The authors have demonstrated the effect of sodium selenite on the hepatotoxicity due to carbon tetrachloride, by observing the distribution and disaggregation of the pyroninophilic granules in the hepatic cell of the mature male albino mice. Each experimental mouse of the selenite and the selenite plus carbon tetrachloride groups was given a single dose of 4 ug. of sodium selenite per kilogram of body weight and that of the control and the carbon tetrachloride groups was given 0.1 ml. of distilled water alone. Six hours after the first administration of distilled water or sodium selenite, the experimental mice of the carbon tetrachloride and the selenite plus carbon tetrachloride groups were given a single dose of l.0 ml. of carbon tetrachloride per kilogram of body weight and those of the selenite groups were given 0.l ml. of paraffin oil alone. Following the 1ast administration of carbon tetrachloride or paraffin oil, the mice were sacrificed by bleeding (cutting the common carotid artery) at the intervals of 2,3,4,6,8, and 12 hours respectively. Histochemical preparations were stained by the methyl-green and pyronin method and oil red 0 method. The hepatotoxicity due to the administration of carbon tetrachoride was evident in the hepatic cells; the pyroninophilic granlues were partly reduced in volume in the hepatic cells of the centrilobular and the intermediate zones as early as the 3 hour-period, and markedly reduced or disappeared in the centrilobular and some part of the intermediate zones associated with hydropic degeneration as well as in the 6 hour-period. Thereafter marked reduction or dissolution of the pyroninophilic granules was found and extended as the periportal zone at the 12 hour-period. However, the pyroninophilic granules in the hepatic cells of selenite plus carbon tetrachbride group showed no significant changes in the hepatic cells of these zones, compared to the histochemical feature of the granules in the hepatic cells of the control and the selenite groups. Consequently it is suggested that the lipid peroxidative decomposition of the microsomal membranes, which is induced with carbon tetrachloride, would be prevented by a previous administration of sodium selenite.
Animal ; Carbon Tetrachloride Poisoning*/pathology ; Cell Nucleus/drug effects ; Cytoplasm/drug effects ; Cytoplasmic Granules ; Lipids ; Liver/drug effects* ; Liver/pathology ; Male ; Mice ; Selenium/pharmacology* ; Vacuoles/drug effects

OBJECTIVETo study the effect of total glucosides of paeony (TGP) on lipopolysaccharides (LPS)-induced nuclear factor-kappaB (NF-kappaB) activation in macrophages.

METHODRat peritoneal macrophages were pre-treated with TGP for 2 h and stimulated with LPS for 20 min or 0.5 h. Inhibitory kappaBalpha (IkappaBalpha) protein in the cytoplasm and NF-kappaB p65 protein in the nuclear were analyzed by western blot. Further, DNA binding activity of NF-kappaB complex was detected.

RESULTTGP enhanced the amounts of IkappaBalpha protein in the cytoplasm and decreased the amounts of NF-kappaB p65 protein in the nuclear of LPS-induced macrophages. TGP also inhibited the LPS-mediated DNA binding activity of NF-kappaB complex in macrophages.

CONCLUSIONTGP can inhibit LPS-induced NF-kappaB activation in macrophages through arresting IKBalpha protein degradation, NF-kappaB p65 protein nuclear translocation and DNA binding activity of NF-kappaB complex.

Animals ; Cell Nucleus ; drug effects ; metabolism ; Cytoplasm ; drug effects ; metabolism ; DNA ; metabolism ; Dose-Response Relationship, Drug ; Glucosides ; pharmacology ; Macrophages, Peritoneal ; cytology ; drug effects ; metabolism ; NF-kappa B ; metabolism ; Paeonia ; chemistry ; Protein Transport ; drug effects ; Rats ; Transcription Factor RelA ; metabolism

The effects of morphine HCI on the rat mesenteric mast cells were studied with the electron microscopy. The materials were prepared for electron microscopy by osmium tetroxide fixation and embedding in Epon. The rat mesenteric mast cells showed no distinct morphological changes due to morphine HCl, but the mast cell granlues were changed in various ways. For instance, they formed dusters, showed granular lysis, and an appearance of electron transparency. Frequently, some granules appeared in the extracellular space and the boundary of the granules was not evident. From the results mentioned above, it was suggested that rat mesenteric mast cell granules were affected by morphine HCl in the shape, the granular matrix, and the granular boundaries.
Animal ; Cell Nucleus/ultrastructure ; Cytoplasm/ultrastructure ; Cytoplasmic Granules/drug effects ; Cytoplasmic Granules/ultrastructure ; Golgi Apparatus/ultrastructure ; Male ; Mast Cells/drug effects ; Mast Cells/ultrastructure* ; Mesentery/drug effects ; Mesentery/ultrastructure* ; Microscopy, Electron ; Mitochondria, Muscle/ultrastructure ; Morphine/pharmacology* ; Rats

AIMTo explore the effects of sinomenine(Sin) on intracellular free calcium ([Ca2+]i) and the activity of PKC (protein kinase C) of the cultured aortic vascular smooth muscle cells (VSMC) during ischemia and hypoxia.

METHODSThe effect of Sin on changes in [Ca2+]i were determined in cultured rabbit VSMC after exposure to high K+, norepinephrine (NE) and caffeine (Caf). Fluorescent Ca2+ -indicater fura-2/AM was used. The effects of Sin were compared with that of verapamil (Ver). The hypoxia model was made, then the activity of PKC was measured by y scintillation counting instrument.

RESULTSSin (10 x 10(-6) mol x L(-1), 3 x 10(-5) mol x L(-1) 10(-4) mol x L(-1)) inhibited the elevation of [Ca2+]i induced by high K+ -depolarization in a concentration dependent manner. In addition, Sin inhibited the elevation of [Ca2+]i induced by NE in the presence of extracellular Ca2+. In the absence of extracellular Ca2+, Sin (3 x 10(-5) mol.L(-1)) also had no blocking effect on the NE-induced [Ca2+]i increase. It was found that the activity of PKC treated with Sin in VSMC cytoplasm and cell membrane in normal condition increased, the activity of PKC in cytoplasm in ischemia and hypoxia situation increased, but the activity of PKC in cell membrane decreased. When VSMC was treated with Sin, the activity of PKC in cytoplasm decreased and that of cell membrane increased.

CONCLUSIONThe results suggest that Sin might decrease the[Ca2+] i of VSMC by blocking both VDC and ROC, could regulate the PKC activities induced by ischemia and hypoxia.

Animals ; Aorta ; cytology ; drug effects ; Calcium ; metabolism ; Cell Hypoxia ; Cells, Cultured ; Cytoplasm ; metabolism ; Morphinans ; pharmacology ; Muscle, Smooth, Vascular ; cytology ; drug effects ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; physiology ; Protein Kinase C ; metabolism ; Rabbits

OBJECTIVETo study whether aristololactam I (AL-I) can enter renal proximal tubular epithelial cells and the situation of intracellular distribution and accumulation.

METHODCultured human renal proximal tubular epithelial cell line (HK-2) was used as the subject. Intracellular fluorescence from AL-I and its distribution are examined by fluorescence microscopy after a treatment with different concentration of AL-I, the intracellular accumulation of AL-I was also investigated by incubated cells in AL-I -free medium for 48 h after washing-out the media containing AL-I.

RESULTAfter treatment of AL-I (concentration from 5 microg x mL(-1) to 20 microg x mL(-1)), glaucous fluorescence could be observed inside renal proximal tubular epithelial cells at 0.5 h, and the fluorescence distributed only in cytoplasm while not be observed in nuclei. Moreover, the fluorescence of AL-I could be kept in cytoplasm for more than 48 h after washing out the media containing AL-I .

CONCLUSIONAL-I is able to enter renal proximal tubular epithelial cells in short time and accumulate in cytoplasm, but not enter nuclei. This property may contribute to the cytotoxic mechanism of renal injury induced by AL-I, which may partially explain the persistent renal toxicity of AAs and its metabolites in the development of aristolochic acid nephropathy.

Animals ; Aristolochic Acids ; metabolism ; toxicity ; Cell Line ; Cell Nucleus ; drug effects ; metabolism ; Cytoplasm ; drug effects ; metabolism ; Epithelial Cells ; cytology ; drug effects ; metabolism ; pathology ; Humans ; Kidney Diseases ; metabolism ; pathology ; Kidney Tubules, Proximal ; cytology ; pathology ; Microscopy, Fluorescence

This research was aimed to study the effect of Emodin gel on the hypertrophic scars of rabbit ears. A total of 18 rabbits were randomly divided into Emodin group (9 rabbits) and control group (9 rabbits) after the successful animal model for hypertrophic scars had been made. The rabbits in the Emodin group were treated with Emodin Gel, while no special treatment was given to those in the control group. The other living conditions were all kept the same in the two groups. The diameter,hardness, and expression of transforming growth factor-beta (TGF-beta) and interleukin-1 (IL-1) of hypertrophic scars were measured after 4 weeks. Transmission electron microscopy was applied to observe the ultra-structure of the fibroblasts of hypertrophic scars. But there was no difference between the two groups in the diameter of hypertrophic scars (P>0.05). The hardness, expression of TGF-beta and IL-1 in hypertrophic scars in the Emodin group decreased, compared to the control group (P<0.05, P<0.01, P<0.05). Transmission electron microscopy showed that the fibroblast and organelle lessened in the cytoplasm and the collagen fibers dissolved obviously. The study showed that Emodin gel decreased the hardness of hypertrophic scars in the rabbit ears, and inhibited the proliferation of fibroblasts in local area. Therefore, Emodin gel treatment would be one of the methods to prevent and treat hypertrophic scars.
Animals ; Cicatrix, Hypertrophic ; drug therapy ; Cytoplasm ; Disease Models, Animal ; Emodin ; pharmacology ; Fibroblasts ; drug effects ; Gels ; Hardness ; Interleukin-1 ; metabolism ; Microscopy, Electron, Transmission ; Rabbits ; Transforming Growth Factor beta ; metabolism

In the present study, the apoptotic mechanism and polyamine transporter recognition of WJH-6, a novel polyamine conjugate, were investigated in K562 and HL-60 cells. The cytotoxicity of WJH-6 was assessed by MTT assay; cell cycle distribution and apoptosis were measured by flow cytometry; the protein expression of Caspase-3, Caspase-8, Caspase-9, Bid and mitochondrial membrane potential (MMP) were evaluated by high content screening (HCS) analysis; the protein expression of cytochrome c was measured by Western blotting. The results showed that WJH-6 could be recognized and transported by polyamine transporter (PAT). Furthermore, WJH-6 was able to inhibit K562 and HL-60 cells proliferation and induce apoptosis. This apoptotic effect was relative to MMP loss, cytochrome c release from mitochondria to cytoplasm and the activation of Caspase-8, Caspase-9, Caspase-3 and Bid. These results suggested that WJH-6-induced K562 and HL-60 cells apoptosis was related with mitochondrial damage.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; BH3 Interacting Domain Death Agonist Protein ; metabolism ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Caspase 9 ; metabolism ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cytochromes c ; metabolism ; Cytoplasm ; metabolism ; Enzyme Activation ; drug effects ; HL-60 Cells ; Humans ; K562 Cells ; Membrane Potential, Mitochondrial ; drug effects ; Mitochondria ; metabolism ; Polyamines ; pharmacology

Amyotrophic lateral sclerosis (ALS) is a fatal disease characterized by the premature loss of motor neurons. While the underlying cellular mechanisms of neuron degeneration are unknown, the cytoplasmic aggregation of several proteins is associated with sporadic and familial forms of the disease. Both wild-type and mutant forms of the RNA-binding proteins FUS and TDP-43 accumulate in cytoplasmic inclusions in the neurons of ALS patients. It is not known if these so-called proteinopathies are due to a loss of function or a gain of toxicity resulting from the formation of cytoplasmic aggregates. Here we present a model of FUS toxicity using the yeast Saccharomyces cerevisiae in which toxicity is associated with greater expression and accumulation of FUS in cytoplasmic aggregates. We find that FUS and TDP-43 have a high propensity for co-aggregation, unlike the aggregation patterns of several other aggregation-prone proteins. Moreover, the biophysical properties of FUS aggregates in yeast are distinctly different from many amyloidogenic proteins, suggesting they are not composed of amyloid.
Amyotrophic Lateral Sclerosis ; metabolism ; pathology ; Cell Proliferation ; drug effects ; Cytoplasm ; drug effects ; metabolism ; DNA-Binding Proteins ; genetics ; metabolism ; Detergents ; pharmacology ; Humans ; Kinetics ; Peptides ; metabolism ; Prions ; chemistry ; metabolism ; Protein Binding ; drug effects ; Protein Multimerization ; drug effects ; Protein Structure, Quaternary ; Protein Transport ; RNA-Binding Protein FUS ; chemistry ; genetics ; metabolism ; Saccharomyces cerevisiae ; cytology ; drug effects ; genetics ; metabolism ; Saccharomyces cerevisiae Proteins ; chemistry ; metabolism

Failure of a glaucoma filtering surgery mainly results from scarring at the filtering wound. Postoperative proliferation of fibroblasts plays an important role in scar tissue formation. Colchicine is a cytoplasmic microtu bule inhibitor capable of inhibiting fibroblast proliferation. The effect of colchicine on fibroblast proliferation at the filtering wound after fiItering surgery was investigated. Posterior lip sclerectomies were performed in each eye of albino rabbits. Under the topical or oral administration of colchicine, intracular pressure, conjunctival fibrosis, histologic finding, and drug toxicity were examined postoperatively. Compared to the untreated groups, reductions of intraocular pressure and conjunctival fibrosis in colchicine-treated groups after filtering surgery were statistically significant(p<0.05), and above changes in the topical administration group were more significant than in the oral administration group(p<0.05). Histologically, reductions of active fibroblasts and collagen fibers at the filltering wound and the subconjunctival area were seen in colchicine-treated eyes. Above findings were more prominent in the topical administration group. There were no ocular and systemic toxicities in both groups. The above results suggest that administration of colchicine, especially topical administration, can increase the success rate of filtering surgery.
Administration, Oral ; Administration, Topical ; Cicatrix ; Colchicine* ; Collagen ; Cytoplasm ; Drug-Related Side Effects and Adverse Reactions ; Fibroblasts* ; Fibrosis ; Filtering Surgery* ; Glaucoma* ; Intraocular Pressure ; Lip ; Rabbits ; Wounds and Injuries

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