We investigated the effect of desferrioxamine (DFO), an iron chelator, on the DNA synthesis and the cell cycle of cultured hepatoma cells. Using Hep 3B cells as the hepatoma cell lines, DNA synthesis was measured by [3H] thymidine incorporation, and the cell cycle analysis was performed by flow cytometry including bivariate DNA/BrdU analysis. [3H] thymidine uptake was decreased by DFO in a dose dependent manner. The proportion of S phase cells increased and that of G0/G1 phase cells decreased after the addition of DFO in the culture media in a dose dependent manner up to 20 micrograms/ml of DFO. The S phase duration of the exponentially proliferating Hep 3B cells was 9.9 hours when cultured without DFO, but it was markedly prolonged (54.1 hours) after the addition of 20 micrograms/ml of DFO. After removal of DFO from the culture media following 24 hours of incubation with 20 micrograms/ml of DFO, a sequential increase from early through mid and late-S to G2/M phase was observed. In conclusion, the antiproliferative effect of DFO on cultured human hepatoma cell lines was caused by the inhibition of DNA synthesis which was related to a block in the early-mid S interface or mid S phase of the cell cycle.
Bromodeoxyuridine/diagnostic use ; Carcinoma, Hepatocellular/*pathology ; Cell Cycle/drug effects ; Cell Division/drug effects ; Deferoxamine/*pharmacology ; Flow Cytometry ; Human ; Liver Neoplasms/*pathology ; Tumor Cells, Cultured/drug effects

BACKGROUND: Posttraumatic stress disorder (PTSD) and depression are highly comorbid. Psilocybin exerts substantial therapeutic effects on depression by promoting neuroplasticity. Fear extinction is a key process in the mechanism of first-line exposure-based therapies for PTSD. We hypothesized that psilocybin would facilitate fear extinction by promoting hippocampal neuroplasticity. METHODS: First, we assessed the effects of psilocybin on percentage of freezing time in an auditory cued fear conditioning (FC) and fear extinction paradigm in mice. Psilocybin was administered 30 min before extinction training. Fear extinction testing was performed on the first day; fear extinction retrieval and fear renewal were tested on the sixth and seventh days, respectively. Furthermore, we verified the effect of psilocybin on hippocampal neuroplasticity using Golgi staining for the dendritic complexity and spine density, Western blotting for the protein levels of brain derived neurotrophic factor (BDNF) and mechanistic target of rapamycin (mTOR), and immunofluorescence staining for the numbers of doublecortin (DCX)- and bromodeoxyuridine (BrdU)-positive cells. RESULTS: A single dose of psilocybin (2.5 mg/kg, i.p.) reduced the increase in the percentage of freezing time induced by FC at 24 h, 6th day and 7th day after administration. In terms of structural neuroplasticity, psilocybin rescued the decrease in hippocampal dendritic complexity and spine density induced by FC; in terms of neuroplasticity related proteins, psilocybin rescued the decrease in the protein levels of hippocampal BDNF and mTOR induced by FC; in terms of neurogenesis, psilocybin rescued the decrease in the numbers of DCX- and BrdU-positive cells in the hippocampal dentate gyrus induced by FC. CONCLUSIONS A single dose of psilocybin facilitated rapid and sustained fear extinction; this effect might be partially mediated by the promotion of hippocampal neuroplasticity. This study indicates that psilocybin may be a useful adjunct to exposure-based therapies for PTSD and other mental disorders characterized by failure of fear extinction.
Humans ; Mice ; Animals ; Psilocybin/metabolism* ; Fear ; Extinction, Psychological ; Brain-Derived Neurotrophic Factor/metabolism* ; Bromodeoxyuridine/pharmacology* ; Hippocampus/metabolism* ; Neuronal Plasticity ; TOR Serine-Threonine Kinases/metabolism*

OBJECTIVEThe literature has shown that cognitive and emotional changes may occur after chronic treatment with glucocorticoids. This might be caused by the suppressive effect of glucocorticoids on hippocampal neurogenesis and cell proliferation. Paroxetine, a selective serotonin reuptake transporter, is a commonly used antidepressant for alleviation of signs and symptoms of clinical depression. It was discovered to promote hippocampal neurogenesis in the past few years and we wanted to investigate its interaction with glucocorticoid in this study.

METHODSAdult rats were given vehicle, corticosterone, paroxetine, or both corticosterone and paroxetine for 14 d. Cell proliferation in the dentate gyrus was quantified using 5-bromo-2-deoxyuridine (BrdU) immunohistochemistry.

RESULTSThe corticosterone treatment suppressed while paroxetine treatment increased hippocampal cell proliferation. More importantly, paroxetine treatment could reverse the suppressive effect of corticosterone on hippocampal cell proliferation.

CONCLUSIONThis may have clinic application in preventing hippocampal damage after glucocorticoid treatment.

Analysis of Variance ; Animals ; Bromodeoxyuridine ; metabolism ; Cell Count ; Cell Proliferation ; drug effects ; Corticosterone ; pharmacology ; Drug Interactions ; Hippocampus ; cytology ; Male ; Neural Inhibition ; drug effects ; Neurons ; drug effects ; Paroxetine ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Serotonin Uptake Inhibitors ; pharmacology

OBJECTIVETo investigate the effect of Buyang Huanwu Decoction (BYHWD) and the different combinations of its ingredients on neurogenesis following ischemic stroke in rats.

METHODSThe model rats of ischemic stroke was established by blocking cerebral media artery with electrocoagulation through craniectomy, and electric stimulation, given from 24 h after blocking, 2 h daily for 15 successive days. They were divided into four groups, Group A treated with saline, Group B treated with BYHWD, Group C treated with BYHWD but earthworm subtracted, and Group D treated with Danggui Buxue Decoction (DGBXD). The expression of 5-bromodeoxyuridine (BrdU) in cerebral tissue was determined by immunohistochemical method.

RESULTSLarge amount of BrdU immunoreactive cells presented in the hippocampal region of rats in Group B and C, densely arranged, partial in cluster, with the figure significantly different to that in Group A (P < 0.01), and the amount in the ischemic side was significantly more than that in the opposite side (P < 0.05). While comparing between Group A and D, the amount of BrdU immunoreactive cells in the hippocampal region showed insignificant difference (P > 0.05).

CONCLUSIONBYHWD has a effect in promoting neurogenesis better than DGBXD.

Animals ; Bromodeoxyuridine ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Hippocampus ; drug effects ; metabolism ; pathology ; Immunohistochemistry ; Infarction, Middle Cerebral Artery ; drug therapy ; Male ; Neurons ; drug effects ; metabolism ; pathology ; Neuroprotective Agents ; pharmacology ; therapeutic use ; Phytotherapy ; Random Allocation ; Rats ; Rats, Wistar

We investigated the effect of low dose radiation on diabetes induced suppression of neurogenesis in the hippocampal dentate gyrus of rat. After 0.01 Gy, 0.1 Gy, 1 Gy and 10 Gy radiation was delivered, the dentate gyrus of hippocampus of streptozotocin (STZ)-induced diabetic rats were evaluated using immunohistochemistry for 5-bromo-2-deoxyuridine (BrdU), caspase-3, and terminal deoxynucleotidyl transferase-mediated nick end-labeling (TUNEL) staining. The number of BrdU positive cells in the non-diabetic rats, diabetic rats without radiation, diabetic rats with 0.01 Gy radiation, diabetic rats with 0.1 Gy radiation, diabetic rats with 1 Gy radiation and diabetic rats with 10 Gy radiation were 55.4+/-8.5/mm2, 33.3+/-6.4/mm2, 67.7+/-10.5/mm2, 66.6+/-10.0/mm2, 23.5+/-6.3/mm2 and 14.3+/-7.2/mm2, respectively. The number of caspase-3 positive cells was 132.6+/-37.4/mm2, 378.6+/-99.1/mm2, 15.0+/-2.8/mm2, 57.1+/-16.9/mm2, 191.8+/-44.8/mm2 and 450.4+/-58.3/mm2, respectively. The number of TUNEL-positive cells was 24.5+/-2.0/mm2, 21.7+/-4.0/mm2, 20.4+/-2.0/mm2, 18.96+/-2.1/mm2, 58.3+/-7.9/mm2, and 106.0+/-9.8/mm2, respectively. These results suggest low doses of radiation paradoxically improved diabetes induced neuronal cell suppression in the hippocampal dentate gyrus of rat.
Rats, Sprague-Dawley ; Rats ; Radiotherapy/methods ; Neurons/*metabolism ; Male ; In Situ Nick-End Labeling ; Hippocampus/*cytology/metabolism/radiation effects ; Diabetes Mellitus, Experimental/radiotherapy ; Dentate Gyrus/drug effects/*radiation effects ; Cell Proliferation ; Caspase 3/metabolism ; Bromodeoxyuridine/pharmacology ; Apoptosis ; Animals

OBJECTIVEThis study aimed to determine whether pentylenetetrazol-induced status epilepticus (SE) can induce dentate granule cell neurogenesis in the developing rat and the effect of MK-801, a noncompetitive antagonism of N-methyl-D-aspartate receptor (NMDAR), on neurogenesis.

METHODSTwo hundred and sixteen postnatal days 7, 14, 21 or 28 Sprague Dawley (SD) rats were involved in this study. Each age group consisted of 54 rats which were randomly assigned into a SE group, a SE + MK-801 group and a Normal control group (n=18 each). SE was induced by intraperitoneal injection of PTZ (80 mg/kg). The SE + MK-801 group was injected intraperitoneally with MK-801 (1 mg/kg) at 1 hr after SE episode. All rats were given 5-bromodeoxyuridene (BrdU) intraperitonealy to label newborn cells at 6, 13 and 27 days after seizures and then were sacrificed 24 hrs after BrdU injection. The immunohistochemistry method was used to measure the expression of BrdU, TuJl (betaIII tubulin), and glial fibrillary acidic protein (GFAP) in the dentate gyrus of hippocampus of rats.

RESULTSThe number of the BrdU positive cells in the SE group was significantly higher than in the age-matched normal controls at 7 and 14 days after SE episode (P <0.05 or 0.01). Approximately 82.5% and 80.3% of BrdU-labeled cells in the SE and the Control groups were co-expressed TuJ1 respectively. MK-801 treatment decreased the BrdU positive cells compared with the SE group at 7 and 14 days after SE seizures (P < 0.01). On the 28th day after SE episode there were no differences among the three groups for the BrdU positive cells.

CONCLUSIONSPTZ-induced SE can increase the dentate granule cell neurogenesis in the developing rat. NMDAR plays an important role in neurogenesis following seizures.

Animals ; Bromodeoxyuridine ; metabolism ; Dentate Gyrus ; drug effects ; physiopathology ; Dizocilpine Maleate ; pharmacology ; Female ; Glial Fibrillary Acidic Protein ; analysis ; Immunohistochemistry ; Male ; Pentylenetetrazole ; Rats ; Rats, Sprague-Dawley ; Status Epilepticus ; chemically induced ; physiopathology

OBJECTIVETo investigate the effects of erythropoietin (EPO) on the neuronal proliferation and apoptosis in neonatal rats after infection-induced brain injury and the neuroprotective mechanism of EPO in neonatal rats with infection-induced brain injury.

METHODSTwenty-six two-day-old neonatal rats were randomly divided into 3 groups: control group (intraperitoneally given an equal volume of normal saline), lipopolysaccharide (LPS) group (intraperitoneally given LPS 0.6 mg/kg), and EPO group (intraperitoneally given LPS 0.6 mg/kg and EPO 5 000 U/kg). These groups were injected with respective drugs for 5 consecutive days. Meanwhile, each group was intraperitoneally injected with 5-bromo-2'-deoxyuridine (BrdU) (50 mg/kg) once a day for 5 consecutive days. The expression of BrdU and cleaved Caspase-3 in the hippocampal dentate gyrus was detected by immunohistochemistry at 24 hours after the last injection.

RESULTSThe number of neuronal cells in the hippocampal dentate gyrus in the LPS and EPO groups was significantly greater than in the control group (P<0.05), but there was no significant difference between the LPS and EPO groups. The EPO group had a significantly higher number of BrdU-positive cells in the subgranular zone of hippocampal dentate gyrus than the LPS group (51±9 vs 29±6; P<0.05), but a significantly lower number of BrdU-positive cells than the control group (51±9 vs 67±12; P<0.05). The EPO group had a significantly lower number of cleaved Caspase-3-positive cells in the subgranular zone of hippocampal dentate gyrus than the LPS group (27.9±1.5 vs 34.0±1.3; P<0.05), but a significantly higher number of cleaved Caspase-3-positive cells than the control group (27.9±1.5 vs 21.0±1.7; P<0.05).

CONCLUSIONSEPO can promote hippocampal neuronal proliferation and reduce neuronal apoptosis in neonatal rats after infection-induced brain injury.

Animals ; Animals, Newborn ; Apoptosis ; drug effects ; Brain Diseases ; drug therapy ; pathology ; Bromodeoxyuridine ; metabolism ; Caspase 3 ; metabolism ; Cell Proliferation ; drug effects ; Erythropoietin ; pharmacology ; therapeutic use ; Hippocampus ; pathology ; Neurons ; pathology ; Rats ; Rats, Sprague-Dawley

p21Cip/WAF1, an important regulator of cell proliferation, is induced by both p53- and extracellular signal regulated kinase (ERK) pathways. The induction of p21Cip/WAF1 occurs by prolonged activation of the ERKs caused by extracellular stimuli, such as zinc. However, not all the cells appeared to respond to ERK pathway dependent p21Cip/WAF1 induction. Here we investigated the cause of such difference using colorectal cancer cells. p21Cip/WAF1 induction and concomitant reduction of bromodeoxyuridine (BrdU) incorporation were observed by zinc treatment within HT-29 and DLD-1. However, HCT-116 cells with high endogenous p21Cip/WAF1 levels did not show any additional increment of p21Cip/WAF1 levels by zinc treatment and did maintain high BrdU incorporation level. The p21Cip/WAF1 induction by zinc depended upon prolonged activation of extracellular signal regulated kinase (ERK) was not observed in HCT-116 cells. The percentage of BrdU positive cells was 50% higher in p21Cip/WAF1 -/- HCT-116 cells compared to p21Cip/WAF1 +/+ HCT- 116 cells, and no cells induced p21Cip/WAF1 incorporated BrdU in its nucleus, yet confirming the importance of p21Cip/WAF1 induction in anti- proliferation. These results again support that p21Cip/WAF1 induction is a determinant in the regulation of colonic proliferation by the ERK pathway.
Bromodeoxyuridine/metabolism ; Cell Cycle Proteins/*metabolism ; Cell Line, Tumor ; Cell Proliferation ; Colorectal Neoplasms/enzymology/*metabolism ; Enzyme Activation ; Extracellular Signal-Regulated MAP Kinases/metabolism/*physiology ; Flavonoids/pharmacology ; Humans ; Protein Kinase Inhibitors/pharmacology ; Research Support, Non-U.S. Gov't ; Signal Transduction/drug effects ; Zinc/pharmacology

PURPOSE: Cell transplantation of myelin-producing exogenous cells is being extensively explored as a means of remyelinating axons in X-linked adrenoleukodystrophy. We determined whether 3,3',5-Triiodo-L-thyronine (T3) overexpresses the ABCD2 gene in the polysialylated (PSA) form of neural cell adhesion molecule (NCAM)-positive cells and promotes cell proliferation and favors oligodendrocyte lineage differentiation. MATERIALS AND METHODS: PSA-NCAM+ cells from newborn Sprague-Dawley rats were grown for five days on uncoated dishes in defined medium with or without supplementation of basic fibroblast growth factor (bFGF) and/or T3. Then, PSA-NCAM+ spheres were prepared in single cells and transferred to polyornithine/fibronectin-coated glass coverslips for five days to determine the fate of the cells according to the supplementation of these molecules. T3 responsiveness of ABCD2 was analyzed using real-time quantitative polymerase chain reaction, the growth and fate of cells were determined using 5-bromo-2-deoxyuridine incorporation and immunocytochemistry, respectively. RESULTS: Results demonstrated that T3 induces overexpression of the ABCD2 gene in PSA-NCAM+ cells, and can enhance PSA-NCAM+ cell growth in the presence of bFGF, favoring an oligodendrocyte fate. CONCLUSION: These results may provide new insights into investigation of PSA-NCAM+ cells for therapeutic application to X-linked adrenoleukodystrophy.
ATP-Binding Cassette Transporters/*metabolism ; Adrenoleukodystrophy/genetics/*therapy ; Animals ; Animals, Newborn ; Bromodeoxyuridine ; Cell Differentiation ; Fibroblast Growth Factor 2/pharmacology ; Fibronectins/metabolism ; Immunohistochemistry ; Neural Cell Adhesion Molecules/*genetics ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; Sialic Acids/metabolism ; Stem Cells ; Thyroid Hormones/*metabolism ; Triiodothyronine/pharmacology

This study is to observe the effect of ilexonin A (IA) on the expression of basic fibroblast growth factor (bFGF) and growth associated protein-43 (GAP-43), and neurogenesis after cerebral ischemia-reperfusion in rats and explore its possible mechanism of protecting neuronal injury. Models of middle cerebral artery occlusion (MCAO) were established in SD rats. Before and after two hours ischemia-reperfusion, IA (20 and 40 mg x kg(-1)) was injected immediately and on 3, 7, 14, and 28 d once a day. The neurological severity was evaluated by neurological severity scores (NSS); neuronal injury in the boundary zone of the infarction area was evaluated by TUNEL and Niss1 staining. The expressions of bFGF and GAP-43 and neurogenesis were evaluated by Western blotting and 5-bromodeoxyuridine (Brdu) fluorescence staining, respectively. After treatment with IA, the NSS of treatment groups were lower than that of the models (3 and 7 d). The number of TUNEL positive neurons decreased and Nissl positive neurons increased at the same time (3 d). The expressions of bFGF and GAP-43 increased significantly in the boundary zone of the infarction area when compared to model group. Moreover, IA markedly enhanced the neurogenesis in the brain after ischemia-reperfusion, which revealed an increase of Brdu/NeuN positive cells in the boundary zone of the infarction area. The possible mechanism of protecting neuronal injury of IA may be related to inhibition on neuronal apoptosis, upregulation of bFGF and GAP-43, and neurogenesis in boundary zone of infarction after cerebral ischemia-reperfusion.
Animals ; Apoptosis ; drug effects ; Brain Ischemia ; etiology ; Bromodeoxyuridine ; metabolism ; Fibroblast Growth Factor 2 ; metabolism ; GAP-43 Protein ; metabolism ; Infarction, Middle Cerebral Artery ; complications ; Male ; Neurogenesis ; drug effects ; Neurons ; pathology ; Neuroprotective Agents ; pharmacology ; Organic Chemicals ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; etiology ; metabolism