Quality Evaluation Of Genus Paris And Spectrum-effect Relationship Of Anti-tumor Activity From P.forrestii

Objective:To explore the differences of chemical constituents among Paris and the effective substances of anti-hepatoma of P.forrestii,so as to provide a reference for the quality standard of Paridis rhizoma.Method:1.Different species of P.were collected and the content of 7 kinds of saponins analyzed by HPLC and HPLC chemical fingerprints were established.2.UPLC-ESI-Q-TOF-MS was used to identify the chemical constituents,the Principal Component Analysis（PCA）and Hierarchical Cluster Analysis（HCA）were applied for the chemical composition differences in P..3.MTT array was used to detect the inhibitory effects of P.forrestii on the Hep G2 cells,The quantitative real-time PCR and Western blot were used to detect the m RNA expressions of signal molecules including PI3K,AKT,m TOR,P70S6K,and 4EBP,as well as related protein expressions,respectively.Partial Least Squares Discriminant Analysis（PLS-DA）and Orthogonal Partial Least Squares Discriminant Analysis（OPLS-DA）were used to utilized the data of the spectrum and efficacy.Results:1.P.polyphylla Smith var.yunnensis,P.daliensis and P.forrestii have higher contents of total saponins in P..The results of fingerprint similarity analysis are shown that the samples with the same growing years and the same batch have high similarity.2.A method for UPLC-MS of P.was established,and 10 species in P.were analysis.72 compounds were identified in P.polyphylla Smith var.yunnanensis,46compounds were identified in P.polyphylla Smith var.Chinensis,75 compounds were identified in P.daliensis,62 compounds were identified in P.polyphylla Smith var.polyphylla,37 compounds were identified in P.thibetica,41 compounds were identified in P.mairei,72 compounds were identified in P.Rugosa,48 compounds were identified in P.polyphylla Smith var.kwantungensis,41 compounds were identified in P.polyphylla var.Stenophylla,81 compounds were identified in P.forrestii.PolyphyllinⅠ,Ⅱ,Ⅴ,Ⅵ,Ⅶcould be detected in all samples.PCA score plot and HCA analysis showed P.polyphylla Smith var.polyphylla,P.polyphylla Smith var.kwantungensis,P.daliensis can be clustered in their own category,and the others cluster in one groups.3.The P.forrestii has the highest inhibitory effect on the Hep G2 cells after treated 72 h,the IC₅₀of P.forrestii was 3.1-131.5μg/m L.P.forrestii significantly down-regulated the m RNA and the protein expression of PI3K,AKT,m TOR and P70S6K,up-regulate the expression of 4EBP.The results of PLS-DA and OPLS-DA analysis showed that a variety of ingredients in P.forrestii with higher resistance to the anti-tumor effect.Conclusion:In this study,the content and chemical composition of 10 species of the genus P.was determined.P.forrestii,P.daliensis and other medicinal materials could be used as potential alternative resources.The different chemical components of samples were preliminaries identified and provided a reference for characteristic markers of the chemical composition difference of Paridis rhizoma.The anti-tumor effect of P.forrestii may inhibit cell activity by regulating the PI3K/AKT/m TOR signaling pathway,which provides an experimental basis for further study on the efficacious material basis of P..