JOURNAL OF MILITflRV PHRRMflCO-MCDICINC N°7-2014
ESTABLISHMENT OF ACUTE RENAL FAILURE MODEL BY ADENINE ON MICE
Ho Anh Son*; Nguyen Thi My Thanh**
Pham Xuan Phong***; Nguyen Linh Toan*
SUMMARY
The models of renal failure are considered as important tools to study mechanism and pre- clinical treatment Adenine-based protocols to induce renal failure are available in rats, but have not been adapted in mice due to their reluctance to consume adenine In this study, we developed a method for induction of acute renal failure through forced adenine ingestion in mice within 2 weeks. Adenine-ingestive mice showed that uremia level is higher than that in control group.
Histopathology showed the lesion on glomerulus and tubules.
* Key words: Adenine; Renal failure; Mice; Model.
INTRODUCTION
Kidney disease is a global health problem.
Animal models of renal failure are important tools to study pathophysiological events in kidney disease that allows translational studies aiming to improve treatment of kidney disease patients. Established techniques for induction of renal failure in mice are mostly dependent on surgical interventions.
It is quite differentiate to kidney lesion in renal failure disease. Creating a renal failure mouse models based on adenine will provide a useful model to investigate the effectiveness of new therapy on this model. The purpose of this study is to establish acute renal failure by adenine administration.
SUBJECTS AND METHODS 1. Subjects.
- 20 Swiss mice, 8 to 10-week old (VMMU).
The animals were housed in laboratory conditions for at least 5 days prior to experiment with food and water available ad bilitum. Animals were divided in two group:
+ Adenine group (12 mice).
+ Control group (8 mice).
- Adenine powder (Sigma, USA) 2. Methods.
- Adenine group was treated with adenine (250 mg/kg of body weight per day) for 2 weeks to induce the chronic renal failure experimentally. Control group received physiological saline. Adenine solution was administered intragastricly.
- Body weight of animals was measured every week.
' Vietnam Military Medical University
** Vinh Medical University
**• Military Institute of Traditional Medicine
Corresponding author: Ho Anh Son ([email protected])
JOURNfll OF MIUTflRV PHflRMflCO-MCDICINC N°7-2014
- in final period of the experiment, mice were deeply anaesthetized with thiopental (100 mg/kg). The kidney was isolated and preserved in 10% fonnalin solution for 24 - lU hoyrs. Then, it was embedded in paraffin, sliced 5 pm thick, dyed with hematoxyline am eosin stain (HE stain); the results were observed under the optical microscope.
- Biochemical analysis was performed on all studied sera animals. Biochemical parameters (serum urea nitrogene, creatinine) were determined.
RESULTS 1. Body weight changes.
Table 1: Body weight during adenine administration (g).
DayO(X ±2SD) D a y 7 ( X ±2SD) Day14{X±2SD}
ADENINE 21.0 ±0.89 20.66 ± 0.52 20.6 ± 0 85
CONTROL 21.1 ±0.96 21.7 ±1.09 22.5 ± 0.93
P
>0.05
<0.06
< 0.005 During administration, adenine-treated group showed significant body weight lower than that in control group at day 7 and endpoint (p < 0.05; p < 0.005).
23.00 22.50 22.00 21.50 21.00 20.50 20.00 19.50
DayO day 14
Figure 1: Adenine-treated loss weight group
In the adenine-treated group, there was a significant decline in body weight during adenine administration.
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JOURNfll OF millTARV PHnRmnCO-MEDKINE N°7-20T4
2. Lethal number of animals during adenine administration.
i 1 4 V-
i 12 I 1 0
Day Day Day Day Day Day Day Day Day Day Day Day Day Day 1 ! 3 t 5 6 7 8 9 10 11 12 13 14
Figure 2: Animal death during adenine administration,
A counting lethal animal presented 50% of animals dead at the end of experiment.
The number of animals declined after 3 days of adenine ingestion and kept stable after 10 days of adenine treatment.
3. Serum biochemistries.
Table 2: Serum urea and creatinine level at endpoint.
Control(X±2SD) Adenine ( X ± 2 S D )
P
URE (mmol/L) 6.1 ±0.61 14.9 ± 6.2
<0.01
CREATINfN (ijmof/L) 25.6 ± 6.1 32.2 ±14.4
< 0 05 At endpoint, there was a significant increase in serum urea and creatinine in adenine- treated animals (p < 0.01; p < 0.05) Among them, 6/6 mice treated with adenine showed that urea level was higher than that in control group; 4/6 animals showed creatinine level was higher than that in control group.
45
JOURNfll OF MIUTflRV PHflRMflCO.MCDKINt N°7-2014 4. Histology.
Figure 3: Lesion on glomerulus and tubules in adenine treated mice.
The kidney histology showed lesion on glomerulus, thicken basement membrane, and enlarge bowman space, tubular area with much of vacuoles.
DISCUSSION
Mouse models of chronic renal failure are important tools for pre-clinical testing in humans. The limitations of existing surgical models, including requirements of surgical skills, demand for animal facilities to support post-operative care, high mortality rate suggested us to develop a non-surgical model of renal dysfunction based on intake of adenine. In this study, we had successful in establishing adenine induced high uremia and glomerular-tubular lesion mice model. These lesions might cause
disorders in secreting and absorbing in tubules, leading to high level of urea and creatinine.
In fact, there were some more common non-surgical options to study uremia including radiation nephropathy and administration of nephrotoxic drugs such as folic acid, cyclosporine [3] and cisplatine [5]. However, these models are non-reversible, strain-dependent and of limited use due to systematic toxicity.
Several different uremic models have also been developed in rats [1, 6]. Advantages 46
JOURNAL OF MIUTHRV PHRRMRCO-MCDICINC N°7-2014 of rat models are that collected blood and
urine volumes are significantly greater, blood samples at intermediate points of time are more easily obtainable.
CONCLUSION
After administration of adenine with dose of 250 mg/kg bw/day within 2 weeks on mice, the results showed:
- Serum urea and creatinine leval were higher than that in control group.
- Lesion on glomerulus and tubules:
thicken basement membrane, and enlarge bowman space, tubular area with much of vacuoles.
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