| ObjectiveTo study the effects of intravenous iron on oxidative stress in rats with adanine-induced chronic kidney disease, and to investigate the role of antioxidative vitamin supplemetation on the oxidative stress induced by intravenous iron in CKD rats, so as to provide experimental basis for guiding iron therapy and nutriton nursing with CKD patients.Methods1. Division of groups: Male Wistar rats (n=48) were divided randomly into six groups, including three control groups (CTL group, CTL+Fe group and CTL+Fe+V_E group) and three CKD groups (CKD group, CKD+Fe group and CKD+Fe+V_E group).Each group had 8 rats.2. Preparation for model: To be induced chronic kidney disease, the rats of three CKD groups were fed adenine 200mg/kg/d for 3 weeks. At the same time, every rat ofcontrol groups were fed pure water 2 ml/d. Then two control groups' (CTL+Fe group and CTL+Fe+VE group) and two CKD groups' (CKD+Fe group and CKD+Fe+VE group) rats were treated with a single intravenous infusion of iron dextran (lOOmg/kg). After intravenous iron therapy, CTL+Fe+VE group and CKD+Fe+Vn group were supplemented vitamin E (50 mg/kg/d) immediately for two weeks. It took 5 weeks to do this experiment.3. Collection of general data on rat model: Diet, activites, color and luster of hair, body weight were observed during the experiment.4. Measurement of renal function parameters: 24-hour urine was collected at the 1st, 3rd week, and urinary volume and protein content were measured. Urinary creatinine (Cr) and urea nitrogen (UN), serum Cr and BUN separately were measured at the end of the 3rd week, and then 24-hour creatinine clearace (CCr) was calculated.5. Measurement of hemogram: Red blood cell (RBC) count and hemoglobin concentration were measured at the end of the 5th week.6. Measurement of iron parameters: The levels of serum iron and ferritin were measured at the end of the 5th week.7. Measurement of antioxidant status: The concentration of plasma glutathione (GSH) and vitamin E (Ve) were measured at the end of the observation.8. Measurement of markers of oxidative stress: The concentration of plasma nitricoxide (NO), and malondialdehyde (MDA) were measured at the end of the experiment.9. Pathological diagnosis: The pathological changes of kidney were obtained at the end of expriment.Results1. Genaral data: During the experiment, the rats of three CKD groups drank more and ate less day by day. And the hair of these rats became seared and moulted. Thebody weights of these rats increased slowly,and got significantly lighter than those of the control groups' rats (P<0. 01) . Two rats separately in the CKD group and CKD+Fe group died at the 4th week.2. Renal function parameters: The 24-hour urinary volume and protien content of three CKD groups were significantly higher than those of three control groups at the end of the 1st and 3rd week (P<0. 01) .In addition, a marked increase in the levels of serum Cr and BUN and a significant reduction in CCr were found in the CKD groups at the end of the 3rd week. (P<0. 01). Both in three CKD groups and in three control groups, no remarkable differences were revealed in the serum Cr, BUN levels and in the 24-hour urinary protien content.3. Hemogram: The RBC count and Hb concentration of the CKD groups were significantly lower than those of the control groups (P<0. 01). Compared with the CKD+Fe group and CKD+Fe+VE group, the RBC count and Hb concentration of the CKD group were significantly lower (P<0. 01). But the RBC count of CKD+Fe+VE group was significantly higher than that of the CKD+Fe group (P<0. 01) .No notable differences in RBC and Hb were found in three control groups.4. Iron parameters: In the iron-treated groups, both the serum iron and ferritin concentration were markedly higher than those in the untreated groups (P<0. 01) .5. Antioxidant status and markers of oxidative stress5.1 Changes of CKD rats: Compared with the CTL group, the plasma GSH and VE concentration of the CKD group were significantly lower (P<0. 05) ; and the plasma NO and MDA concentration were significantly higher (P<0. 05) at the end of the 5th week.5.2 Changes of control and CKD rats receiving intravenous iron therapy: At the end of the 5th week,the plasma GSH of the CKD+Fe group were significantly lower than those of CTL+Fe group (P<0.01) .At the same time, the plasma NO and MDAconcentration of the CKD+Fe group were remarkablely higher than those of the CTL +Fe group and CKD group (P<0.05). Except the plasma VE concentration, there were no significant diferences in the plasma GSH, NO and MDA concentration between the CTL and CTL+ Fe groups . The plasma VE concentration of the CTL+ Fe group was lower than that of the CTL group (P<0. 05) .5.3 Changes of the VE-treated rats receiving intravenous iron therapy: At the end of the 5th week, compared with the CKD+Fe group's ,the CKD+Fe+VE group's plasma GSH and Ve concentration were significantly higher (P<0.01), and the plasma NO and MDA concentration were significantly lower (P<0.05) .Compared with the CTL+Fe+VE group's , the CKD+Fe+VE group's plasma MDA concentration was higher , but the Ve concentration was also significantly higher (P<0. 01). There were no remarkable differences in the NO and MDA concentration between CKD+Fe+VE group and CTL+Fe+VE group. Except the plasma Ve concentration, no notable differences existed in the plasma GSH, NO and MDA concentration between the VE-treated CTL group and two untreated CTL groups. The plasma Ve concentration of CTL+Fe+VE group was significantly higher than that of CTL and CTL+Fe group (P<0.05) .6. Pathological diagnosis: All three CKD groups had pathological changes in the kidneys at the end of experiment. The pathological findings implied the main pathological changes were in renal tubular and interstitial region. Renal tubules dilatation and atrophy, minor interstitial fibrosis was found. The pathological changes in the glomeruli were not severe. And there were no pathological changes in the kidneys of three control groups.Conclusion1. Feeding rats adenine 200mg/kg/d for 3 weeks will increase the levels of serumCr ,BUN and urinary protien content, and decrease CCr. All of these symptoms are associated with CKD.So this method can be used to make CKD animal model. Furthermore, this animal modal will appear anemia at the end of the 5th week.2. Oxidative stress exists in rats with adenine-induced chronic kidney disease.3. A single intravenous infusion of iron may result in an increase of serum iron and ferritin levels, improve anemia, and aggravate pre-existing oxidative stress in rats with adenine-induced CKD.4. Antioxidative vitamin (Ve ) supplementation can improve anemiabetter ,enhance antioxidative capacity, and attenuate oxidative stress in CKD rats with a single intravenous iron therapy.
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