Study Of Toxic Theory Of The Peptides Toxins From Amantia Fuliginea And Pilot Study In Alexipharmic For These Toxins

The toxic theory of amanitatoxins(including alpha-amanitin) and alexipharmic for these toxins in liver of murine was studied.The result as follows:1 Though a known system and technology of isolation , purification and analysis of amantoxins,the main peptides toxins of Amantia fuliginea Hongo were produced. The body fluid of sufferer due to Amantia fuliginea were studied and the minim toxins were detected.2.The phenomenon was found with HE dyeing that the nuclear chromatin condenses and marginates below nuclear envelope,apoptoic bodis are formed and the cellur junctions are almost lose in liver cells in mouse after injected the crude amantoxins or alpha-amanitin.The number of necrosis cells increase parallel with the treated time by these toxins.3.Treated with fluorescence dye as AO(0.1 %),Hoechst 33258 and EB, the condensed nuclear chromatin was observed and the apoptosis index(AI) can calculate from visual field of fluorescence microscope. AI is correlate with the treated time of toxins.4.From the figure of ultrastructure of liver cells in toxic mouse , the pyknotic chromation which likes lune , integrated organelle in cell such as mitochondrion(Mt),endoplasmic reticulum(ER)and nuclear envelope and vacuolation of cytoplasm were observed ,but the cristae of Mt cann't observed distinctly.5.The DNA-fragment by agarose gel electrophoresis (1.5%, 5V/cm, 60min) demonstrated the typical ladder pattern on the liver cells directlyexposed to the crude amantoxins and alpha-amanitin,and it was correlate with the treated time and dose of these toxins in mouse.6.Used immunohistochemistry assay-SABC, the positive expressions of Bcl-2,Bax proteins in liver tissues in murine which treated with crude amantoxins and alpha-amanitin were detected, but the positive signal of Bcl-2 was larger than one of Bax in forepart.7.The cell cycle and the relative content of DNA of hepatocytes in murine were investigated with flow cytometry (FCM).alpha-amanitin can induce the Grstagnation in cycle and reduced content of DNA,but the obvious apoptosis apex wasn't found in maps of FCM when the dose was 0.6 g/g (avoirdupois of murine).8.The liquid distilled from Tuhuanlian(Sphenomeris chinensis) cann't free the sufferer from the poison of the peptide toxins of Amantia fuliginea ,but the ones from Ganoderma japonicum can serve as a effective alexipharmic for this poison to liver in murine.Conclusiomthe the peptide toxins of Amantia fuliginea can induce the apoptosis of hepatocytes in murine.alpha-amanitin can induce apoptosis or the the G1-stagnation in cell cycle in different experiment.