Pentoxifylline Attenuates Arsenic Trioxide-Induced Cardiac Oxidative Damage in Mice
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Hindawi Oxidative Medicine and Cellular Longevity Volume 2021, Article ID 6406318, 10 pages https://doi.org/10.1155/2021/6406318 Research Article Pentoxifylline Attenuates Arsenic Trioxide-Induced Cardiac Oxidative Damage in Mice Atefeh Gholami ,1,2 Sara Ataei ,3 Davoud Ahmadimoghaddam ,1,2 Navid Omidifar ,4 and Amir Nili-Ahmadabadi 1,2 1Medicinal Plants and Natural Products Research Center, Hamadan University of Medical Sciences, Hamadan, Iran 2Department of Pharmacology and Toxicology, School of Pharmacy, Hamadan University of Medical Sciences, P.O. Box 8678-3- 65178, Hamadan, Iran 3Department of Clinical Pharmacy, School of Pharmacy, Hamadan University of Medical Sciences, Hamadan, Iran 4Clinical Education Research Center, Department of Pathology, School of Medicine, and Biotechnology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran Correspondence should be addressed to Amir Nili-Ahmadabadi; [email protected] Received 14 June 2020; Revised 9 December 2020; Accepted 22 December 2020; Published 8 January 2021 Academic Editor: Paula Felippe Martinez Copyright © 2021 Atefeh Gholami et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. This study was undertaken to evaluate the therapeutic potential effect of pentoxifylline (PTX) against arsenic trioxide (ATO)- induced cardiac oxidative damage in mice. Thirty-six male albino mice were divided into six groups and treated intraperitoneally with normal saline (group 1), ATO (5 mg/kg; group 2), PTX (100 mg/kg; group 3), and different doses of PTX (25, 50, and 100 mg/kg; groups 4, 5, and 6, respectively) with ATO. After four weeks, the blood sample was collected for biochemical experiments. In addition, cardiac tissue was removed for assessment of oxidative stress markers and histopathological changes (such as hemorrhage, necrosis, infiltration of inflammatory cells, and myocardial degeneration). The findings showed that ATO caused a significant raise in serum biochemical markers such as lactate dehydrogenase (LDH), creatine phosphokinase (CPK) and troponin-I (cTnI), glucose, total cholesterol (TC), and triglyceride (TG) levels. In addition to histopathological changes in cardiac tissue, ATO led to the significant increase in cardiac lipid peroxidation (LPO) and nitric oxide (NO); remarkable decrease in the activity of cardiac antioxidant enzymes such as catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx); and the depletion of the total antioxidant capacity (TAC) and total thiol groups (TTGs). PTX was able to reduce the increased levels of serum cardiac markers (LDH, CPK, cTnI, TC, and TG), cardiac LPO, and improve antioxidant markers (TAC, TTGs, CAT, SOD, and GPx) alongside histopathologic changes. However, no significant changes were observed in elevated serum glucose and cardiac NO levels. In conclusion, the current study showed the potential therapeutic effect of PTX in the prevention of ATO-induced cardiotoxicity via reversing the oxidative stress. 1. Introduction (APL), but its usage has been limited because of cardiovascu- lar side effects, such as ventricular tachycardia, QT prolonga- Arsenic is an environmental contaminant that is widely tion, torsade de pointes, and sudden cardiac death [6, 7]. widespread in water, soil, and air due to its industrial and These side effects can be caused through mitochondrial dys- agricultural applications [1]. The epidemiologic evidence function and excess generation of reactive oxygen species showed that high-chronic arsenic exposure has been associ- (ROS) [8], functional changes of ion channels, and disrupted ated with hepatorenal failure and cardiovascular disorders balance of intracellular and extracellular ions [9]. [2–4]. However, arsenic compounds have been used to treat Phosphodiesterase inhibitors block one or more subtypes various diseases from the past to the present [5]. of the phosphodiesterase enzymes (PDEs), thereby prevent- Arsenic trioxide (ATO) is an effective chemotherapeutic ing the inactivation of the cAMP and/or cGMP in various drug used in the treatment of acute promyelocytic leukemia cells. In recent years, the antioxidant and anti-inflammatory 2 Oxidative Medicine and Cellular Longevity properties of phosphodiesterase inhibitors have been consid- Group 3: the mice received PTX (100 mg/kg/day) ered in several studies [10–12]. For instance, Mohammadi Group 4: the mice received ATO (5 mg/kg/day) + PTX et al. (2011) showed that selective phosphodiesterase inhibi- (25 mg/kg/day) tors could increase survival of Langerhans islets by prevent- Group 5: the mice received ATO (5 mg/kg/day) + PTX ing free radical formation [13]. Moreover, sildenafil, as (50 mg/kg/day) phosphodiesterase 5-selective inhibitor, can have beneficial Group 6: the mice received ATO (5 mg/kg/day) + PTX role in improvement of toxicities caused via cadmium [14] (100 mg/kg/day) and lead acetate [15]. It should be noted that groups 4-6 were treated with dif- Pentoxifylline (PTX), as a methyl xanthine derivative and ferent doses of PTX 1 h before ATO administration. In addi- nonselective PDE, is commonly used to treat intermittent tion, the highest dose of PTX (100 mg/kg) was considered to claudication and peripheral vascular diseases, reducing plate- show its safety in group 3. Twenty-four hours after the com- let aggregation and improving red blood cell deformability pletion of treatment, each animal was weighed and anesthe- [16]. Recent evidence showed that PTX inhibits ROS genera- tized by ketamine (50 mg/kg) and xylazine (10 mg/kg), and tion and improves capillary circulation and tissue oxygena- its blood sample was taken through cardiac puncture. Then, tion in various organs. For instance, Yao et al. (2016) blood sample was centrifuged (at 3000 g, 10 min), and its showed that PTX could prevent intermittent hypobaric hyp- serum was kept at -20°C for the biochemical analysis. Fur- oxia induced-oxidative stress in testicular tissue by maintain- thermore, the heart was removed for preparation of tissue ing redox homeostasis [17]. Zhang et al. (2005) reported that homogenate (10%, w/v). Briefly, half of the heart tissue was PTX might be beneficial in reducing hydrogen peroxide homogenized with phosphate-buffered saline (50 mM, induced embryo injury and improve in vitro fertilization pH 7.3) and centrifuged at 3000 g, 10 min at 4°C. Finally, its (IVF) outcome [18]. Additionally, the findings of Egin et al. supernatant was removed for the biochemical experiments. (2016) indicate the effective effects of PTX on oxidative stress Another part of tissue was fixed in 10% formaldehyde solu- reduction in the abdominal compartment syndrome animal tion for histopathological analysis. model [19]. Despite the antioxidant properties of PTX, there is no 2.2. Determination of Glucose and Total Triglyceride and evidence of the therapeutic potential of this drug on ATO- Cholesterol. Glucose, total cholesterol, and triglyceride serum induced cardiotoxicity. Therefore, the current study was levels were determined using commercial kits (Pars Azmoon, designed to assess the PTX effects on the oxidative damage Tehran kit, Iran). induced by ATO in the heart tissue of mice. 2.3. Lactate Dehydrogenase Assay. Lactate dehydrogenase 2. Materials and Methods (LDH) activity in serum sample was measured by determin- ing the rate of oxidation of NADH by an enzymatic colori- Pentoxifylline, 2,4,6-tripyridyl-s-triazine (TPTZ), 1,1,3,3-tet- metric kit (Pars Azmoon Co., Tehran, Iran). The ramethoxypropane, bovine serum albumin (BSA), sulfanil- ′ absorbance change per minute was detected at 340 nm using amide, 5,5 dithiobis-2-nitro benzoic acid (DTNB), 2- spectrophotometric instrument (Analytik Jena Specord 50 thiobarbituric acid (TBA), and N-(1-naphthyl) ethylenedia- Plus), and its results were expressed as U/L. mine dihydrochloride were obtained from Sigma-Aldrich Chemical Company (St. Louis, MO, USA). Arsenic trioxide 2.4. Creatine Phosphokinase Assay. The activity of serum cre- powder was purchased from Merck (Darmstadt, Germany). atine phosphokinase (CPK) was assayed by an enzymatic col- orimetric kit (Pars Azmoon Co., Tehran, Iran). Based on the 2.1. Animals and Experimental Protocol. Thirty-six male ’ albino mice (25 ± 2:5g) ranging from 1 to 2 months in age kit s procedure, creatine kinase converts creatine into ADP were obtained from the animal house of Hamadan University and phosphocreatine. The absorbance change per minute of Medical Sciences (HUMS). The animals were kept in stan- was detected at 340 nm, and its data were expressed as U/L. dard cages at suitable temperature (23 ± 2°C), 12/12 h light/- dark cycle, and relative humidity 50% and received a 2.5. Troponin-I Assay. Cardiac troponin-I (cTnI) levels in standard diet and water ad libitum. The ethical concerns of serum samples were assayed by Enzyme Linked-Immuno- animals’ experiments were considered carefully, and its pro- Sorbent Assay (ELISA) kit, according to the manufacturer’s tocol was approved by the HUMS ethics review board (Ethi- instructions (Shanghai Crystal Day Biotech Co., LTD, China). cal code number: IR.UMSHA.REC.1397.463). In this study, the toxic dose of ATO 5 mg/kg/day was 2.6. Lipid Peroxidation Assay. Cardiac lipid peroxidation was used based on the animal model proposed by Li et al. measured via the reaction of TBA with active-aldehyde inter- (2002) [20]. In addition, based on pilot studies, the dosage mediates such as MDA. Briefly, heart homogenate superna- range of PTX was considered 25-100 mg/kg/day. tant (100 μl) was mixed with 500 μl reagent containing TBA Accordingly, the mice were divided randomly into six (0.2%) in H2SO4 (0.05 M) and subsequently heated for groups of six each and treated for four consecutive weeks 30 min at 100°C in boiling water bath [21, 22]. The peak by intraperitoneal (i.p.) injection as follows: absorbance was detected at 532 nm against different concen- Group 1: the mice received normal saline (control group) tration of MDA as the standard, and its results reported as Group 2: the mice received ATO (5 mg/kg/day) nmol/mg protein.