ABSTRACT
Objective To establish acute hepatotoxic model induced by Amanita exitialis and to study the characteristics of acute toxic liver failure induced by mushrooms containing peptide toxins,in hope for providing some help to experimental research on poisoning induced by mushrooms containing peptide toxins.Methods UPLC-MS/MS (Ultra performance liquid chromatography-tandem mass spectrometry) method was used to detect peptide toxins in Amanita exitialis.To establish acute toxic liver hepatic failure model,the beagles were fed with 60 mg/kg of lyophilized powder of Amanita exitialis fungus which encapsulated in starch capsules.Toxic sighs were observed,coagulation function,hepatic and renal function,liver histopathological morphology,peptide toxin concentration in plasma and urine were detected during the experiment.Results Total peptide toxins in Amanita exitialis was (3 482.6 ± 124.94) mg/ kg.All the beagles had toxic signs including vomiting and diarrhea in 12-48 h after ingestion.On 24 h after ingestion,the beagles' ALT,AST,TBIL,ALP,PT and APTT levels increased obviously.On 36 h after ingestion,the beagles' ALT,AST,PT and APTT values reached their peaks (ALT:283.2 ± 112.9 Kallmann unit;AST:223.9 ±93.8 Kallmann units;PT:132.9 ± 152.6 s;APTT:131.4 ± 153.9 s).On 48 h after ingestion,the beagles' TBIL and ALP levels reached their peaks (TBIL:23.3 ± 14.6 mol/L;ALP:274.5 ± 115.5 U/L).The beagles' TBIL,TP and APTT returned to normal 1 week after ingestion,their ALT,AST and ALP levels returned to normal 3 weeks after ingestion.Three dogs died during 24-72 h after ingestion.Liver histopathological morphology study showed hemorrhagic necrosis of hepatocytes.Peptide toxins can be detected in plasma within 24 h after ingestion.Peptide toxins can be detected in urine within 96 h after ingestion.Conclusion Amanita peptide toxins can cause hemorrhagic necrosis of liver cells and lead to acute liver failure.This model is consistent with clinical pathophysiological process of acute toxic liver failure induced by mushrooms containing peptide toxins,and it can be applied to the study of diagnosis and treatment of poisoning induced by mushrooms containing peptide toxins.
ABSTRACT
Although cadmium is a well known heavy metal which has an influence testis and brings about male infertility, the mechanism of action in the testis is still fully unknown. In these experiment, cadmium chloride 4 mg/kg of body weight administered intraperitoneally to the rat (Sprague-Dawley) and sacrificed after 1 week, and morphological changes were observed by LM and TEM. In addition, electrophoresis, immunoprecipitation and Western blotting and N-terminal analysis performed to reveal the protein changes. 1. Major findings under light microscope were hemorrhagic necrosis and death of all the spermatogenic cells and supporting cells within the seminiferous tubules, and decreased volume of ECM, many apoptotic bodies, and death of interstitial cells and fibroblasts within interstitium. 2. The EM findings were disruption of nuclear membrane and disappearance of cell organelles of spermatogenic cells and supporting cells within seminiferous tubules, and decreased filopodia, increased inclusion bodies, vacuolation and apoptotic changes of the interstitial cells and fibroblastic cells, many short electron-dense collagen fibers in the extracellular matrix of interstitium. 3. Two proteins of molecular weight 42 kDa and 21 kDa which disappeared after cadmium treatment were rat collagen type I alpha 2. According to the above results, it is considered that cadmium degrades the collagen of the wall of small blood vessels within seminiferous tubules and interstitium and disrupts vascular walls, which results hemorrhagic necrosis, death of all the spermatogenic cells, and the death of interstitial cells and fibroblastic cells.