jujube polysaccharides has not yet been in-depth.Interestingly,recent studies have shown that a variety of non-digestible sugars can improve the bioavailability of flavonoids,but its mechanism is still unclear.Ziziphus jujube cv.Goutouzao,as a kind of high-quality red jujube in Yan'an,Shaanxi Province,is rich in a variety of bioactive ingredients.This study took the Ziziphus jujube cv.Goutouzao polysaccharides(GZSP)as the research object to explore the various biological activities of GZSP in cells level,mainly including the antioxidant activity,antitumor activity,and its absorption promotion effect on genistein.The main research contents and results of this experiment are as follows:1.Extraction,physical and chemical properties detection,and chemical composition analysis of polysaccharides in Ziziphus jujube cv.Goutouzao:the water-soluble polysaccharides in Ziziphus jujube cv.Goutouzao(GZSP)are extracted by the water-extraction and alcohol-precipitation method.The sulfuric acid-phenol method and 3,5-dinitrosalicylic acid method was used to determine the total sugar content and reducing sugar content of GZSP.The results showed that the contents of total sugar and reducing sugar in GZSP were 76.5%and 2.1%,respectively.The microstructure and chemical composition of GZSP were analyzed by scanning electron microscopy,infrared spectroscopy,and high-performance liquid chromatography.The results show that the microstructure of jujube polysaccharides is irregular flaky particles,with typical polysaccharide functional groups,mainly composed of glucose,arabinose,galactose,glucuronic acid.2.Polysaccharides in Ziziphus jujube cv.Goutouzao has significant antioxidant and antitumor activities:The antioxidant test results showed that GZSP has significant scavenging ability on DPPH·,O2·-,OH· and reduction effect on Fe3+,indicating that GZSP has good antioxidant activity.The effect of GZSP on the viability of colorectal carcinoma LoVo cells and macrophage RAW 264.7 cells was detected by MTT method.The results showed that GZSP has no direct effect on the viability of LoVo cells,but GZSP can be significant promoting the proliferation of macrophages RAW 264.7 cells.After GZSP are co-cultured with RAW 264.7 cells,the supernatant of the culture medium interferes with LoVo cells,which can significantly inhibit the proliferation of LoVo cells.And LDH method to detect cytotoxicity also confirmed this.Flow cytometry was used to detect cell cycle,cell apoptosis,and intracellular ROS levels.The results showed that the indirect intervention of GZSP blocked the cycle progress of LoVo cells in the G0/G1 phase,increased the intracellular ROS and induce cell apoptosis.The ELISA method was used to detect the TNF-? content of cells produced by RAW 264.7 under the indirect intervention of GZSP.The results showed that the content of TNF-? was positively correlated with the concentration of GZSP.It can be seen that GZSP indirectly inhibit the growth of tumor cells by enhancing immunity.3.Jujube polysaccharides can promote the absorption of genistein:The effect of GZSP,genistein and the combination of them on viability of Caco-2 cells was detected by MTT method.The results show that 10-40 ?M genistein and 100-400 ?g/mL GZSP are safe concentration range.UPLC-qTOF/MS was used to detect the changes in the content of genistein in Caco-2 cells by the intervention of different concentrations of GZSP and same concentration of genistein.The results showed that GZSP can significantly increase the content of genistein in cells.The expression levels of phase II metabolic enzymes(UGT and SULT)and efflux transporters(MRP1,MRP2 and P-gp)in Caco-2 cells were detected by ELISA.The results showed that GZSP can significantly reduce the expression of UGT,SULT and MRP2,but had no significant effect on the expression levels of MRP1 and p-gp.To sum up,this study discovered that polysaccharides in Ziziphus jujube cv.Goutouzao(GZSP)can indirectly inhibit the growth of colorectal carcinoma cells through immune activation at the cellular level.At the same time,it was discovered that GZSP can enhance the absorption of genistein in cells by reducing the expression of phase ? metabolic enzymes(UGT and SULT)and efflux transporters(MRP2).These results provide a reference for the development of jujube polysaccharide-assisted antitumor functional foods and compound nutritional health foods.