Rheumatoid arthritis (RA) and its animal model adjuvant arthritis (AA) are inflammatory diseases characterized by chronic inflammation, systemic oxidative stress and disturbed mitochondrial bioenergetics of skeletal muscle. The present study aimed to evaluate the effects of coenzyme Q10 - CoQ10 (100 mg/kg b.w.), omega-3-polyunsaturated fatty acids - omega-3-PUFA (400 mg/kg b.w.) and their combined treatment in AA on impaired skeletal muscle mitochondrial bioenergetics, inflammation and changes in levels CoQ9 and CoQ10 in plasma. Markers of inflammation (C-reactive protein, monocyte-chemotactic protein-1), antioxidant capacity of plasma, respiratory chain parameters of skeletal muscle mitochondria and concentrations of CoQ9 and CoQ10 in plasma and in muscle tissue were estimated. Treatment of the arthritic rats with CoQ10, omega-3-PUFA alone and in combination partially reduced markers of inflammation and increased antioxidant capacity of plasma, significantly increased concentrations of coenzyme Q in mitochondria and improved mitochondrial function in the skeletal muscle. Combined treatment has similar effect on the mitochondrial function as monotherapies; however, it has affected inflammation and antioxidant status more intensively than monotherapies. Long-term supplementary administration of coenzyme Q10 and omega-3-PUFA and especially their combination is able to restore the impaired mitochondrial bioenergetics and antioxidant status in AA.

• MeSH
• Antioxidants metabolism   MeSH
• Arthritis, Experimental blood   diet therapy   MeSH
• C-Reactive Protein metabolism   MeSH
• Chemokine CCL2 blood   MeSH
• Rats MeSH
• Fatty Acids, Omega-3 therapeutic use   MeSH
• Rats, Inbred Lew MeSH
• Dietary Supplements MeSH
• Arthritis, Rheumatoid blood   diet therapy   MeSH
• Mitochondria, Muscle metabolism   MeSH
• Ubiquinone analogs & derivatives   metabolism   therapeutic use   MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Male MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Evaluation Study MeSH
• Names of Substances
• Antioxidants MeSH
• C-Reactive Protein MeSH
• Chemokine CCL2 MeSH
• coenzyme Q10 MeSH Browser
• Fatty Acids, Omega-3 MeSH
• Ubiquinone MeSH
• ubiquinone 9 MeSH Browser

Nitrotyrosine formation is caused by presence of reactive oxygen and nitrogen species. Nitration is a very selective process leading to specific modification of only a few tyrosines in protein molecule. 2D electrophoresis and western blotting techniques coupled with mass spectrometry are common methods used in analysis of proteome. Here we describe protocol for analysis of peroxynitrite-induced protein nitration in isolated mitochondria. Mitochondrial proteins are separated by 2D electrophoresis and transferred to nitrocellulose membrane. Membranes are then incubated with antibodies against nitrotyrosine. Positive spots are compared with corresponding Coomassie-stained gels, and protein nitration is confirmed with mass spectrometry techniques.

• Keywords
• 2D electrophoresis, Immunoblotting, Mass spectrometry, Mitochondria, Nitrotyrosine,
• MeSH
• Electrophoresis, Gel, Two-Dimensional methods   MeSH
• Mass Spectrometry methods   MeSH
• Immunoblotting methods   MeSH
• Peroxynitrous Acid chemistry   MeSH
• Mitochondrial Proteins analysis   metabolism   MeSH
• Cattle MeSH
• Mitochondria, Heart chemistry   metabolism   MeSH
• Tyrosine analogs & derivatives   analysis   metabolism   MeSH
• Animals MeSH
• Check Tag
• Cattle MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• 3-nitrotyrosine MeSH Browser
• Peroxynitrous Acid MeSH
• Mitochondrial Proteins MeSH
• Tyrosine MeSH