Effect of Adenosine Triphosphate on Carbamazepine and Paracetamol Interaction Mediated Liver Injury in Female Swiss Albino Mice

Abstract

Aim and Objective: Liver being the main site of drug and toxicant metabolism has become a prime target for drug induced injury. It is the leading cause for acute liver failure and liver transplantation in western countries. It is expected that Paracetamol (PCM) when used along with Carbamazepine (CBZ) results in enhanced liver toxicity. Recently in our lab we have studied Paracetamol and Carbamazepine interaction causing the hepatotoxicity in female Swiss albino mice. From the studies it was concluded that whenever necrosis occurs in liver, the cells which are in the Sphase will start to divide and provides new cells, so that normal lobular structure of liver is restored and progression of injury is inhibited. But due to decline in the concentration of ATP, the restoration process is critically affected. Some studies have shown that ATP administration during the early phase of injury can restore normal liver function and tissue healing mechanisms in animal model. Based on this concept we have formulated the hypothesis whether supplementation of external energy in the form of ATP will stimulate the tissue repair and rescue the animals from the hepatotoxicity and lethal effects of CBZ and PCM combination. So the aim of the present study was to check the effect of ATP on bio-activation mechanism and tissue repair response [mitosis] in CBZ and PCM interaction toxicity. Materials and Methods: Female Swiss albino mice were divided into three groups- Control, CBZ+PCM treated and CBZ+PCM+ATP treated. In the present study, CBZ+PCM group received CBZ in corn oil (300mg/kg, p.o.) for 4 days and on 5th day PCM challenge was given (300mg/kg, p.o. in 0.25 % tragacanth solution). Whereas the CBZ+PCM+ATP group of animals were given CBZ (300mg/kg, orally) for five days immediately followed a single dose of PCM (300mg/kg in 0.25% tragacanth, orally) along with ATP (14mg/20gm mouse, s.c at -1, +1, 3, 5, & 12hr after CBZ and PCM administration on the 5th day). The normal control animals were given only vehicle (10ml/kg, p.o.). Both blood and liver tissues were collected at different time-intervals for assessing liver injury and tissue repair response. After 5th day of dosing, both blood and liver samples were collected at different time intervals like- 0, 12, 24, 36, 48, and 96 hrs from CBZ+PCM and CBZ+PCM +ATP treated animals. Whereas in control animals, the blood and liver samples were collected at only three different time intervals like – 0, 36 and 96 hrs. The blood samples collected were utilized for various serum biochemical estimations such as SGOT, SGPT, Bilirubin and Glucose. Whereas the liver samples were used for estimation of glycogen, GSH, SOD, Catalase, MDA and histopathological evaluations. Results: Serum estimation showed elevated levels of SGPT, SGOT and bilirubin in CBZ+PCM treated mice at 0 hr (5th day) and increased upto 36 hours. Thereafter, it declined gradually by 48 hours indicating liver injury is reversing nearly normal by 96 hrs. CBZ+PCM+ATP treated animals showed comparatively lower elevation in SGPT, SGOT and bilirubin levels indicating that progression of injury is slower in ATP treated animals. The histopathological analysis of CBZ+PCM treated group showed a remarkable centrilobular necrosis and prompt and robust mitotic cell division at 24 and 36 hours respectively. The CBZ+PCM+ATP treated animals showed lesser necrosis in comparison to CBZ+PCM treated groups. In CBZ+PCM treated groups serum glucose levels were elevated upto 24 hours and decreased gradually. Supporting liver glycogen declined gradually indicating liver regeneration process demands energy in terms of glucose. Serum glycogen levels decreased upto 36 hours indicating energy consumption for cell division process and reverted slowly after 36 hrs and normal by 96 hrs. ATP supplementation in CBZ+PCM+ATP treated animals there was increase in the glucose levels and also no drastic fall in glycogen levels was observed. The unexpected mortality during the course of the study in CBZ+PCM+ATP treated animals is suspected due to higher glucose levels leading to hyperglycemic condition in the animals. The lipid peroxidation in CBZ+PCM treated groups was evident from higher MDA levels which were comparatively lower in CBZ+PCM+ATP treated groups. Also the decline in GSH levels with the progression of injury in CBZ+PCM treated groups indicated the elevated levels of toxic metabolites which were normalized by 96 hr as the tissue was getting repaired. Whereas in case of CBZ+PCM+ATP group the levels were comparatively higher than CBZ+PCM treated groups indicating the slower progression of injury at each time points. In the same way due to higher oxidative stress in CBZ+PCM treated groups, the SOD levels kept on declining until the process of tissue repair came into action. But the animals on ATP treatment on account of slower progression of injury did had decline in SOD levels, but in comparison to CBZ+PCM treated groups, the SOD levels were higher in ATP treated groups. Catalase activity was found to be significantly decreased after CBZ+PCM dose upto 36 hour and then was recovering back to normal by 96 hour. The ATP treated groups had comparatively lower decline in catalase levels in comparison to CBZ+PCM treated groups. Conclusions: Based upon my present data, the liver injury was significantly higher in CBZ+PCM group. In ATP treatment group the progression of injury was comparatively lower as evident from liver injury, energy and oxidative stress parameters. Rare mitotic cells were seen in CBZ+PCM+ATP treated animals, because, these animals suffer from less liver injury compared to CBZ+PCM treated animals. Further, histopathology technique may not be able to capture mitotic cells in these animals on account of slower progression of injury in comparison to CBZ+PCM treated animals. The unexpected mortality during the course of the study in CBZ+PCM+ATP treated animals is suspected due to elevated serum glucose levels leading to hyperglycemia like condition. Further studies needed to establish and understand the exact reason behind the lethality of animals.