Comparative Efficacy of Folic Acid and Adenine in Inducing Renal Anemia in Rat Models

Abstract

Abstract:Chronic kidney disease occurs when a disease or condition impairs kidney function, causing kidney damage to worsen over several months or years. Anemia is an important complication of chronic kidney disease (CKD), with increasing prevalence in the more advanced stages of the disease. The etiology of anemia in CKD is multifactorial, and the key mechanisms involve relative deficiency of erythropoietin (EPO), iron deficiency and maldistribution, and shortened erythrocyte life span.

Objective:  To determine the most effective method for inducing chronic kidney disease (CKD) with anemia in male rats by administering adenine and folic acid and comparing the outcomes of four different models.

First, folic acid was intraperitoneally injected into rats at a dose of 250mg/kg per week. Second, folic acid was also intraperitoneally injected into rats at a dose of 250mg/kg every 2 weeks. Third, adenine was intraperitoneally injected into rats at a dose of 250mg/kg/ per week. Finally, adenine was intraperitoneally injected into rats at a dose of 250mg/kg every 2 weeks.

 Methods:  Thirty male Wistar albino rats were divided into five groups, with n=6 in each group. Group I, the healthy control group, received a weekly intraperitoneal injection of normal saline for four weeks. Group II, the Adenine model group, received a weekly intraperitoneal injection of adenine at 250 mg/kg. Group III, also part of the adenine model group, received an intraperitoneal injection of adenine at a dose of 250 mg/kg every two weeks for four weeks. Group IV, the Folic acid model group, received a weekly intraperitoneal injection of folic acid at 250 mg/kg for four weeks. Lastly, Group V, also part of the folic acid model group, received an intraperitoneal injection of folic acid at 250 mg/kg every two weeks for four weeks. After a 28-day induction period, the rats were sedated and euthanized. Blood samples were obtained, and the serum was collected for further biomarker testing using ELISA kits.

Results:  Upon comparing the rats' body weight at the start and end of the experiment, no group showed a statistically significant change. The dosage of folic acid per week was the most important factor in the increase of relative kidney weight, while there was a significant variation in relative kidney weight across the analyzed groups. After measuring kidney function tests (urea, creatinine), hematological parameters (hematocrit, ferritin), it was found that the folic acid model (250 mg/kg/wk, IP for 4 weeks) was significantly the best to induce renal anemia in male rats, it caused a critical increase in the levels of urea and creatinine, which indicates the occurrence of chronic kidney failure. The aforementioned model also caused a decrease in the hematocrit rate and a reduction in the level of ferritin in the blood, which gives the impression of renal anemia. The adenine model, in the same dose and duration, came second in ranking of the optimal induction model.

Conclusion: Compared to the other rat models, the folic acid model group (250 mg/Kg/wk) is more effective in producing anemia associated with chronic renal disease. Due to the observed alterations in many parameters including kidney function, oxidative stress markers, and inflammatory markers, the weekly dosage of folic acid demonstrated its superiority in the context of renal anemia. This will be useful in the future for discovering innovative drugs and understanding the renal anaemia process.