Biological activities of the natural imidazole-containing peptidomimetics n-acetylcarnosine, carcinine and L-carnosine in ophthalmic and skin care products

Apart from genetically programmed cell aging, different external aggressors related to oxidative stress and lipid peroxidation (LPO) can accelerate the skin aging phenomenon. Oxidative stress associated with the formation of lipid peroxides is suggested to contribute to pathological processes in aging and systemic diseases known as the risk factors for cataract. Despite the fact that L-carnosine-related peptidomimetics N-acetylcarnosine (N-acetyl-beta-alanyl-L-histidine) (NAC) and carcinine (beta-alanylhistamine) are metabolically related to L-carnosine and have been demonstrated to occur in tissues of many vertebrates, including humans, these compounds were shown resistant toward enzymatic hydrolysis. A series of related biocompatible imidazole-containing peptidomimetics were synthesized in order to confer resistance to enzymatic hydrolysis and ex vivo improvement of protective antioxidative properties related to L-carnosine. The included findings revealed a greater role of N-acetylcarnosine (NAC) and carcinine ex vivo in the prolongation and potentiation of physiological responses to the therapeutical and cosmetics treatments with L-carnosine as antioxidant. 3-D molecular conformation studies proposed the antioxidant activity of peptidomimetics (carcinine, L-prolylhistamine, N-acetylcarnosine, L-carnosine) for metal ion binding, quenching of a number free radicals, and binding of hydroperoxide or aldehyde (including dialdehyde LPO products) in an imidazole-peroxide adducts. NAC can act as a time release (carrier) stable version of L-carnosine during application in ophthalmic pharmaceutical and cosmetics formulations which include lubricants. Carcinine, L-prolylhistamine show efficient deactivation of lipid hydroperoxides monitored by HPLC and protection of membrane phospholipids and water soluble proteins from the lipid peroxides-induced damages. This activity is superior over the lipophilic antioxidant vitamin E. The biologically significant applications of carnosine mimetics were patented by Dr. Babizhayev and the alliance Groups (WO 2004/028536 A1; WO 94/19325; WO 95/12581; WO 2004/064866 A1).     

The membrane-stabilizing action of zinc carnosine (Z-103) in stress-induced gastric ulceration in rats

Zinc compounds have been shown to antagonize various types of gastric ulceration in rats. Zinc carnosine (Z-103), a newly developed agent was, therefore, examined for its antiulcer effect in stress-induced ulceration and also its membrane stabilizing action in rat stomachs. Cold-restraint (restrained at 4 degrees C for 2 h) stress induced severe hemorrhagic lesions together with increased mast cell degranulation and beta-glucuronidase release in the gastric glandular mucosa. Z-103 pretreatment with a single oral dose (3, 10 or 30 mg/kg) reversed these actions in a dose-dependent manner. When the compound was incubated in concentrations of 10(-7, 10(-6), 10(-5) or 10(-4) M, with isolated hepatic lysosomes, it significantly reduced the spontaneous release of beta-glucuronidase in the medium. The present study not only demonstrates the antiulcer effect of Z-103 but also indicates that the protective action is likely to be mediated by its membrane-stabilizing action on mast cells and lysosomes in the gastric glandular mucosa.     

Lithospermic acid as a novel xanthine oxidase inhibitor has anti-inflammatory and hypouricemic effects in rats

Lithospermic acid (LSA) was originally isolated from the roots of Salvia mitiorrhiza, a common herb of oriental medicine. Previous studies demonstrated that LSA has antioxidant effects. In this study, we investigated the in vitro xanthine oxidase (XO) inhibitory activity, and in vivo hypouricemic and anti-inflammatory effects of rats. XO activity was detected by measuring the formation of uric acid or superoxide radicals in the xanthine/xanthine oxidase system. The results showed that LSA inhibited the formation of uric acid and superoxide radicals significantly with an IC50 5.2 and 1.08 microg/ml, respectively, and exhibited competitive inhibition. It was also found that LSA scavenged superoxide radicals directly in the system beta-NADH/PMS and inhibited the production of superoxide in human neutrophils stimulated by PMA and fMLP. LSA was also found to have hypouricemic activity on oxonate-pretreated rats in vivo and have anti-inflammatory effects in a model of gouty arthritis. These results suggested that LSA is a competitive inhibitor of XO, able to directly scavenge superoxide and inhibit superoxide production in vitro, and presents hypouricemic and anti-inflammatory actions in vivo.     

Antioxidative Activity of 5,6,7,8-Tetrahydrobiopterin and its Inhibitory Effect on Paraquat-Induced Cell Toxicity in Cultured Rat Hepatocytes

The in vitro antioxidative activity of 5,6,7,8-tetrahydrobiopterin (BPH4) was measured and the ability of BPH4 to prevent paraquat-induced cell damage was examined in cultured hepatocytes. The scavenging activity of BPH4 against superoxide anion radicals was assayed in two systems, i.e., xanthine/xanthine oxidase (X/XOD) and rat macrophage/phorbol myristate acetate (MξPMA) radical-generating systems. BPH4 showed an extremely strong superoxide anion radical-scavenging activity in both assay systems. Biopterin (BP) itself did not show any activity in the X/XOD system, but was effective in the MξPMA system. The antioxidative activities of BPH4 against both superoxide anion and hydroxyl radicals were confirmed by spin trapping-ESR spectrometry. BPH4 also protected rat brain homogenate against auto-oxidation. We further examined the effect of BPH4 on paraquat-induced cell toxicity in cultured rat hepatocytes. The paraquat-induced elevation of the release of lactate dehydrogenase (LDH), a marker enzyme for cytotoxicity from cultured hepatocytes was suppressed by BPH4 in a dose-dependent manner. The elevation of lipid peroxides simultaneously induced by paraquat was also inhibited by BPH4 in the same manner. These results suggest that BPH4 might be useful in the treatment of various diseases whose pathogenesis is active oxygen-related.     

Influence of Lanthanides on the Antioxidative Defense System in Maize Seedlings Under Cold Stress

The influence of LaCl₃, CeCl₃, and NdCl₃ on the antioxidative defense system in maize seedlings under cold stress was investigated. It was found that maize seedlings cultivated in cold stress developed distinct cold symptoms, and the plant growth was significantly inhibited as expected, while Ln-treated seedling growth was improved. Cold stress in maize seedlings also increased the permeability of plasma membrane, malondialdehyde as a degradation product of lipid peroxidation, and reactive oxygen species such as superoxide radicals and hydrogen peroxide, and decreased activities of the antioxidant enzymes such as superoxide dismutase, catalase, ascorbate peroxidase, and glutathione content; however, Ln treatments cultivated in cold stress decreased the permeability of plasma membrane, malondialdehyde, and reactive oxygen species, and increased activities of the antioxidative defense system. It implied that Ln could increase oxidative-stress resistance under cold stress. On the other hand, the assay of physiological and biochemical parameters demonstrated that Ce relieving chilling injury of maize seedlings caused by cold stress was most significant, medium in the Nd treatment, and last in the La treatment. These results suggested that the increase of cold resistance of maize seedlings caused by Ln might be closely related to its properties of 4f electron shell and variable valence.     

Protective effect of Aquilegia vulgaris (L.) on carbon tetrachloride-induced oxidative stress in rats

The ethyl ether extract of A. vulgaris inhibited in vitro microsomal lipid peroxidation (IC50 58.8 microg/ml) and showed moderate ability to scavenge superoxide radicals and to chelate iron ions. The extract (100 mg/kg body weight, po) decreased uninduced and enzymatic microsomal lipid peroxidation in the liver of male rats pretreated with CCl4 (1 ml/kg body weight) by 27 and 40%, respectively. Activity of antioxidant and related enzymes (catalase and glucose-6-phosphate dehydrogenase) inhibited by CCl4 was significantly restored after administration of the extract. The extract itself significantly enhanced superoxide dismutase activity. There was no effect of the extract on hepatic glutathione level and cytochrome P450 content, both were decreased by CCl4. Neither CCl4 nor the tested extract affected activities of NADPH-cytochrome P450 reductase and two monooxygenases, aniline hydroxylase and aminopyrine n-demethylase. It can be concluded that the protective effect of the A. vulgaris extract in CCl4-induced liver injury is mediated by inhibition of microsomal lipid peroxidation and restoring activity of some antioxidant and related enzymes.     

Antioxidative effect of α-tocopherol incorporation into lecithin liposomes on ascorbic acid-Fe2+-induced lipid peroxidation

The antioxidative effect of α-tocopherol incorporated into lecithin liposomes was studied. Lipid peroxidation of liposome membranes, assayed as malondialdehyde production, was catalyzed by ascorbic acid and Fe²⁺. The peroxidation reaction, which did not involve the formation of singlet oxygen, superoxide, hydrogen peroxide, or a hydroxyl radical, was inhibited by α-tocopherol and a model compound of α-tocopherol, 2,2,5,7,8-pentamethyl-6-hydroxy-chroman (TMC), but not by phytol, α-tocopherylquinone, or α-tocopheryl acetate. One mole of α-tocopherol completely prevented peroxidation of about 100 moles of polyunsaturated fatty acid. Decrease in membrane fluidity by lipid peroxidation, estimated as increase of fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) embedded in the membrane, was also inhibited by α-tocopherol and TMC, reflecting their antioxidant functions. Cholesterol did not act as an antioxidant, even when incorporated in large amount into the liposome membranes, but it increased the antioxidative efficiency of α-tocopherol. When a mixture of liposomes with and without α-tocopherol was incubated with Fe²⁺ and ascorbic acid, α-tocopherol did not protect the liposomes not containing α-tocopherol from peroxidation. However, preincubation of the mixture, or addition of Triton X-100 allowed the α-tocopherol to prevent peroxidation of the liposomes not containing α-tocopherol. In contrast, in similar experiments, liposomes containing TMC prevented peroxidation of those without TMC without preincubation. Tocopherol in an amount so small as to exhibit only a slight antioxidative effect was oxidized when incorporated in egg lecithin liposomes, but it mostly remained unoxidized when incorporated in dipalmitoyllecithin liposomes, indicating that oxygen activated by ascorbic acid-Fe²⁺ does not oxidize α-tocopherol directly. Thus, decomposition of α-tocopherol may be caused by its interaction with peroxy and/or alkoxyl radicals generated in the process of lipid peroxidation catalyzed by Fe²⁺ and ascorbic acid.     

Protective effects of polysaccharide from Euphorbia kansui (Euphorbiaceae) on the swimming exercise-induced oxidative stress in mice

The present study examined the effects of derivatives of galactosides and glucosides in a polysaccharide extract from Euphorbia kansui (Euphorbiaceae) on exercise-induced oxidative stress in mice. Exhaustive swimming exercise significantly increases the degree of lipid peroxidation in terms of malondialdehyde content and reduces the antioxidant activities of superoxide dismutase (SOD) and glutathione peroxidase (GPx). Our findings revealed that chronic oral treatment with the extract elevates enzymatic activities of SOD and GPx accompanied by a corresponding decrease in malondialdehyde. The antioxidative activities of these compounds against exercise-induced oxidative stress are correlated with various activities such as reducing the production of superoxide and hydroxyl radicals, inhibiting lipid peroxidation, enhancing antioxidative defenses, and increasing the production of SOD and GPx activity and expression in different tissues. These compounds may be involved in glycogen metabolism to meet the requirement of working skeletal muscles and act as antioxidants by terminating the chain reaction of lipid peroxidation to maintain the morphological stability of mitochondria in spinal motor neurons. These observations suggest that E. kansui has antioxidative and antifatigue properties and can be given as prophylactic and (or) therapeutic supplements for increasing antioxidant enzyme activities and preventing lipid peroxidation during strenuous exercise.     

Effect of ethanol extract of Piper betle Linn leaf on healing of NSAID-induced experimental ulcer--a novel role of free radical scavenging action

Treatment with ethanol extract of leaf of P. betle at a dose of 150 mg/kg body weight daily for 10 days, after induction of peptic ulcer by NSAID in albino rats, produced significant healing effect. During healing process, on treatment with the extractive, antioxidative factor, e.g. superoxide dismutase and catalase activity, mucus and total gastric tissue sulfhydryl group were increased. In contrast, oxidised lipid and oxidatively modified proteins were reduced to near normalcy, within 7 to 10 days, however, change in the untreated group was not significant. The extract also showed significant in vitro free radical scavenging action. The results suggest that the antioxidant or free radical scavenging activity of the plant extract, may be responsible for its healing action.     

Antioxidant activity of carotenoid lutein in vitro and in vivo

Carotenoid lutein was evaluated for its antioxidant potential both in vitro and in vivo. Lutein was found to scavenge superoxide radicals, hydroxyl radicals and inhibited in vitro lipid peroxidation. Concentrations needed for 50% inhibition (IC50) were 21, 1.75 and 2.2 microg/mL respectively. It scavenged 2,2-diphenyl-1-picryl hydrazyl (IC50 35 microg/mL) and nitric oxide radicals (IC50 3.8 microg/mL) while 2,2-azobis-3-ethylbenzthiozoline-6-sulfonic acid radicals were inhibited at higher concentration. Ferric reducing power (50%) of lutein was found to be equal 0.3 micromols/mL of FeSO4.7H2O. Its oral administration inhibited superoxide generation in macrophages in vivo by 34.18, 64.32 and 70.22% at doses of 50, 100 and 250 mg/kg body weight. The oral administration of lutein in mice for 1 month significantly increased the activity of catalase, superoxide dismutase, glutathione reductase and glutathione in blood and liver while the activity of glutathione peroxidase and glutathione-S-transferase were found to be increased in the liver tissue. Implication of these results in terms of its role in reducing degenerative diseases is discussed.     

Effect of antiperoxidative drugs on gastric damage induced by ethanol in rats

Lesion formation due to oral administration of absolute ethanol could be prevented by parenteral pretreatment with antiperoxidative drugs such as butylated hydroxytoluene (BHT), quercetin and quinacrine. Also effective were allopurinol and oxypurinol, inhibitors of xanthine oxidase, but not superoxide dismutase (SOD) and hydroxyl radical scavengers, such as sodium benzoate and dimethyl sulfoxide (DMSO). BHT, quercetin, quinacrine and sulfhydryl compounds such as reduced glutathione and cysteamine which offer gastroprotection in vivo against ethanol inhibited lipid peroxidation induced in vitro by ferrous ion in porcine gastric mucosal homogenate, but SOD, sodium benzoate, DMSO, allopurinol and oxypurinol did not. These results suggest the possibility that an active species, probably derived from free iron mobilized by the xanthine oxidase system, other than oxygen radicals such as hydroxyl radicals, contributes to lipid peroxidation and lesion formation in the gastric mucosa after absolute ethanol administration.     

Spermine: an anti-oxidant and anti-inflammatory agent.

This work demonstrates that spermine is a natural antioxidant and anti-inflammatory agent. It is found that: (1) Spermine inhibits the cytochrome C reduction initiated by FMLP- or PMA-stimulated human granulocytes. (2) Spermine inhibits the Fe(III)/xanthine oxidase stimulated lipid peroxidation of brain phospholipid liposomes. The antioxidative effect disappears at high Fe(III) concentrations. (3) Spermine forms a complex with Fe(II). (4) Spermine inhibits the Fe(II)-induced depolymerization of hyaluronic acid, and EDTA abolishes this effect. (5) Spermine or spermine-Fe(II) has no superoxide mimetic effect. These findings suggest that spermine has at least two antioxidative mechanisms of action: (I) Spermine inhibits the generation of the transport of superoxide radicals from stimulated granulocytes, and (II) Spermine inhibits the Haber-Weiss reaction by forming an unreactive chelate with Fe. Spermine thus prevents generation of destructive hydroxyl radicals.     

Design and biological activity of imidazole-containing peptidomimetics with a broad-spectrum antioxidant activity

Histidine-containing natural dipeptides, such as L-carnosine, were reported to be effective against different oxygen-derived free radicals, and also lipoperoxyl radicals. However, L-carnosine appeared to be uneffective in vivo, due to the presence of ubiquitous specific or semi-specific hydrolytic enzymes. Therefore, a series of peptidomimetics were synthesized in order to confer resistance to enzymatic hydrolysis. Some structural modifications were also done in an attempt to improve the antioxidant power of the molecule, for example, in relation to binding of ferrous ions. The following methods were used for the evaluation of peptidomimetics:– The resistance to enzymatic deactivation was determined using either a purified specific enzyme or a multi-enzymatic system.– The free radical scavenging potential was measured by different in vitro methods involving different free radical species and biological targets.– Deactivation of lipid hydroperoxides was monitored by HPLC and protection of membrane phospholipids and proteins was demonstrated.– The effectiveness of peptidomimetics in vivo was evaluated.It was shown that decarboxylation of L-carnosine results in an important improvement of the resistance towards hydrolytic enzymes. In vitro experiments have demonstrated, for a series of peptidomimetics, free radical scavenging and lipid hydroperoxide deactivating properties similar to or even better than the natural peptide (depending on the experimental design). In addition, the two peptidomimetics -alanylhistamine and L-prolylhistamine proved to be far superior in inhibiting the lipid hydroperoxide-mediated cross-linking of a representative protein. Finally, -alanylhistamine was able to protect skin enzymes from UV-induced degradation in vivo.     

Antioxidant activity of liver growth factor,a bilirubin covalently bound to albumin

We previously reported that treatment of spontaneously hypertensive rats (SHR) with liver growth factor (LGF), an albumin–bilirubin complex with a covalent bond, reduces blood pressure, improves nitric oxide (NO)-dependent vasodilatation, and exerts vascular antifibrotic actions. Because bilirubin, albumin, and albumin-bound bilirubins have antioxidant properties, we hypothesize that LGF might exert its cardiovascular actions through an antioxidant mechanism. We have tested in vitro the capacity of LGF to scavenge ABTS cation and peroxyl and hydroxyl radicals and to protect vascular NO from degradation by superoxide anion. We have also compared the antioxidant capacity of LGF with that of its molecular components albumin and bilirubin and the reference antioxidant trolox. LGF exhibited antioxidant capacity against all free radicals tested at lower concentrations than albumin, bilirubin, and trolox. LGF, bilirubin, and albumin were also able to protect endothelial NO from superoxide anion degradation in a fashion similar to that of superoxide dismutase or tiron, but at much lower concentrations. These data, together with our previous results in SHR, suggest that LGF might exert its cardiovascular regenerative actions, at least in part, through an antioxidant mechanism and that LGF could be a relevant circulating antioxidant in situations of oxidative stress.     

Antioxidant effect of zinc and zinc-metallothionein in the acute cytotoxicity of hydrogen peroxide in Ehrlich ascites tumour cells

This study was concerned with the role of zinc (Zn) and zinc-metallothionein (Zn-MT) in oxidative stress. Hydrogen peroxide-induced oxidative injury was examined in Ehrlich ascites tumour cells isolated from control host mice, mice pretreated with 10 mg/kg ZnSO4 (i.p.) to increase cellular Zn/Zn-MT levels, and mice exposed to Zn-deficient diet to reduce the cellular Zn/Zn-MT levels. The results of the present study showed that Ehrlich cells with seven-fold differences in Zn-MT concentrations could be obtained by manipulating the Zn status of host mice and that high Zn and Zn-MT levels can make Ehrlich cells more resistant to H2O2-induced oxidative injury (cell viability, lipid peroxidation, [Ca2+]i) while cells with reduced Zn/Zn-MT levels were more susceptible to this treatment. H2O2 treatment resulted in oxidation of MT thiolate groups and loss of its metal binding capacity with translocation of Zn released from oxidized MT to other cellular sites. Preincubation of Ehrlich cells with ZnSO4 in vitro also conferred some degree of resistance to H2O2 toxicity, suggesting the inherent antioxidative property of Zn ions. These data suggested that Zn-MT can be considered as an antioxidant by virtue of its thiolate groups and its Zn ions that are released in the presence of oxidative stress.     

Mechanisms involved in gastric protection of melatonin against oxidant stress by ischemia-reperfusion in rats

The generation of oxygen-derived free radicals has been suggested to be significantly responsible for ischemia-reperfusion injury in gastrointestinal tissues. Biochemical mechanisms include the xanthine-oxidase-derived oxidants mainly the superoxide anion. Both in vitro and in vivo studies have demonstrated that the pineal hormone melatonin possesses free radical scavenging and antioxidant properties. The indolamine has been effective in reducing the induced-oxidative damage in several tissues and biological systems. The aim of this study was to elucidate additional antioxidant mechanisms responsible for the gastroprotection afforded by the indolamine in ischemia-reperfusion gastric injury. Therefore, changes of related enzymes such as xanthine-oxidase, superoxide dismutase, glutathione reductase and total glutathione were investigated. Our results showed that treatment with 5, 10 or 20 mg kg(-1) of melatonin, administered i.p., clearly diminished the percentage of damage to 49.56 +/- 17.20, 37.54 +/- 11.40 and 26.70 +/- 8.12 respectively. Histologically there was a reduction of exfoliation of superficial cells and blood cell infiltration. These protective effects were related to a significant reduction of xanthine-oxidase activity (2.23 +/- 0.38 U/mg prot x 10(-4) with the highest tested dose of melatonin) and significant increases in superoxide dismutase reaching a value of 6.20 +/- 0.56 U/mg prot with 25 mg/Kg of melatonin and glutation reductase activities (417.44 +/- 29.72 and 649.43 +/- 81.11 nmol/min/mg prot with 10 and 20 mg/Kg of melatonin). We conclude that the free radical scavenger properties of melatonin mainly of the superoxide anion, probably derived via the xanthine-oxidase pathway, and the increase of antioxidative enzymes significantly contributes to mediating the protection by the hormone against ischemia-reperfusion gastric injury.     

Effects of metal ions on the antioxidant enzyme activities, protein contents and lipid peroxidation of carp tissues

1. Studies were performed regarding the effects of CuSO4 in concentrations of 5, 10, 25 and 50 ppm and ZnSO4 in concentrations of 10 and 100 ppm on the antioxidant enzyme activities, lipid peroxidation and protein contents of tissues of common carp (Cyprinus carpio morpha L.) exposed to these pollutants for 24 hr. 2. The results demonstrated that CuSO4 was more toxic than ZnSO4 and that both treatments brought about significant changes in these parameters in carp hepatopancreas (liver), gill and white muscle. 3. An increase of the CuSO4 concentration led to significant decreases in the antioxidant enzyme activities, except that of glutathione peroxidase, which was increased significantly, and significant increases in the lipid peroxidation and protein contents. 4. An increase of the ZnSO4 concentration led to slight changes in the antioxidant enzyme activities, lipid peroxidation and protein contents of carp tissues.     

Protection against oxygen radicals: an important defence mechanism studied in transgenic plants

Free radicals and other active derivatives of oxygen are inevitable by-products of biological redox reactions. Reduced oxygen species, such as hydrogen peroxide, the superoxide radical anion and hydroxyl radicals, inactivate enzymes and damage important cellular components. In addition, singlet oxygen, produced via formation of triplet state chlorophyll, is highly destructive. This oxygen species initiates lipid peroxidation, and produces lipid peroxy radicals and lipid hydroperoxides that are also very reactive. The increased production of toxic oxygen derivatives is considered to be a universal or common feature of stress conditions. Plants and other organisms have evolved a wide range of mechanisms to contend with this problem. The antioxidant defence system of the plant comprises a variety of antioxidant molecules and enzymes. Considerable interest has been focused on the ascorbate-glutathione cycle because it has a central role in protecting the chloroplasts and other cellular compartments from oxidative damage. It is clear that the capacity and activity of the antioxidative defence systems are important in limiting photo-oxidative damage and in destroying active oxygen species that are produced in excess of those normally required for signal transduction or metabolism. In our studies on this system, we became aware that the answers to many unresolved questions concerning the nature and regulation of the antioxidative defence system could not be obtained easily by either a purely physiological or purely biochemical approach. Transgenic plants offered us a means by which to achieve a more complete understanding of the roles of the enzymes involved in protection against stress of many types: environmental and man-made. The ability to engineer plants which express introduced genes at high levels provides an opportunity to manipulate the levels of these enzymes, and hence metabolism in vivo. Studies on transformed plants expressing increased activities of single enzymes of the antioxidative defence system indicate that it is possible to confer a degree of tolerence to stress by this means. However, attempts to increase stress resistance by simply increasing the activity of one of the antioxidant enzymes have not always been successful presumably because of the need for a balanced interaction of protective enzymes. The study of these transformed plants has allowed a more complete understanding of the roles of individual enzymes in metabolism. Protection against oxidative stress has become a feasible objective through the application of molecular genetic techniques in conjunction with a biochemical and physiological approach.     

Lipid peroxidation of skeletal muscle: an in vitro study

The process of lipid peroxidation of skeletal muscle has been examined in vitro by monitoring the autoxidation of skeletal-muscle homogenates. Skeletal-muscle tissue has been shown to have considerable capacity for autoxidation and the process has been found to be initiated by a free-radical-mediated mechanism, critically dependent on the presence of free iron in the homogenate. The initiating radicals have not been firmly identified, but the results suggest that neither superoxide or hydroxyl radicals are involved. An in vitro technique for assessment of the antioxidant capacity of muscle tissue has also been developed which is capable of demonstrating differences between muscle tissues with differing vitamin E contents.     

Antioxidant and antitumor activities of the polysaccharide from seed cake of Camellia oleifera Abel

To explore biomedical potential of the polysaccharide from seed cake of Camellia oleifera Abel, we investigated antioxidant and antitumor capacities of the polymer. The results showed that the polysaccharide is capable of scavenging both superoxide anion and hydroxyl radicals in vitro. The highest scavenging rate of superoxide anion and hydroxyl radicals is 85% and 76%, respectively. Using the model animal, Caenorhabditis elegans, we further show that the polysaccharide can increase antioxidant enzyme activity, decrease lipid peroxidation level, and reduce paraquat-induced oxidative damage at a polysaccharide concentration more than 50mg/l. We also revealed that the polysaccharide has some ferric chelating ability and strong in vivo antitumor activity. The antitumor rate against Sarcoma180 solid tumor grown in BALB/C mice reached 85.6% at the highest dose of 40×20mg/kgdays.