ABSTRACT
About half of the world population carries at least one allele of the Ala92-DIO2, which slows down the activity of the type 2 deiodinase (D2), the enzyme that activates T4 to T3. Carrying the Ala92-DIO2 allele has been associated with increased body mass index and insulin resistance, but this has not been reproduced in all populations. To test if the genetic background affects the impact of this polymorphism, here we studied the genetically distant C57Bl/6J (B6) and FVB/N (FVB) mice carrying the Ala92-Dio2 allele as compared to control mice carrying the Thr92-Dio2 allele. Whereas B6-Ala92-Dio2 and B6-Thr92-Dio2 mice-fed chow or high-fat diet-behaved metabolically similar in studies using indirect calorimetry, glucose- and insulin tolerance tests, and measuring white adipose tissue (WAT) weight and liver steatosis, major differences were observed between FVB-Ala92-Dio2 and FVB-Thr92-Dio2 mice: carrying the Ala92-Dio2 allele (on a chow diet) resulted in hypercholesterolemia, smaller WAT pads, hepatomegaly, steatosis, and transcriptome changes in the interscapular brown adipose tissue (iBAT) typical of ER stress and apoptosis. Acclimatization at thermoneutrality (30 °C) eliminated most of the metabolic phenotype, indicating that impaired adaptive (BAT) thermogenesis can be involved. In conclusion, the metabolic impact of carrying the Ala92-Dio2 allele depends greatly on the genetic background of the mouse, varying from no phenotype in B6 mice to a major phenotype in FVB mice. These results will help the planning of future clinical trials studying the Thr92Ala-DIO2 polymorphism and may explain why some clinical studies performed in different populations across the globe have obtained inconsistent results.
Subject(s)
Iodide Peroxidase , Iodothyronine Deiodinase Type II , Mice, Inbred C57BL , Animals , Male , Iodide Peroxidase/genetics , Mice , Diet, High-Fat , Genetic Background , Adipose Tissue, White/metabolism , Adipose Tissue, Brown/metabolism , Polymorphism, Genetic , Insulin Resistance/genetics , Fatty Liver/geneticsABSTRACT
Levothyroxine (LT4) is a form of thyroid hormone used to treat hypothyroidism. In the brain, T4 is converted to the active form T3 by type 2 deiodinase (D2). Thus, it is intriguing that carriers of the Thr92Ala polymorphism in the D2 gene (DIO2) exhibit clinical improvement when liothyronine (LT3) is added to LT4 therapy. Here, we report that D2 is a cargo protein in ER Golgi intermediary compartment (ERGIC) vesicles, recycling between ER and Golgi. The Thr92-to-Ala substitution (Ala92-D2) caused ER stress and activated the unfolded protein response (UPR). Ala92-D2 accumulated in the trans-Golgi and generated less T3, which was restored by eliminating ER stress with the chemical chaperone 4-phenyl butyric acid (4-PBA). An Ala92-Dio2 polymorphism-carrying mouse exhibited UPR and hypothyroidism in distinct brain areas. The mouse refrained from physical activity, slept more, and required additional time to memorize objects. Enhancing T3 signaling in the brain with LT3 improved cognition, whereas restoring proteostasis with 4-PBA eliminated the Ala92-Dio2 phenotype. In contrast, primary hypothyroidism intensified the Ala92-Dio2 phenotype, with only partial response to LT4 therapy. Disruption of cellular proteostasis and reduced Ala92-D2 activity may explain the failure of LT4 therapy in carriers of Thr92Ala-DIO2.
Subject(s)
Brain , Endoplasmic Reticulum Stress , Hypothyroidism , Iodide Peroxidase , Polymorphism, Genetic , Unfolded Protein Response , Amino Acid Substitution , Animals , Brain/enzymology , Brain/pathology , Endoplasmic Reticulum/enzymology , Endoplasmic Reticulum/genetics , Golgi Apparatus/enzymology , Golgi Apparatus/genetics , HEK293 Cells , Humans , Hypothyroidism/drug therapy , Hypothyroidism/enzymology , Hypothyroidism/genetics , Hypothyroidism/pathology , Iodide Peroxidase/genetics , Iodide Peroxidase/metabolism , Mice , Mice, Transgenic , Mutation, Missense , Thyroxine/therapeutic use , Triiodothyronine/therapeutic use , Iodothyronine Deiodinase Type IIABSTRACT
PURPOSE: The aim of the present study was to evaluate the in vitro effect of carotenoid astaxanthin (ASTA) on the phagocytic and microbicidal capacities, cytokine release, and reactive oxygen species production in human neutrophils. METHODS: The following parameters were evaluated: cytotoxic effect of ASTA on human neutrophils viability, phagocytic and microbicidal capacities of neutrophils by using Candida albicans assay, intracellular calcium mobilization (Fura 2-AM fluorescent probe), superoxide anion (lucigenin and DHE probes), hydrogen peroxide (H2O2, phenol red), and nitric oxide (NO·) (Griess reagent) production, activities of antioxidant enzymes (total/Mn-SOD, CAT, GPx, and GR), oxidative damages in biomolecules (TBARS assay and carbonyl groups), and cytokine (IL-6 and TNF-alpha) release. RESULTS: Astaxanthin significantly improves neutrophil phagocytic and microbicidal capacity, and increases the intracellular calcium concentration and NO· production. Both functional parameters were accompanied by a decrease in superoxide anion and hydrogen peroxide and IL-6 and TNF-α production. Oxidative damages in lipids and proteins were significantly decreased after ASTA-treatment. CONCLUSIONS: Taken together our results are supportive to a beneficial effect of astaxanthin-treatment on human neutrophils function as demonstrated by increased phagocytic and fungicide capacity as well as by the reduced superoxide anion and hydrogen peroxide production, however, without affecting neutrophils capacity to kill C. albicans. This process appears to be mediated by calcium released from intracellular storages as well as nitric oxide production.
Subject(s)
Antioxidants/pharmacology , Neutrophils/drug effects , Phagocytosis/drug effects , Antioxidants/toxicity , Calcium/metabolism , Candida albicans/drug effects , Catalase/metabolism , Cell Survival/drug effects , Cells, Cultured , Cytokines/metabolism , Dimethyl Sulfoxide/pharmacology , Dimethyl Sulfoxide/toxicity , Glutathione Peroxidase/metabolism , Glutathione Reductase/metabolism , Humans , Immune System/drug effects , Neutrophils/metabolism , Neutrophils/physiology , Nitrogen Oxides/metabolism , Oxidative Stress , Reactive Oxygen Species/metabolism , Superoxide Dismutase/metabolism , Xanthophylls/pharmacology , Xanthophylls/toxicityABSTRACT
Upon mitogen sensitization, lymphocytes undergo proliferation by oxyradical-based mechanisms. Through continuous resting-restimulation cycles, lymphocytes accumulate auto-induced oxidative lesions which lead to cell dysfunction and limit their viability. Astaxanthin (ASTA) is a nutritional carotenoid that shows notable antioxidant properties. This study aims to evaluate whether the in vitro ASTA treatment can limit oxyradical production and auto-oxidative injury in human lymphocytes. Activated lymphocytes treated with 5 microM ASTA showed immediate lower rates of O(2)(*-) /H(2)O(2) production whilst NO* and intracellular Ca(2+) levels were concomitantly enhanced (