ABSTRACT
Skeletal muscle weakness in patients suffering from rheumatoid arthritis (RA) adds to their impaired working abilities and reduced quality of life. However, little molecular insight is available on muscle weakness associated with RA. Oxidative stress has been implicated in the disease pathogenesis of RA. Here we show that oxidative post-translational modifications of the contractile machinery targeted to actin result in impaired actin polymerization and reduced force production. Using mass spectrometry, we identified the actin residues targeted by oxidative 3-nitrotyrosine (3-NT) or malondialdehyde adduct (MDA) modifications in weakened skeletal muscle from mice with arthritis and patients afflicted by RA. The residues were primarily located to three distinct regions positioned at matching surface areas of the skeletal muscle actin molecule from arthritis mice and RA patients. Moreover, molecular dynamic simulations revealed that these areas, here coined "hotspots", are important for the stability of the actin molecule and its capacity to generate filaments and interact with myosin. Together, these data demonstrate how oxidative modifications on actin promote muscle weakness in RA patients and provide novel leads for targeted therapeutic treatment to improve muscle function.
Subject(s)
Actins/metabolism , Arthritis, Rheumatoid/metabolism , Muscle Weakness/metabolism , Muscle, Skeletal/metabolism , Oxidative Stress , Actins/chemistry , Animals , Arthritis, Rheumatoid/complications , Disease Models, Animal , Female , Humans , Malondialdehyde , Mice , Mice, Inbred C57BL , Molecular Dynamics Simulation , Muscle Contraction/physiology , Muscle Weakness/etiology , Muscle Weakness/physiopathology , Muscle, Skeletal/physiopathology , Myosins/chemistry , Myosins/metabolism , Polymerization , Protein Processing, Post-Translational , Tyrosine/analogs & derivativesABSTRACT
PURPOSE: To quantify and compare the time-course and potency of the analgesic and antipyretic effects of naproxen in conjunction with the inhibition of PGE(2) and TXB(2). METHODS: Analgesia was investigated in a rat model with carrageenan-induced arthritis using a gait analysis method. Antipyretics were studied in a yeast-induced fever model using telemetrically recorded body temperature. Inhibition of TXB(2) and PGE(2) synthesis was determined ex vivo. Pharmacokinetic profiles were obtained in satellite animals. Population PKPD modeling was used to analyze the data. RESULTS: The IC(50) values (95% CI) of naproxen for analgesia (27 (0-130) µM), antipyretics (40 (30-65) µM) and inhibition of PGE(2) (13 (6-45) µM) were in similar range, whereas inhibition of TXB(2) (5 (4-8) µM) was observed at lower concentrations. Variability in the behavioral measurement of analgesia was larger than for the other endpoints. The inhibition of fever by naproxen was followed by an increased rebound body temperature. CONCLUSION: Due to better sensitivity and similar drug-induced inhibition, the biomarker PGE(2) and the antipyretic effect would be suitable alternative endpoints to the analgesic effects for characterization and comparisons of potency and time-courses of drug candidates affecting the COX-2 pathway and to support human dose projections.
