What Should Ethylene Glycol Induced Urolithiasis Model
Male albino rats weighing about (180 – 220g) were housed in polypropylene cages (47 x 34 x 18cm3) with saw dust (renewed after every 48hrs).The animals were acclimatized to standard laboratory conditions (temperature 25 ± 20 C) and maintained on 12h light and 12 h dark cycle for one month before the start of the experiment. The animals were provided with regular rat chow and drinking water ad libitum. The experiment were approved by the institutional animal ethical committee (IAEC) of CPCSEA (Committee for the purpose of control and supervision of experiment on animals), protocol number IAEC/APCAS/01/2013/02. ii. Ethylene glycol induced urolithiasis model Twenty four animals were randomly divided into four groups, Group I, Group II, Group
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Collection and analysis of urine At the end of the experiment, all animals were places in individual metabolic cages (Nalgane, NalgeNunc Int., Ltd) and twenty –four hour urine samples was collected in a 20µl of 20% sodium azide as a preservative. All the animals had free access to drinking water during the urine collection period. After measuring urinary volume, urine was analysed for calcium, oxalate, phosphate and magnesium.
4. Body weight of rats The body weights of each rat in various groups were estimated once before the treatment and after the end of the treatment for the respective doses. The relative body weight (RBW) of each animal was then calculated as follows. RBW = ABT (g) / IB (g) x 100, Where ABT is the absolute body weight at one time interval and IB is the body weight of rest on the beginning of the treatment.
5. Serum analysis On day 29, the rats were anaesthetized with diethyl ether and sacrificed by decapitation after 24h of above treatment. Blood was collected from each animal, into a centrifuge tube without a coagulant and allowed to clot at room temperature to collect serum. The serum was separated by centrifugation at 10,000 x g for 10min in refrigerated research centrifuge. The collected serum sample was used for the estimation of Creatinine, Urea and Uric
Collection and analysis of urine At the end of the experiment, all animals were places in individual metabolic cages (Nalgane, NalgeNunc Int., Ltd) and twenty –four hour urine samples was collected in a 20µl of 20% sodium azide as a preservative. All the animals had free access to drinking water during the urine collection period. After measuring urinary volume, urine was analysed for calcium, oxalate, phosphate and magnesium.
4. Body weight of rats The body weights of each rat in various groups were estimated once before the treatment and after the end of the treatment for the respective doses. The relative body weight (RBW) of each animal was then calculated as follows. RBW = ABT (g) / IB (g) x 100, Where ABT is the absolute body weight at one time interval and IB is the body weight of rest on the beginning of the treatment.
5. Serum analysis On day 29, the rats were anaesthetized with diethyl ether and sacrificed by decapitation after 24h of above treatment. Blood was collected from each animal, into a centrifuge tube without a coagulant and allowed to clot at room temperature to collect serum. The serum was separated by centrifugation at 10,000 x g for 10min in refrigerated research centrifuge. The collected serum sample was used for the estimation of Creatinine, Urea and Uric