The Effects Of Fresh Garlic Extract On Acetaminophen Essays

The Effects Of Fresh Garlic Extract On Acetaminophen Essays The Effects Of Fresh Garlic Extract On Acetaminophen Essay The Effects Of Fresh Garlic Extract On Acetaminophen Essay Introduction Oxidative emphasis and lipid peroxidation play cardinal functions in the pathogenesis and patterned advance of several upsets. Cancer, ageing, coronary artery disease, and inflammatory procedures have all been linked to the coevals of reactive O species and toxic metabolites of lipid peroxidation reactions. 1, 2, 3 In many theoretical accounts, depletion of liver glutathione shops and other antioxidant molecules constitute an of import mechanism for the initiation of oxidative emphasis and the attendant harm to biological molecules such as proteins and nucleic acids, and the activation of atomic written text factors that may be of import in the coevals of proinflammatory cytokines. Several anti-oxidants have been used in the intervention of oxidative stress-mediated diseases, including vitamins ( C and E ) , carotenoids, and minerals such as Se. 9, 10, 11,12 Besides, ethnomedical patterns have relied on the usage of works merchandises which are now known to incorporate antioxidant secondary metabolites.13 Garlic and garlic merchandises have been employed in medical pattern since antiquity. Assorted pharmacological surveies have besides reported on the benefits of its infusions and merchandises on critical physiological maps including their antioxidant, 14 cardioprotective, 15 hepatoprotective, 16 anticancer 17 and anti-inflammatory effects. 18 However, most of these surveies focused on the usage of aged garlic infusion ( AGE ) or other commercial merchandises. Here we report on the anti-oxidant and anti-lipid peroxidative belongingss of fresh ethanolic infusion of local Ugandan cultivars of Allium sativum in mice theoretical accounts of Datril induced lipid per oxidation and oxidative emphasis. We hypothesize that regular ingestion of fresh Allium sativum could forestall oxidative emphasis and protect against diseases associated with oxidative emphasis and lipid peroxidation reactions. MATERIALS AND METHODS 1. Collection, Identification, and Processing of Garlic Bulbs. Bulbs of a local assortment of garlic ( Allium sativum L. ) were obtained from Ishaka Town in Western Uganda, and identified by a qualified taxonomer. Cold extraction of the Allium sativum was carried out at room temperature ( 18-22 O C ) as follows: Fresh Allium sativum bulbs were land to a all right paste utilizing a mechanical bomber and 50 g of the paste was put in a 250 milliliter conelike flask and covered with 100 milliliters of 80 % ethyl alcohol, stoppered with cotton wool, and allowed to stand in the dark at room temperature for 48 hours. The ethanolic infusion was filtered off with a Whatman no. paper into pre-weighed evaporating dishes, while the residue in the flask was washed with a farther 100 milliliter of 80 % ethyl alcohol and added to the infusions in the evaporating dishes. The filtrates were so evaporated to a syrupy residue utilizing a rotary extractor at 40 O C. The dishes were so weighed once more on a ternary beam balance and the per centum output was calcula ted as follows: Weight of extract = weight of vaporizing dish after vaporization weight of dish before add-on of infusion ; Percentage output = entire weight of extract ? weight of paste used ( 50 g ) A- 100. The infusions were pooled together into an airtight container and stored refrigerated ( at -4 oC ) until required for usage. For usage, a part of the infusion was weighed and dissolved in normal saline solution. Fresh readyings were made on each twenty-four hours of the experiment. The resulting solutions were injected intraperitonially into the mice. 2. Lab Animals Swiss mice 6-8 hebdomads old weighing 18-32 g were obtained from the Pharmacology Department of the Mbarara University of Science and Technology in Uganda. They were maintained and habituated in plastic coops in the carnal house of the School of Health Sciences, Kampala International University, Western Campus for one hebdomad, and so after used for the surveies. The mice had free entree to H2O and were fed standard gnawer pellets ( purchased from a local commercial provider ) ad libitum. Addiction conditions were 12 hour dark/light rhythms, and mean environmental temperature of 20 o C. 3. Acute Toxicity Test and Determination of LD50 The LD50 of the infusion was determined in the mice by the process described by Bernas et Al. ( 2004 ) .19 The assurance interval of the LD50 was estimated by the Litchfield Wilcoxon method utilizing a computing machine software.20 4. Experimental Design Thirty Swiss mice of both sexes were used for the experimental survey. The animate beings were grouped indiscriminately into 6 groups of 5 each and administered with the drugs/extracts as follows: Group I received physiological saline i.p. merely ; group II received acetaminophen 250 mg/kg i.p. individual dosage merely ; group III was given garlic infusion 250 mg/kg for 5 yearss before a individual i.p. dosage of acetaminophen 250 mg/kg ; group IV received 500 mg/kg garlic infusion for 5 yearss before 250 mg/kg Datril ; group V were given 750 mg/kg garlic infusion for 5 yearss before 250 mg/kg Datril ; group VI received 25 mg/kg silymarin for 5 yearss before a individual i.p dosage of acetaminophen 250 mg/kg. The infusion was administered as a individual one time day-to-day dosages, while Datril was administered after 12 hours fast. 5. Sample Collection The mice were sacrificed under quintessence anesthesia, and their livers were obtained from the mice washed with ice cold normal saline, followed by 0.15 M Tris-buffer ( pH 7.4 ) , blotted and weighed. The liver was so homogenized in 0.15 M Tris buffer to a concentration of 10 g per 100ml of homogenate and used for TBARS, glutathione, catalase, and SOD checks. 6. Biochemical Assays Thiobarbituric acid reactive substances ( TBARS ) in the liver homogenates were estimated by the method of Ohkawa et al 21 as a step of lipid peroxidation reactions. Catalase activities in the homogenates were estimated by the method of Johansson and Borg, 22 ( which depended on the reaction between methyl alcohol and catalase in the presence of H peroxide ) with kits obtained from Calbiochem USA. Superoxide dismutase check was estimated by the method of Kakkar et Al, 23 utilizing kits obtained from Calbiochem. The NWLSS GSH spectrophotometric assay kit was used for the appraisal of glutathione in the homogenates ( Northwest Life Sciences Specialties LLC, USA ) . In this method, 5-5 dithiobis ( 2-Nitrobenzoic acid ) DTNB, reacts with glutathione to organize 5-thionitrobenzoic acid ( TNB ) which has optimum soaking up at a wavelength of 412 nanometers. The maker s protocol was purely followed. 7. Datas Analysis Datas were presented as average AÂ ± criterion mistake of the mean. Statistical analysis was by the one manner analysis of discrepancy ( ANOVA ) utilizing the SPSS version 10 package, and a P value lt ; 0.05 was considered important. Consequence Administration of toxic doses of Datril produced pronounced depletion of the liver glutathione shops and the antioxidant enzymes, superoxide dismutase, and catalase, and important lift of lipid peroxidation merchandises estimated as thiobarbituric acid reactive substances ( TBARS ) . Liver glutathione degree in group II was significantly lower than in the negative control ( p lt ; 0.005 ) as are SOD ( P lt ; 0.001 ) and catalase ( p lt ; 0.05 ) . The liver TBARS degree in group II was significantly higher than in group I ( P lt ; 0.005 ) . The disposal of fresh Allium sativa infusion and silymarin protected against these alterations in a dose dependent mode and brought the values to degrees comparable to those of the negative controls ( P gt ; 0.01 ) as shown in table 1 and in figure 1. Table 1:Liver TBARS, GSH, SOD, and CAT of mice in the six groups Group Treatment TBARS ( mM/Kg ) GSH ( ug/mg protein ) Turf ( U/g liver ) CAT ( U/g liver ) I. NEG CONTROL 0.5 Master of Library Science Normal saline i.p. 11.5 Â ± 2.5 48Â ± 4.6 85Â ±6.8 85Â ±4.4 II. POS CONTROL 250 mg/Kg APAP i.p. 26.2 Â ± 1.8 P lt ; 0.005 12Â ±2.4 P lt ; 0.001 14Â ±3.6 P lt ; 0.001 50Â ± 3.9 P lt ; 0.05 III. 250 mg/Kg APAP + 250 mg/Kg garlic infusion 20 Â ±1.2 P lt ; 0.01 27Â ±4.1 P lt ; 0.01 californium. group I P lt ; 0.005 californium. group II 38Â ±2.1 P lt ; 0.001 californium. group I P lt ; 0.005 californium. group II 65Â ± 2.0 P lt ; 0.01 californium. group I P lt ; 0.05 californium. group II Four 250mg/Kg APAP + 500 mg/Kg garlic infusion 15.1 Â ±0.8 P gt ; 0.05 californium. group I ; p lt ; 0.01 californium. group II 32Â ±3.1 P lt ; 0.05 californium. group I P lt ; 0.001 californium. group II 44Â ±1.8 P lt ; 0.01 californium. group I P lt ; 0.0001 californium. group II 74Â ± 1.8 P lt ; 0.05 californium. group I P lt ; 0.005 californium. group II Volt 250 mg/kg APAP + 750 mg/Kg garlic infusion 12.2 Â ± 0.6 P gt ; 0.1 californium. group I ; P lt ; 0.001 californium. group II 38Â ±2.8 P lt ; 0.05 californium. group I P lt ; 0.001 californium. group II 62Â ±2.5 P lt ; 0.05 californium. group I ; P lt ; 0.001 californium. group II 82Â ± 2.4 P gt ; 0.1 californium. group I ; P lt ; 0.01 californium. group II Six 250 mg/Kg APAP + 25 mg/Kg silymarin 10.8 Â ±0.8 P gt ; 0.1 californium. group I ; P lt ; 0.005 californium. group II 45Â ±2.9 P gt ; 0.1 californium. group I ; P lt ; 0.0001 californium. group II 76Â ±4.8 P gt ; 0.1 californium. group I ; P lt ; 0.005 californium. group II 78Â ±2.5 P gt ; 0.1 californium. group I ; P lt ; 0.000 californium. group II Discussion Natural antioxidants play important functions in the bar and intervention of many organic and inflammatory diseases associated with oxidative stress.24 Polyphenols and flavonoids that are present in plant-derived merchandises are widely reported to exercise important influences on the remotion of reactive O and N species and have been utile in such diseases as diabetes mellitus and artherosclerosis.25 This survey demonstrated that fresh Allium sativa infusion exerted important protection against oxidative emphasis and lipid peroxidation induced by Datril overdose. It besides showed that fresh Allium sativa preserved liver GSH, and up-regulated superoxide dismutase and catalase activities in the liver. These observations are consistent with the ascertained effects of infusions from other workss in continuing liver GSH 26, and more so agrees with the study of Sabaya and others 27 in the relation to the action of Allium sativa infusion on valproic acid induced hepatotoxicity. In this re gard, Allium sativa mimics the activities of cysteine prodrugs such as N-acetyl cysteine ( NAC ) and S-adenosyl methionine ( SAM ) , which are known to continue liver GSH degrees in Datril hepatotoxicity 28, 29. It is besides possible that the infusion prevented GSH depletion by forestalling NAPQI formation in Datril overdose. The mechanism here could be suppression of enzymes of stage I metabolism, notably CYP2E1 and CYP3A, which are the primary enzymes responsible for acetaminophen biotransformation into NAPQI. Greenbaltt et al30 have shown that certain H2O soluble components of aged Allium sativum can suppress CYP3A in normal human liver microsomes. It has been suggested that drugs which can cut down cytochrome P450 mediated NAPQI formation such as Co chloride, Tagamet, and piperonyl butoxide could protect the liver against acetaminophen hepatotoxicity 31, 32. Several studies have besides shown that isothiocyanate and allyl sulphide compounds of Allium sativa inhibited cytochrome P450 enzymes such as CYP2E1 that act in stage I metamorphosis of acetaminophen 33, 34 Besides, several other surveies have reported that Allium sativa and Allium cepa ( onion ) organic sulfides are capable of hei ghtening glutathione S transferase activity in the liver,35 and isothiocyanate is a really powerful inducer of stage II metabolizing enzymes such as quinone reductase and glutathione -S transferase.36, 37 Allium sativa may besides speed up NAPQI elimination by supplying substrates that are required for its junction. Such substrates may include thiol ( organosulphure ) compounds, aminic acids, and sulphate ions. It may besides accomplish this by increasing NAPQI binding to glucuronic acid.38 Investigation of these possibilities requires surveies of the pharmacokinetics of NAPQI in animate beings having Allium sativa infusion, and the effects of Allium sativa infusion on cytochrome P450 enzymes responsible for NAPQI metamorphosis. GSH saving could ensue from the supply of substrates for GSH biogenesis by the Allium sativa infusion. Allium sativa is known to incorporate organic sulfides such as S-allyl cysteine ( SAC ) and S-allyl mercaptocysteine ( SAMC ) which could be utilized for GSH biogenesis ( 221,222 ) . 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