|Year : 2018 | Volume
| Issue : 1 | Page : 38-42
Hepatoprotective effects of hexane root extract of Alchornea laxiflora in sodium arsenate toxicity in wistar albino rats
Esosa Samuel Uhunmwangho, Nurudeen Olajide Rasaq, Iyanuoluwa Olubukola Osikoya
Department of Biochemistry, University of Medical Sciences, Ondo City, Ondo State, Nigeria
|Date of Web Publication||12-Jan-2018|
Nurudeen Olajide Rasaq
Department of Biochemistry, University of Medical Sciences, P.M.B. 536, Ondo City, Ondo State
Source of Support: None, Conflict of Interest: None
Background: Medicinal plants are have been used in the treatment of myriad disease conditions, Alchornea laxiflora is one of such medicinal plant. Aim: To investigate the hepatoprotective effect of hexane root extract of Alchornea laxiflora against sodium arsenate induced liver damage in wistar male rats. Setting and Design: Extraction and administration of bioactive extract. Materials and Methods: Extraction of air-dried ground root of Alchornea laxiflora was done by extracting 500 g with 500 cm of 95% hexane for 72 hrs.The animals (120-150 g) were pre-treated with the extract at varying doses (0.1, 0.5, 1.0, 10, 50 and 100 mg/kg body weight) for seven days orally prior to the intra-peritoneal administration of the toxicant (sodium arsenate) at a dose of 2 mg/kg body weight at the eight day. Hepatoprotective activity of the extracts was evaluated by studying the Cytochrome b5content, Glutathione-S-transferase and 4- Nitroanisole-o-demethylase activities in the liver, Plasma levels of Alanine aminotransferase (AST), Alanine aminotransferase (ALT), Alkaline phosphatase (ALP) and the levels of Total protein, Albumin, Globulin and Glutathione of the various groups. Statistical Analysis: The data were analysed with Microsoft excels (for windows 2007) and Student t-test and ANOVA. Result and Conclusion: The results suggest that pre-treatment of rats with hexane root extract of Alchornea laxiflora for seven days reduced the elevated levels of liver enzymes, reduced the levels of induced liver metabolizing enzymes and the levels of Total protein, Albumin, Globulin and Glutathione that was increased by the toxicity of sodium arsenate in rats and as such possess hepatoprotective effect against sodium arsenate.
Keywords: Alchornea laxiflora, hepatoprotective, liver damage, sodium arsenate
|How to cite this article:|
Uhunmwangho ES, Rasaq NO, Osikoya IO. Hepatoprotective effects of hexane root extract of Alchornea laxiflora in sodium arsenate toxicity in wistar albino rats. CHRISMED J Health Res 2018;5:38-42
|How to cite this URL:|
Uhunmwangho ES, Rasaq NO, Osikoya IO. Hepatoprotective effects of hexane root extract of Alchornea laxiflora in sodium arsenate toxicity in wistar albino rats. CHRISMED J Health Res [serial online] 2018 [cited 2021 Jul 24];5:38-42. Available from: https://www.cjhr.org/text.asp?2018/5/1/38/223116
| Introduction|| |
A large section of the world's population relies on conventional remedies to treat a plethora of diseases. Medicinal herbs are an indispensable part of the world healthcare due to low cost, easy access, and ancestral experience. A number of evidence has been accounted to demonstrate promising potentials of medicinal plants. Alchornea laxiflora (beth) pax and Hoffman is of the family Euphorbiacea. It is a shrub spread throughout tropical Africa. Decoctions of leaves are used in the treatment of inflammation and infectious diseases. The decoction of the leaves is also used in the treatment and management of inflammatory and infectious diseases, as well as an important component of herbal anti-malaria formulations. Earlier reports on A. laxiflora revealed the presence of alkaloids, cardiolides, saponins, and phenolic compounds such as quercetin, with high concentration residing in the leaves than the roots. The presence of terpenoids compounds was discovered in the root samples of A. laxiflora.
Arsenate, an extremely toxic heavy metal is a common environmental pollutant, with sodium arsenate (NaASO2) shown in several studies to be the most toxic of all arsenic compounds. Arsenics with genotoxic, hepatic, tumorigenic, and carcinogenic potential have been severally implicated in the outset of chromosomal aberration, micronucleus and sister chromatids exchange in mammalian cells., In view of the above background information, this study aims to investigate the hepatoprotective effect of hexane root extracts of A. laxiflora on sodium arsenate-induced toxicity in albino Wistar rats.
| Materials and Methods|| |
This study was approved by the University of Medical Sciences and the National Research and Ethics Committee.
Fresh roots of A. laxiflora were obtained from local gardens in Ondo City and Authenticated at the Department of Botany, University of Medical Sciences, Ondo City, Nigeria. The roots were washed with distilled water, air-dried and 500 g was extracted with 500 cm of hexane 95% (Sigma, Chemical Co. London) Utilizing a giant Soxhlet apparatus for continuous extraction at 80°C for 72 h. In each case, the crude extracts were collected and concentrated to low volumes with rotatory evaporator and the concentrated extracts were weighed and transferred to labeled sample tubes.
Forty adult albino rats of Wistar strain weighing between 150 and 200 g were obtained from the animal house in the Department of Biochemistry, University of Medical Sciences, Ondo City, Nigeria. The animals were kept in clean disinfected cages and were allowed to acclimatize to laboratory conditions for 2 weeks before the commencement of the study. They were feed on standard rat pellets from Pfizer feeds, Nigeria and were allowed free access to water and food. The animals were randomized into eight groups with each group having five animals each, the sample size was determined by power analysis using PASS software 15 (NCSS statistical software).
Animals were pretreated with hexane root extract of A. laxiflora for 7 days and at day 8 and 9, the toxicant (sodium arsenate) was administered (intraperitoneally) at a dose of 2 mg/kg body weight. The plant extract was administered (orally) to all groups except the positive control, whereas the negative control received only 2 mg/kg of toxicant for 1 day all through the course of the experiment. The plant extract was administered (orally) to all groups except the positive control (Group 1) which received only water, whereas the negative control (Group 2) received only 2 mg/kg of toxicant for 1 day all through the course of the experiment. Group 3–8 received graded doses of the plant extracts (0.1, 0.5, 1.0, 10, 50, and 100) mg/kg body weight, respectively, after receiving the toxicant for a day.
Tissue preparation and sample collection
The animals were sacrificed 24 h after the last administration by cervical decapitation. Livers were isolated from sacrificed animals, washed in ice cold 1.15% kcl and homogenized in homogenizing buffer (50 mM Tris.HCl, 1.15% KCl, pH 7.4). The homogenate was centrifuged at 9000 g for 20 min and the resulting supernatant further centrifuged at 105,000 g for 1 h at 4°C to obtain the microsomal fraction. This fraction was re-suspended in 0.25 M sucrose and stored frozen. The blood obtained from the heart was also separated with the clear supernatant (serum) separated from the pellets, the homogenate and the serum were then used for the estimation of the liver function parameters and liver metabolizing enzymes.
Liver Marker Enzymes: serum and liver homogenate were analyzed for the following parameters: aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP). These were carried out using standard reagent and kits from Randox Laboratory limited, U.K. Total protein, albumin, and globulin levels were also carried out using standard kits from Randox Laboratory limited, U.K. 4-nitroanisole demethylase activity was carried out according to the method of Shigematsu et al., Glutathione-S-transferase by Meyer et al. and Cytochrome b5 levels by Gan et al.
All analyses were carried out in triplicates. The data were recorded as a mean ± standard deviation and analyzed by Microsoft excels (for windows 2007). Student's t-test and ANOVA was carried out to test any significant differences between their means. Values of P < 0.05 were considered to be statistically significant.
| Results|| |
Effects of hexane root extract of Alchornea laxiflora on liver marker enzymes
Sodium arsenate administration significantly increased (P< 0.05) the activities of liver marker enzymes (AST, ALT and ALP) compared to the positive control. However, administration of hexane root extract of A. laxiflora significantly reduced the activities in a dose-dependent manner [Table 1].
|Table 1: Effects of hexane root extract of Alchornea laxiflora on liver marker enzymes|
Click here to view
Effects of hexane root extract of Alchornea laxiflora on some liver metabolizing enzymes
Administration of sodium arsenate significantly (P< 0.05) increased the activities of liver metabolizing enzymes (4-Nitroanisole demethylase, Glutathione–S-transferase, and Cytb5) compared to the positive control, however, administration of hexane root extract of A. laxiflora significantly reduced the activities in a dose-dependent manner [Table 2].
|Table 2: Effects of hexane root extract of Alchornea laxiflora on some liver metabolizing enzymes|
Click here to view
Effects of hexane root extract of Alchornea laxiflora on some liver biochemical parameters
Administration of sodium arsenate significantly increased (P< 0.05) the total protein, albumin level, and globulin levels compared to the positive control, however, administration of hexane root extract of A. laxiflora significantly reduced the levels of total protein, albumin, and globulin levels in a dose-dependent manner [Table 3].
|Table 3: Effects of hexane root extract of Alchornea laxiflora on some liver biochemical parameters|
Click here to view
| Discussion|| |
The liver is a selfless organ strategically located in the body of mammals. In metabolism, the liver plays a major function of supplying nutrients to other organs of the body. Due to this and other functions, all other organs are referred to as extra-hepatic in comparison to the liver. The liver is also known for its role in the metabolism of foreign compounds (xenobiotics) where the liver converts toxic and nonpolar compounds to nontoxic and polar ones, the form in which they are easily excreted out of the body using an array of endogenous molecules and phase 1 and phase 2 metabolizing enzymes. Even though the liver has natural abilities to regenerate and sustain itself after a damage (occasioned by the metabolism of foreign compounds), given the myriads of roles the liver plays and the severity of liver diseases (a major health concern) it is desirable that natural hepatoprotective herbal/medicinal plants which can help the liver in the metabolism of foreign compounds/toxicants be used to protect the integrity of the liver. A. laxiflora is one such medicinal plant, the possible reversal potentials of its hexane leaf extract against sodium arsenate induced liver toxicity was reported by Uhunmwangho et al. In the present study, the hepatoprotective effect of hexane root extract of A. laxiflora was verified on sodium arsenate induced hepatotoxicity in Wistar rats. From the result obtained in [Table 1] shows the levels of AST, ALT, and ALP of test groups (0.1–100 mg/kg) and the controls (positive and negative). It was obvious that the administration of 2 mg/kg body weight of sodium arsenate for 1 day significantly increased (P< 0.05) the levels of AST, ALT, and ALP of the positive control (H20 only) compared to the negative control (NaASO2). This was to confirm that the administration of NaASO2 indeed caused liver damage and consequently the increased levels of the intracellular enzymes. That liver damage results in the increased activities of these liver enzymes has been confirmed by several reports spanning decades of research on hepatotoxicity.,,, However, prior administration of hexane root extract of A. laxiflora significantly reduced (P< 0.05) the levels of these markers enzymes in a dose-dependent manner with the 100 mg/kg bringing the levels closer to that observed for the positive control and far from the negative control. This is to attest to the hepatoprotective ability of the extract. It was able to heal the liver, reseal the damaged cell membranes and the liver ultrastructure this is not strange given the phytochemicals constituent of A. laxiflora. The medicinal properties of herbal plants is tied to their contents of secondary metabolites.
Also from the result obtained in [Table 2] shows the effects of prior administration of hexane root extract of A. laxiflora on liver metabolizing enzymes during sodium arsenate induced hepatotoxicity in male Wistar rats. As earlier mentioned, the liver is also known for its major role in the detoxification of toxic/foreign compounds and as confirmed by the results from this study, administration of sodium arsenate 2 mg/kg in a day significantly (P< 0.05) increased the activities of the liver metabolizing enzymes (4-nitroanisole demethylase, Glutathione-S-transferase and Cyt-b5) of the negative control compared to the positive control. This is not surprising given that these enzymes are inducible, that is, the administration of toxicants will increase the levels of these enzymes and which would in-turn ensure that noxious substances (which are often non polar) are metabolized to polar and nontoxic forms that are readily excreted out of the body. This is a confirmed physiological/biochemical response of the liver to toxicants as reported by several scholarly works spanning decades of research in hepatotoxicity ,, to mention but a few. However, prior administration of hexane root extract of A. laxiflora before sodium arsenate intoxication significantly reduced (P< 0.05) the levels of these liver metabolizing enzymes to a level closer to that observed in the positive control. This shows that the extract possesses some hepatoprotective properties. It is plausible to assert that the extract possesses enormous hepatoprotective ability which may be related to its antioxidant potentials, i.e., it is able to combat free radicals. Ajith et al. in their study reported that the major route by which toxicants induce damage is through free radical generation or through an intermediate that generates free radicals which if not abated will almost always lead to oxidative damage and hence, it is plausible to assert that the free radical scavenging potentials of medicinal plants can combat these free radicals (generated during the metabolism of foreign compounds) and thereby protecting the liver during toxicant administration. The free radical scavenging properties of hexane root extract of A. laxiflora has been reported by Uhunmwangho et al. Hence, it is safe to say that the hepatoprotective ability of this extract is tied to its antioxidant potentials. [Table 3] shows the effects of the extract on total protein, albumin and globulin levels. Total protein, albumin, and globulin levels are also used as indicators of liver damage. Total protein levels are generally increased during liver metabolism of noxious chemicals as a result of the induction of liver metabolizing enzymes, also the levels of albumin and globulin which are increased during hepatotoxicity. The same trend is also observed in this study where the administration of 2 mg/kg sodium arsenate for 1 day significantly (P< 0.05) increased the levels of these parameters. However, the administration of hexane root extract of A. laxiflora significantly reduced (P< 0.05) the levels of TP, AL, and GB in a dose-dependent manner with the 100 mg/kg of the extract showing the most reduction. This may be due to the ability of the extract to combat free radicals generated during toxicant metabolism and as such the levels of total protein, albumin, and globulin which were initially increased by the toxicant were significantly brought (P< 0.05) closer to the positive control. This shows that the extract could protect the liver against hepatotoxicants. In conclusion, it is safe to conclude that the administration of hexane root extract of A. laxiflora at 0.1–100 mg/kg has hepatoprotective effects on the liver against sodium arsenate hepatotoxicity in rats. This may due to the already established free radical scavenging potentials of the extract. However, more research is needed to confirm if the same is obtainable in man.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| References|| |
Olaleye MT, Adegboye OO, Akindahunsi AA. Alchornea cordifolia
extract protects wistar albino rats against acetaminophen – Induced liver damage. Afr J Biotechnol 2006;5:2439-45.
Burkill HM. The useful plants of West tropical, Africa, royal botanic gardens. Kew 1994;2:144-50.
Adewale AA. Personal Communication with Local Traditional Medical Practitioner in Ibadan, Nigeria. 1993.
Meyer DJ, Coles B, Pemble SE, Gilmore KS, Fraser GM, Ketterer B, et al.
Theta, a new class of glutathione transferases purified from rat and man. Biochem J 1991;274(Pt 2):409-14.
Farombi EO, Ogundipe OO, Samuel Uhunwangho E, Adeyanju MA, Olarenwaju Moody J. Antioxidant properties of extracts from Alchornea laxiflora
(Benth) Pax and Hoffman. Phytother Res 2003;17:713-6.
Domingo JL, Bosque MA, Llobet JM, Corbella J. Amelioration by BAL (2,3-dimercapto-1-propanol) and DMPS (sodium 2,3-dimercapto-1-propanesulfonic acid) of arsenite developmental toxicity in mice. Ecotoxicol Environ Saf 1992;23:274-81.
Patrick L. Toxic metals and antioxidants: Part II. The role of antioxidants in arsenic and cadmium toxicity. Altern Med Rev 2003;8:106-28.
Yamanaka K, Mizol M, Kato K, Hasegawa A, Nakano M, Okada S, et al.
Oral administration of dimethylarsinic acid, a main metabolite of inorganic arsenic, in mice promotes skin tumorigenesis initiated by dimethylbenz(a)anthracene with or without ultraviolet B as a promoter. Biol Pharm Bull 2001;24:510-4.
Uhunmwangho SE, Omage K, Erifeta OG, Josiah JS, Nwangwu CO. Possible reversal of sodium arsenate-induced liver toxicity by hexane leaf extract of Alchornea laxiflora
. Asian J Med Sci 2013;5:3-8.
Reitman S, Frankel S. A colorimetric method for the determination of serum glutamic oxalacetic and glutamic pyruvic transaminases. Am J Clin Pathol 1957;28:56-63.
Kochmar JF, Moss DW. Fundamentals of clinical chemistry. In: Tietz NW, editor. Fundamentals of Clinical Chemistry. Philadelphia, PA: WB Saunders, and Co.; 1976. p. 604.
Shigematsu H, Yamano S, Yoshimura H. NADH-dependent O-deethylation of p-nitrophenetole with rabbit liver microsomes. Arch Biochem Biophys 1976;173:178-86.
Gan L, von Moltke LL, Trepanier LA, Harmatz JS, Greenblatt DJ, Court MH, et al.
Role of NADPH-cytochrome P450 reductase and cytochrome-b5/NADH-b5 reductase in variability of CYP3A activity in human liver microsomes. Drug Metab Dispos 2009;37:90-6.
Barrett KE, Barman SM, Boitano S, Brooks HL. Transport and metabolic functions of liver. In: Barrett KE, Barman SM, Boitano S, Brooks HL, editors. Ganong's Review of Medical Physiology. 23rd
ed. New Delhi: McGraw-Hill; 2010. p. 479-87.
Sallie R, Tredger JM, Williams R. Drugs and the liver. Part 1: Testing liver function. Biopharm Drug Dispos 1991;12:251-9.
Ulicná O, Greksák M, Vancová O, Zlatos L, Galbavý S, Bozek P, et al.
Hepatoprotective effect of rooibos tea (Aspalathus linearis
) on CCl4-induced liver damage in rats. Physiol Res 2003;52:461-6.
Porchezhian E, Ansari SH. Hepatoprotective activity of Abutilon indicum
on experimental liver damage in rats. Phytomedicine 2005;12:62-4.
Işeri S, Ercan F, Gedik N, Yüksel M, Alican I. Simvastatin attenuates cisplatin-induced kidney and liver damage in rats. Toxicology 2007;230:256-64.
Croteau R, Kutcher TM, Lewis NG. Natural products (secondary metabolite). Biochemistry and molecular biology of plants. American Society of plant Physiologists, Rock ville; 2002. p. 1250-318.
Park BK, Kitteringham NR, Pirmohamed M, Tucker GT. Relevance of induction of human drug-metabolizing enzymes: Pharmacological and toxicological implications. Br J Clin Pharmacol 1996;41:477-91.
Manson MM, Ball HW, Barrett MC, Clark HL, Judah DJ, Williamson G, et al.
Mechanism of action of dietary chemoprotective agents in rat liver: Induction of phase I and II drug metabolizing enzymes and aflatoxin B1 metabolism. Carcinogenesis 1997;18:1729-38.
Hodgson E, Goldstein JA. Metabolism of toxicants: Phase I reactions and pharmacogenetics. In: Hodgson E, Smart RC, editors. Introduction to Biochemical Toxicology. 3rd
ed. New York: Wiley; 2001.
Rose RL, Hodgson E. Adaptation to toxicants. In: Hodgson E, Smart RC, editors. Introduction to Biochemical Toxicology. 3rd
ed. New York: Wiley; 2001.
Ajith TA, Hema U, Aswathy MS. Zingiber officinale
roscoe prevents acetaminophen-induced acute hepatotoxicity by enhancing hepatic antioxidant status. Food Chem Toxicol 2007;45:2267-72.
[Table 1], [Table 2], [Table 3]