ABSTRACT
Hesperetin, an abundant bioactive component of citrus fruits, is poorly water-soluble, resulting in low oral bioavailability. We developed new formulations to improve the water solubility, antioxidant activity, and oral absorption of hesperetin. Two nano-based formulations were developed, namely hesperetin-TPGS (D-α-tocopheryl polyethylene glycol 1000 succinate) micelles and hesperetin-phosphatidylcholine (PC) complexes. These two formulations were prepared by a simple technique called solvent dispersion, using US Food and Drug Administration (FDA)-approved excipients for drugs. Differential scanning calorimetry (DSC) and dynamic light scattering (DLS) were used to characterize the formulations' physical properties. Cytotoxicity analysis, cellular antioxidant activity assay, and a pharmacokinetic study were performed to evaluate the biological properties of these two formulations. The final weight ratios of both hesperetin to TPGS and hesperetin to PC were 1:12 based on their water solubility, which increased to 21.5- and 20.7-fold, respectively. The hesperetin-TPGS micelles had a small particle size of 26.19 nm, whereas the hesperetin-PC complexes exhibited a larger particle size of 219.15 nm. In addition, the cellular antioxidant activity assay indicated that both hesperetin-TPGS micelles and hesperetin-PC complexes increased the antioxidant activity of hesperetin to 4.2- and 3.9-fold, respectively. Importantly, the in vivo oral absorption study on rats indicated that the micelles and complexes significantly increased the peak plasma concentration (Cmax) from 2.64 μg/mL to 20.67 and 33.09 μg/mL and also increased the area under the concentration-time curve of hesperetin after oral administration to 16.2- and 18.0-fold, respectively. The micelles and complexes increased the solubility and remarkably improved the in vitro antioxidant activity and in vivo oral absorption of hesperetin, indicating these formulations' potential applications in drugs and healthcare products.
Subject(s)
Animals , Dogs , Female , Humans , Rats , Administration, Oral , Antioxidants/chemistry , Biological Availability , Calorimetry, Differential Scanning , Dose-Response Relationship, Drug , Drug Carriers , Hep G2 Cells , Hesperidin/chemistry , Light , Madin Darby Canine Kidney Cells , Micelles , Phosphatidylcholines/chemistry , Polyethylene Glycols/chemistry , Rats, Sprague-Dawley , Scattering, Radiation , Solubility , Solvents , Vitamin E/chemistry , Water/chemistry , alpha-Tocopherol/chemistryABSTRACT
The biological significance of singlet oxygen (1O2), an electronically excited species of oxygen, has been realized only in the last two decades. This was mainly due to the lack of proper methodology to generate this reactive oxygen species (ROS) in pure form and its reactions with biological molecules. Recent studies, using newly developed detection methods, show that 1O2 being generated in many biological systems, can significantly and quite often adversely alter several crucial biomolecules including DNA, proteins and lipids with undesirable consequences including cytotoxicity and/or disesase development. The reactions of 1O2 with the biological molecules are rather specific, as compared to other ROS. There are various compounds, mainly derived from natural sources that offer protection against damage induced by 1O2. Among the antioxidants carotenoids are the most effective singlet oxygen quenchers followed by tocopherols and others. The same reactive species if generated specifically in diseased states such as cancer can lead to the cure of the disease, and this principle is utilized in the newly developing modality of cancer treatment namely photodynamic therapy. Singlet oxygen, in low concentrations can also act as signaling molecule with several biological implications. This review clearly brings out the biological significance of 1O2.
Subject(s)
Animals , Antioxidants/chemistry , Carotenoids/chemistry , DNA Damage , Humans , Lipid Metabolism , Mutagenesis , Neoplasms/metabolism , Photochemistry , Photochemotherapy , Proteins/metabolism , Singlet Oxygen/adverse effects , Vitamin E/chemistryABSTRACT
1. Hepatotoxicity is the most common finding in patients with iron overload since the liver is the major recipient of iron excess, even though the kidney could be a target of iron toxicity. The effect of iron overload was studied in the early stages after iron-dextran injection inrats, as a model for secondary hemocromatosis. 2. Total hepatic and kidney iron content was markedly over control values 20h after the iron administration. Plasma GOT, GPT and LDH activities were not affected, suggesting that liver cell permeability was not affected by necrosis. 3. Spontaneous liver chemiluminescence was measured as an indicator of oxidative stress and lipid peroxidation. Light emission was increased four-fold 6 h after iron supplementation. 4. Increases in the generation of thiobarbituric acid reactive substances (TBARS) in liver and kidney homogenates were detected after iron administration. 5. The activities of catalase, SOD and glutathione peroxidase were determined. Enzymatic activities declined in liver homogenates by 25, 36 and 32 por cento, respectively, 20 h after iron injection. These activities were not affected in kidney as compared to control values, except for SOD activity that was decreased by 26 por cento. 6. The content of alfa-tocopherol was decreased by 31 por cento in whole kidney homogenates and by 40 por cento in plasma. 7. Our data indicate that lipid peroxidation occurs after mild iron overload both in liver and kidney. Enzymatic antioxidantes are consumed significantly in liver and alfa-tocopherol content decreases in kidney, suggesting an organ-specific antioxidant effect