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Whiplash Rove Beetle Dermatitis Among Students at University Sains International Journal of Pharmacy Teaching & Practices 2015, Vol.6, Issue 1, 1603-1608. Hepatoprotective Activity of Ficus Pseudopalma Blanco against Acetaminophen-Induced Liver Toxicity in Sprague-Dawley Rats *1,2,3librado A. Santiago, 2joy M. Buccat, 2mary Rose T. Domalanta 1,3anna Beatriz R. Mayor 1research Center For The Natural And Applied Sciences, 2department Of Biochemistry-Faculty Of Pharmacy, And 3the Graduate School University Of Santo Tomas Introduction Research Article Hepatotoxicity and drug-induced injury account for a big number of death, hospital admission and acute *1,2,3 2 2 Please cite this paper as librado A. Santiago, joy M. Buccat, mary liver failure (ALF) worldwide. Acetaminophen is 1,3 Rose T. Domalanta, anna Beatriz R. Mayor. Hepatoprotective among the commonly available non-prescription Activity of Ficus Pseudopalma Blanco against Acetaminophen-Induced analgesics in the Philippines. This over-the-counter Liver Toxicity in Sprague-Dawley Rat. IJPTP, 2015, 6(1), 1603-1608. (OTC) drug serves as an active ingredient in most Corresponding Author: popular pain medications and muscle relaxants that Librado A. Santiago are readily consumed in therapeutic doses over time Faculty of Pharmacy, or even too much in committing suicide without University Of Santo Tomas realizing that the drug may cause subclinical damage Email: [email protected] to the liver. Acetaminophen produces a potentially fatal, hepatic centrilobular necrosis when taken in overdose [1]. Acetaminophen overdose, publicized to trigger the most number of ALF cases in the United States, leads to mitochondrial dysfunction ABSTRACT and nuclear DNA fragmentation, resulting in necrotic The hepatoprotective activity of F. pseudopalma was cell death [2]. Acetaminophen is metabolically evaluated against acetaminophen-induced liver toxicity and activated by a group of enzymes located in the was compared with N-acetyl cysteine (NAC) and Lupeol. endoplasmic reticulum by cytochrome P450 in the Acetaminophen (500mg/kg BW) was induced daily to the rats liver to a reactive metabolite identified to be N- for seven days and the plant extract (200mg/kg and acetyl-p-benzoquinone imine (NAPQI) [3,4] 400mg/kg), NAC (100mg/kg) and lupeol (10mg/kg) were instigating glutathione (GSH) depletion that led to administered four hours after acetaminophen induction. The the development of the currently used antidote N- liver index, biomarkers of liver damage (ALT, AST, ALP and acetylcysteine (NAC) in the market for albumin) and antioxidant enzyme levels were (GSH and acetaminophen overdose. catalase) all determined. Histopathological analysis of the liver samples was performed to further assess the protective ability Previous study showed that F. pseudopalma Blanco of F. pseudopalma. Following the 7-day oral administration of of the Moraceae family, an endemic medicinal plant F. pesudopalma at 200mg/kg and 400 mg/kg BW ameliorated widely cultivated in the country, is a potent the toxic effects brought by the overdose of acetaminophen. antioxidant and scavenger of various free radicals The liver enzymes in the sera were normalized, the liver index such as DPPH, nitric oxide and lipid peroxides [5,6] in improved, glutathione and catalase levels restored and lipid which lupeol, a pentacyclic triterpene, was found to peroxides remarkably reduced in the liver homogenate be one of the major bioactive compounds [7]. although albumin levels remained unaltered. Toxicity study also revealed that F. pseudopalma has Histopathological examination indicated an improved no toxic effects towards Sprague-Dawley rats at condition of the hepatocytes though not completely healed or 2000mg/kg dose [8]. In addition to that, the plant repaired as result of the 7-day continued acetaminophen- was also studied for its anti-urolithiatic activity and induced injury in the liver. These data suggest that was shown to effectively lessen the effect of administration of antioxidants such as F. pseudopalma, lupeol ethylene glycol-induced kidney stone formation in and NAC may be important in the repair of damage brought by rats [9]. Lastly, F. pseudpalma was also acetaminophen-induced hepatotoxicity. demonstrated to have a good cytotoxic activity towards hepatocellular carcinoma (HpeG2) and prostate (PRST2) cancer cell lines [10,11]. However, Keywords: Ficus pseudopalma, N-acetylsyteine, Lupeol, F. pseudopalma has not been investigated in liver- hepatoprotective, acetaminophen, antioxidant damaged animal model. This study aimed to demonstrate whether the administration of this 1603 International Journal of Pharmacy Teaching & Practices 2015, Vol.6, Issue 1, 1603-1608. plant can ameliorate the effects of acetaminophen-induced with potassium phosphate buffer (pH 7.4) and were hepatotoxicity in rats by studying the levels of biomarkers of used for the antioxidant assays (GSH, lipid liver damage compared to lupeol and NAC. peroxidation). All colorimetric assays were done using Corona micro plate reader SH-1000 (Hitachi, Materials and Method Japan). Experimental Animals and Induction of Liver Damage using Acetaminophen Assessment of the Biochemical Parameters and The crude ethanolic leaf extract of F. pseudopalma was Antioxidant Enzyme Levels evaluated for its hepatoprotective activity using female Colorimetric analysis of the biochemical parameters Sprague-Dawley rats induced with acetaminophen. were performed using commercially available test The animal experiment was approved by the University of kits. These included the measurement of alanine Santo Tomas Institute of Animal Care and Use Committee aminotransferase (ALT), aspartate aminotransferase (UST-IACUC) and certified by the Bureau of Animal Industry (AST), alkaline phosphatase (ALP) and albumin. In (BAI) and the experimental animals were purchased from the addition to that, levels of antioxidant enzymes such Food and Drug Administration. as the reduced glutathione (GSH), lipid peroxides The rats were numbered and randomly grouped into six (6), and catalase were also estimated. consisting of six (6) rats each: Statistical Analysis Group 1-Normal Control (2% Tween 80) Results were expressed as mean± SD and statistical Group 2-Acetaminophen (500mg/kg BW) analysis was performed using ANOVA, to determine Group 3- Acetaminophen + F. pseudopalma (200mg/kg BW) the significant differences between the groups with Group 4- Acetaminophen + F. pseudopalma (400mg/kg BW) p<0.05 implied significant. Group 5- Acetaminophen + Lupeol (10mg/kg BW) Group 6- Acetaminophen + N-acetylcysteine (100mg/kg BW) Results & Discussion It has long been established that NAPQI-mediated All treatments were done orally for seven (7) days following a mitochondrial injury of hepatocytes, an important previously described protocol with some modification [12]. reactive intermediate and a big source of initating Acetaminophen was administered four (4) hours before reactive oxygen species (ROS) and reactive nitrogen administration of the F. pseudoaplma extract, lupeol and NAC. species, play important role in acetaminophen- Single overdose of acetaminophen or repeated and excessive induced hepatotoxicity. ingestion of therapeutic doses may lead to its accumulation in the body that further lead to hepatoxicity. Acetaminophen toxicity can be explained by its metabolism inside the body, which involves its oxidation to form N-acetyl-p- benzoquinoneimine (NAPQI) [13]. NAPQI is a highly reactive intermediate alkylating metabolite that is when produced in higher amounts (due to acetaminophen overdose) bonds covalently with various cell proteins forming inactive conjugates [14]. These inactive conjugates contribute to the irreversible hepatocyte injury and necrosis. Also during this certain period, levels of reduced glutathione (GSH) and other antioxidant enzymes such as catalase is lower compared to Figure 1. Liver index (%) of the different treatment that of lipid peroxides, which are produced via the groups. Values (n=6) represent mean±SEM, p<0.05). augmentation of oxidative stress [15]. To understand the mechanism of action of F. pseudopalma this Liver Index study was undertaken. In this experiment, the initial weight The weight of each rats were noted prior to of each rats were noted and blood was collected in order to cervical dislocation. The livers were obtained and determine the initial ALT, AST, albumin and alkaline weighed to determine the liver index. The liver phosphatase levels in the serum. The rats were then treated index is an indicator of liver toxicity and was daily with the specified doses of the chemicals and extract evaluated by obtaining the ratio of the liver weight using an oral gavage and were allowed to feed with ordinary to the body weight of each rats [17]. Figure 1 rat chow and drinking water. Blood serum was again collected presents the mean liver index for each group, where on 8th day and the final weight of each rats was also noted. Group 2 (Acetaminophen [500mg/kg] control) had The rats were killed via cervical dislocation and the livers were the highest liver index followed by Group 3 collected. Two livers from each group were placed in separate (200mg/kg F. paseudopalma). The liver index for container with 10% buffered formalin and were sent to Hi- Group 6 (100mg/kg NAC) was closely similar to that Precision Diagnostics for histopathological analysis. The other of Group 1 (Normal control). four livers from each group were placed in a plastic container 1604 International Journal of Pharmacy Teaching & Practices 2015, Vol.6, Issue 1, 1603-1608. group (Group
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