Effects of Oxymetholone on Kidney Tissue of One-Day Old Offspring of Pregnant Rats
Research Article
Effects of oxymetholone on kidney tissue of one-day old offspring of pregnant rats
Hojjatollah Karimi Jashni*1, Shamsi Bandak2, Amir Ashkan Mahjoor2
Received: 03/17/2011 Revised: 05/10/2011 Accepted: 05/19/2011
1. Dept. of Anatomy, School of Medicine, Jahrom University of Medical Sciences, Jahrom, Iran 2. Dept. of Biology, Islamic Azad University, Jahrom Branch, Jahrom, Iran
Journal of Jahrom University of Medical Sciences, Vol. 9, No. 3, Fall 2011
Abstract:
Introduction: Oxymetholone is a drug used by some athletes as energy booster. However, its overuse may result in disorders such as lung cancer, ovarian cycle irregularities, liver cancer and adenoma of renal tubules. The aim of this study is to investigate the effects of oxymetholone on kidney tissue of one-day offspring of pregnant rats. Material and Methods: In this experimental study, 30 pregnant rats were divided into control, sham and experiment groups. The experiment group and the sham group were respectively administered 10 mg oxymetholone and 10 mg dimethyl sulfo oxide intraperitoneally for 15 days. On first day after delivery, the neonates were weighed and dissected. Kidney weight, length, width, and thickness were measured. Subsequently, tissue sections were studied microscopically. Data were analyzed using one-way analysis of variance with SPSS software. Results: The findings indicate that fetal weight, kidney weight, length, width and thickness, glomerular diameter, and cortical and medullary thickness in the experiment group were significantly greater compared to the control group (P<0.05), while the diameter of the interglomerular space was diminished. Moreover, certain pathological changes, such as glomerular cell proliferation, necrosis of cortical tubules and cast formation inside tubules, were observed. Conclusion: Oxymetholone, at the dosage used in our study, induces pathological changes in kidney tissue, as well as increased body and kidney weight in rat neonates.
Keywords: Oxymetholone, Kidney, Tissue Downloaded from jmj.jums.ac.ir at 0:51 +0330 on Thursday January 21st 2021 [ DOI: 10.29252/jmj.9.3.2 ] Introduction: hydroxymethyl group is placed in the Oxymetholone is an anabolic drug position of carbon atom number 2 (1). synthesized by Ringold in 1959. It is a 17- Anabolic-androgenic drugs, particularly alpha-alkamine and a synthetic derivative of oxymetholone, are used at small doses (1-5 testosterone, synthesized by methylation of mg/Kg) for treating anemia, failure to thrive 17-alpha carbon and saturation of 5-alpha in children, and heart failure (2). These carbon in testosterone. In addition, a drugs increase erythropoietin production and * Corresponding Author, Address: Department of Anatomy, School of Medicine, Jahrom University of Medical Sciences, Ostad Motahari Avenue, Jahrom Tel: 0791 3340406-10 Email: [email protected] 8
Effects of oxymetholone on kidney tissue
probably affect bone marrow directly to in vivarium. The male rats were used for increase hemoglobin and red blood cell (3). fertilization of female rats. Conception and In addition, oxymetholone promotes pregnancy were confirmed with observation nitrogen retention, protein synthesis and of vaginal plaque, after which the male and calcium deposition on bones (4). female rats were separated. The females Furthermore, it increases sodium, potassium were assigned to control, sham and and chlorine salts, protecting athletes from experiment groups. During 15 days of injuries during intense exercises (5). pregnancy, rats in the experiment group Anabolic-androgenic steroids must be used were injected intraperitoneally with 10 at 10-100 times their normal (therapeutic) mg/Kg/day oxymetholone, and rats in the doses to assume anabolic properties, and sham group were injected intraperitoneally their adverse reactions occur at these high with 10 mg/Kg/day dimethyl sulfo oxide. doses (6). Prior to 1936, it was unknown that Rats in the control group were preserved anabolic steroids boost muscle tissue. From under normal conditions of diet and that time on, anabolic steroids began to be temperature. After delivery, the neonates used as performance enhancers (7). were isolated from their mothers and Anabolic steroids were abused by Olympics weighed on scales. 5 neonates from each weight lifters in 1954, and their abuse has mother rat in the experiment group, and 3 gradually spread to other fields of sports (8). neonates from each mother rat in the sham Nowadays, abusing anabolic-androgenic and control groups were anesthetized with steroids is a hot issue throughout the world. ether and dissected. In order to extract the According to reports issued by the National kidneys, the rat was fixed onto Styrofoam Olympics Committee, half of doping cases with pins, its abdomen was incised, and the pertains to abusing anabolic agents (9). kidneys were carefully removed using Anabolic drugs, especially oxymetholone, pincers and scissors with fine blades. The are used by athletes for enhancing muscle kidneys were then irrigated with physiologic growth and force. These substances delay serum, their dimensions were measured cell death, but cause complications such as using a vernier caliper, and weighed using hepatic cancer, acne, premature alopecia, precise analytical scales. The specimens hirsutism in women, increased serum lipid were then fixed, sectioned and stained with (LDL) and cardiac diseases (9, 10). They Hematoxylin-Eosin stain to be studied may also result in behavioral complications histologically with light microscopy. We such as aggression, anger, and violent used graticule and Dinolit software to behaviors (11, 12). In the present study, we measure cortex thickness, medulla thickness, investigate the tissue and pathological glomerular diameter, and interglomerular effects of oxymetholone on kidney tissue at space and assess them for pathologies. higher than physiological dosages in one- Subsequently, microscopic images were Downloaded from jmj.jums.ac.ir at 0:51 +0330 on Thursday January 21st 2021 [ DOI: 10.29252/jmj.9.3.2 ] day old rat neonates. taken using a photomicroscope. Since Clark (1997) used 12 mg/Kg oxymetholone, which Material and Methods: is almost 1 million times the normal dose of For the present study, we obtained 30 female testosterone in rat’s body, to investigate and 10 male healthy adult Wistar rats from ovarian cycle, we used a close dose of 10 Razi Immunization Institute, Shiraz. Female mg/Kg in the present study. rats were kept in trios in special cages and The resulting data were analyzed using one- the male rats were kept in separate plastic way analysis of variance (ANOVA) with cages in proximity of the cage of female rats
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Effects of oxymetholone on kidney tissue
SPSS software. P values < 0.05 were Bowman’s capsule space were significantly considered significant. different from the control group (p<0.05). In other words, all these parameters, except for Results: the diameter of Bowman’s capsule space, Our findings indicate that in the experiment were significantly greater in the experiment group, the weight of one-day old neonates, group compared to controls. The diameter of as well as kidney weight, length, width and Bowman’s capsule space was significantly thickness, cortical and medullary thickness, smaller in the experiment group compared to glomerular diameter, and diameter of the controls (Tables 1, 2).
Table 1: Body weight, kidney weight, length, width and thickness in one-day old rats Observation Body Weight (g) Kidney Weight (g) Kidney Length (mm) Kidney Width (mm) Kidney Thickness (mm) Control Group 5.8800 ± 0.12497 0.0353 ± 0.00157 5.4867 ± 0.10111 3.300 ± 0.04819 2.3933 ± 0.010427 Sham Group 5.8867 ± 0.11868 0.0353 ± 0.00115 5.4867 ± 0.10134 3.2867 ± 0.04254 2.3733 ± 0.07013 Experiment Group 6.6423 ± 0.15347 0.041 ± 0.00182 5.9000 ± 0.12031 3.5525 ± 0.07604 2.8358 ± 0.07927 P Value <0.001 <0.05 <0.05 <0.05 <0.001
Table 2: Cortical thickness, medullary thickness, glomerular diameter and interglomerular space diameter in one-day old rats Observation Cortical Thickness (μ) Medullary Thickness (μ) Glomerular Diameter (μ) Interglomerular Space Diameter (μ) Control Group 393.24 ± 11.8873 496.40 ± 15.0934 22.1300 ± 0.31690 4.6933 ± 0.13746 Sham Group 397.87 ± 14.9239 495.90 ± 11.6060 22.2333 ± 0.42765 4.7267 ± 0.11773 Experiment Group 465.10 ± 7.4565 535.76 ± 10.4765 23.4075 ± 0.37417 4.3475 ± 0.08251 P Value <0.001 <0.05 <0.05 <0.05
On pathological investigations, we observe outside the capillaries resulting in thickening proliferation of glomerular cells, a lesion of capillaries (Figure 1), necrosis of tubules with homogenous involvement of glomeruli in cortex, increased eosinophility of tubule and enlargement, hyperplasia and a clover- of tubular epithelial cells, tubular destruction leaf appearance of glomeruli. Moreover, it is (Figure 2), and cast formation inside tubules obvious that glomeruli have lost their cluster (Figure 3). form, mesangium is dispersed annularly
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Figure 1: Section of a one-day rat’s kidney from the experiment group (magnified x400; H&E staining)
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Effects of oxymetholone on kidney tissue
Figure 2: Section of a one-day rat’s kidney from the experiment group (magnified x400; H&E staining)
Cast
Figure 3: Section of a one-day rat’s kidney from the experiment group (magnified x400; H&E staining). Arrow marks the cast
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Figure 4: Section of a one-day rat’s kidney from the experiment group (magnified x400; H&E staining)
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Conclusion: Considering the structural similarities of In this study, oxymetholone induced an nandrolone and oxymetholone, the increase increase in body weight, kidney weight, in kidney volume and weight, as observed in length, width and thickness, cortical and our study, may be attributed to medullary thickness, and glomerular oxymetholone effects. diameter in one-day rats, as well as a Androgens may induce renal disorders decrease in diameter of Bowman’s capsule through altering the amounts of cytokinases space. Steroid hormones enter cells easily and growth factors. It is believed that due to their lipid core and exert their effect transforming growth factor plays a pivotal by acting directly on nucleus. They promote role in nephropathy by inducing the DNA transcription and increase intracellular proliferation of mesangial cells (17). The mRNA production, which results in proliferation of glomerular cells observed in increased synthesis of proteins necessary for our study is probably the result of the hormone’s response. This response oxymetholone effects. promotes muscle and bone growth through A study by Deshmukh et al. reported that synthesis of structural proteins, or modifies anabolic-androgenic steroids are excreted as body physiology and function (13). glucoroid compounds which may create Oxymetholone is an oral anabolic energy casts in the urinary tubules (18). This may booster synthesized from 17-alpha account for the observation of cast in renal alkylation of testosterone. Compared to tubules in our study. Previous studies testosterone, oxymetholone has high suggest that oxymetholone crosses placenta anabolic and low androgenic properties. Its readily and affects the fetus (19). Thus, the anabolic effects are mediated through variations and lesions observed in the increased nitrogen retention and experiment group of our study may be consequently increased protein synthesis attributed to oxymetholone effects. (14, 15). Nandrolone is another anabolic medication. Conclusion: In general, it may be stated that It has been reported that nandrolone at the dosage used in our study, increases kidney weight and volume, oxymetholone induces pathological changes cortical volume, and volume of the proximal in kidney tissue of one-day old rat neonates, and distal convoluted tubules in the as well increasing body and kidney weight. experiment group compared to controls (16).
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