BACKGROUND: This study was carried out to evaluate the efficacy of desferrioxamine as a chelating agent in iron overloaded patients with severe aplastic anemia due to multiple transfusion. METHODS AND MATERIALS: From Oct. 1995 to Aug. 1996, 15 patients with aplastic anemia, diagnosed from May 1995 to Jan. 1996 at St. Mary's Hospital, who had a transfusional hemosiderosis were included in this study. They received 19 courses of high-dose desfer-rioxamine therapy for 6 days(20 to 30 mg/kg daily as a 24-hour intravenous infusion) . Before and after treatment, we measured serum ferritin, iron, TIBC, 24-hour urinary excretion of iron. RESULTS: 1) The range of iron load before treatment was between 4.5 and 20.0 gram. 2) Because of limit of detection(1,800 microgram/L), it was difficult to compare the changes of serum ferritin level after therapy to those of before therapy. 3) There was no significant differences between the levels of serum iron before and after therapy(214.3+/-62.8 vs 220.0+/-53.3). And there was no significant differences between TIBC before and after therapy(235.8+/-64.6 vs 259.4+/-60.1). 4) Iron/TIBC ratios were significantly deceased after desferrioxamine treatment compared to those of before therapy(0.90+/-0.04 vs 0.85+/-0.04, P<0.001) and mean urinary excretions of iron were increased by high-dose desferrioxamine compared to those by test dose(6.5+/-7.6 vs 29.1+/-14.3, P<0.001) CONCLUSION: High-dose desferrioxamine therapy is very effective for chelating and excretion of iron in iron overloaded patients with severe aplastic anemia due to multiple transfusion. A repeat administration of desferrioxamine is necessary for the iron overloaded patient to eliminate the risk of a transfusional hemosidersis.
Anemia, Aplastic* ; Deferoxamine* ; Ferritins ; Hemosiderosis* ; Humans ; Iron ; Iron Overload

There are several factors concerning to anemia in chronic renal failure patients. But when rHuEPO is used, most of these factors can be overcome, and the levels of hemoglobin are increased, However, about 10% of the renal failure patients represent rHuEPO-resistant anemia eventhough high dosage of rHuEPO. For these cases, desferrioxamine can be applied to correct rHuEPO resistnacy, and many mechanism og DFO are arguing. So we are going to know whether DFO can applied to correct anemia of the such patients, how long its effect can continued. The seven patients as experimental group(DFO+EPO) who represent refractoriness to rHuEPO and the other seven patients as control group(EPO) were included. Experimental group has lower than 9 g/dL of hemoglobin levels despite high rHuEPO dosage (more than 4000U/Wk) and showed normochromic anemia. There were no definitive causes of anemia such as hemorrhage or iron deficiency. Control group patients has similar characteristics in age, mean dialysis duration but showed adequate response to rHuEPO. DFO was administered to experimental group for 8 weeks along with rHuEPO(the rHuEPO individual mean dosage had been determined by mean dosage of the previous 6 months. Total mean dosage; 123.5 U/Kg/Wk). After 8 weeks of DFO administration, the hemoglobin and rHuEPO dosage levels were checked for 15 consecutive months. It should be noted that the patients determined their own rHuEPO dosage levels according to hemoglobin levels and economic status. In control group, rHuEPO was administered by the same method used in experimental group without DFO through the same period. Fifteen months of ovservation period after DFO trial were divided as Time I(7 months after DFO trial) and Times II(8 months after Time I). The results are as follows: Before DFO trial, mean hemoglobin level of experimental group was 7.8 g/dL, which is similar level(p>0.05) to control group(mean Hb; 8.2 g/dL). But in experimental group, significantly(p<0.05) higher dosages of rHuEPO(mean; 123.5 U/Kg/Wk) than control group (mean;41.6 U/Kg/Wk) had been used. It means resistancy to rHuEPO of experimental group. But after DFO trial, the hemoglobin levels of the experimental group were increased significantly(p<0.05), and these effect were continued to II.(Time I; mean 8.6g/dL, Time II; mean 8.6g/dL) The effects of DFO to hemoglobin were continued for 15 months after DFO trial with simiral degree through Time I, Time II. Also, rHuEPO dosage used in the experimental group were decreased to simiral levels of the control group after DFO trial and these effect were also continued for 15 months(Time I; mean 48.1 U/Kg/Wk. Time II; mean 51.8 U/Kg/Wk). In the same period, hemoglobin levels and rHuEPO dosages used in the control group were not changed significantly. Notibly, hemoglobin increment and rHuEPO usage decrement in experimental group were showed maxilly in the 1st month after DFO trial. That is, after the use of DFO, erythropoiesis was enhanced with a reduced rHuEPO dosage. So we think rHuEPO reisistancy can be overcome by DFO therapy. In conclusion, the DFO can improve the anemia caused by chronic renal failure at least over 1 year, and hence, can reduce the dosage of rHuEPO for anemia correction. Additional studies in order to determined the mechanism of DFO on erythropoiesis and careful attention to potential side effects DFO will be needed.
Anemia* ; Deferoxamine* ; Dialysis ; Erythropoiesis ; Hemorrhage ; Humans ; Iron ; Kidney Failure, Chronic ; Renal Dialysis* ; Renal Insufficiency

OBJECTIVE: To evaluate endothelin-1 (ET-1) expression in the villous explants from normal and preeclamptic (PE) placentae under hypoxic condition. METHODS: Villous explants from normal (n=5) and PE (n=4) placentae were obtained. To obtain hypoxic culture condition, villous explants were cultured in hypoxic chamber or treated with deferoxamine (DFO). ET-1 mRNA expressions in villous explants were evaluated by RT-PCR following 0, 24, and 48 h of culture in hypoxic chamber, and 0, 2, 4, 6 h following DFO treatment. ET-1 protein levels in media were measured by enzyme immunoassay. RESULTS: After 24 and 48 hours of incubation of villous explants from normal and PE placentae in hypoxic chamber, ET-1 mRNA and protein levels were increased in both groups, however, ET-1 production seemed to be more exaggerated in the villous explants from PE placentae. During 6 h of DFO exposure, ET-1 mRNA level was increased in the villous explants from PE placenta comparing to those from normal placentae (p<0.05). Interestingly, the increase of ET-1 mRNA expression in the villous explants from PE placentae was more exaggerated than those from normal placentae. Concordantly, increments of protein level between 0 to 2 h and 2 to 4 h were significantly higher in villous explants from PE placentae (p<0.05). CONCLUSION: ET-1 mRNA and protein were increased in villous explants from PE placentae compared to those from normal placentae under hypoxic condition. Furthermore, villous explants from PE placentae showed upregulated ET-1 expression upon hypoxic stimulation. This enhanced sensitivity to hypoxia may contribute to ET-1 overexpression in PE placenta in vivo and it needs further investigation for clarification.
Anoxia ; Deferoxamine ; Endothelin-1* ; Immunoenzyme Techniques ; Placenta ; Pre-Eclampsia ; Pregnancy* ; RNA, Messenger

PURPOSE: It has been suggested in our previous study that the serum level of xanthine oxidise(XO) activity, glutathione(GSH), malonyldialdehyde(MDA) could be used as marker of oxidant stress in association with renal ischemia/reperfusion(I/R) injury. The present study was undertaken to establish the early marker of renal 1/R injury and to investigate the effect of deferoxamine on renal 1/R injury. MATERIALS AND METHOD: In Sprague-Dawley rats(male, 200-250gm, n=60), bilateral renal arteries were clamped for 60mins after pretreatment with deferoxamine(group A) or saline(group B). After 30min of bilateral renal recirculation, left nephrectomy and blood sampling in inferior vena cava were performed for in-vitro spectrophotometric study. Control animals(group C) did not undergo I/R operation. In-vivo renal function studies were performed in both group A and B with measurement of creatinine clearance rate(Ccr) at 7th day of experiments a%or renal ischmia for 60min. RESULTS: The levels of XO activity and XO type conversion ratio in renal tissue (RT) and serum(5) were measured. These levels were significantly high in group B, but were lower in group A compared to those of control group. The values of GSH(micrometer/g tissue), a scavenger of OFR, were decreased in group A (RT:0.183+/-0.019,5:0.201+/-0.029) and greatly decreased in group B(RT:0.159+/-0.009,5:0.164+/-0.022) compared to control group(RT:0.201+/-0.006,5:0.224+/-0.031). The values of MDA(nM/g tissue), a stable end product of lipid peroxidation, were increased in group A(RT:0.149+/-0.003, 5:0.058+/-0.004) compared to control group(RT:0.128+/-0.013, 5;0.055+/-0.005), but the values were significantly lower in group A compared to group B(RT:0.171+/-0.005, 5:0.070+/-0.003). Subsequent investigation was focused on the established renal function study after 1/R, which was determined using Ccr(ml/min). The Ccr in group A(2.06+/-0.03) was significantly higher compared to that of group 8(1.48+/-0.18), although it was slightly lower than in control group(2.18+/-0.05). CONCLUSIONS: From these results, it is suggested that renal I/R injury is highly correlated with the production of OFR. The levels of GSH and MDA in renal tissue and serum seem to be probable markers of oxidant stress in association with renal I/R injury. Furthermore, deferoxamine could reduce the degree of renal damage resulting from ameliorating the production of OFR following renal I/R injury.
Animals ; Creatinine ; Deferoxamine* ; Lipid Peroxidation ; Nephrectomy ; Rats* ; Rats, Sprague-Dawley ; Renal Artery ; Vena Cava, Inferior ; Xanthine

OBJECTIVES: This study was designed to evaluate cytotoxicity of TAFMAG, which is a trade name of natural mineral fiber mined and produced in China. METHODS: The cytotoxicity of TAFMAG was evaluated by measuring iron content, lipid peroxidation, erythrocyte hemolysis, and cytotoxicity in vitro. These results were compared with the data of chrystotile and wollastonite as a positive and negative control, respectively. RESULTS: There was significant increase of Fenton activity in TAFMAG and chrysotile with dose-response pattern. The iron chelating agent, desferrioxamine, significantly decreased Fenton activity of the particulates except wollastonite. TAFMAG and chrysotile fibers significantly increased malondialdehyde concentration from lipid peroxidation of the red blood cell membrane. In erythrocyte hemolysis test, TAFMAG & chrysotile had stronger effect on erythrocyte hemolysis than wollastonite with the concentration of 1,000g/ml. Furthermore, TAFMAG was more hemolytic than chrysotile with the concentration of 5.000 g/ml. There was a significant cytotoxic effect in TAFMAG and chrysotile on RAW cell compared with wollastonite. CONCLUSIONS: In vitro study suggested that TAFMAG may have a similar health hazard as usual asbestos.
Asbestos ; Asbestos, Serpentine ; China ; Deferoxamine ; Erythrocytes ; Hemolysis ; Iron ; Lipid Peroxidation ; Malondialdehyde ; Membranes ; Mineral Fibers

Objective: To investigate the effects of deferoxamine on macrophage polarization and wound healing in mice with deep tissue injury (DTI) and its mechanism. Methods: The experimental research methods were adopted. Fifty-four male C57BL/6J mice of 6-8 weeks old were divided into DTI control group, 2 mg/mL deferoxamine group, and 20 mg/mL deferoxamine group according to random number table, with 18 mice in each group. DTI was established on the back of mice by magnet compression method. From post injury day (PID) 1, mice were injected subcutaneously with 100 µL normal saline or the corresponding mass concentration of deferoxamine solution every other day at the wound edge until the samples were collected. Another 6 mice without any treatment were selected as normal control group. Six mice in each of the three DTI groups were collected on PID 3, 7, and 14 to observe the wound changes and calculate the wound healing rate. Normal skin tissue of mice in normal control group was collected on PID 3 in other groups (the same below) and wound tissue of mice in the other three groups on PID 7 and 14 was collected for hematoxylin-eosin (HE) staining to observe the tissue morphology. Normal skin tissue of mice in normal control group and wound tissue of mice in the other three groups on PID 7 were collected, and the percentages of CD206 and CD11c positive area were observed and measured by immunohistochemical staining, and the mRNA and protein expressions of CD206, CD11c, and inducible nitric oxide synthase (iNOS) were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction and Western blotting, respectively. Normal skin tissue of mice in normal control group and wound tissue of mice in DTI control group and 20 mg/mL deferoxamine group were collected on PID 3, 7, and 14, and the protein expressions of signal transducer and activator of transcription 3 (STAT3) and interleukin-10 (IL-10) were detected by Western blotting. The sample number in each group at each time point in the above experiments. The RAW264.7 cells were divided into 50 μmol/L deferoxamine group, 100 μmol/L deferoxamine group, 200 μmol/L deferoxamine group, and blank control group, which were treated correspondingly, with 3 wells in each group. The positive cell percentages of CD206 and CD86 after 48 h of culture were detected by flow cytometry. Data were statistically analyzed with analysis of variance for repeated measurement, one-way analysis of variance, and least significant difference test. Results: On PID 7, the wound healing rates of mice in 2 mg/mL and 20 mg/mL deferoamine groups were (17.7±3.7)% and (21.5±5.0)%, respectively, which were significantly higher than (5.1±2.3)% in DTI control group (P<0.01). On PID 14, the wound healing rates of mice in 2 mg/mL and 20 mg/mL deferoamine groups were (51.1±3.8)% and (57.4±4.4)%, respectively, which were significantly higher than (25.2±3.8)% in DTI control group (P<0.01). HE staining showed that the normal skin tissue layer of mice in normal control group was clear, the epidermis thickness was uniform, and skin appendages such as hair follicles and sweat glands were visible in the dermis. On PID 7, inflammation in wound tissue was obvious, the epidermis was incomplete, and blood vessels and skin appendages were rare in mice in DTI control group; inflammatory cells in wound tissue were reduced in mice in 2 mg/mL and 20 mg/mL deferoxamine groups, and a few of blood vessels and skin appendages could be seen. On PID 14, inflammation was significantly alleviated and blood vessels and skin appendages were increased in wound tissue of mice in 2 mg/mL and 20 mg/mL deferoxamine groups compared with those in DTI control group. On PID 7, the percentages of CD206 positive area in wound tissue of mice in 2 mg/mL and 20 mg/mL deferoxamine groups were significantly higher than that in DTI control group (P<0.01), the percentage of CD206 positive area in wound tissue of mice in DTI control group was significantly lower than that in normal skin tissue of mice in normal control group (P<0.01), the percentage of CD206 positive area in wound tissue of mice in 20 mg/mL deferoxamine group was significantly higher than that in normal skin tissue of mice in normal control group (P<0.01). The percentages of CD11c positive area in wound tissue of mice in 2 mg/mL and 20 mg/mL deferoxamine groups were significantly lower than those in DTI control group and normal skin tissue in normal control group (P<0.05 or P<0.01), and the percentage of CD11c positive area in normal skin tissue of mice in normal control group was significantly higher than that in DTI control group (P<0.05). On PID 7, the CD206 mRNA expressions in the wound tissue of mice in 2 mg/mL and 20 mg/mL deferoxamine groups were significantly higher than that in DTI control group (P<0.01), but significantly lower than that in normal skin tissue in normal control group (P<0.01); the CD206 mRNA expression in wound tissue of mice in DTI control group was significantly lower than that in normal skin tissue in normal control group (P<0.01). The mRNA expressions of CD11c and iNOS in wound tissue of mice in 2 mg/mL and 20 mg/mL deferoamine groups were significantly lower than those in DTI control group (P<0.01). The mRNA expressions of CD11c in the wound tissue of mice in DTI control group, 2 mg/mL and 20 mg/mL deferoamine groups were significantly higher than that in normal skin tissue in normal control group (P<0.01). Compared with that in normal skin tissue in normal control group, the mRNA expressions of iNOS in wound tissue of mice in 2 mg/mL and 20 mg/mL deferoamine groups were significantly decreased (P<0.01), and the mRNA expression of iNOS in wound tissue of mice in DTI control group was significantly increased (P<0.01). On PID 7, the protein expressions of CD206 in the wound tissue of mice in 2 mg/mL and 20 mg/mL deferoamine groups were significantly higher than those in DTI control group and normal skin tissue in normal control group (P<0.01), and the protein expression of CD206 in wound tissue of mice in DTI control group was significantly lower than that in normal skin tissue in normal control group (P<0.01). The protein expressions of CD11c and iNOS in wound tissue of mice in 2 mg/mL and 20 mg/mL deferoamine groups were significantly lower than those in DTI control group (P<0.01). The protein expressions of CD11c and iNOS in wound tissue of mice in DTI control group were significantly higher than those in normal skin tissue in normal control group (P<0.01). The CD11c protein expressions in wound tissue of mice in 2 mg/mL and 20 mg/mL deferoamine groups were significantly higher than those in normal skin tissue in normal control group (P<0.05 or P<0.01). The protein expression of iNOS in wound tissue of mice in 2 mg/mL deferoamine group was significantly lower than that in 20 mg/mL deferoamine group and normal skin tissue in normal control group (P<0.05). On PID 3, 7, and 14, the protein expressions of STAT3 and IL-10 in wound tissue of mice in 20 mg/mL deferoxamine group were significantly higher than those in DTI control group (P<0.05 or P<0.01), and the protein expressions of STAT3 were significantly higher than those in normal skin tissue in normal control group (P<0.05 or P<0.01). On PID 7 and 14, the protein expressions of IL-10 in wound tissue of mice in 20 mg/mL deferoxamine group were significantly higher than those in normal skin tissue in normal control group (P<0.01). On PID 3, 7, and 14, the protein expressions of IL-10 in wound tissue of mice in DTI control group were significantly lower than those in normal skin tissue in normal control group (P<0.05 or P<0.01). After 48 h of culture, compared with those in blank control group, the CD206 positive cell percentages in 100 μmol/L and 200 μmol/L deferoamine groups were significantly increased (P<0.01), while the CD86 positive cell percentages in 100 μmol/L and 200 μmol/L deferoamine groups were significantly decreased (P<0.01). Conclusions: Deferoxamine can promote the polarization of macrophages toward the anti-inflammatory M2 phenotype and improve wound healing by enhancing the STAT3/IL-10 signaling pathway in DTI mice.
Animals ; Deferoxamine/pharmacology* ; Inflammation ; Interleukin-10 ; Macrophages ; Male ; Mice ; Mice, Inbred C57BL ; Wound Healing

OBJECTIVE: To investigate the effect of local administration of deferoxamine mesylate (DFO) on vascularization and osteogenesis and its ability to maintain the activity of hypoxia inducible factor-1α (HIF-1α), by constantly observing early changes of vessel-like structures and bone tissues during bone defects healing. METHODS: Skull critical bone defect models were constructed on a total of thirty male SD rats (6-8 weeks old). The rats were randomly divided into experimental group (DFO group) or control group (normal saline group). 300 μL 200 μmol/L DFO solution or normal saline was locally injected on the 4th day after the defect was made. On the 5th, 7th, 10th, 14th, and 28th days after surgery, three rats in each group were sacrificed respectively. HE staining and Masson staining were performed to observe new bone formation and mineralization. HIF-1α immunohistochemistry staining was performed to examine relative expression of protein. Qualitative analysis and comparation were performed by t-tests on relative expression of HIF-1α, numbers of blood vessels and percentages of mineralization tissues of new bone areas. RESULTS: On the 5th, 7th, 10th, 14th and 28th days after surgery, the average numbers of blood vessels were 30.40±12.15, 62.00±17.87, 73.43±15.63, 40.00±7.84, 48.71±11.64 in the DFO group, and 18.75±6.63, 19.13±2.80, 51.35±16.21, 27.18±7.32, 30.88±13.43 in the control group. The number of blood vessels in the DFO group was significantly higher than that of the control group at each time point (P < 0.05). The mass of new bone in the DFO group was higher than that in the control group on the 14th and 28th days after surgery. The percentage of mineralization tissues of new bone area on the 14th and 28th days after injection were (27.73±5.93)% and (46.53±3.66)% in the DFO group, and (11.99±2.02)% and (31.98±4.22)% in the control group. The percentage of mineralization tissues in the DFO group was significantly higher than that of the control group at each time point (P < 0.001). The relative expression of HIF-1α in the DFO group compared with the control group was 2.86±0.48, 1.32±0.26, 1.32±0.32, 1.28±0.38 and 1.05±0.34 on the 5th, 7th, 10th, 14th and 28th days, with significant expression difference on the 5th day (P < 0.01). CONCLUSION Use of DFO in bone defects promotes vascularization and osteogenesis in the defect area, and maintains the protein activity of HIF-1α temporarily.
Animals ; Bone Regeneration ; Deferoxamine/therapeutic use* ; Male ; Rats ; Rats, Sprague-Dawley ; Skull

Adult ; Deferoxamine ; adverse effects ; therapeutic use ; Humans ; Lung Diseases, Interstitial ; chemically induced ; Male ; Myelodysplastic Syndromes ; drug therapy

OBJECTIVETo study the molecular mechanism of apoptosis of leukemic cells (K562 cells) induced by iron chelating agent deferoxamine (DFO).

METHODSThe exponentially growing K562 cells were used (1×10(6)/mL) in this study. The K562 cells were treated with different concentrations of DFO (10, 50 and 100 mmol/L), DFO+FeCl3 (10 μmol/L each) or normal saline (blank control). The cellular labile iron pool was measured with a fluorimetric assay using the metalsensitive probe calcein-AM. The viable count and cell viability were determined by typanblue assay. Cell apoptosis was determined by morphological study and flow cytometry assay. Caspase-3 activity in K562 cells was detected by colorimetry.

RESULTSAfter DFO treatment, the cellular labile iron pool and the viability of K562 cells were reduced and the cell apoptosis increased in a time- and dose-dependent manner compared with the blank control group. The apoptosis rate of K562 cells in the DFO+FeCl3 treatment group was not significantly different from that in the blank control group. The caspase-3 activity in K562 cells increased significantly 24 hrs after 50 and 100 μmmol DFO treatment when compared with the blank control group (P<0.01). There was a negative correlation between cellular labile iron pool and caspase-3 activity of K562 cells (r=-0.894, P<0.05).

CONCLUSIONSDFO induces apoptosis of leukemic cells possibly through decreasing cellular labile iron pool and increasing caspase-3 activity of the cells.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Deferoxamine ; pharmacology ; Flow Cytometry ; Humans ; Iron Chelating Agents ; pharmacology ; K562 Cells

Ascorbic acid (AA) and dehydroascorbic acid (DHA) are known to have protective effects in experimental central nerve system disorder models such as stroke, ischemia, and epileptic seizures. The present study was conducted to examine the protective effect of AA and DHA on kainic acid (KA) neurotoxicity using organotypic hippocampal slice cultures (OHSC). Protective effects of AA and DHA on KA-induced cell death were evaluated by analyzing caspase-3. In addition, to determine if the prooxidant effect of AA is related to iron, the effect of AA on cell death was examined using desferrioxamine (DFO), an iron chelator. After 12h-KA treatment, significant delayed neuronal death was detected in CA3 region, but not in CA1. The AA (500 micrometer) and DHA (100 and 500 micrometer) pretreatments significantly prevented cell death by inhibiting caspase-3 activation in CA3 region. In the concentration of 1,000 micrometer, however, AA pretreatment might have prooxidant effect, but AA-induced oxidative reaction is mainly not related to transition metal ions. These data showed that the pretreatments of intermediate-dose AA and DHA protected KA-induced neuronal damage in OHSCs and co-pretreatment of AA and DFO did not affect cell death except for a few cases. These data suggest that both AA and DHA pretreatment have antioxidant or prooxidant effect depending on doses treated on KA-induced neuronal injury and the possible prooxidant effect of AA may not depend on the Fenton reaction.
Ascorbic Acid ; Caspase 3 ; Cell Death ; Deferoxamine ; Dehydroascorbic Acid ; Epilepsy ; Ions ; Iron ; Ischemia ; Kainic Acid ; Neurons ; Stroke