Abstract
BackgroundSida cordata, a member of Family Malvaceae is used in folk medicine for various ailments including liver diseases. In this study we investigated, its flavonoid constituents, in vitro antioxidant potential against different free radicals and hepatoprotection against carbon tetrachloride (CCl4)-induced liver damage in rat.MethodsDried powder of S. cordata whole plant was extracted with methanol and the resultant (SCME) obtained was fractionated with escalating polarity to obtain n-hexane fraction (SCHE), ethyl acetate fraction (SCEE), n-butanol fraction (SCBE) and the remaining soluble portion as aqueous fraction (SCAE). Diverse in vitro antioxidants assays such as DPPH, H2O2, •OH, ABTS, β-carotene bleaching assay, superoxide radical, lipid peroxidation, reducing power, and total antioxidant capacity were studied to assess scavenging potential of methanol extract and its derived fractions. On account of marked scavenging activity SCEE was selected to investigate the hepatoprotective potential against CCl4 induced toxicity in Sprague–Dawley male rats by assessing the level of serum markers (alkaline phosphatase, alanine transaminase, aspartate transaminase, lactate dehydrogenase, bilirubin, and γ-glutamyltransferase) and of liver antioxidant enzymes such as catalase (CAT), superoxide dismutase (SOD), peroxidase (POD), glutathione-S-transfers (GST), glutathione reductase (GSR), glutathione peroxidase (GSH-Px), and reduced glutathione (GSH) and lipid peroxidation (TBARS). Histology of the liver was performed to study alteration in histoarchitecture. Existence of active flavonoids was established by thin layer chromatographic studies.ResultsConsiderable amount of flavonoid and phenolic contents were recorded in the methanol extract and its derived fractions. Although the extract and all its derived fractions exhibited good antioxidant activities however, the most distinguished scavenging potential was observed for SCEE. Treatment of SCEE decreased the elevated level of serum marker enzymes induced with CCl4 administration whereas increased the activity of hepatic antioxidant enzymes (CAT, SOD, POD, GST, GSR and GSH-Px). Hepatic concentration of GSH was increased while lipid peroxidation was decreased with SCEE administration in CCl4 intoxicated rats. Presence of apigenin with some unknown compounds was observed in SCEE by using thin layer chromatography.ConclusionsThese results revealed the presence of some bioactive compound in the ethyl acetate fraction, confirming the utility of S. cordata against liver diseases in folk medicine.
Highlights
Sida cordata, a member of Family Malvaceae is used in folk medicine for various ailments including liver diseases
There has been a mounting interest in the medical implications of free radicals. Free radicals such as superoxide radical (O2-), hydroxyl radical (OH), hydrogen peroxide (H2O2) and lipid peroxides (LOOH) are becoming of enormous concern in human diseases. They are recognized for DNA damage, lipid peroxidation and protein breakdown along with role in the pathogenesis of many clinical disorders such as inflammatory diseases, cardiac diseases, asthma, Alzheimer’s, Parkinson’s and aging [2,3]
Extract and fraction yield S. cordata crude methanol extract gave a yield of 18 percent (w/w), proceeded to further fractionation by using different organic solvents based on a difference of polarity index
Summary
Plant collection and preparation of extract The whole plant was collected from the campus of Quaidi-Azam University, Islamabad, Pakistan and recognized by their local names and confirmed by Prof. Antioxidant assays Samples preparation S. cordata extract, fractions and positive standards (ascorbic acid, butylated hydroxytoluene, catechin and gallic acid) 200 μg were dissolved in 1 ml analytical methanol. These solutions were further serially diluted to 100 μg/ml and 50 μg/ml. Each sample (100 μl) at varying concentrations (200, 100, 50 μg/ml in methanol) and 500 μl of yolk homogenate were pipetted into eppendorfs and volume was made up to 1 ml with distilled water It was mixed with 1.5 ml of acetic acid (20%, pH 3.5) and TBA (0.8%, w/v) in sodium dodecyl sulphate (1.1%, w/v). Correlation between IC50 values of different assays with total flavonoids and total phenolics was calculated by Pearson’s correlation coefficient with a significance level of P
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