Browsing by Author "Hughes, R.D."

Now showing 1 - 9 of 9

Antioxidant activity of Obseckia aspera
(1998) Thabrew, M.I.; Hughes, R.D.; McFarlane, I.G.
Plants of the Osbeckia family have been shown to possess hepatoprotective properties, which could be due to the presence of antioxidant compounds. The plant extract was shown to inhibit significantly in a dose-dependent manner, the activities of the DPPH(1,1-diphenyl-2-picrylhydrazyl) free radical (EC50 of 27.5 g/mL), xanthine oxidase (EC50 of 1.16 mg/mL) and demonstrate a scavenging effect on hydroxyl radical mediated damage to deoxyribose (EC50 of 140 g/mL). The plant extract possessed some prooxidant activity from the effect on bleomycin-induced DNA damage, but this was less than that shown by comparable concentrations of (+)-catechin or silymarin
Assay to detect inhibitory substances in serum of patients with acute liver failure
(Wichtig Editore, 1999) Anderson, C.; Thabrew, M.I.; Hughes, R.D.
Patients with acute liver failure accumulate toxic substances in the circulation which may impair recovery of hepatic function. The aim of this study was to test an in vitro assay to detect inhibitory substances in the serum of patients with acute liver failure. Human liver-derived HepG2 cells were incubated for 24h in 96 well plates (30,000 cells/well) with sera (10%) from 24 patients with acute liver failure due to paracetamol overdose or NANB hepatitis and 11 normal controls. DNA synthesis was determined from the incorporation of 3H-thymidine and cell viability by the metabolism of the tetrazolium dye MTS. HepG2 cells exposed to acute liver failure sera incorporated significantly less 3H-thymidine (median 30% of control, range 0.2-169%) than normal sera (100%, 76-133%, p=0.002). Cell viability was also reduced (75%, 33-112% vs 100%, 96-105%, p<0.001). There was no correlation between these values and patient outcome or levels of plasma TNF-alpha or serum interferon-gamma. The assay detected inhibitory substances in sera of patients with acute liver failure and could be used to monitor the use of liver support systems.
Cytotoxic effects of a decoction of Nigella sativa, Hemidesmus indicus and Smilax glabra on human hepatoma HepG2 cells
(Elsevier, 2005) Thabrew, M.I.; Mitry, R.R.; Morsy, M.A.; Hughes, R.D.
A decoction of Nigella sativa seeds, Hemidesmus indicus root and Smilax glabra rhizome is used by traditional medical practitioners in Sri Lanka to treat cancer and has been shown to prevent chemically induced carcinogenesis in rats. The cytotoxicity of the decoction and the individual plant extracts were tested on the human hepatoma HepG2 cell line. The effects of 24 h incubation with different concentrations (0--50 mg/ml) of the extracts on HepG2 cells were determined. Results from MTT and SRB assays, and [(14)C]-leucine and [(3)H]-thymidine uptake demonstrated that the decoction had a strong dose-dependent cytotoxic activity. The greatest inhibitory effects were observed on DNA synthesis with both the decoction (91+/-S.E. 3.7% inhibition) and N. sativa plant extract (88+/-3.8%) even at low concentrations (5 mg/ml). The three individual plant extracts were cytotoxic in the order of potency N. sativa>H. indicus>S. glabra. Flow cytometric analysis using Annexin V and propidium iodide staining showed that after 24 h exposure to the decoction, cells were in the late stage of apoptosis and/or necrosis. Further experiments are worthwhile to determine the anticancer potential of this plant decoction and its components.
Phenolic and terpenoid constituents from the Sri Lankan medicinal plant Osbeckia aspera
(Informa Healthcare, 2008) Grayer, R.J.; Thabrew, M.I.; Hughes, R.D.; Bretherton, S.; Lever, A.; Veitch, N.C.; Kite, G.C.; Lelli, R.
A crude aqueous acetone extract of Osbeckia aspera. Blume (Melastomataceae), a plant from Sri Lanka used traditionally to treat liver disease, was fractionated by column and preparative paper chromatography, and the fractions were analyzed by high-performance liquid chromatography (HPLC) using diode array and mass spectrometric detection. Phenolic acids (gallic, protocatechuic, and ellagic acid), flavonol glycosides [quercetin 3-O.-β-galactopyranoside, quercetin 3-O.-β-glucopyranoside, kaempferol 3-O.-β-glucopyranoside, and kaempferol 3-O.-(6″-O.-p.-coumaroyl-β-glucopyranoside) (tiliroside)] and flavonol aglycones (quercetin and kaempferol) were identified by comparison of their retention times, UV and MS spectra with those of authentic standards. Five compounds from a methanol extract were identified by NMR spectroscopy as the flavonol glycosides, quercetin 3-O.-(3″-O.-acetyl-β-galactopyranoside) and kaempferol 3-O.-[2″,6″-di-O.-(E.,E.)-p.-coumaroyl-β-glucopyranoside], and the norsesquiterpenoids 6,9-dihydroxy-4,7-megastig-madien-3-one, 9-hydroxy-4,7-megastigmadien-3-one and 9-hydroxy-4-megastigmen-3-one. A crude water extract, 50% acetone extract and fractions from this extract, a 100% methanol extract, and three of the phenolic acids in the fractions were tested for in vitro. hepatoprotective activity against bromobenzene and 2,6-dimethyl-N.-acetyl p.-quinoneimine toxicity to HepG2 liver-derived cells. The crude water extract showed protective activity against both liver toxins, whereas the fractions and compounds were more protective against 2,6-dimethyl-N.-acetyl p.-quinoneimine than bromobenzene. Of the three phenolic acids present in the extracts that were tested, gallic and protocatechuic acids were more active at protecting the liver cells from the two toxic compounds than ellagic acid
Phytogenic agents in the therapy of the Liver disease
(1996) Thabrew, M.I.; Hughes, R.D.
Plant extracts have been used by traditional medical practitioners for the treatment of liver disorders for centuries. This article reviews the clinical trials carried out with thirteen plants and their constituents in patients with liver disease, including acute viral hepatitis, chronic viral hepatitis, chronic cholecystitis, alcoholic liver disease and mushroom poisoning. There is considerable scientific evidence that phytogenic agents can have significant beneficial effects on liver dysfunction and the course of liver disease. At present, silymarin has the most proven overall clinical hepatoprotective effects, although glycyrrhizin appears to be more beneficial in chronic viral hepatitis. With the high worldwide incidence of viral hepatitis, further study of isolated phytochemicals is important in relation to their potential antiviral activity against the different hepatitis viruses.
Protection against galactosamine and test-butyl hydroperoxide induced hepatocyte damage by Melothria maderaspatana extract
(Wiley, 1995) Thabrew, M.I.; Gove, C.D.; Hughes, R.D.; McFarlane, I.G.; Williams, R.
The aqueous extract of Melothria maderaspatana is used by traditional medical practitioners to treat jaundice in man. The effect of Melothria maderaspatana extract on damage induced in freshly isolated rat hepatocytes by D-galactosamine and tert-butyl hydroperoxide (TBH) has been investigated. On incubation of hepatocytes with galactosamine or TBH in the presence of the plant extract, a significant dose-dependent protection against hepatocyte damage was observed, with maximum protection at a concentration of 500 g/mL. At this concentration the galactosamine-induced release of lactate dehydrogenase (LDH) and aspartate amino-transferase (AST) were reduced by 40.7 percent + or - 4.2 percent and 37.7 percent + or - 6.1 percent respectively, compared with control incubations. The TBH-induced lipid peroxidation (estimated from malondialdehyde production) was decreased by 26.0 + or - 3.7 percent together with a 38.4 percent + or - 4.4 percent and 40.8 + or - 7.6 percent reduction in the release of cellular LDH and AST respectively into the incubation medium. On post-treatment with the plant extract the protective activity was found to decrease with increase in time of exposure of the cells to either of the toxins. The direct protective effects of Melothria extract on hepatocytes support the use of this plant as a herbal remedy.
Protective effects of Osbeckia octandra against galactosamine and tert-butyl hydroperoxide induced hepatocyte damage
(Elsevier, 1995) Thabrew, M.I.; Gove, C.D.; Hughes, R.D.; McFarlane, I.G.; Williams, R.
Ayurvedic and other 'traditional' medical practitioners in Sri Lanka use the mature leaves of the plant Osbeckia octandra for its hepatoprotective properties. In this study the effects of an aqueous extract of Osbeckia octandra against injury induced by D-galactosamine and tert-butyl hydroperoxide (TBH) were investigated in freshly isolated rat hepatocytes. The plant extract (500 micrograms/ml) significantly reduced the inhibition of protein synthesis (as assessed by the incorporation of 14C-leucine into protein) in hepatocytes incubated for 1 h with 10 mM galactosamine by a mean of 25.6 +/- 3.6% and decreased the release of cellular lactate dehydrogenase (LDH) and aspartate aminotransferase (AST) enzyme activities into the medium by 55.3% and 32.8%, respectively. With TBH, the plant extract decreased lipid peroxidation (estimated from malondialdehyde formation) by a mean of 29.9 +/- 1.1% together with a 46.8% and 54.7% decrease in the release of LDH and AST, respectively into the incubation medium. Significant protection was also obtained when the Osbeckia extract was added to the incubation medium up to 30 min after pre-exposure of the hepatocytes to either galactosamine or, to a lesser extent, TBH. The results support the use of Osbeckia as a hepatoprotective agent.
Protective effects of Osbeckia octandra against paracetamol-induced liver injury
(Informa Healthcare, 1995) Thabrew, M.I.; Hughes, R.D.; Gove, C.D.; Portmann, B.; Williams, R.; McFarlane, I.G.
Osbeckia octandra is a plant used in traditional medicine to treat jaundice and other liver disorders. In this study, the effects of Osbeckia leaf extract on paracetamol-induced liver injury were investigated both in vivo in mice and in rat hepatocytes in vitro. 2. Oral administration of Osbeckiaextract (330 mg/kg) at the same time as paracetamol (450 mg/kg) to mice, resulted in a significant protection (p < 0.05) against liver damage, as assessed by improvements in the blood Normotest (39.1 +/- 1.9 versus 46.3 +/- 2.0 s), total liver glutathione (730 +/- 39 versus 574 +/- 27 micrograms/250 mg liver), plasma aspartate aminotransferase level (916 +/- 225 versus 1965 +/- 291 iu/l), and liver histopathology at 24 h after paracetamol administration. 3. In experiments to assess the direct effects of Osbeckia extract, significant protection was also found in freshly isolated rat hepatocytes against damage induced by 185 microM 2,6-dimethyl N-acetyl p-quinoneimine (2,6-diMeNAPQI, an analogue of NAPQI, the toxic metabolite of paracetamol) in vitro. When Osbeckia extract (500 micrograms/ml) was added to the incubation medium at the same time as 2,6-diMeNAPQI significant changes in cell viability (78.4 +/- 3.3 versus 47.2 +/- 5.8% of control, p < 0.001), cell reduced glutathione (GSH) level (35.0 +/- 3.1 versus 23.8 +/- 1.5%, p = 0.009), and reduced release of lactate dehydrogenase (129.9 +/- 6.6 versus 224.6 +/- 12.1%, p < 0.001) were demonstrated after 1 h incubation as compared with 2,6-diMeNAPQI alone. 4. Significant protection was still obtained against 2,6-diMeNAPQI in vitro when addition of Osbeckia extract was delayed by 20 min. These results indicate that Osbeckia extract can protect against paracetamol-induced liver injury
Screening of hepatoprotective plant components using a HepG2 cell cytotoxicity assay
(Wiley, 1997) Thabrew, M.I.; Hughes, R.D.; Mcfarlane, I.G.
Identification of the active components of plants with hepatoprotective properties requires screening of large numbers of samples during fractionation and purification. A screening assay has been developed based on protection of human liver-derived HepG2 cells against toxic damage. Various hepatotoxins were incubated with HepG2 cells in 96-well microtitre plates (30,000 cells well-1) for 1 h and viability was determined by metabolism of the tetrazolium dye 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxy phenyl)-2-(4-sulphophenyl)-2H-tetrazolium (MTS). Bromobenzene (10 mM) and 2,6-dimethyl-N-acetyl-p-quinoneimine (2,6-diMeNAPQI, 200 mM) had greater toxic effects than tert-butyl hydroperoxide (1.8 mM) or galactosamine (10 mM), reducing mean viability to 44.6 +/- 1.2% (s.e.m.) and 56.1 +/- 2.1% of control, respectively. Protection against toxic damage by these agents was tested using a crude extract of a known hepatoprotective Sri Lankan plant, Osbeckia aspera, and two pure established hepatoprotective plant compounds, (+)-catechin and silymarin (1 mg mL-1). Viability was significantly improved by Osbeckia (by 37.7 +/- 2.4%, P < 0.05, and 36.5 +/- 2.1%, P < 0.05, for bromobenzene and 2,6-diMeNAPQI toxicity, respectively). Comparable values for (+)-catechin were 68.6 +/- 2.9% and 63.5 +/- 1.1%, and for silymarin 24.9 +/- 1.4% and 25.0 +/- 1.6%. This rapid and reproducible assay should prove useful for the isolation and identification of active hepatoprotective compounds in crude plant extracts.