Ascorbic Acid-induced Lipid Peroxidation Research Articles

Protective effect of black raspberry seed containing anthocyanins against oxidative damage to DNA, protein, and lipid.

This study aimed to determine bioactive components and radical scavenging capacity of black raspberry seed extracts as byproducts obtaining during the juice (FSE) and wine (WSE) making process. Cyanidin-3-O-rutinoside was identified as a major anthocyanin and the total anthocyanin contents of fresh and wine seed were 78.24 and 41.61mg/100g of dry weight, respectively. The total phenolic and flavonoid contents of FSE and WSE were 2.31g gallic acid equivalent (GAE) and 360.95mg catechin equivalent (CE), and 2.44g GAE and 379.54mg CE per 100g dry weight, respectively. The oxygen radical absorbance capacity (ORAC) values were 1041.9μM TE/g for FSE and 1060.4μM TE/g for WSE. Pretreatment of the FSE and WSE inhibited the generation of intracellular reactive oxygen species (ROS), DNA and protein damage induced by hydroxyl radicals, and Fe(3+)/ascorbic acid-induced lipid peroxidation in a dose dependent manner. WSE more effectively protected from oxidative damage than FSE. Results from the current study suggest that black raspberry seeds as byproducts from juice and wine processing could be potential sources for natural antioxidants.

Combination of counterpropagation artificial neural networks and antioxidant activities for comprehensive evaluation of associated-extraction efficiency of various cyclodextrins in the traditional Chinese formula Xue-Zhi-Ning

Xue-Zhi-Ning (XZN) is a widely used traditional Chinese medicine formula to treat hyperlipidemia. Recently, cyclodextrins (CDs) have been extensively used to minimize problems relative to medicine bioavailability, such as low solubility and poor stability. The objective of this study was to determine the associated-extraction efficiency of various CDs in XZN. Three various type CDs were evaluated, including native CDs (α-CD, β-CD), hydrophilic CD derivatives (HP-β-CD and Me-β-CD), and ionic CD derivatives (SBE-β-CD and CM-β-CD). An ultra high-performance liquid chromatography (UHPLC) fingerprint was applied to determine the components in CD extracts and original aqueous extract (OAE). A counterpropagation artificial neural network (CP-ANN) was used to analyze the components in different extracts and compare the selective extraction of various CDs. Extraction efficiencies of the various CDs in terms of extracted components follow the ranking, ionic CD derivatives>hydrophilic CD derivatives>native CDs>OAE. Besides, different types of CDs have their own selective extraction and ionic CD derivatives present the strongest associated-extraction efficiency. Antioxidant potentials of various extracts were evaluated by determining the inhibition of spontaneous, H2O2-induced, CCl4-induced and Fe2+/ascorbic acid-induced lipid peroxidation (LPO) and analyzing the scavenging capacity for DPPH and hydroxyl radicals. The order of extraction efficiencies of the various CDs relative to antioxidant activities is as follows: SBE-β-CD>CM-β-CD>HP-β-CD>Me-β-CD>β-CD>α-CD. It can be demonstrated that all of the CDs studied increase the extraction efficiency and that ionic CD derivatives (SBE-β-CD and CM-β-CD) present the highest extraction capability in terms of amount extracted and antioxidant activities of extracts.

In vitro antioxidant and cytotoxic activities of Arnebia benthamii (Wall ex. G. Don): a critically endangered medicinal plant of Kashmir Valley.

Arnebia benthamii is a major ingredient of the commercial drug available under the name Gaozaban, which has antibacterial, antifungal, anti-inflammatory, and wound-healing properties. In the present study, in vitro antioxidant and anticancer activity of different extracts of Arnebia benthamii were investigated. Antioxidant potential of plant extracts was evaluated by means of total phenolics, DPPH, reducing power, microsomal lipid peroxidation, and hydroxyl radical scavenging activity. The highest phenolic content (TPC) of 780 mg GAE/g was observed in ethyl acetate, while the lowest TPC of 462 mg GAE/g was achieved in aqueous extract. At concentration of 700 µg/mL, DPPH radical scavenging activity was found to be highest in ethyl acetate extract (87.99%) and lowest in aqueous extract (73%). The reducing power of extracts increased in a concentration dependent manner. We also observed its inhibition on Fe2+/ascorbic acid-induced lipid peroxidation (LPO) on rat liver microsomes in vitro. In addition, Arnebia benthamii extracts exhibited antioxidant effects on Calf thymus DNA damage induced by Fenton reaction. Cytotoxicity of the extracts (10–100 µg/mL) was tested on five human cancer cell lines (lung, prostate, leukemia, colon, and pancreatic cell lines) using the Sulphorhodamine B assay.

Antioxidant and hepatoprotective effects of Crataegus songarica methanol extract.

The protective activity of the methanolic extract of the Crataegus songarica leaves was investigated against CCl4- and paracetamol-induced liver damage. On folklore levels, this plant is popularly used to treat various toxicological diseases. We evaluated both in vitro and ex vivo antioxidant activity of C. songarica. At higher concentration of plant extract (700 µg/ml), 88.106% inhibition on DPPH radical scavenging activity was observed and reducing power of extract was increased in a concentration-dependent manner. We also observed its inhibition on Fe2+/ascorbic acid-induced lipid peroxidation on rat liver microsomes in vitro. In addition, C. songarica extract exhibited antioxidant effects on calf thymus DNA damage induced by Fenton reaction. Hepatotoxicity was induced by challenging the animals with CCl4 (1 ml/kg body weight, i.p.) and paracetamol (500 mg/kg body weight) and the extract was administered at three concentrations (100, 200, and 300 mg/kg body weight). Hepatoprotection was evaluated by determining the activities of liver function marker enzymes and antioxidant status of liver. Administration of CCl4 elevated the levels of liver function enzymes, SGOT, SGPT, and LDH. We also observed a dramatic increase in ALT, AST, bilirubin, and alkaline phosphatase levels in rats administered 500 mg/kg body weight of paracetamol. Decreased antioxidant defense system as glutathione (GSH), catalase (CAT), glutathione peroxidase (GPX), glutathione reductase (GR), glutathione-S-transferase (GST), and superoxide dismutase (SOD) were observed in rats treated with CCl4 and paracetamol. Pretreatment with the extract decreased the elevated serum GOT, GPT, LDH, bilirubin, and alkaline phosphatase activities and increased the antioxidant enzymes in a dose-dependent manner. Therefore, C. songarica methanol extract may be an effective hepatic protective agent and viable candidate for treating hepatic disorders and other oxidative stress-related diseases.

IN VITRO LIPID PEROXIDATION INHIBITORY AND ANTI-ARTHRITIC ACTIVITIES OF SOME INDIAN MEDICINAL PLANTS

Anti-lipid peroxidation effect of aqueous extracts of Ocimum sanctum, Tinospora cordifolia and Withania somnifera was evaluated against Fe2+-ascorbic acid-induced lipid peroxidation using rat liver mitochondria as model system, whereas, anti-arthritic activity was evaluated by proteinase inhibitory assay. O. sanctum showed potent anti-lipid peroxidation and anti-arthritic activities. T. cordifolia exhibited moderate anti-lipid peroxidation activity, but considerable anti-arthritic activity, whereas, W. somnifera revealed least lipid peroxidation inhibitory activity and considerable anti-arthritic activity. Besides, Folin-Ciocalteu reagent in terms of gallic acid equivalent achieved the total phenolic content and the trend was found to be O. sanctum > T. cordifolia > W. somnifera.

High molecular weight persimmon tannin is a potent antioxidant both ex vivo and in vivo

The antioxidant activities of high molecular weight persimmon condensed tannin (HMWPT) were evaluated in an ex vivo tissue system and in vivo. Addition of HMWPT to mouse liver homogenate protected the samples against auto-peroxidation or H2O2- or Fe2+/ascorbic acid-induced lipid peroxidation. The IC50 values for HMWPT were 4.32±0.20μg/mL (auto-oxidation), 1.36±0.40μg/mL (H2O2-induced peroxidation) or 0.20±0.09mg/mL (Fe2+/ascorbic acid-induced peroxidation). Mice were oxidatively stressed with bromobenzene to test the antioxidant activity of HMWPT in vivo. An oral dose of HMWPT at 200 or 400mg/kg HMWPT significantly (P<0.01) prevented the bromobenzene-induced decrease in serum and liver superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities, and decreased liver malondialdehyde levels in bromobenzene-treated mice (P<0.01). The results suggest that dietary HMWPT may provide protection from oxidative damage both ex vivo and in vivo.

Anti-hypercholesterolaemia, antioxidant activity and free radical scavenger effects of traditional Chinese medicine prescriptions used for stroke.

The generation of oxygen radicals and lipid peroxidation may be factors in the cerebral damage secondary to ischaemia of the cerebrovascular disease, as in stroke. Five traditional Chinese medicinal prescriptions were investigated for their antioxidant activity: Shiee Fuu Jwu Iu Tang (TCMP1), Oh Yaw Shuen Chin Saan (TCMP2), Buu Yang Hwan Wuu Tang (TCMP3), Sheau Shiuh Ming Tang (TCMP4), and Chir Hwu Jia Long Guu Muu Lih Tang (TCMP5). Anti-lipid peroxidation, anti-superoxide formation and free radical scavenger activity were determined by the FeCl2--ascorbic acid-induced lipid peroxidation effects on lipids in-vitro, xanthine oxidase inhibition, cytochrome C system and an electron spin resonance spectrometer, respectively. The results showed that TCMP5 had greater anti-lipid peroxidation and anti-superoxide formation activity than the other prescriptions. TCMP4 had the greatest free radical scavenging effect, TCMP5 showed the greatest superoxide radical scavenger activity and TCMP3 showed the greatest hydroxyl radical scavenger activity. Tests were also performed to evaluate the effects of the five prescriptions on blood lipid in-vivo. The test showed that the prescriptions decreased the level of total cholesterol and LDL-cholesterol in serum in high cholesterol-fed rats. From these results, it seems probable that these prescriptions may be effective in the prevention and therapy of stroke and ischaemia.

Effects of Tryptophan and 5-Hydroxytryptophan on the Hepatic Cell Membrane Rigidity Due to Oxidative Stress

The ability of several indoleamines to scavenge free radicals is well documented. Our aim was to evaluate the ability of 0.01-3 mM tryptophan (Trp) and 0.1-5 mM 5-hydroxytryptophan (5-OH-Trp) to protect hepatic cell membranes against 0.1 mM FeCl(3) plus 0.1 mM ascorbic acid-induced lipid peroxidation and increases in membrane rigidity. Membrane fluidity was evaluated using fluorescence spectroscopy. Lipid and protein oxidation were estimated by quantifying malondialdehyde (MDA) plus 4-hydroxyalkenals (4-HDA) concentrations and carbonyl group content, respectively. Exposure to FeCl(3) plus ascorbic acid increased hepatic cell membrane rigidity, MDA + 4-HDA and carbonyl content. The presence of 5-OH-Trp, but not Trp, attenuated these changes. In the absence of oxidative stress, neither indoleamine modified fluidity, MDA + 4-HDA or carbonylation. These results suggest that C5 hydroxylation determines the ability of Trp to preserve membrane fluidity in the presence of oxidative stress.

Evaluation of antioxidant profile and activity of amalaki (Emblica officinalis), spirulina and wheat grass

Aqueous and alcoholic extracts of amalki (Emblica officinalis), spirulina and wheatgrass were prepared and analyzed for antioxidant vitamin content (vitamin C and E), total phenolic compounds. Antioxidant status, reducing power and effect on glutathione S-transferase (GST) activity were evaluated in vitro. Vitamin C content of crude amalaki powder was found to be 5.38 mg/g, while very less amount 0.22 mg/g was detected in wheat grass. Amalki was rich in vitamin E like activity, total phenolic content, reducing power and antioxidant activity. Total antioxidant activity of aqueous extract of amalki, spirulina and wheat grass at 1mg/ml concentration were 7.78, 1.33 and 0.278 mmol/l respectively. At similar concentrations the total antioxidant activity of alcoholic extract of amalaki, spirulina and wheat grass was 6.67, 1.73 and 0.380 mmol/l respectively. Amalki was also found to be rich source of phenolic compounds (241mg/g gallic acid equivalent). Alcoholic extract of wheat grass showed 50 % inhibition in FeCl(2)- ascorbic acid induced lipid peroxidation of rat liver homogenates in vitro. Both aqueous and alcoholic extracts of amalaki inhibited activity of rat liver glutathione S-transferase (GST) in vitro in dose dependant manner. Since GST acts as powerful drug metabolizing enzyme its inhibition by amalaki offers possibility of its use for lowering therapeutic dose of herbal preparations. The aqueous extracts of both amalki and spirulina also showed protection against t-BOOH induced cytotoxicity and production of ROS in cultured C(6) glial cells.

Antidiabetic Properties of 2,5-Dihydroxy-4,3′-Di(β-D-Glucopyranosyloxy)-trans-Stilbene from Mulberry (Morus bombycisKoidzumi) Root in Streptozotocin-Induced Diabetic Rats

We investigated the antidiabetic properties of 2,5-dihydroxy-4,3-di(beta-D-glucopyranosyloxy)-trans-stilbene (DGTS) isolated from Morus bombycis Koidzumi in streptozotocin (STZ)-induced diabetic rats. The DGTS prevented the increase in aspartate aminotransferase, alanine aminotransferase, and blood urea nitrogen levels in serum of diabetic rats. At doses of 200-800 mg/kg, DGTS improved hyperglycemia in the rats, and the hypoglycemic effect of DGTS was comparable to that of tolbutamide. The histological observations showed that DGTS prevented atrophy of pancreatic beta-cells and vascular degenerative changes in the islets. DGTS reversed STZ-induced diabetes and had antioxidant activity in assays of FeCl(2)/ascorbic acid-induced lipid peroxidation in the rats. Levels of cytochrome P450 2E1 mRNA, as measured by reverse transcription-polymerase chain reaction, were lower in the livers of the DGTS-treated rats than those of the control group. These results suggest that DGTS might be beneficial in the treatment of type 1 diabetes.

Hepatoprotective and antioxidant effects of Morus bombycis Koidzumi on CCl 4-induced liver damage

The antioxidant activity and liver protective effect of Morus bombycis Koidzumi were investigated. Aqueous extracts of M. bombycis Koidzumi had higher superoxide radical scavenging activity than other types of extracts. The aqueous extract at a dose of 100 mg/kg showed significant hepatoprotective activity when compared with that of a standard agent. The biochemical results were confirmed by histological observations indicating that M. bombycis Koidzumi extract together with CCl 4 treatment decreased ballooning degeneration. The water extract recovered the CCl 4-induced liver injury and showed antioxidant effects in assays of FeCl 2–ascorbic acid-induced lipid peroxidation in rats. Based on these results, we suggest that the hepatoprotective effect of the M. bombycis Koidzumi extract is related to its antioxidative activity.

Free radical scavenging and hepatoprotective actions of Quercus aliena acorn extract against CCl4-induced liver

In the present study, we investigated the protective effect of Quercus aliena acorn extracts against CCl4-induced hepatotoxicity in rats, and the mechanism underlying the protective effects. Aqueous extracts of Quercus aliena acorn had higher superoxide radical scavenging activity than other types of extracts. The Quercus aliena acorn extracts displayed dose-dependent superoxide radical scavenging activity (IC50 = 4.92 μg/ml), as assayed by the electron spin resonance (ESR) spin-trapping technique. Pretreatment with Quercus aliena acorn extracts reduced the increase in serum aspartate aminotransferase (AST) and serum alanine aminotransferase (ALT) levels. The hepatoprotective action was confirmed by histological observation. The aqueous extracts reversed CCl4-induced liver injury and had an antioxidant action in assays of FeCl2- ascorbic acid induced lipid peroxidation in rats. Expression of cytochrome P450 2E1 (CYP2E1) mRNA, as measured by RT-PCR, was significantly decreased in the livers of Quercus aliena acorn-pretreated rats compared with the livers of the control group. These results suggest that the hepatoprotective effects of Quercus aliena acorn extract are related to its antioxidative activity and effect on the expression of CYP2E1.

Antioxidant activity of Cinnamomum cassia.

The antioxidant activities of hot water extracts (HWECC) and ethanol extracts (EECC) from the dry bark of Cinnamomum cassia Presl were evaluated in this study. Results showed that at 1.0 mg/mL, the ethanol extracts of C. cassia (96.30%) exhibited a greater inhibition than the alpha-tocopherol (93.74%) on FeCl(2)-ascorbic acid induced lipid peroxidation of rat liver homogenate in vitro. From 0.05 to 1.0 mg/mL, the EECC demonstrated the highest superoxide anions scavenging activity and the strongest anti-superoxide formation activity (p < 0.05). The same extract also showed an excellent antioxidant activity in enzymatic and nonenzymatic liver tissue oxidative systems. EECC revealed the strongest antioxidant activity followed by alpha-tocopherol and HWECC. Compared to alpha-tocopherol, the IC(50) values of EECC were found to be lower in thiobarbituric acid test (IC(50) = 0.24 mg/mL vs 0.37 mg/mL), in cytochrome c test (IC(50) = 0.16 mg/mL vs 0.27 mg/mL) and in xanthine oxidase inhibition test (IC(50) = 0.09 mg/mL vs 0.19 mg/mL). The present study concludes that EECC could be used as a good source of antioxidant in the dietary supplement.

Antioxidant activity of medicinal plants used in phytotherapy

Oxygen free radicals play an important role in the development of different disorders like inflammatory-immune injury, carcinogenesis, hepatic toxicity and artherosclerosis. The antioxydant role of a wide spectrum of natural products has been established. Flavonoids and other phenolic compounds (proanthocyanidins, rosmarinic acid, hydroxicinnamic derivatives, catechines, etc.) of plant origin have been reported as scavengers and inhibitors of lipid peroxidation.&#x0D; We have studied the antioxidant activity as well as content and composition of natural phenolics in a series of medicinal plants with phytotherapeutical significance. Thus we determined the total phenol contents and studied the composition of flavonoids, polyphenols, phenolic acids of different vegetative and reproductive organs of medicinal plants: Anthriscus cerefolium (L.) Hoffm., Petroselinum crispum L., Cichorium intybus L., Helichrysum arenarium D.C.„cempervivum tectorum L., Taravacum officinale Web.&#x0D; Characteristic constituents in the various crude drugs were determined by chromatographic (TLC, HPLC) and spectroscopic (UV, UV-VIS) methods. The non specific scavenger activities of the medicinal plant extracts were studied by the chemiluminometric technique. The changes of chemiluminescence intensity of the H,G,•0H-luminol system at increasing concentrations of the H702/ -OH were measured. Inhibitory effects of selected standardized fractions from plants were tested on ascorbic acid induced lipid peroxidation in rat liver and homogenates.&#x0D; The best correlation were established with total phenolics in some medicinal plants (S. tectorum, T. officinale) while activities in other cases seem to be influenced by flavonoids (P. crispum, H. arenarium, A. cerefolium) and by hydroxicinnamic derivatives (C. intybus).&#x0D;

Evaluation of the Antioxidant Activity of Legumes

The anti-lipid peroxidation activity, free radical scavenger activity and anti-superoxide formation of hot water extracts of legumes (HWEL), such as mung bean (Phaseolus radiatus L.), adzuki bean (Phaseolus aureus Roxb.), black bean (Glycine max (L.) Merr.) and rice bean (Phaseolus calcaratus Roxb.), were evaluated to test antioxidant activity. The results showed that all HWEL exhibited remarkable inhibition of FeCl 2 -ascorbic acid induced lipid peroxidation of mouse liver homogenate. All extracts showed anti-lipid peroxidation activities. Both mung bean and adzuki bean extracts demonstrated the strongest anti-lipid peroxidation activity and the highest superoxide anion scavenging activity. All extracts of legumes showed anti-superoxide formation activity. Treatment with black bean extracts exhibited the most powerful activity. Therefore, a component of the diet, HWEL, is a potential antioxidant.

Prooxidant role of histidine in hypoxic stressed mice and Fe3+-induced lipid peroxidation

An attempt was made to study the effect of histidine on reactive oxygen species in a rodent model of hypoxic stress and in Fe(3+)-ascorbic acid-induced lipid peroxidation in mouse brain homogenates. The latency for onset of hypoxic stress-induced convulsions was decreased in histidine-treated animals with a concomitant rise in brain lipid peroxidation levels. In vitro, histidine potentiated Fe(3+)-ascorbic acid-induced lipid peroxidation in mouse brain homogenates while other antioxidants like B-HT and U-74500A inhibited the same. Moreover, Fe(3+)-histidine-induced lipid peroxidation could not be inhibited by preincubation of the system with high concentrations of ascorbic acid. Thus, it is concluded that histidine acts as a strong prooxidant potentiating the genesis of reactive oxygen species during hypoxic stress as well as during Fe(3+)-ascorbic acid-induced lipid peroxidation.

Free radical scavenging and membrane protective effects of methanol extracts from Anthriscus cerefolium L. (Hoffm.) and Petroselinum crispum(Mill.) nym. ex A.W. Hill.

The antilipoperoxidant activity of Anthriscus cerefolium L. (Hoffm.), chervil, Petroselinum crispum (Mill.) Nym. ex A. W. Hill., parsley extracts were evaluated with ascorbic acid induced lipid peroxidation on rat brain homogenates. These results are completed by the antiradical potential of these extracts against a solution of OH. radical. In all cases luteolin-7-O-glucoside was used as a reference material.

Triterpene antioxidants from ganoderma lucidum.

Ganoderma lucidum was studied for its antioxidative activity by bioassay guided isolation in conjunction with in vitro tests. The powdered crude drug was treated with boiling water and the aqueous extract (Ex1) was further separated to obtain terpene and polysaccharide fractions. The two fractions and Ex1 were screened for their antioxidative effect against pyrogallol induced erythrocyte membrane oxidation and Fe (II)-ascorbic acid induced lipid peroxidation. All tested samples showed antioxidative activities in a dose dependent manner and the terpene fraction was found to possess the highest effect compared with the others. Chemical isolation of the terpene fraction resulted in the detection of ganoderic acids A, B, C and D, lucidenic acid B and ganodermanontriol as major ingredients.

Articular chondrocytes and synoviocytes in a co-culture system: influence on reactive oxygen species-induced cytotoxicity and lipid peroxidation.

A new co-culture system of rat articular chondrocytes and synoviocytes (HIG-82; cell line) was incubated with phorbol myristate acetate (PMA), H2O2 or a combination of Fe2+ and ascorbic acid to simulate inflammation-like radical attacks in articular joints. Chondrocytes were characterized by immunocytochemistry against collagen type II, transmission electron (TEM) and light microscopy. Lipid peroxidation was investigated by measuring thiobarbituric-acid-reactive material in the supernatants, cytotoxicity by determining release of lactate dehydrogenase and proliferation by measuring [3H]thymidine incorporation, culture protein and DNA. PMA or Fe2+ and ascorbic acid induced lipid peroxidation in chondrocytes and synoviocytes that was decreased significantly in co-cultures. PMA and H2O2 dose dependently induced release of lactate dehydrogenase in chondrocytes, which was lowered in co-cultures or in previously co-cultured chondrocytes to a nearly basal level. In contrast, conditioned media of synoviocyte cultures showed no lowering effect on the radical-induced toxicity. Protection against H2O2-induced damage of cellular membranes by co-culturing was also shown by TEM. Synoviocytes released chondrocyte-stimulating growth factors spontaneously without previous interaction. Chondrocytes establish protective mechanisms against reactive oxygen species via an interaction with synoviocytes. Our co-culture model presents a possible way to study mechanisms of inflammation in articular joints under defined conditions.

Antioxidative activity of synthetic melanins. Cardiolipin liposome model

The inhibiting effect of melanin synthesized from dihydroxyphenylalanine (DOPA), dopamine, adrenaline and adrenolutin on the ultraviolet- or the Fe 2+-ascorbic acid-induced lipid peroxidation. Antioxidative activity of melanins enhances in the order: dopamine-melanin<melanin synthesized from dopamine in the presence of Cu 2+ < DOPA - melanin < melanin synthetized from adrenaline in the presence of Cu 2+ ⋟ adrenolutin-melanin , and correlates with thei ability to scavenge superoxide anion radical. The optical screening effect of the investigated melanins in the inhibition of lipid peroxidation was not higher than 15% for the most active melanins.