The effects of hesperidin on sodium arsenite‐induced different organ toxicity in rats on metabolic enzymes as antidiabetic and anticholinergics potentials: A biochemical approach
Abstract
In our work, it was purposed to investigate the effects of sodium arsenite (SA) and hesperidin (HSP) administered to rats on some metabolic enzymes including carbonic anhydrase (CA), aldose reductase (AR), paraoxonase‐1 (PON1), α‐glycosidase (α‐Gly), butyrylcholine esterase (BChE), acetylcholine esterase (AChE) enzymes activities in the brain, heart, liver, testis, and kidney tissues of rats. CA activities were signifi‐ cantly decreased in testis, liver, and heart tissues of rats given HSP, SA, SA+HSP‐100, and SA+HSP‐200 compared to control (p < 0.05). In liver tissue, AChE and BChE en‐ zymes activities were significantly reduced given in all groups. In all tissues, α‐Gly activity was reduced given in all groups. In the current study, aldose reductase en‐ zyme activity was reduced significantly in testis, brain, and heart tissues of all groups compared to standard (p < 0.05). PON1 enzyme activity was increased significantly in kidney and liver tissues of rats HSP groups and decreased SA groups compared to control. α‐Glycosidase is the key enzyme involved in the digestion of the carbohydrate. Another enzyme α‐amylase hydrolyzes the α‐linked polysaccharide derivatives into oligosaccharide molecules, and α‐glycosidase enzymes, which are characterized in small intestine, catalyze the final stage in the digestive mechanism of carbohydrate molecule to release absorbable monosaccharides like glucose. Conforming to the cholinergic hypothesis, impairment of the cholinergic pathways plays a good role in the development of neurodegenerative diseases like depression, schizophrenia, Alzheimer’s disease (AD) problems with the regulation of traumatic brain injury and sleep. The AD is the main reason for dementia disease, and mild to moderate cases are generally treated with AChE inhibitors. Human CA inhibitor compounds are clini‐ cally used for more than 70 years as antiglaucoma and diuretics drugs.
1| INTRODUC TION
Arsenic, one of the most harmful metalloids, is widely found every‐ where in the earth’s crust and biosphere (Das et al., 2010). Besides the natural sources, use of arsenic‐contaminating insecticides, her‐ bicides, rodenticides, and by‐products of fossil fuels are also potent sources of arsenic toxicity (Gupta, Kannan, Sharma, & Flora, 2005). Arsenic contamination can lead to a wide variety of diseases such as cancers, liver and kidney diseases, neurologic disorder, diabetes mel‐ litus, anemia, hypertension, cardiovascular disorders, male and fe‐ male reproductive abnormalities (Dash et al., 2018; Momeni, Oryan, & Eskandari, 2012; Yousef, El‐Demerdash, & Radwan, 2008). In ad‐ dition, arsenic targets various enzymatic reactions; it affects almost all organ systems in humans and other animals (Yousef et al., 2008). Paraoxonase‐1 (PON1), a family of mammalian enzymes, is a significant antioxidant enzyme. Its lactonase and esterase activ‐ ities have also been observed. Also, the physiologically relevant substrates for these enzymes are unknown. Besides that, it is re‐ sponsible for inhibition of the oxidation of low‐density lipoproteins cholesterol (LDL‐C) and protects against the atherogenic activity of oxidized phospholipids. It is related to HDL and has an important role (Demir & Beydemir, 2015). This enzyme is showed protective impacts against oxidative damage on oxidized lipid production from LDL and to arterial endothelial cells. Therefore, reduction or inhibi‐ tion of PON results in endothelial cell apoptosis and cardiovascular disease (Beydemir & Demir, 2016).
Aldose reductase belongs to keto‐aldo reductase superfamily and plays an important role in the metabolic pathway (Demir, Isık, Gulcin, & Beydemir, 2017b). In diabetic condition, the much level of glucose enters the polyol pathway more actively than glycolysis that gives rise to accumulation of sorbitol in different tissues. Sorbitol is a polar molecule and creates osmotic imbalance. It is very hard to dif‐ fuse through cell membrane. This leads to tissue damage by osmot‐ ics welling. It was reported that a number of studies reported that inhibition of AR could be the effective drug target for prevention of diabetic complications (Aslan & Beydemir, 2017).Carbonic anhydrase (CA; carbonate hydrolyase, E.C.4.2.1.1) en‐ zymes are metabolic enzymes, which catalyze the rapid conversion of CO2 to a proton (H+) and bicarbonate (HCO3–). This mechanism iscommon for most organisms pending the evolvement of life sevengenetically various families of this enzyme are α‐, ζ‐, β‐, δ‐, γ‐, η‐, and θ‐CAs. Especially, CA inhibitors (CAIs) have clinically usage for al‐ most 60 years as antiglaucoma, diuretics or for the treat of epilepsy, obesity, convulsants, glaucoma, and more recently cancer (Caglayan & Gulcin, 2018; Gül, Kucukoglu, et al., 2016; Ozmen Ozgun et al., 2016; Taslimi, Gulcin, et al., 2016; Taslimi, Gülçin, et al., 2016). Hence, the interaction of CA isozymes with different types of novel‐ synthesized derivatives had a great importance (Behcet et al., 2018). Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) are significant enzymes that catalyze the breakdown of acetylcho‐ line (ACh) and butyrylcholine (BCh) that function as neurotransmit‐ ter molecules, which are recorded as drug purposes for Alzheimer’sdisease (AD) (Gül, Tuğrak, et al., 2016; Özbey et al., 2016; Sujayev et al., 2016; Turan et al., 2016).α‐Glycosidase can create glucose compound by opening and hydrolyzing linear and branched isomaltose oligosaccharide mole‐ cules, resulting in hyperglycemia.
Indeed, identifying and charac‐ terizing the inhibitor compounds of α‐glycosidase enzyme that can be therapeutically utilized is significant. Commercial α‐glycosidase inhibitor compounds, like acarbose and voglibose have been uti‐ lized to treatment action of diabetes, but they have side effects, including flatulence, liver disturbances, renal tumors, abdominal discomfort, acute hepatitis, hepatic hurt, diarrhea, and abdomi‐ nal fullness (Hiroyuki, Tomohide, & Kazunori, 2001; Zhang et al., 2011).Flavonoids are ubiquitous compounds and are polyphenol compounds found in many vegetable, fruit, and herbal dietary sup‐ plements (Gülçin, 2012; Kandemir, Kucukler, Eldutar, Caglayan, & Gülçin, 2017; Kuzu et al., 2018). These polyphenolic compounds may also act as physiological regulators, chemical messengers, and cell cycle inhibitors (Köksal et al., 2017). Hesperidin (HSP) is abundant in fruits and vegetables like sweet orange lemon and grapes, in citrus fruits (Kandemir, Ozkaraca, Küçükler, Caglayan, & Hanedan, 2017; Liu et al., 2015). It exhibits pharmacological and biological properties like antioxidant, antiapoptotic, chemopre‐ ventive activity, and anti‐inflammatory (Ahmad et al., 2012; Turk et al., 2018).After an extensive literature review, there were no studies re‐ lated to the effects of HSP on SA‐induced heart, brain, kidney, testis, and liver tissues of rats. Therefore, the activities of CA, PON1, α‐gly‐ cosidase, AR, AChE, and BChE enzymes in the brain, heart, testis, kidney, and liver tissues were investigated in male rats in this study.
2| E XPERIMENTAL
The HSP and sodium arsenite (SA, NaAsO2) were provided from Sigma‐Aldrich (USA). Other chemical materials, which were used in the paper, were of highest purity and were purchased from Sigma and Merck. The HSP and SA were based on the formerly searches by Ahmad et al. (2012) and Dash et al. (2018), respectively.In this experiment, 35 male Sprague dawley rats between 220–250 g (10 weeks old) were used. The rats were provided from Medical Experimental and Application Research Center, Ataturk University. Animals were allowed to adapt the environment at a temperature of 24°C ± 2°C, 45% ± 5% humidity and for 1 week before the experi‐ ment with 12 hr light/dark cycle. During the process, rats were fed with water ad libitum and standard rat diet. This paper was designed conforming to ethical norms approved by the Ataturk University Ethical Committee for Animal Experiments.Sprague dawley rats were randomly divided into five classes as seven rats in each class.Twenty‐four hours after the study period, rats were sacrificed under mild sevoflurane anesthesia. The kidney, testis, brain, liver, and heart tissues were isolated immediately from the animals. These tissues were washed and stored at −20°C for biochemical enzyme analysis.The rat heart, liver, brain, kidney, and testis tissues were homog‐ enized in a homogenizer device (Ultra Turrax‐T25) using Tris buffer (pH 7.4, 20 mM) to obtain a (1/10 weight/volume) homogenate. It was then centrifuged for 30 min at 15,000 × g at 4°C. The upper su‐ pernatant was transferred to another clean tube for enzyme analysis.
PON1 activity was p‐nitrophenyl phosphate) as substrat (1 mM) in 50 mM glycine/NaOH (pH 10.5) including 1 mM CaCl2 at 412obtained using paraoxon compound (diethyl nm (Alim & Beydemir, 2016).Aldose reductase enzyme activity was obtained conforming to previ‐ ous articles and measured with using DL‐glyceraldehyde substrate re‐ duce of NADPH at 340 nm spectrophotometrically (Cerelli et al., 1986).In CA enzyme section, changes in absorbance were determined pending 3 min at 348 nm using PNA was utilized as a substrate and performed conforming to Verport method and previous studies (Aksu et al., 2016; Garibov et al., 2016; Gul, Mete, Taslimi, Gulcin, & Supuran, 2017; Taslimi, Sujayev, et al., 2017).butyrylcholinesterase activities assaysIn this procedure, Ellman method was used and performed con‐ forming to previous studies (Gül, Demirtas, Ucar, Taslimi, & Gülçin,2017, Taslimi & Gulçin, 2018, Taslimi, Osmanova, et al., 2018; Taslimi, Caglayan, et al., 2018).α‐Glycosidase inhibition assay was performed using p‐NPG as the sub‐ strate, conforming to the method of Tao, Zhang, Cheng, and Wang (2013) and performed conforming to previous studies (Taslimi & Gulçin, 2017; Taslimi, Akıncıoğlu, & Gulçin, 2017; Taslimi, Caglayan, & Gulçin, 2017).Values were analyzed with one‐way ANOVA using the SPSS statisti‐ cal program. Tukey multiple comparision test was used to compare the studied parameters between the groups. The data are presented as the mean ± standard error of means (SEM). Differences were con‐ sidered significant when the p < 0.05.
3| RESULTS
CA enzyme activities were decreased significantly in testis, liver, and heart tissues of rats given HSP, SA, SA+HSP‐100, and SA+HSP‐200 compared to control (p < 0.05). Against the cytosolic CA, HSP, SA, SA+HSP‐100, and SA+HSP‐200 behaves as good inhibitors. While HSP, SA, SA+HSP‐100, and SA+HSP‐200 were not statistically sig‐ nificant difference compared to control group in brain and kidney tis‐ sues (Figure 1). For example, EU values for liver tissue: SA+HSP‐200 (72.83 ± 0.79)