Study On Gastrointestinal Toxicity Component Of Euphorbia Kansui

Euphorbia kansui T.N.Liou ex T.P.Wang?Euphorbiaceae?is abundantly distributed in the northwestern China.The dried roots of Euphorbia kansui are known as "Kansui" in Chinese medicine."Kansui" was recorded in Sheng Nung' s Herbal as a low grade drug and has been used as a herbal remedy for edema,ascites,asthma and cancer.Previous research of E.kansui has discovered that it contained plentiful ingenane-type diterpenes and euphane/tirucallane-type triterpenes.Kansui is only for external use when it is raw since it's thrill and toxicity,and must be processed if it is to be taken orally,in order to enture the security of the clinical application.12 compounds were isolated with silica gel,HPLC,and TLC as the main method.Among them,9 compounds were elucidated on the basis of chemical and spectral methods.one jatrophane-type diterpenes were named as kansuinin B?EK-01?,two ingenane-type diterpenes were named as 3-O-?2,3-dimethylbutanoyl?-13-O-dodecanoyl-ingenol?EK-02?,5-benzoyl-20-deoxying enol?EK-03?,one triterpenes such as euphol?EK-04?,five others such as Isoscopletin?EK-05?,Adenosine?EK-06?,hydroxytyrosol?EK-07?,2,4,7-3 hydroxy isoflavones?EK-08?,Emodin?EK-09?.Determined by MTT method is used to evaluate the euphorbia kansui isolated part of the monomer compounds of rat intestinal epithelial cell toxicity,IEC-6,the results show that huge ji alkanes type 2 terpenoids EK-03 cytotoxicity.False white lam alkanes type 2 terpenoids EK₀1?Kansuinine B?and triterpene compounds EK-04?euphorbia alcohol?did not show good cytotoxic effect,EK-09?emodin?on intestinal epithelial cells ICE-6 has better inhibition.In order to investigat the mechanism of toxicity attenuation on intestinal epithelial cell line IEC-06 cells of EK-03.1?This section has studied the toxicity on intestinal epithelial cell IEC-06 of EK-03 by MTT assay.The results showed that EK-03 significantly inhibited the proliferation and morphological changes of IEC-06 cells.The results indicated that EK-03 can significantly induce hepatotoxicity?P<0.01?and in concentration-dependent manner.2?Using flow cytometry and HCS and other methods to detect the cell cycle and apoptosis rate;Results show that the EK-03 can induce intestinal epithelial cells?IEC 6?apoptosis of rats?P<0.01?,and in concentration-dependent manner.3?The HCS to detect apoptosis pathways,Real-time PCR and Western blot method to detect cell apoptosis gene mRNA expression and protein expression.EK-03 can significantly reduce the IEC-6 Hoechst nucleus fluorescence intensity of fluorescence intensity,mitochondrial membrane potential,cytochrome C fluorescence intensity?P<0.05?,and a certain dose of correlation;Can significantly increase the cell membrane permeability of the fluorescence intensity?P<0.01?,and a certain dose of correlation;Real-timePCR results show EK-03 can significantly increase of rat intestinal epithelial cell IEC 6 caspase 3,8,9 mRNAexpression?P<0.01?,and a certain dose of correlation;Western blot analysis results show that the EK-03 can significantly increase of rat intestinal epithelial cell IEC-06 cytc cytosol,Beta actin,Bax,AIF,Apaf-1 protein expression?P<0.01?,and a certain dose of correlation;Can significantly cut of rat intestinal epithelial cell IEC-6 cytc mitochondria,the Bcl 2 protein expression?P<0.01?,and a certain dose of relevance.Results showed that the EK-03 passed the mitochondrial pathway showed intestinal toxicity.