Reducing Ability Assay Research Articles

Protection by sildenafil and theophylline of lead acetate-induced oxidative stress in rat submandibular gland and saliva.

The role of oxidative stress in lead toxicity has been proposed in many organs, however, no study has been performed in the salivary glands, which are important parts of the gastrointestinal tract with a high implication in health of the whole body. Recently, it has been proposed that increasing the levels of cGMP and cAMP in the cells may protect from the neurotoxicity of lead. The objective of this study was to determine the ability of lead acetate to produce oxidative stress in rat submandibular as the main salivary gland of the body and to study the role of pretreatment by specific phosphodiesterase inhibitors in the prevention of oxidative stress. Lead acetate (100 mg/kg), alone or in combination with theophylline (25 mg/kg) and sildenafil (5 mg/kg), was administered intraperitoneally to rats. After 2 hours and under general anaesthesia, the submandibular gland ducts were cannulated intraorally using microcannula, and pure saliva was collected for 30 min using pilocarpine (8 mg/kg) as a secretagogue. The submandibular glands were then isolated free under surgery. Oxidative stress in the gland and pure saliva were evaluated measuring lipid peroxidation (thiobarbituric acid reactive substances assay), total thiol groups content and total antioxidant capacity (the ferric reducing ability assay). Results showed significant oxidative stress in the gland and secretions as indicated by increased lipid peroxidation, decreased total antioxidant capacity and thiol group levels. The use of cAMP and cGMP phosodiesterase inhibitors, theophylline and sildenafil, prevented lead-induced increased lipid peroxidation and also protected from decreased thiol groups content and total antioxidant power of the gland and secretions. The same trend of effects was observed in gland and saliva. It is concluded that lead toxicity is mediated through oxidative stress in salivary glands, while increasing intracellular cAMP and cGMP levels may prevent lead-induced oxidative stress.

Anthocyanins are potent antioxidants in model systems but do not reduce endogenous oxidative DNA damage in human colon cells

Anthocyanins are common colored plant flavonoids, occurring as glycosides of the respective anthocyanidin-chromophores. Like other flavonoids, anthocyanidins are also expected to have antioxidative and anti-mutagenic properties in vivo, although only few data are available. To gain more knowledge on possible protective mechanisms in mammalian cells, we have compared their extracellular and intracellular antioxidative potential in vitro and in human colon tumor cells. We used Aronia melanocarpa Elliot anthocyanin (AA) concentrates, fractions thereof, concentrates from Elderberry, Macqui, and Tintorera fruits, as well as pure compounds. In vitro, antioxidative properties of the samples were studied with the ferric reducing ability assay (FRA assay). As a measure of intracellular oxidative/antioxidative effects, H2O2-induced strand breaks as well as oxidized DNA bases were determined in human tumor HT29 clone 19A cells using a microgelelectrophoresis assay (comet test). Major results were that isolated compounds (aglycons and glycosides) and complex plant samples are powerful antioxidants in vitro. In fact their activities by far exceeded those of Trolox and vitamin C in the FRA assay. Also, H2O2-induced DNA strand breaks were reduced in cells treated with the complex plant extracts. In contrast, endogenous generation of oxidized DNA bases was not prevented. In summary, the intracellular steady state of oxidized DNA bases is not altered by anthocyanins or anthocyanidins. This finding raises questions with respect to the cancer preventive potential of anthocyanidins within specific tissues, such as the colon. Extracellularly, however, the compounds are potent antioxidants. This points to their potential for providing systemic protection in vivo, e.g., by scavenging oxidants in the blood stream and in the colon. Notably, both aglycons and glycosides have equally strong antioxidant activity.