Preparation Of Ginsenoside Rg5 And Its Anti-gastric Cancer And Anti-breast Cancer Effects

Cancer(such as gastric cancer,breast cancer,etc.)is a malignant tumor.In recent years,the incidence of cancer has continued to rise,causing great harm to human health and quality of life,and also bringing great burden to the medical industry and the national economy.Chemotherapy is commonly used for the treatment of cancer,but chemotherapeutic drugs generally have the disadvantages of high price and side effects.Long-term use of chemotherapeutic drugs will lead to drug resistance,which seriously hinders the process of cancer treatment.Therefore,it is of great significance to find safe,effective,widely sourced and few side effects anticancer drugs for the treatment of gastric cancer and breast cancer.In this study,ginsenoside Rg5 was extracted from ginseng powder,and then transformed and purified.The effect and molecular mechanism of ginsenoside Rg5 on gastric cancer and breast cancer were investigated in vitro and in vivo,respectively.It will provide research basis for clinical treatment of gastric cancer or breast cancer with ginsenoside Rg5.The main contents of this paper are as follows:(1)Preparation of ginsenoside Rg5 Total ginsenosides were separated and purified from ginseng powder by methanol extraction and gradient elution with D101-macroporous resin.The content of total ginsenosides was 93%.The pure total ginsenosides were converted into rare ginsenosides by cellulase conversion method combined with citric acid hydrolysis method.The conversion process of Rg5 was optimized by single factor analysis and response surface analysis.The optimum conditions of cellulase conversion method were obtained as follows: cellulase concentration 40 mg/mL,pH 5.5,temperature 45 ℃,conversion time 30 h;and the optimum conditions of citric acid hydrolysis method were obtained as follows: acid concentration 1.50 mol/L,temperature was 100 ℃,and preparation time 6 h.Under these conditions,the yield of ginsenoside Rg5 was 27.02%.Then,ginsenoside Rg5 was separated and purified by preparative liquid chromatography,and analysed by liquid chromatography.The purity of final monomer of ginsenoside Rg5 was 98.9%.(2)Anti-gastric cancer activity of ginsenoside Rg5 To study the mechanism of apoptosis of gastric cancer cells,MTT assay,flow cytometry,AO/EB double staining,JC-10 assay and western blot assay were used to verify that Rg5 could inhibit the growth of gastric cancer cells,induce G2/M cycle arrest and apoptosis of gastric cancer cells through mitochondria and death receptors.Transmission electron microscopy and western blotting showed that Rg5 could induce autophagy in gastric cancer cells.At the same time,Rg5-induced apoptosis and autophagy of gastric cancer cells promoted each other.The effects of Rg5 on the upstream signaling pathways of cell cycle,apoptosis and autophagy in gastric cancer cells were further studied.It was found that Rg5 could induce ROS and activate MAPK pathways(P38,JNK,ERK)in gastric cancer cells,and then induce apoptosis,autophagy and cell cycle arrest.Finally,we constructed a tumor-bearing nude mice model of gastric cancer,and found that Rg5 had obvious anti-tumor activity in vivo with few side effects.The results of western blotting and immunohistochemistry showed that Rg5 could regulate the protein expression of apoptosis,autophagy and MAPK pathway in gastric cancer,which was consistent with the results of cell experiments in vitro.These results suggested that Rg5 could achieve anti-gastric cancer effect through a variety of anti-cancer molecular mechanisms.It was expected to become a new anti-gastric cancer drug and provide reference for clinical application.(3)Anti-breast cancer activity of ginsenoside Rg5 In this paper,a series of in vitro cell experiments,staining experiments and western blotting experiments proved that Rg5 could significantly inhibit the growth and migration of breast cancer cells,and induce cell cycle arrest,apoptosis and autophagy.For further investigate the upstream signaling pathway of Rg5 on breast cancer,the molecular mechanism of which against breast cancer in vivo and in vitro was studied.The expression levels of PI3 K protein,AKT protein,mTOR protein,Bad protein and their phosphorylation were determined by immunohistochemistry and western blotting.It was found that Rg5 could inhibit the growth of breast cancer by inhibiting PI3K/AKT signaling pathway.Rg5 regulated apoptosis and expression of autophagy-related proteins in breast cancer.According to the tumor-bearing nude mice model of breast cancer,Rg5 prepared in this paper could also significantly inhibit the growth of breast cancer transplantation in nude mice.It had the same efficacy as docetaxel,the first-line chemotherapeutic drug for breast cancer.However,Rg5 has the advantage of few side effects,which is expected to become a new generation of anti-breast cancer drugs.In conclusion,this study prepared high purity ginsenoside Rg5 from natural ginseng powder by extraction,transformation,isolation and purification,and then studied its anti-tumor effect on gastric cancer and breast cancer in vitro and in vivo.In vivo and in vitro experiments showed that Rg5 could effectively inhibit the growth of gastric cancer by activating the ROS-mediated MAPK signaling pathway to induce cell cycle arrest,cell apoptosis and cell autophagy.At the same time,Rg5 could induce apoptosis and autophagy of breast cancer cells by inhibiting PI3K/AKT signaling pathway,and then inhibited the proliferation of breast cancer cells.In addition,in the tumor-bearing nude mice model of gastric cancer and breast cancer,ginsenoside Rg5 had remarkable anti-tumor effect and few side effects,which provided an effective theoretical basis and important research basis for the clinical treatment of gastric cancer and breast cancer.