Amifostine ; pharmacology ; Animals ; Benzene ; toxicity ; Blood ; drug effects ; Blood Cell Count ; Male ; Mice ; Random Allocation

Objective of this study was to perform bioinformatics analysis of the characteristics of gene expression profiling regulated by amifostine and predict its novel potential biological function to provide a direction for further exploring pharmacological actions of amifostine and study methods. Amifostine was used as a key word to search internet-based free gene expression database including GEO, affymetrix gene chip database, GenBank, SAGE, GeneCard, InterPro, ProtoNet, UniProt and BLOCKS and the sifted amifostine-regulated gene expression profiling data was subjected to validity testing, gene expression difference analysis and functional clustering and gene annotation. The results showed that only one data of gene expression profiling regulated by amifostine was sifted from GEO database (accession: GSE3212). Through validity testing and gene expression difference analysis, significant difference (p < 0.01) was only found in 2.14% of the whole genome (460/192000). Gene annotation analysis showed that 139 out of 460 genes were known genes, in which 77 genes were up-regulated and 62 genes were down-regulated. 13 out of 139 genes were newly expressed following amifostine treatment of K562 cells, however expression of 5 genes was completely inhibited. Functional clustering displayed that 139 genes were divided into 11 categories and their biological function was involved in hematopoietic and immunologic regulation, apoptosis and cell cycle. It is concluded that bioinformatics method can be applied to analysis of gene expression profiling regulated by amifostine. Amifostine has a regulatory effect on human gene expression profiling and this action is mainly presented in biological processes including hematopoiesis, immunologic regulation, apoptosis and cell cycle and so on. The effect of amifostine on human gene expression need to be further testified in experimental condition.
Amifostine ; pharmacology ; Computational Biology ; Gene Expression ; drug effects ; Gene Expression Profiling ; methods ; Humans ; Microarray Analysis ; Molecular Sequence Annotation

Amifostine ; pharmacology ; Anemia, Aplastic ; chemically induced ; pathology ; prevention & control ; Animals ; Benzene ; toxicity ; Dose-Response Relationship, Drug ; Male ; Mice

This study was designed to assess the feasibility of calcium phosphate cement/amifostine complex as a new material for filling the bone defect caused by tumor resection. Mixed-molding method was used, the mass ratios of 0%, 0.1%, 0.5%, 1%, 2% of amifostine/calcium phosphate cement complex being adopted. The curing time, mechanical strength, porosity, scanning electron micrograph, osteosarcoma cells' vitality and vascular endothelial cells' vitality relevant to the complex in vitro were observed. Calcium phosphate cement being loaded with 0.1% and 0.5% amifostine did not affect the curing time, strength, pore size and porosity of calcium phosphate bone cement. In addition proliferation and differentiation of osteosarcoma cells and vascular endothelial cells were not affected. These data suggest that phosphate cement containing 0.1% and 0.5% amifostine be of significance in the treatment regimen as bone defect filling materials..
Amifostine ; pharmacology ; Bone Cements ; pharmacology ; Calcium Phosphates ; pharmacology ; Cell Line, Tumor ; Cell Survival ; drug effects ; Cells, Cultured ; Chemical Phenomena ; Endothelial Cells ; cytology ; drug effects ; Humans ; Osteosarcoma ; pathology

To study the effects of S-2-(3-aminopropylamino) ethyl phosphorothioic acid (WR-2721, amifostine) on proliferation inhibition and apoptosis of HL-60 human leukemia cell line, the cell apoptosis rate of HL-60 was determined by annexin V/PI double staining method. Cell proliferation and chemotherapy sensitivity were analyzed with XTT assay, and the changes of cell cycle were observed through flow cytometry. The results showed that WR-2721 could significantly inhibit HL-60 cell proliferation. After treatment (30 min, 37 degrees C) with WR-2721, the sensitivity of HL-60 cells to VP16 was enhanced, and the IC(50) descended from 52.5 micro g/ml to 40.5 microg/ml. After 72 hours treatment of HL-60 cells with WR-2721, the early apoptotic cells (annexin V-FITC positive/PI negative) were increased from (5.5 +/- 1.9)% to (48.5 +/- 8.4)% (P < 0.001), late apoptotic cells (annexin V-FITC positive/PI positive) were increased from (1.2 +/- 0.5)% to (39.0 +/- 4.0)% (P < 0.001), and HL-60 cells were arrested in G(2)-M phase. In conclusion, WR-2721 treatment can enhance HL-60 cell chemotherapy sensitivity to VP16, inhibit proliferation, induce apoptosis and accumulation of cells in G(2)-M phase.
Amifostine ; pharmacology ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Drug Synergism ; Etoposide ; pharmacology ; HL-60 Cells ; Humans

OBJECTIVETo evaluate the protective effect of amifostine on hydroquinone-induced apoptosis of bone marrow mononuclear cells in vitro.

METHODSThe mononuclear cells were separated and divided into four groups: blank control, amifostine group, hydroquinone group, amifostine + hydroquinone group. The cell apoptotic rate was examined in separated group at different time point, and apoptosis was detected by HT stain, then cell morphology was observed under fluorescent microscope and DNA fragments was tested by agarose gel electrophoresis. In addition, apoptotic and necrotic rate was detected by flow cytometer.

RESULTSAfter 10 hour culture, DNA ladder was detected in the hydroquinone group, but not in other groups. The apoptotic rate was not significantly different between amifostine group and blank control group at different culture time (P > 0.05). After 8 - 12 hour culture, the apoptotic rate in amifostine + hydroquinone group was significantly lower than that in the group of hydroquinone alone (P < 0.01). After 18 - 48 hour culture, the necrotic rate in amifostine + hydroquinone group was lower than that in the group of hydroquinone alone (P < 0.05).

CONCLUSIONAmifostine can protect cell from hydroguinone-induced bone marrow damage through inhibition on cell apoptosis, and decrease in cell necrosis.

Amifostine ; pharmacology ; Apoptosis ; drug effects ; Bone Marrow Cells ; cytology ; Cells, Cultured ; Humans ; Hydroquinones ; antagonists & inhibitors ; Leukocytes, Mononuclear ; cytology ; Protective Agents ; pharmacology

The aim was to study the protective effects of amifostine (AMF) on normal hematopoietic stem/progenitor cells against the chemotherapeutic damage from etoposide (VP-16). The cord blood mononuclear cells (CBMNC), fresh and frozen peripheral blood stem cells (PBSC), and HL-60 cells were divided into AMF, AMF + VP-16, VP-16 and control groups, each group cell viability was determined by using trypan blue exclusion test, the CFU-GM culture was used to count cells, the apoptosis was detected by flow cytometry. The results showed that in CBMNC, fresh and frozen PBSC samples, cell viability and the number of CFU-GM in AMF + VP-16 group were all significantly higher than those in VP-16 group (P < 0.05); the CFU-GM incidence in AMF + VP-16 group was higher than that in VP-16 group, and the GFU-GM life in AMF + VP-16 group was also longer than that of latter, in CBMNC samples, the number of CFU-GM in AMF groups was higher than that in control group, but there was no statistical significance between the two groups (P > 0.05), in HL-60 cell apoptotic rate in AMF + VP-16 group was little higher than that in VP-16 group, but no statistical significance between these two groups (P > 0.05). It is concluded that AMF can significantly protect normal hematopoietic stem/progenitor cells against the damage from VP-16. Moreover, AMF does not affect cytotoxity of VP-16 on HL-60 cells, and can not stimulate the growth and differentiation of cord hematopoietic stem/progenitor cells directly.
Amifostine ; pharmacology ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Etoposide ; pharmacology ; Flow Cytometry ; HL-60 Cells ; Hematopoietic Stem Cells ; cytology ; drug effects ; Humans ; Leukocytes, Mononuclear ; cytology ; drug effects ; Protective Agents ; pharmacology

The aim of this study was to investigate the effect of WR2721(amifostine) against bone marrow hematopoietic damage of mice exposed to 6.5 Gy of (60)Co-γ ray. A total of 60 C57/BL6J mice was divided into 3 groups:normal group (mice were injected with physiological salt solution), irradiation group (mice were injected with physiologic salt solution before irradiation) and WR2721 group (mice were injected with WR2721 before irradiation). The WBC, neutrophil (Neut), Plt and RBC levels in peripheral blood of 3 group mice were counted within 60 days after irradiation; the bone marrow nuclear cells (BMNC) were counted at 2 and 24 hours after irradiation; the hematopoietic stem/progenitor cell (LK/LSK) level and colony formation capability were detected by flow cytometry at 2 and 24 hours after irradiation. The results indicated that the counts of WBC and neut at 4 and 18 days, Plt at 7-18 days and RBC at 10-30 day after irradiation in WR2721 group were higher than those in irradiation group (P < 0.05); the BMNC, LSK and LK levels obviously increased at 24 hours after irradiation (P < 0.05), the CFU-GEMM, CFU-GM, CFU-MK BFU-E and CFU-E all significantly increased at 2 and 24 hours after irradiation (P < 0.01), as compared with irradiation group. It is concluded that WR2721 can effectively alleviate early hematopoietic damage and promote the fast recovery of peripheral blood cells in mice exposed to γ-ray, suggesting that the WR2721 has significant radioprotective effect on hematopoietic system.
Amifostine ; pharmacology ; Animals ; Blood Cell Count ; Bone Marrow Cells ; cytology ; drug effects ; radiation effects ; Gamma Rays ; Hematopoietic Stem Cells ; cytology ; drug effects ; radiation effects ; Male ; Mice ; Mice, Inbred C57BL ; Radiation-Protective Agents ; pharmacology ; Whole-Body Irradiation

This study was purposed to investigate the protective effects of lipoprotein HS-6101(6101) on rhesus monkey total body irradiated with 7.0 Gy ⁶⁰Coγ-ray. A total of 30 health adult rhesus monkeys were randomly divided into symptomatic therapy (ST), WR2721 and HS-6101 30, 90 and 270 mg/kg groups (n = 6), the rhesus monkeys of each groups were injected with physiological saline 0.3 ml/kg, WR-2721 30 mg/kg, or HS-6101 30, 90 and 270 µg/kg, respectively. All agents were once intramuscularly injected at 1 hr prior irradiation. General observation, peripheral blood cell counts, colony forming unite assay of bone marrow hemopoietic progenitor cells, and histopathological examination were performed. The results showed that animals in symptomatic therapy group begin to die on the 13(th) day and 4 animals died within 24 days, the average survival time was 18.2 ± 4.3 days; 2 animals in WR-2717 groups died on day 15.8 and day 18.5 post irradiation respectively. 1 animal in HS-6101 270 mg/kg group died on day 35.8, all other animals survived. Nadirs of peripheral blood white blood cells, neutrophils and platelets of animals in HS-6101 treatment groups were significantly higher than those in other 2 groups including ST and WR-2721 groups, and the hemopoietic recovery were also significantly speeding up(P < 0.05 and 0.01). In vitro results showed that HS-6101 obviously promoted 7.0 Gy ⁶⁰Coγ irradiated monkey's bone marrow mononuclear cells to form various hematopoietic progenitor cell colonies (P < 0.05 and 0.01) . Compared with symptomatic therapy and WR-2717 groups, bone marrow histopathological changes in HS-6101 treatment groups showed more active hemopoietic cell proliferation and higher density structure. It is concluded that HS-6101 90 µg/kg treatment can promote the bone marrow recovery of 7.0 Gy ⁶⁰Coγ irradiated monkey, alleviate their animal symptom, simplify the treatment measures and improve the animal survival rate. The HS-6101 shows remarkable radioprotective effects as compared with the currently internationally acknowledged radioprotectant of WR-2721.
Amifostine ; Animals ; Blood Cell Count ; Blood Platelets ; Bone Marrow ; Bone Marrow Cells ; Hematopoietic Stem Cells ; Hematopoietic System ; drug effects ; radiation effects ; Lipoproteins ; pharmacology ; Macaca mulatta ; Radiation Injuries ; drug therapy ; Survival Rate

OBJECTIVETo investigate the sites and pattern of renal toxicity in rats treated with cisplatin and the protective effect of amifostine, and to understand whether Fas/FasL system is involved in cisplatin-induced nephrotoxicity.

METHODSForty-eight Sprague-Dawley rats were randomly divided into 3 groups: control group (0.9% saline solution), cisplatin group (6 mg/kg) and amifostine group (cisplatin 6 mg/kg + amifostine 200 mg/kg). Serum BUN and creatinine were measured by automatic biochemiscal analysis. Renal histopathological lesions were examined by light microscopy. TUNEL method was used for counting apoptotic cells. Immunohistochemistry and image analysis system were used for observing the expression of Fas/FasL system in renal tissues.

RESULTSCompared with control group and amifostine group, serum BUN and creatinine were significantly elevated on day 3 (P < 0.05) and day 5 (P < 0.01 and P < 0.05, respectively), and recovered to normal on day 10. Severe necrosis and apoptosis of renal proximal tubular cells were revealed by elevated number of positively staining apoptotic cells examined by TUNEL method. Increased immunostaining intensity of Fas/FasL system in renal tissues in cisplatin-treated group was detected by immunohistochemistry and image analysis system.

CONCLUSIONAmifostine can reduce cisplatin-induced nephrotoxicity and its mechanism is probably associated with the suppression of Fas/FasL expression in renal tissues.

Amifostine ; pharmacology ; Animals ; Antineoplastic Agents ; adverse effects ; Apoptosis ; drug effects ; Blood Urea Nitrogen ; Cisplatin ; adverse effects ; Creatinine ; blood ; Fas Ligand Protein ; metabolism ; Kidney Tubules, Proximal ; metabolism ; pathology ; Male ; Necrosis ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; fas Receptor ; metabolism