| Aralia elata is a plant which belongs to the genus of Aralia and the family of Araliaceae.The tast is bitter and acrid,and the nature is neutral.It has the effect of anti-inflammatory,anti-fatigue,hepatoprotection and so on.Domestic and foreign researches on chemical constituents and pharmacologic actions mainly concentrate on the root bark and tender shoot.Not only the output is very low and the resource is limited,but it is a tremendous destruction for the resources of A.elata,so it is harmful for continuing exploitation and the applications are limited.Liver cancer is a common malignant tumour of the serious harm to people’s health,and the treatment effect is not optimistic,because of it has already lead to lower quality of life after the ill,lower survival rate.So,The development of an effective liver cancer drugs,relieve the suffering of the patients with liver cancer,delay the course of the disease,improve the quality of life,improve the survival rate is the focus in the field of cancer research.The great amount of leaves of A.elata run its course,it is very wasty.The latest researches show that ETS and monomer saponin can significantly inhibit tumorcell.Our team take the reproducibility of resources into consideration and our prophase experiments show that the leaves of A.elata contain a great amount of saponins,ETS and monomer saponin can inhibit many kinds of tumor cell on different degrees through inhibiting synthesis of DNA,RNA and protein in tumor cell and show structure-activity relationship as well.In order to clarify the mnaterial basis of antitumor activity,develop an antitumor new drug coming from leaves of A.elata and institute the quality standard,we carry out further studies on chemical constituent in its effective part based on our early work.Objective:In this study,we examined the anti-tumor activity of total saponins of Aralia elata(Miq.)Seem leaves(ETS)using both in vitro and in vivo experiments,and investigated the underlying molecular mechanisms.We also evaluated the safety of total saponins of Aralia elata(Miq.)Seem(ETS)in SD ratsMethod:1.Systems refer to relevant Aralia elata and liver cancer related ancient and modern literature.The literatures from Aralia elata,chemical composition and clinical curative effect;Liver disease,etiology,syndrome differentiation,treatment and other respects of Aralia elata and the research progress of traditional Chinese medicine and modern medicine understanding of liver disease.2.By establishing the model of liver cancer in mice,and observe the change of the tumor-burdened general status in mice,viscera index,blood cell,tumor inhibition rate and survival rate,serum IL-2,IL-6,VEGF,TNF-α、IFN-γ content,total saponins by western blot method for active ctive caspase-9,active caspase-3,Bax,Bcl-2,Cyt-C、MMP-2、MMP-9 protein expression;Tunel staining method to detect the cell apoptosis in each dose group of tumor tissue;Immunohistochemical staining method to detect the drug group of VEGF protein expression in tumor tissue.Delve into the mechanism of actiontreatment of ETS in treatment for the liver diseases.3.Choose common typical cell lines such as human liver Bel 7402,human lung adenocarcinoma A549,human oral epithelial carcinoma KB,human ovarian cancer A2780 and rat high metastasis of melanoma B16BL6 strains,colon cancer HCT-8,Use these cells to research ETS and part of the monomer compounds which resistance of tumor cells in vitro killing effect and their structure-activity relationship.4.By acute toxicity test and long toxicity experiments to investigate the security of ETS,record the changes of toxic symptoms,deaths,behavioral change,weight,food consumption and temperature.Detect the changes of the ecg index,index of urine,hematology index,serum biochemical indicators and viscera weight,At the same time,each viscera of ophthalmologic examination and histopathologic examination.Provide the safety evaluation index of ETS.Results1.Literature research:Aralia elata application early in our country,its also has very deeply research in the composition and pharmacological activity.Aralia elata’s root,leaf,fruit can be medicine,With its effect of the duodenal ulcer,chronic gastritis,hepatitis,diabetes,rheumatoid arthritis,edema.Its chemical composition mainly contains saponins,diterpene,triterpenoid,brass,lignin compounds,organic acids,alkaloids,volatile oil and so on,In clinical treatment of neurasthenia,rheumatoid arthritis,diabetes,stomach cramps,constipation,trauma,hemorrhage,Yang deficiency,gas of the weak and the shortage of kidney disease.Of traditional Chinese medicine to the understanding of the origin of liver cancer early,its etiology is the human body,joint outcome of external factors,the phase comparison method of surgical treatment of western medicine,TCM treatment for liver cancer also has obtained the good curative effect2.The result on the study of resistance mechanism primary carcinoma of liver with ETS,it can obviously relieve the weight loss in patients with liver cancer,significantly reduced number of white blood cells in mice tumor and improve tumor mice condition,at the same time significantly extend the survival days of mice a tumor-burdened,ETS has obvious antitumor effect in the body,and had no obvious effect on the immune organs.3.ETS can significantly increase the level of IL-2、IL-6、IFN-γ、TNF-α in aserum,reduce the level of the VEGF.The development of tumor is excessive cellular proliferation and apoptosis abnormal function,ETS has obvious effect in the role of liver cancer in mice,it mechanism because of the inhibition of MMP-2 in tumor tissue,MMP-9,VEGF protein expression and inhibiting tumor angiogenesis related organization.4.Through experiment in the detection of tumor tissue Bax,Bcl-2 protein expression,The results showed that,ETS can reduce the inhibition of protein expression of Bcl-2,enhance the expression of Bax promote protein,a djust the values of Bel-2/Bax to promote apoptosis of tumor cells,At the same time it can inhibit the Caspase 3,Caspase-9,increase the expression of Cyt-C.5.Antitumor activity has a close relationship with the structure type,saponins of sugar chain number,the number of saponins of sugar and sugar and sugar in saponin sugar chain connection.6.The results of the rat acute toxicity experiment showed that the single lavage of the ETS on female rats LD50 is 3.16 g·kg-1,the single lavage of the ETS on male rats LD50 is 5.84 g·kg-1.7.90 days of repeated dose harm dose not observed(NOAEL)is 540mg·kg-1 on SD rats.8.During the treatment period,with the dose in 60,180,540 mg/kg,each viscera organization and body weight were significantly decreased in high dose group.Four weeks after the drug was stopped,there was no any pathological change in organs.The no-observed adverse effect level(NOAEL)of the ETS was 540 mg/kg.Conclusion1.ETS can oinhibit tumor occurrence and development from several aspects.It can act on the death receptor pathway and endoplasmic reticulum Netcom.To inducing tumor cell apoptosis.2.Antitumor activity has a close relationship with the structure type,saponins of sugar chain number,the number of saponins of sugar and sugar and sugar in saponin sugar chain connection.3.The results of the rat acute toxicity experiment showed that the single lavage of the ETS on female rats LD50 is 3.16 g·kg-1,the single lavage of the ETS on male rats LD50 is 5.84 g·kg-1.5.During the treatment period,each viscera organization and body weight were significantly decreased in high dose group.Four weeks after the drug was stopped,there was no any pathological change in organs. |
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