sign in sign up
<<
Home , Corchorus, Corchorus olitorius

Original Article Effects of olitorius Extract on Certain and Biochemical Indices in Sodium Arsenite Exposed Rats Olaniyi T. Adedosu*1, Olufemi E. Akanni2, Olusegun K. Afolabi1 and Adebayo L. Adedeji1

1Department of Biochemistry Ladoke Akintola University of Technology, Ogbomoso. Oyo state Nigeria 2Department of Biomedical Sciences, College of Health Sciences, Ladoke Akintola University of Technology, Osogbo. Nigeria

ABSTRACT Objective: is a household leafy with

several medicinal uses. The anti-oxidative and certain biochemical indices of the methanol extract in an in-vivo and in-vitro experiments were studied. Method: Phenols, Flavonoids were determined by Folin Ciocalteu and Aluminium chloride assays while Hydroxyl, 1-1-diphenyl-2- picrylhydraxyl (DPPH) radicals and Malondialdehyde levels measured using deoxyribose assay, DPPH and the reaction of Malondialdehyde with Thiobarbituric acid. Cholesterol, reduced glutathione (GSH) and enzyme assays were carried out using standardized methods. Results: Extract showed activities as it scavenged hydroxyl, DPPH radicals and inhibited lipid peroxidation respectively in-vitro. Animals fed with extract for 21 days at 50, 100, 150 and 200 mg/kg body weight significantly (p<0.05) decrease MDA in a concentration- dependent manner by 2.02, 1.34, 0.76

and 0.02 mg/ml respectively. However, significant increases (p<0.05) in the tissue level of GSH was observed (52.80, 68.7, 81.80 and Address for 88.80 mg/ml). Furthermore, Extract at 100 mg/kg modulate sodium arsenite induced hepatotoxicity in rats, as combined treatment Correspondence significantly (p<0.05) decreases cholesterol and marker enzymes Department of activities (Alanine aminotransferase, Aspartate aminotransferase and Biochemistry Ladoke Gamma glutamyl transferase) compared with intoxicated animals. Akintola University of Extract contained various amount of phenol and flavonoids from Technology, (2.250-3.500 mg/g) and (1.650-1.880 mg/g) respectively. Ogbomoso. Oyo state Conclusions: Extract shows anti oxidative properties reduces MDA Nigeria. and cholesterol level, enhance GSH increase and exhibit E-mail: hepatoprotective ability, suggestive of their bioactive polyphenol [email protected] components which may be explored as therapeutic agent for management, prevention and treatment of age – related diseases.

American Journal of Phytomedicine and Clinical Therapeutics www.ajpct.org Adedosu et al______ISSN 2321 – 2748

Keywords: Anti-oxidative, Anti-lipid peroxidation, Corchorus Olitorius, Hepatoprotective, Polyphenols.

INTRODUCTION Despite decades of research, the However nature remains the roles of free radicals-induced organ injury is primary source for cures to mans ailments still a matter of debate and currently a and diseases as antioxidants of natural serious issue in diagnosis of many diseases. source have been identified in many It is well known that free radicals cause cell medicinal which has raised various damage through mechanism of covalent interest in this field7. Numerous indigenous binding and lipid peroxidation leading to in Africa were consumed injury of the cell membranes1. Common free innocently without information on their - 8 radicals includes, superoxide anion (O2 ) dietary and health benefits , while many - Nitric oxide (NO), Peroxyl (RO2 ), Hydroxyl medicinal plants are currently gaining (OH), Lipid Peroxyl (LOO-), Alkyl (R-) and approval in the traditional treatment of alkoxy (RO-) radicals which are involved in diseases such as malaria9, about 800 plants various catalysis of enzymes, transition are currently documented with beneficial metals or biological systems2. However non- effects for treatment of diabetes10. free radicals such as Hydrogen Interestingly, most suspected bioactive -1 Peroxide (H2O2), Singlet Oxygen (O2 ), components of these plants are believed to Ozone (O3), Lipid Peroxide (LOOH) are have important roles with natural other forms of activated oxygen generated antioxidant properties which are presumed from aerobic organisms capable of reacting to be safe since they occur in plants hence with most biological molecules including the need for laboratory based experiments proteins, lipids, lipoprotein and DNA3. The and investigations to validate these claims reactive ability of free radicals have been which forms the rationale for this study. considered responsible for a series of Corchorus olitorius (), is a undesired processes such as aging, material native to both tropical and subtropical degradation, food deterioration, oxidative regions throughout the world with mallow- stress and many pathophysiological leaves commonly consumed as a leafy disorders such as arthritis, diabetes, vegetable11,12. The leaves are used in inflammation, neuro-degeneration and ethnomedical practices to treat ache and cancer2-4. pain, dysentery, malaria, enteritis, fever, Interestingly, antioxidants have been gonorrhea, pectoral pains and tumors13. The found to terminate or retard oxidative present study was aimed at evaluating process by scavenging free radicals and as certain antioxidative and biochemical protective agents in humans reduces cellular indices of the extract of Corchorus olitorius oxidative damage and thus retard the leaves via in vivo and in vitro experiments. progress of many chronic diseases as well as lipid peroxidation5. In addition, some MATERIALS AND METHODS antioxidants exert their effect by the levels of endogenous antioxidant defenses up Reagents regulating the expression of the genes Some of the reagents in this study encoding superoxide dismutase (SOD), includes; methanol, folin ciocalteu, gallic catalase, or glutathione peroxidase6. acid, quercetin, sodium carbonate,

AJPCT[3][03][2015] 245-256 Adedosu et al______ISSN 2321 – 2748

thiobarbituric acid, trichloroacetic acid, 2- extracts were administered at 100mg/kg deoxyribose and 1, 1-dipheny-2- body weight for this group. picrylhydrazyl radical (DPPH), all of which were products of Sigma chemical company Collection of blood sample and preparation USA. of liver homogenate Rats were sacrificed by cervical Experimental animals dislocation. Blood was collected directly Twelve weeks old male wistar rats from the heart into plain and well-labelled weighing between 120-140g obtained from sample bottles and centrifuged at 4000rpm the animal house of Biochemistry for 5 minutes to obtain serum for analysis of Department were maintained on commercial biochemical parameters. The hepatic tissues feed for about two weeks before the were homogenized in KCl [10 mM] commencement of the experiment. The phosphate buffer (1.15%) with ethylene- animals were randomly divided into four diamine tetra acetic acid (EDTA, pH 7.4) groups with six (6) animals per group in a and centrifuged at 12, 000 rpm for 60 battery cage and another set of four groups minutes. The supernatant was used to assay with six animals each for the first and for GSH and MDA concentrations. second in vivo study respectively. They were fed ad libitum and handled based on Phytochemical study the recommendations and guide lines of our Phenols and Flavonoids were institutional animal care and ethical measured spectrophotometrically using committee for laboratory animals. Folin Ciocalteu and Aluminium chloride assays by14,15, using gallic acid and Plants material and extract preparation quercetin as standards. The plants were collected from the teaching and research farm and Antioxidants activity authenticated by the botany unit. Fresh Radicals scavenging effects were healthy leaves of the plant were dried and determined using the methods of16,17 for the powdered while 10 grams of this was hydroxyl and DPPH radicals respectively. extracted in 100ml of methanol in the cold MDA levels and GSH determination were for 72 hours. carried out by the methods of18,19 while Inhibition of lipid peroxidation was Administration of test substances to animals determined by the method of20 with ascorbic 48 animals were divided randomly acid as standard. into two major groups with the first group made up of 24 rats divided into four groups Biochemical indices with 6 animals each and were treated with Protein and cholesterol concen- various doses (50, 100, 150 and 200mg/kg trations were determined by the procedure body weight) respectively for 21 days with of21,22. Enzyme assays, ALT, AST and γGT the extracts while the other groups were were determined using the method of23-25 made of 24 rats divided into four groups based on the standardized methods by the namely group A (control), B (sodium International Federation of Clinical arsenite intoxicated rats), C (sodium arsenite Chemistry. and extract) and D (extract only) respectively. Arsenite intoxication was carried out at 2.5 mg/kg body weight while

AJPCT[3][03][2015] 245-256 Adedosu et al______ISSN 2321 – 2748

Statistical analysis free radicals intracellularly30,31 which in turn Statistical analysis were based on the induces oxidative damage to tissues, thus in Duncan`s experimental analysis with mean this study, sodium arsenate was used to standard deviation of sample analysed using induced hepatoxicity in the experimental the student T test and p< 0.05 i.e 95% level animals. Sodium arsenite intoxication of significance26. caused increase in serum protein, cholesterol, malondialdehyde level and RESULTS AND DISCUSSION activities of alanine aminotransferases, aspartate aminotransferases and gamma Considerable evidence indicates that glutamyltransferase (Fig. 1, 2, 3, 4, 5 and 6). increased oxidative damage is associated Elevated levels of these serum enzymes with, and may contribute to the development 27 have been studied and could be indicative of of all major neuro-degenerative diseases . an early liver damage as well as an It is also well known that free assessment of its functions32,33. However radicals cause cell damage through results obtained showed that rats treated mechanism of covalent binding and lipid 1 with sodium arsenite and the extract peroxidation with subsequent tissue injury . ameliorated these effects as these parameters Interestingly, the interaction of were restored nearly to their control levels. oxygen radicals and other oxidants with This then suggest the possible modulatory proteins, lipids, biological membranes and role of the extract for a possible even DNA have been linked with oxidative hepatoprotective ability34. The decreased stress, lipid peroxidation, DNA. Single- cholesterol level may be an indication of the stranded–breaks, changes in structures or possibility of anti-hyper lipidemic property loss of bases and DNA-DNA or DNA- 28 of the extract which may be employed in Protein cross-linkages . treating cholesterol disorders35. Results from However foods, , beverages and the arsenic intoxication can be inferred that other plant products are found to be rich in Corchorus olitorius methanol extract polyphenolic compounds with, free radical possesses free radical scavenging activity scavenging and protective anti oxidant 29 which could have a beneficial effect. defenses in the body , due to their natural Further assessment of the extract source. shows that phenols and flavonoids were Hence in the present study, rats expressed between (2.250-3.500 mg/g) and orally exposed to the methanol extract of (1.650-1.880 mg/g) Garlic acid and Corchorus olitorius for 21days at various quercetin equivalent respectively. (Table 2). doses (50, 100, 150 & 200mg/kg body The extract also scavenged hydroxyl weight) showed significant (P<0.05) and DPPH radical maximally by, 60.01% decrease in the level of liver MDA and a and 93.48% while Fe2+-induced lipid significant increase (P<0.05) in the peroxidation was inhibited maximally by concentration of reduced glutathione (GSH) 60.04% respectively (Table 3). in a dose-dependent manner (Table 1). The overall results of this study from Observation from table 1, is an the antioxidants and biochemical indices indication of the ability of the extract to investigated clearly suggest that extracts of inhibit lipid peroxidation, maintain Corchorus olitorius were hepatoprotective membrane integrity and boost the body’s had antioxidative and anti-lipiperoxidative antioxidant status by increasing GSH levels. effects which are linked to the main Also, studies have shown that repository of appreciable amount of Sodium arsenite is capable of generating

AJPCT[3][03][2015] 245-256 Adedosu et al______ISSN 2321 – 2748

phenolics, flavonoids and other antioxidant enzymes in growing rice antioxidative compounds found in this seedlings. 1997; P. plants. 7. Osawa T, Kato Y. Protective role of antioxidative food factor in oxidative stress CONCLUSION caused by hyperglycemia. Ann NY Acad Sci. 2005; 1043: 440-51. Conclusively, data and information 8. Adedosu OT, Badmus JA, Adeleke GE, obtained from this study are indication of Afolabi OK, Adekunle AS, Fatoki JO, Corchorus olitorius potential to enhance Fakunle PB. Oxidative damage and dietary antioxidants: The roles of extract and nutrition and health as a rich source of fractions of Solanium aethiopicum leaves: dietary antioxidants for biological systems Canadian journal of Pure and Applied susceptible to free radicals-mediated Sciences. 2015; 9: (1); 3185-3192. reactions leading to oxidative stress in 9. Adedosu OT, Akanni EO, Adedosu AN, diseases. Adedeji AL, Yakubu O and Ayinde FA. Comparative antioxidants status of leaves ACKNOWLEDGEMENT extracts of some common anti-malarial plants in . American Journal of The author wishes to appreciate the Phytomedicine and clinical therapeutics. efforts of Dr. A. Akintola of the Botany Unit 2014; 2: (12); 1480-1488. for information on the plant. 10. Patil A, Jadhav V, Arvindekar A and More T. Anti-diabetic activity of Maesa Indica (Roxb) Authors disclosure statement stem back in streptozotocin induced diabetic “No conflict of interests exist”. rats. American Journal of Phytomedicine and clinical therapeutics. 2014; 2: (8); 957-962. 11. Danny OC. “Saluyot now a popular vegetable Funding source declaration worldwide.” Inquirer. February 15, 2010; Funding for this work was provided Retrieved August 7, 2011. by private contributions from the authors. 12. Stewart RH. “The Corchorus () Pages” Malvaceae Info. http://www.malvaceae. REFERENCES info/Genera/Corchorus/Corchorus.html. 2011; Retrieved September 10, 2011. 1. Brattin WJ, Glenda EA. J free radical biol. 13. Yoshikawa M, Shimada H, Saka M, and Med 1985; 1: 27. Matsuda H. Medicinal foodstuffs. Absolute 2. Halliwell B, and Gutteridge JMC. Free stereostructures of corchoionosides A, B, C, radicals, other reactive species and disease. In. histamine inhibitors from the leaves of Free Radicals in Biology and Medicines, Vietnamese Cochorius olitoros L (), Clarendon Press, Oxford. 1999; Pp: 617-783. Chem Pharm Bull 1997; 45: 464 -469. 3. Gulein. Antioxidant and antiradical activities 14. McDonald S, Prenzler PD, Autolovich M, of L-carnitine. Life Sci., 2006; 78: 803-811. Robards K. Phenolic content and antioxidant 4. Kourounakis AP, Galanakis DK. Synthesis activity of olive extracts. Food Chem. 2001; and pharmacological evaluation of novel 73: 73. derivatives of anti-inflammatory activities. 15. Chang CC, Yang MH, Wen HM, and Chern Drug Dev Res 1999; 47: 9-16. JC. Estimation of total Flavonoid content of 5. Lai LS, Chou ST. Studies on the antioxidative propolis by two complimentary colorimetric activities of Hsiantsao (Mesona procumbens. methods. J. Food drug analysis 2002; 10: Hems.). J. Agric. Food Chem. 2001; 49: 963- 117- 182. 968. 16. Halliwell Β. Free radicals and metal ions in 6. Cotelle JM, Verina RS, and Kumar S. health and disease. Proc. Nutr. Soc.1987; 46: Superoxide anions generation and activities of 13-26.

AJPCT[3][03][2015] 245-256 Adedosu et al______ISSN 2321 – 2748

17. Mensor LL, Menezes FS, Leitao GG, Reis 27. Halliwel B. “Dietary polyphenols: good, bad, AS, dos Santos TC, Coube CS, and Leitao or indifferent for your health?” Cardiovasc. SG. Screening of Brazilian plant extracts for Res 2007; 73: (2), 341-7. antioxidant activity by the use of DPPH free 28. Lim YY, Lim TT, Tee JJ. Antioxidant radical method. Phytother. Res. 2001; 15: property of several tropical : a 127-130. Comparative study. Food Chemistry 2007; 18. Varshney R, and Kale RK. Effects of 103: 1003-1008. calmodulin antagonist on radiation induced 29. Shobha RI, Rajeshwari CU, Andallu B. Anti- lipid peroxidation in microsomes. Int. J. peroxidative and anti-diabetic activities of Ani Radiation. Biol. 1990; 58: (5); 733-743. (Pinpinella anisumi L) and 19. Beutler EO, Duran and B.M Kelly. Improved identification of bio-active compounds. method for the determination of blood American Journal of Phytomedicine and glutathione. Lab .Clin. Med. 1963, 61; 882- clinical therapeutics. 2013; 1: (5); 516-527. 888. 30. Pei-chung L, Chiong IH, and Te-chang L. 20. Ruberto G, Baratta MT, Deans SG, and Oxidative stress mediates sodium arsenite Dorman HJD. Analysis of chemical induced expression of heme oxygenase composition and bioactive property monocyte chemoattractant, protein and evaluation. Asian Pac J. 2000; 31: 3-15. interleukin in vascular smooth muscular cells. 21. Lowry OH, Rosenbrough NJ, Fair AI, Toxi Sci 2005; 85: (1); 541-550. Ramdall RJ. Protein measurement with folin – 31. Ramos O, Carrizales I, Leticing Y, Jesus M, phenol reagent J. Biol. Chem 1951; 193: 265- Liha B. Arsenic increased lipid peroxidation 275. in rat tissue by a mechanism independent of 22. Trinder P. Report of the national cholesterol glutathione levels. Environmental Health education programme. Anu. Clin. Biochem. 6: Persp. 1995; 103: 85-88. 24; 4. Expert panel on detection, evaluation 32. Wooton ID. Microanalysis in medical and treatment of high blood cholesterol in Biochemistry. 4th edition Churchill London. adults. Arch. Intern. Med. 1969; 148: 36-39. 1964. 23. Wroblewski F, La Due JS. Serum and Plasma 33. Varley E. Pratical clinical biochemistry 4th Alkaline phosphatase determination. Ann. ed. Heineman. London 1969; 275-308. Intern. Med. 1956; 45: 801. 34. Dajas F, Arrendondo F, Echeverry C, Ferreira 24. Karmen A. Serum and Plasma Aspartate M, Morquio A and Rivera F. Flavonoids and aminotransferase determination. J. Clin. the brain. Evidences and mechanisms for a Invest 1995; 24: 126. protective capacity Carr. Neuro. Pharma. 25. Szasz G. A kinetic photometric method for 2005; 3: (3), 1-2. serum gamma glutamyl transterase. Clin. 35. Laskar S, Majumder SG, Basak B, Dey CD. Chem. 1969; 15: 124-136. Influence of jute (Cochorus olitorius) 26. Bliss CI. Statistics in biology. Statistical protein enriched dieton some enzymes and methods for research in the Natural sciences, liver lipids of Albino rats (Rattus novergicus). Mc Graw Hill Book company N.Y. 1967; 1: Physiol. Bohemoslov 1986; 35: 86-89. 558.

AJPCT[3][03][2015] 245-256 Adedosu et al______ISSN 2321 – 2748

Table 1. Malondialdehyde (MDA) and reduced glutathione (GSH) concentrations in the liver of various treated groups with extract for 21days

Malondialdehyde (MDA) Reduced glutathione (GSH) Treatment group concentration (mg/ml protein) concentration (mg/ml) A (control) 2.89 ± 0.109 47.00 ± 1.655 B (50mg/kg body weight) 2.02 ± 0.023 52.80 ± 6.870 C (100mg/kg body weight) 1.34 ± 0.250 68.75 ± 7.190 D (150mg/kg body weight) 0.76 ± 0.280 81.80 ± 6.990 E (200mg/kg body weight) 0.02 ± 0.001 88.80 ± 4.945

Values are given as mean and standard deviation of six determinations.

Table 2. Total phenolic and flavonoids contents of the methanol extracts of Corchorus Olitorius leaves

Total phenolic content (mg/g) Total flavonoids content Concentration (mg/ml) Gallic acid Equivalent (mg/g) Quercetin Equivalent 100 2.250 ± 0.004 1.650 ± 0.008 200 2.250 ±0.001 1.800 ± 0.039 300 3.000 ±0.004 1.810 ± 0.014 400 3.500 ±0.008 1.880 ± 0.011

Mean value ± standard deviation of three replicates.

Table 3. Percentage inhibition of lipid peroxidation, hydroxyl radical and DPPH scavenging effects at various concentrations by the extract

% inhibition of lipid % hydroxyl radical scavenging % DPPH radical scavenging peroxidation effects effects 41.10 (250.00mg/ml) 15.98 (25.00mg/ml) 51.820 (100.00 mg/ml) 47.65 (300.00 mg/ml) 20.03 (50.00 mg/ml) 80.340 (200.00 mg/ml) 50.31 (350.00 mg/ml) 24.67 (75.00 mg/ml) 90.580 (300.00 mg/ml) 54.40 (400.00 mg/ml) 32.23 (100.00 mg/ml) 92.610 (400.00 mg/ml) 58.08 (450.00 mg/ml) 44.63 (125.00 mg/ml) 92.900 (500.00 mg/ml) 60.74 (500.00 mg/ml) 60.01 (150.00 mg/ml) 93.480 (600.00 mg/ml)

Mean values ± standard deviation of three replicate.

AJPCT[3][03][2015] 245-256 Adedosu et al______ISSN 2321 – 2748

Figure 1. ALT activity in the serum of the various treatment groups

Values are given as mean and standard deviation of six determinations. Values differ significantly from control (P≤0.05).

AJPCT[3][03][2015] 245-256 Adedosu et al______ISSN 2321 – 2748

Figure 2. AST activity in the serum of the various treatment groups

Values are given as mean and standard deviation of six determinations. Values differ significantly from control (P≤0.05).

AJPCT[3][03][2015] 245-256 Adedosu et al______ISSN 2321 – 2748

Figure 3. GGT activity in the liver of various treated groups

Values are given as mean and standard deviation of six determinations. Values differ significantly from control (P≤0.05).

AJPCT[3][03][2015] 245-256 Adedosu et al______ISSN 2321 – 2748

Figure 4. Cholesterol concentrations in the serum of various treatment groups

Values are given as mean and standard deviation of six determinations. Values differ significantly from control (P≤0.05).

AJPCT[3][03][2015] 245-256 Adedosu et al______ISSN 2321 – 2748

Figure 5. MDA concentrations in the liver of each treated groups

Values are given as mean and standard deviation of six determinations. Values differ significantly from control (P≤0.05).

AJPCT[3][03][2015] 245-256