Volume 8, Issue 6, December Issue - 2020, Pages:839-848 |
Authors: Emeka Hillary Ugwuanyi, Chukwuneke Samuel Udem, Ifeanyi Innocent Madubuinyi |
Abstract: This study was aimed to investigate the antioxidant potentials of methanol and petroleum ether leaf extracts of Asystasia vogeliana against paracetamol-induced liver injury in rats. For estimation of antioxidant potentials, in vitro radical scavenging assays were carried out using DPPH, FRAP, and ABTS. For in vivo study, twenty-five male Wistar rats weighing 100-120 g were randomized and assigned into 5 groups (I-V, n=5). Further, Paracetamol (PCM) at 2 g/kg was used to induce acute hepatotoxicity orally. Rats in group I received distilled water (10 ml/kg) only. While, the rats of groups II, III, and IV received MLEAV (200 mg/kg), PLEAV (200 mg/kg), and a standard hepatoprotective reference drug silymarin (25 mg/kg) respectively for 5 days before PCM induction. Rats in group V received distilled water for 5 days before PCM induction. Blood and liver samples were collected for hematology, serum biochemistry, and histopathology analyses using standard procedures. In vitro assays revealed that MLEAV showed significant (P < 0.05) increases in antioxidant activity compared with PLEAV. Further, significant (P < 0.05) reductions in the activities of ALT and ALP while a significant (P < 0.05) increases in the activity of antioxidant enzymes (CAT, SOD, and GPx) were reported in the group II and III compared with group V. There were also no observable lesions in their hepatocytes. Results of the study can be concluded that MLEAV elicited more in vitro and in vivo antioxidant activities than PLEAV, thus it protects the liver of rat from PCM-induced hepatotoxicity. Therefore, MLEAV could be used as a hepatoprotective agent for the clinical management of liver damage. |
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Full Text: 1 Introduction Asystasia vogeliana (Benth) is a green straggling under-shrub, which belongs to the family Acanthaceae. The leaves of the tree are simple, opposite, ovate, and decussate without stipules. The flowers are zygomorphic and usually associated with colored bract (Popoola et al., 2017). A. vogeliana is widely used in traditional medicine as a remedy for hepatitis in Nsukka, Enugu State, Nigeria. It is commonly known as ‘‘ogwu iba ocha n’anya’’ by Nsukka people which literarily meaning is a drug for treating hepatitis (Okoli, 2015). Indigenous use of A. vogeliana revealed that its infusion with the leaves of other plants such as Cassia alata, Cymbopogon citrus, and fruit juice of Citrus aurantifolia showed higher fidelity level in the management of malaria, chronic fever, gonorrhea, and leprosy (Popoola et al., 2017). Oxygen is vital for aerobic life processes. However, about 5% or more of the inhaled O2 is converted to reactive oxygen species (ROS), (Phaniendra et al., 2015). When the generation of ROS overtakes the antioxidant defense of the cells, the free radicals start attacking various biomolecules such as proteins (Stadtman & Levine, 2000), lipids (Yla-Herttuala, 1999), and DNA (Marnett, 2000), thus leading to several physiological disorders (Phaniendra et al., 2015). Antioxidants are chemical substances that biotransform free radicals to harmless molecules by donating their electrons to free radicals. Antioxidants have gained increasing interest due to their protective roles in pharmaceutical and food industries against oxidative damage, and in animals against oxidative stress damage in biomolecules (Gulcin, 2020). Thus, antioxidants carry out their actions either by reducing the energy of the free radical or suppressing radical formation or breaking chain propagation or repairing damaged and reconstituting membrane (Sahu et al., 2009; Gulcin, 2020). Phenolics compounds have been associated with antioxidant activity due to their high free radical scavenging activities (Rezaie et al., 2015). Acetaminophen (paracetamol) induced hepatotoxicity is a frequently used model for liver damage study, for screening hepatoprotective activity of natural medicines (Ezzat et al., 2012). Acute liver injury is a common consequence of taking an overdose of acetaminophen (Liu et al., 2019). The application of herbal drugs in managing liver diseases has increased globally, because of the belief that herbal drugs are less toxic and have minimum side effects. Also, due to the undesirable therapeutic effects associated with orthodox drugs, the application of traditional medicine including herbal remedies has gained favor as better alternative management (Ali et al., 2009; Girish et al., 2009; Ali et al., 2019). There is a dearth of information on the antioxidant capacity of the A. vogeliana and its hepatoprotective effects on animals. Therefore, the current study was aimed to investigate the in vitro and in vivo antioxidant potentials of the A. vogeliana leaf extracts. The study also evaluated the hepatoprotective effects of the leaf extracts on paracetamol-induced hepatotoxicity in Albino Wistar rats. 2 Materials and Methods 2.1 Plant collection The study was carried out in 2017, for this A vogeliana leaves were collected from Nsukka, Enugu State, Nigeria. The collected A. vogeliana samples were identified by a plant taxonomist, Department of Pharmacognosy and Phytotherapy, University of Nigeria, Nsukka. The plant specimen (Px UNN / 0015) was kept in the department’s herbarium for ease of reference. 2.2 Experimental animals The study was carried out using 25 healthy male Albino Wistar rats weighing between 100-120 g. These experimental animals were obtained from Diamond Research Farm, Nsukka. The animals were housed in the department’s laboratory animal units. Two weeks of acclimatization was allowed to the animals before the experiment started. During this period, these experimental animals were fed with commercial pelletized feed (Vital feed) ® and water ad libitum. The rats were managed according to the Guide for the Use and Care of Laboratory Animals of the National Research Council (National Research Council, 2010). Ethical approval was taken from the Ethical Committee, University of Nigeria, (no. ECUN/15/78756). 2.3 Chemicals and drugs Methanol and petroleum ether were purchased from Sigma Aldrich Company Missouri, USA. Silymarin (Silybon-70®) was purchased from Micro Labs Limited, India. Paracetamol was purchased from Emzor Pharmaceutical Company Limited, Nigeria. The reagents used were of analytical quality. 2.4 Extraction of plant material A. vogeliana leaves were air-dried on a laboratory bench at ambient temperature (26?C - 28?C). The leaves were pulverized into a coarse powder using a hammer mill (JS-390, Japan), and 200g of the powdered leaves was soaked in 1000 ml (80 % v/v) petroleum ether for 48 h and shaken at 2 h interval. The extract was refined using filter paper (Whatman No.1). The extract was later concentrated in vacuo by a rotatory evaporator. The extract was labeled as petroleum ether leaf extract of A. vogeliana (PLEAV) and then kept at 4°C in the refrigerator till the time of use. The marc was air-dried, reweighed, and cold macerated in 1000 ml (80% v/v) methanol for 3 days at ambient temperature (26?C - 28?C). The extract was obtained similarly as described in petroleum ether. The extract was labeled as methanol leaf extract of A. vogeliana (MLEAV). 2.5 In vitro antioxidant assays 2.5.1 DPPH radical scavenging activity 1, 1 – diphenyl – 2 – picrylhydrazyl (DPPH) radical scavenging activity of the two leaf extracts MLEAV and PLEAV were determined by adopting the procedure described by (Sahu et al., 2013), and as modified by (Iqbal et al., 2015). Each of the extracts was dissolved in a suitable volume of methanol to give final concentrations which ranges from 10 to 160 µg / ml. Ascorbic acid solutions which served as the standard solution was also prepared in methanol using the same concentration range. Two milliliters (2 ml) of each solution were put into a test tube and mixed with 4 ml of 0.3 Mmol / L DPPH. A control solution was prepared by adding methanol (2 ml) to 0.3 Mmol / L DPPH. The experiment was carried out in triplicate. The solutions were mixed well and left in the dark for 30 min. After 30 min, the solutions were analyzed on a UV-VIS spectrophotometer (JenWay®, UK) at 517 nm. The antioxidant activity was evaluated using the formula: |
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