Study On The Biological Activity And Mechanism Of Action Of Steroids In Simao Vine

In this work,Epigynum aruitum is the research object.The steroidal saponin components were enriched from E.aruitum and evaluated of immunosuppressive activity.Futhermore pregnane glycosides were screened the immunosuppressive activities and its mechanism was studied.In addition,the extracts from E.aruitum by different solvents,which were used to study the cell proliferation inhibitory activity of T acute lymphoblastic leukemia(T-ALL).Next,androstanes were screened for T-ALL cell proliferation inhibitory activity and its mechanism was also studied.The main results were concluded as follows:(1)The steroidal saponins were enriched by different extraction methods,and extracts were screened by LC-MS/MS assay,saponins detection and evaluated for Con A-induced spleen lymphocyte immune proliferation inhibitory activity.The results showed that the extracts with higher proportion of steroidal saponins had stronger immunosuppressive activity.Therefore,we believe that pregnane glycosides have immunosuppressive activity potential.(2)Pregnane glycosides 1-9 have been isolated from E.aruitum,and pregnane glycosides were used for screening immunosuppressive activity.The results showed that the immunosuppressive effect of epigycoside E(2)was comparable as the positive control DXM.Further studies showed that epigycoside E(2)caused G0/G1 arrest,and inhibited TNF-? and IL-2 production.(3)E.aruitum was extracted with different solvents,Polarities of PE,EA,n-Bu and W were screened the proliferation inhibitory activity of T-ALL cells.EA has the best inhibitory activity and 3-O-ethyl-androst-3?,12?-triol-4,14-diene-16-trione is the main components of EA by LC-MS/MS analysis.Further,EA was divided into different polar segments,E20%-2 and E80% have strong proliferation inhibition effects of T-ALL cells.Interestingly,mass spectrometry analysis,the main component of the extract E80% was androstanes.According to previous research,androstanes have strong immunosuppressive effects.Moreover,autoimmune diseases and lymphoproliferative tumors are closely related to T or B lymphocytes.Therefore,we tentatively consider that androstanes have the potential to inhibit the proliferation of T-ALL cells.(4)In order to prove the inhibitory effect of androstanes on the proliferation of T-ALL cells,androstanes 10-19 were screened for activity.Compound 13 had a strong inhibitory effect on CCRF-CEM cells and compound 14 had a strong inhibitory effect on proliferation of MOLT-4 cells.Both of them caused G0/G1 cell arrest;induced cell apoptosis,Bax/Bcl-2 ratio was increased,elevated p53 protein level;and increased intracellular ROS content to synergistically inhibited the proliferation of T-ALL cells.