To investigate toxicity of aflatoxin Gl and its mechanism, light microscopic, histochemical, electron microscopic and autoradiographic studies were done on the rat liver at various time intervals after the administration of aflatoxin Gl. Light microscopic alteration was first observed at 6 hours and necrosis of periportal hepatic cells was found at 18 hours. However, reduction of Feulgen positivity of the nucleus and pyroninophilia of cytoplasm was observed as early as 1 hour. Ultrastructural changes were noted at 6 hours and were advanced at l8 hours. Early changes consisted of nucleolar segregation, dilatation of rough endoplasmic reticulum, swelling of mitochondria and detachment of membrane bound ribosomes followed later by disruption of cytoplasmic organellae and focal necrosis. These changes were most marked at periportal region. Autoradiographic studies showed inhibition of H3-uridine incorporation into the nucleus at 1 hour, was most marked at 6 hours, and showed some recovery at 18 hours. H3-uridine labeling in the cytoplasm was also inhibited and the most marked inhibition was noted at 1 hour after the aflatoxin administration. These data indicate aflatoxin Gi has a hepatotoxic effect, particulary at the periportal region. This toxic effect is likely due to inhibition of nuclear RNA synthesis which leads to inhibition of ribosomal RNA and eventually protein synthesis. The DNA synthesis is also inhibited, as shown by reduction of Feulgen reaction in the nucleus.
Aflatoxins/toxicity* ; Animal ; Autoradiography ; Cell Nucleus/ultrastructure ; Cytoplasm/ultrastructure ; Histocytochemistry ; Liver/ultrastructure* ; Microscopy, Electron ; Mitochondria, Liver/drug effects ; Rats ; Uridine/metabolism

BACKGROUND/AIMS: In spite of increasing interests about nonalcoholic steatohepatitis (NASH), there are few reports about the ultrastructure of hepatocyte in this disease. The aim of this study was to clarify abnormal electron microscopic (EM) findings and related factors in NASH. METHODS: Total of fourteen patients who underwent liver biopsy due to steatohepatitis were included. Precise personal history was taken and variable blood tests such as liver function test, lipid profile, and serum iron study were done. Pathologic examination with light and electron microscopy was done by single pathologist. RESULTS: Eleven men and three women were included and mean age was 33.7+/-12.8 years. Nine patients drinking less than 40 g/week was grouped as "NASH group" and other 5 patients drinking more than 40 g/week and body mass index less than 25 was grouped as "ASH (Alcoholic Steatohepatitis) group". Polymorphism of mitochondria such as megamitochondria or loss of cristae was major abnormal EM findings and was more common in "NASH group" than "ASH group" (p=0.027). There was no significant clinical or pathological factors related with the presence of these abnormal EM findings. CONCLUSIONS: Polymorphism of mitochondria is major abnormal EM finding of steatohepatitis and is more common in NASH than ASH. And there is no significant clinical or pathological factors which could predict the presence of these abnormal EM findings.
Adolescent ; Adult ; Fatty Liver/*pathology ; Female ; Hepatocytes/*ultrastructure ; Humans ; Liver Diseases, Alcoholic/*pathology ; Male ; Microscopy, Electron, Transmission ; Middle Aged ; Mitochondria, Liver/ultrastructure

OBJECTIVETo investigate the clinicopathological features of infantile cytomegalovirus hepatitis.

METHODLiver biopsies from 30 cases of infantile cytomegalovirus hepatitis were observed under optical microscope and electronic microscope.

RESULTThe main clinical manifestations were jaundice, splenohepatomegaly and hypohepatia. Laboratory test showed dysfunction of liver, high level of CMV DNA, and high titer of anti-CMV antibody. Imaging examination demonstrated hepatomegaly. The histological changes were hepatocellular degeneration, necrosis, apoptosis, and fibrosis. The histological characteristics of cytomegalovirus hepatitis, including intranuclear inclusions in multinucleated giant cells and pseudo-lumens, were also observed under optical microscope. In addition, virion was observed in the nuclei and cytoplasm of hepatocytes under electronic microscope.

CONCLUSIONThe viral DNA and serological tests have limited utility for the diagnosis of infantile cytomegalovirus hepatitis, and the final diagnosis depends on histopathology.

Biopsy, Needle ; Cytomegalovirus Infections ; pathology ; Female ; Hepatitis, Viral, Human ; pathology ; Hepatocytes ; pathology ; ultrastructure ; Humans ; Inclusion Bodies, Viral ; pathology ; Infant ; Infant, Newborn ; Liver ; pathology ; Male ; Mitochondria, Liver ; pathology ; ultrastructure

OBJECTIVETo study the effect of Renshen Yangrong decoction (RYD) in protecting morphology and structure of mitochondria in brain and liver of D-galactose induced subacute senile mice.

METHODSForty ICR mice were randomly divided into the youth control group, the model group, the high, middle and low-dose RYD groups. The ultra-structure changes of mitochondria were observed by electronic microscope, the specific surface area, body density, numerical density on area, deltammit area and ridge count of mitochondria were quantitatively analyzed with medical image analyzer.

RESULTSIn the model group, the numbers of mitochondria decreased, mitochondria swelled and significantly expanded, ridge was fragmented, malformed and dissolved even vanished. Those manifestations were coincided with the morphological changes of mitochondria in senile mice. In the different dose of RYD groups, the numbers of mitochondrion increased, mitochondrion tumid were significantly improved, senile degenerative changes of ridge and intima were obviously alleviated in a dose-dependent manner.

CONCLUSIONRYD can anti-age by improving the changes of mitochondria structure of liver and brain accompanied with ageing.

Aging ; drug effects ; Animals ; Brain ; drug effects ; ultrastructure ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Liver ; drug effects ; ultrastructure ; Mice ; Mice, Inbred ICR ; Mitochondria ; drug effects ; ultrastructure ; Panax ; Protective Agents ; pharmacology ; therapeutic use

OBJECTIVETo investigate the protective effect of ischemic postconditioning (IPC) against hepatic ischemia-reperfusion injury.

METHODSTwenty-four normal male Wistar rats were randomly divided into sham-operated group, ischemia-reperfusion group (IR) and IPC group, and in the latter two groups, the rats were subjected to acute hepatic ischemia-reperfusion. IPC was achieved by several brief pre-reperfusion and withdrawn before persistent reperfusion. The concentration of malondialdehyde (MDA) and activity of several antioxidant enzymes in the hepatic tissue were measured. The cell apoptosis was detected by TdT-mediated dUTP-biotin nick end labeling (TUNEL) and the expression of Bcl-2 protein measured by immunohistochemistry. The mitochondrial ultrastructural and morphological changes of the hepatic cells were observed by electron microscopy.

RESULTSCompared with IR group, IPC group showed significantly reduced concentration of MDA and the hepatocellular apoptotic index (P<0.05) with markedly enhanced activity of the antioxidant enzymes and Bcl-2 protein expression (P<0.05).The mitochondrial ultrastructural damage was also relieved obviously in IPC group.

CONCLUSIONIPC can reduce the hepatocellular apoptosis after reperfusion and offers protection against hepatic IR injury.

Animals ; Apoptosis ; Ischemic Postconditioning ; methods ; Liver ; blood supply ; metabolism ; pathology ; Male ; Malondialdehyde ; metabolism ; Mitochondria, Liver ; ultrastructure ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Reperfusion Injury ; prevention & control

OBJECTIVETo study the protective effect of Shaoganduogan (SGDG) on serum transaminase, liver pathology and hepatocyte mitochondria in rat with subacute liver injury induced by carbon tetrachloride.

METHODSubacute liver injury of rats were induced by carbon tetrachloride, and cured by different doses of SGDG through intragastric administration. The activity of serum ALT, AST, liver pathology and ultrastructure, activity of ATPase, SOD and content of MDA of hepatocyte mitochondria were observed.

RESULTSGDG can remarkably reduce the transaminase, alleviate the degeneration and necrosis of liver cells ,enhance activity of Na+ -K+ ATPase, Ca2+ ATPase, SOD, reduce content of MDA of mitochondria, alleviate ultrastructure change of mitochondria, reduce section area, perimeter equivalent diameter and average optical density perimeter of liver cells.

CONCLUSIONSGDG has obvious effect of liver protection, the mechanisms are related with alleviating mitochondria injury.

Animals ; Carbon Tetrachloride ; adverse effects ; Chemical and Drug Induced Liver Injury ; pathology ; Drugs, Chinese Herbal ; pharmacology ; Hepatocytes ; drug effects ; pathology ; Male ; Malondialdehyde ; metabolism ; Mitochondria ; drug effects ; enzymology ; metabolism ; ultrastructure ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo determine the roles of mitochondrial apoptosis and energy metabolism in hepatocytes during the pathogenic process of acute renal failure (ALF) by assessing disease-related differential activities of several key mitochondrial enzymes, including citrate synthase (CS), carnitine palmitoyltransferase-1 (CPT-1) and cytochrome c oxidase (COX).

METHODSThirty-two male Sprague Dawley rats were given D-galactosamine followed by and lipopolysaccharide (LPS) to induce acute liver failure and sacrificed after 4 (4 h group), 8 (8 h group) 12 (12 h group) and 24 hours (24 h group) of treatment. Eight unmodeled rats served as controls. Effects related to apoptosis were evaluated by pathological analysis of hepatic tissues and TUNEL staining. Ultrastructural changes in mitochondria were assessed by electron microscopy. The activity and expression of CS, CPT-1 and COX were measured.

RESULTSHepatocyte apoptosis was present in the 4 h treatment group and was increased obviously in the 8 h treatment group. Hepatocyte necrosis was first observed in the 12 h treatment group and was significantly higher in the 24 h treatment group, with inflammatory cell invasion. Ultrastructural changes in mitochondria were present in the 4 h treatment group, and the 24 h treatment group showed mitochondria with completely destroyed outer membranes, which resulted in mitochondrial collapse. Activity and protein expression of CS, CPT-1 and COX were increased in the 4 h group (vs. controls), were at their peak in the 8 h group (CS:t =1.481, P less than 0.01; CPT-1:t =2.619, P less than 0.05; COX:t =1.014, P less than 0.01) and showed a decreasing trend in the 12 h group. In addition, the activities of CS, CPT-1 and COX were enhanced at the stage of hepatocyte apoptosis, suggesting that these enzymes were involved in the initiation and development of ALF.

CONCLUSIONEnergy metabolism plays an important role in hepatocyte apoptosis during ALF.

Animals ; Apoptosis ; Carnitine O-Palmitoyltransferase ; metabolism ; Citrate (si)-Synthase ; metabolism ; Disease Models, Animal ; Electron Transport Complex IV ; metabolism ; Hepatocytes ; cytology ; enzymology ; Liver Failure, Acute ; metabolism ; pathology ; Male ; Mitochondria ; ultrastructure ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the anti-cancer effects of crude extract from Melia toosendan Sieb. et Zucc and its possible molecular mechanisms in vitro and in vivo.

METHODSTransonic alcohol-chloroform extraction method was used to extract toosendanin from the bark of Melia toosendan Sieb. et Zucc, and the content of toosendanin in the crude extract was measured by high performance liquid chromatography (HPLC). Anti-cancer effects of crude extract from Melia toosendan Sieb. et Zucc were investigated in in vivo and in vitro studies. In the in vitro experiment, human hepatocellular carcinoma cell lines SMMC-7721 and Hep3B were co-incubated with toosendanin crude extract of different concentrations, respectively. In the in vivo experiment, BALB/c mice were subcutaneously inoculated with mouse hepatocellular carcinoma H22 cells and treated with crude extract.

RESULTSHPLC revealed the content of toosendanin was about 15%. Crude extract from Melia toosendan Sieb. et Zucc inhibited cancer cells growth in a dose- and time-dependent manner. The 50% inhibitory concentration (IC50, 72 h) was 0.6 mg/L for SMMC-7721 cells and 0.8 mg/L for Hep3B cells. Both high-dose [0.69 mg/(kg d)] and low-dose [0.138 mg/(kg d)] crude extract could markedly suppress cancer growth, and the inhibition rate was greater than 50%. Hematoxylin and eosin staining showed necrotic area in cancers and transmission electron microscopy displayed necrotic and apoptotic cancer cells with apoptotic bodies. Immunohistochemistry showed that the expression of Bax and Fas increased and the expression of Bcl-2 reduced.

CONCLUSIONSToosendanin extract has potent anti-cancer effects via suppressing proliferation and inducing apoptosis of cancer cells in vivo and in vitro. The mechanism of apoptosis involves in mitochondrial pathway and death receptor pathway.

Animals ; Antineoplastic Agents ; pharmacology ; therapeutic use ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; drug therapy ; pathology ; ultrastructure ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; therapeutic use ; Female ; Immunohistochemistry ; Liver Neoplasms ; drug therapy ; pathology ; ultrastructure ; Male ; Melia ; chemistry ; Mice, Inbred BALB C ; Mitochondria ; drug effects ; metabolism ; Neoplasm Transplantation ; Plant Extracts ; therapeutic use ; Reference Standards ; bcl-2-Associated X Protein ; metabolism ; fas Receptor ; metabolism

OBJECTIVETo investigate the effect of berbamine on human hepatoma cell line SMMC7721.

METHODSThe effects of 24 h and 48 h incubation with different concentrations (0 to approximately 64 microg/ml) of the berbamine on SMMC7721 cells were evaluated using 3-4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT) assay. Hoechst 33258 staining was conducted to distinguish the apoptotic cell, and the appearance of sub-G1 stage was determined by PI (propidium iodide) staining, the percentage of apoptotic cell was determined by flow cytometry following annexin V/PI staining. Flow cytometry was performed to analyze the cell cycle distribution and the mitochondrial membrane potential (psi(m)); the expression of activated caspase3 and caspase9 was analyzed by Western-blot.

RESULTSThe proliferation of SMMC7721 was decreased after treatment with berbamine in a dose- and time-dependent manner. Berbamine could induce apoptosis in SMMC7721 cells and could cause cell cycle arrest in G0/G1 phase, to induce loss of mitochondrial membrane potential (psi(m)) and activate caspase3 and caspase9. Berbamine-induced apoptosis could be blocked by the broad caspase inhibitor z-VAD-fmk.

CONCLUSIONBerbamine exerts antiproliferative effects on human hepatocellular carcinoma SMMC7721 cells. The anticancer activity of berbamine could be attributed partly to its inhibition of cell proliferation and induction of apoptosis in cancer cells through loss in mitochondrial transmembrane potential and caspase activation.

Alkaloids ; pharmacology ; Apoptosis ; drug effects ; Benzylisoquinolines ; pharmacology ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Enzyme Activation ; drug effects ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Medicine, Chinese Traditional ; Membrane Potentials ; drug effects ; Mitochondria, Liver ; drug effects ; metabolism ; ultrastructure