Effect of hepcidin antagonists on anemia during inflammatory disorders.

Iron is an essential element for the mammalian body however, its homeostasis must be regulated accurately for appropriate physiological functioning. Alterations in physiological iron levels can lead to moderate to severe iron disorders like chronic and acute iron deficiency (anemia) or iron overload. Hepcidin plays an important role in regulating homeostasis between circulating iron and stored iron in the cells as well as the absorption of dietary iron in the intestine. Inflammatory disorders restrict iron absorption from food due to increased circulating levels of hepcidin. Increased production of hepcidin causes ubiquitination of ferroportin (FPN) leading to its degradation, thereby retaining iron in the spleen, duodenal enterocytes, macrophages, and hepatocytes. Hepcidin inhibitors and antagonists play a consequential role to ameliorate inflammation-associated anemia. Many natural and synthesized compounds, able to reduce hepcidin expression during inflammation have been identified in recent years. Few of which are currently at various phases of clinical trial. This article comprises a comprehensive review of therapeutic approaches for the efficient treatment of anemia associated with inflammation. Many strategies have been developed targeting the hepcidin-FPN axis to rectify iron disorders. Hepcidin modulation with siRNAs, antibodies, chemical compounds, and plant extracts provides new insights for developing advanced therapeutics for iron-related disorders. Hepcidin antagonist's treatment has a high potential to improve iron status in patients with iron disorders, but their clinical success needs further recognition along with the identification and application of new therapeutic approaches.

Novel Liposome Eencapsulated Guanosine Di Phosphate based Therapeutic Target against Anemia of Inflammation.

Hepcidin, master regulator of iron homeostasis, causes anemia under infectious and inflammatory conditions by reducing intestinal absorption of iron with decreased release of iron from macrophages and liver despite adequate iron stores leading to Anemia of Inflammation (AI). Many therapeutic trials have been carried out but none have been effective due to its adverse effects. In present study, we discover that Guanosine 5'-diphosphate (GDP) encapsulated in lipid vesicle (NH+) was found to inhibit NF-ҝB activation by limiting phosphorylation and degradation of IҝBα, thus, attenuating IL-6 secretion from macrophage cells. Moreover, the suppressed IL-6 levels down regulated JAK2/STAT3 pathway with decrease inflammation-mediated Hamp mRNA transcription (HepG2) and increase iron absorption (Caco2) in HepG2/Caco2 co-culture model. Analogous results were obtained in acute and chronic AI mice model thus, correcting haemoglobin level. These results proved NH + GDP as novel therapeutic agent to overcome limitations and suggests it as potential drug to ameliorate AI.

Identification of Guanosine 5'-diphosphate as Potential Iron Mobilizer: Preventing the Hepcidin-Ferroportin Interaction and Modulating the Interleukin-6/Stat-3 Pathway.

Hepcidin, a peptide hormone, is a key regulator in mammalian iron homeostasis. Increased level of hepcidin due to inflammatory conditions stimulates the ferroportin (FPN) transporter internalization, impairing the iron absorption; clinically manifested as anemia of inflammation (AI). Inhibiting hepcidin-mediated FPN degradation is proposed as an important strategy to combat AI. A systematic approach involving in silico, in vitro, ex vivo and in vivo studies is employed to identify hepcidin-binding agents. The virtual screening of 68,752 natural compounds via molecular docking resulted into identification of guanosine 5'-diphosphate (GDP) as a promising hepcidin-binding agent. The molecular dynamics simulations helped to identify the important hepcidin residues involved in stabilization of hepcidin-GDP complex. The results gave a preliminary indication that GDP may possibly inhibit the hepcidin-FPN interactions. The in vitro studies revealed that GDP caused FPN stabilization (FPN-GFP cell lines) and increased the FPN-mediated cellular iron efflux (HepG2 and Caco-2 cells). Interestingly, the co-administration of GDP and ferrous sulphate (FeSO4) ameliorated the turpentine-induced AI in mice (indicated by increased haemoglobin level, serum iron, FPN expression and decreased ferritin level). These results suggest that GDP a promising natural small-molecule inhibitor that targets Hepcidin-FPN complex may be incorporated with iron supplement regimens to ameliorate AI.

Metallic/bimetallic magnetic nanoparticle functionalization for immobilization of α-amylase for enhanced reusability in bio-catalytic processes.

Novel magnetic nanoparticles coated with silica and gold were synthesized for immobilization of α-amylase enzyme and characterized with Fourier transform infrared spectroscopy, transmission electron microscopy. Effect of various limiting factors such as substrate concentration, temperature, and pH on the catalytic activity of enzyme was investigated. The optimum pH for free and immobilized enzyme was found unaffected (7.0), whereas optimum temperature for the enzyme activity was increased from 60°C for free enzyme to 80°C for immobilized counterpart. The gains in catalytic attributes concomitant to ease of recovery of the enzyme reflect the potential of the approach and the product to be useful for the enzymatic bioprocessing. The Michaelis-Menten constant (Km) value of the immobilized α-amylase was higher than that of free α-amylase, whereas maximum velocity (Vmax), and turn over number (Kcat), values were almost similar. Immobilized α-amylase maintained 60% of the enzyme activity even after recycling ten times.

Potential of Alginate Encapsulated Ferric Saccharate Microemulsions to Ameliorate Iron Deficiency in Mice.

Iron deficiency is one of the most prominent mineral deficiencies around the world, which especially affects large population of women and children. Development of new technologies to combat iron deficiency is on high demand. Therefore, we developed alginate microcapsule with encapsulated iron that had better oral iron bioavailability. Microcapsules containing iron with varying ratios of sodium alginate ferric(III)-saccharide were prepared using emulsification method. In vitro studies with Caco-2 cells suggested that newly synthesized microemulsions had better iron bioavailability as compared to commercially available iron dextran formulations. Ferrozine in vitro assay showed that alginate-encapsulated ferric galactose microemulsion (AFGM) had highest iron bioavailability in comparison to other four ferric saccharate microemulsions, namely AFGlM, AFMM, AFSM, and AFFM synthesized in our laboratory. Mice studies also suggested that AFGM showed higher iron absorption as indicated by increased serum iron, hemoglobin, and other hematopoietic measures with almost no toxicity at tested doses. Development of iron-loaded microemulsions leads to higher bioavailability of iron and can provide alternative strategies to treat iron deficiency.

Preventive Role of Vitamin D in Silica-Induced Skin Fibrosis: A Study in Relation to Oxidative Stress and Pro-Inflammatory Cytokines.

The protective effects of vitamin D analogue calcipotriol in silica-induced skin fibrosis were studied in the present study. Male BALB / c mice were divided into four groups; Control, Vitamin D, Silica and Silica+Vitamin D. Silica was administered as a single intradermal injection (40 µg / µL, dissolved in normal saline; particle size 1 - 5 µm) in the hind limbs of animals in Silica & Silica+Vitamin D group. Vitamin D group animals received topical application of 100µL of vitamin D solution (10-7M in Ethanol) daily for 12 weeks. Silica+Vitamin D group animals received co-treatment of silica and vitamin D as described for other groups. After 12 weeks of treatment, dermal thickness and hydroxyproline content of treated sections were measured. The TNF-α and IL-6 levels were measured in serum of all treated animals. The silica-induced oxidative stress was measured in terms of lipid peroxidation in skin tissue. Antioxidant defence system was assessed in terms of levels of reduced glutathione, superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase. A significant increase in the dermal thickness and hydroxyproline content was observed after silica treatment (931 ± 57.98 to 1804.61 ± 146.20 µm)(p < 0.05). Vitamin D co-treatment reduced dermal thickness and hydroxyproline content compared to Silica group (p < 0.05). Similarly a decrease in TNF-α and IL-6 levels were also observed after vitamin D treatment. A significant reduction in oxidative stress in terms of lipid peroxidation (4.92 ± 0.70 to 2.40 ± 0.31 nmol / mg protein). Therefore, present study suggested that vitamin D could be an effective agent against silica-induced skin fibrosis and oxidative stress.