Establishment Of Experimental Pig Model Of Amanita Exitialis Poisoning

Objective: To establish a standard model of experimental pigs with Amanita exitialis poisoning,and to lay a foundation for further study on the mechanism of peptide toxicity,toxin detection,and new drug development.Methods:(1)morphology,molecular biology and toxin detection were used to identify the Amanita exitialis.(2)the toxin content of Amanita exitialis was determined by Reverse Phase High Performance Liquid Chromatograph(RP-HPLC).(3)eight Banna miniature pigs were randomly divided into blank control group(n=3)and experimental group(n=5).the experimental group was given 2mg/kg Amanita exitialis extract orally.clinical manifestations,liver function,coagulation function(ALT,AST,TBIL,MYO,CK,CK-MB,LDH,APTT,PT),histopathological and urine toxin test results changes of the two groups were observed after exposure.Results:(1)manifestation: the experimental group died within 75 hours,and the earliest time of death was 42 h after exposure,and the latest time of death was 75 h after exposure.during 6h to 52 h,the experimental group showed different degrees of digestive tract and mental symptoms,such as nausea,vomiting,diarrhea,decreased appetite and fatigue,there was no death and no gastrointestinal symptoms in the control group.(2)Blood biochemical and coagulation functions:ALT increased at the 28 th hour and reached a peak of 234.00±112.30U/L(P<0.01)at the 52 th hour after exposure.AST began to increase at the 6th hour and reached the peak of 13127.75±3506.19U/L(P<0.01)at the 52 th hour.TBIL increased at the 12 th hour and reached a peak of130.9mmol/L(P<0.01)at the 72 th hour after exposure.MYO increased significantly at the 28 th hour and reached to 7898ng/m L at the 72 th hour(P>0.05).LDH increased significantly at the 28 th hour and reached the peak value of 6492±1436.23U/L(P<0.05).CK reached a peak of 37572.00U/L at the 72 th hour(P>0.05).the value of CK-MB reached a peak of 3263±2488.31U/L at the 52 th hour,(P<0.05).the value of APTT reached a peak of 44.6±17.68S(P<0.05)at the 39 th hour,and PT reached a peak of 78.15±17.75S(P<0.05)at the 39 th hour.(3)Pathological changes:(1)the liver: the liver of the experimental group was normal in appearance,reddish-brown,with no dilatation of hepatic artery,hepatobiliary duct,portal vein,and gallbladder enlargement.the texture was soft,with varying degrees of focal or patchy bleeding on the diaphragmatic and visceral surfaces of the liver.under light microscope: at low magnification,the structure of hepatic lobules was destroyed to varying degrees.at high magnification,hepatic cord structure destruction,hepatic cell swelling,nuclear pyknosis,nuclear rupture,nucleolysis and other cell necrosis manifestations were observed.some hepatic cells showed fatty degeneration and bile acid crystallization,while portal area and central vein showed no obvious changes.(2)The heart:in the naked eye,the heart was normal in appearance,tough in quality,with normal vascular shape on the surface and no bleeding.the heart was flattened along the ventricular septal section,and the endocardial surface was observed,the bleeding focus in the area of the endocardial membrane,and the second and tricuspid outflow tract can be seen.under light microscope: at low magnification,there is patchy hemorrhage in myocardial interstitium.at high magnification,there are normal myocardial cells,accompanied by a large number of red blood cells and some infiltration of lymphocytes.(3)The kidney: normal appearance,reddish-brown,tough,no bleeding,cut along the coronal plane,normal shape of renal pelvis and calyces.under light microscope: at low magnification,the structure of renal corpuscle was complete,while at high magnification,the structure of glomerulus was complete,the renal tubule cells were slightly swollen,some proximal convolution tubule cells were necrotic,lumen collapsed.(4)Urine toxin detection: on the 6th hour after exposure,α-Amanitin was detected in the urine of the experimental group,which was 15.73±3.79μg/m L,but not detected after24 h.Conclusion: by morphological and molecular biology identification,we identified the species and genus of the Amanita exitialis extract.The types and contents of peptide toxins in Amanita exitialis were determined by RP-HPLC.combined with the clinical manifestations,laboratory examination,toxin detection and histopathological characteristics of experimental animals,we preliminarily established the liver failure model of Banna miniature pig with Amanita exitialis poisoning.