Antitumor Activity Study Of Ginsenoside H

Backgroud and Objective:Ginseng is one of the most popular Chinese herbal medicines in the world,chich consists mainly of three key ingredients,saponins,polysaccharides,and phenols.Ginsenosides are the most widely studied in tumor chemical prevention and treatment.Ginsenosides are considered as the most effective anti-tumor component of ginseng.Ginsenoside Rh2 is presumed to have very strong antitumor activity in ginsenoside based on the structure-activity relationship of ginsenoside and the results of the existing studies.However,the clinical development of Rh2 is limited attribute to the low bioavailability of ginsenoside Rh2 and first-pass effect.Ginsenoside H emulsion injection developed by Shandong Luye Pharma Co.,Ltd.is mainly composed of 20（S）-ginsenoside Rh2.The improvement of ginsenoside H emulsion injection through the formulation increases drug exposure in vivo and overcomes the low bioavailability of Rh2.Method:1.Proliferation inhibition effect of ginsenoside H emulsion injection was determined using MTT assay on A549,Bel-7402,BGC-823,CaEs-17,HCT-8,HO8910,MCF-7,PC-3M,SMMC-7721 and U251 cell lines.2.Anti-tumor activity and effective dose range were explored using mouse tumor model of U14 cervical cancer.Ginsenoside H emulsion injection was administered intravenously at doses of 40,20 and 10 mg/kg once daily for 10 consecutive days.3.The synergistic anti-tumor effect of ginseng glucoside H emulsion injection combined with cyclophosphamide was determined using mouse model of cervical cancer U14.In addition,the effect of ginseng glucoside H emulsion injection on leukopenia induced by cyclophosphamide was investigated in the mouse cervical cancer model.4.The effect of ginsenoside H emulsion injection on the immune system of mouse with U14 cervical cancer was explored.5.The pharmacological mechanism of ginsenoside Rh2,an active ingredient of ginseng glucoside H emulsion injection,was invetigated by bioinformatics method.Results:1.The in vitro result demonstrated that ginsenoside H had antiproliferative effect on cell lines A549,Bel-7402,BGC-823,CaEs-17,HCT-8,HO8910,MCF-7,PC-3M,SMMC-7721 and U251.The half maximal inhibitory concentration(IC₅₀)was 37.18μg/ml,33.15μg/ml,31.02μg/ml,30.17μg/ml,30.14μg/ml,31.27μg/ml,35.21μg/ml,32.47μg/ml,33.93μg/ml,and 26.57μg/ml respectively.The IC₅₀ of of various tumor cell lins was in the same order of magnitude for liver cancer,gastric cancer,prostate cancer,colon cancer,esophageal cancer and neuroglioma.2.The in vivo results of U14 animal model showed that ginsenoside H emulsion injection could significantly inhibit the growth of cervical cancer in mice at 20 mg/kg and above.3.Although 2.5¹0 mg/kg showed a certain inhibitory effect on tumor,the effect was not obvious.But the inhibitory effect was significant in combination with cyclophosphamide.The ginsenoside H emulsion injection also showed an improvement in leukopenia caused by cyclophosphamide.4.In the mouse U14 model,we also found that ginsenoside H emulsion injection promoted secretion of IL-2,thereby activating NK cells,promoting T cell proliferation,and enhancing the killing effect of the immune system on the tumor.Co-administration of ginsenoside H with chemotherapeutic agent could correct immunosuppressive effects caused by chemotherapeutic agent.5.Other studies indicated that action target of Rh2 was Annexin A2,which interacted with Fibronectin Type III Domain Containing 3B（FNDC3B）.We found a significant negative correlation between FNDC3B and the prognosis of cervical cancer using bioinformatics data mining.Co-expression gene analysis of FNDC3B suggested that FNDC3B could be associated with endoplasmic reticulum stress.We explored the antitumor effect of ginsenoside H emulsion injection and explored the mechanism of action of ginsenoside H by bioinformatics in the present study.Conclusion:Combined with the results of the experiments conducted,the bioinformatics data mining results,and the available literature,the antitumor effect of ginsenodise H emulsion injection and proposed a novel mechanism of ginsenoside Rh2.We belevied that ginsenoside Rh2 could be directly combined with Annexin A2,and then affected the correlation between Annexin A2 and FNDC3B.In turn,it disrupted the homeostasis of the endoplasmic reticulum,causeed stress of the endoplasmic reticulum,and thus influenced the proliferation,migration,and invasion of tumor cells,as well as the immune system,and finally produced antitumor activity.Highlights:1.In the present study,we firstly confirmed the efficacy of the novel formulation of 20（S）-ginsenoside Rh2.We further analyzed the mechanism of action of ginsenoside Rh2 by bioinformatics.2.We proposed a new mechanism of ginsenoside Rh2.Rh2 can interact with Annexin A2 directly and affect Annexin A2 and FNDC3B interaction,consequently result in endoplasmic reticulum stress and induce tumor inhibition.