Background Ageing is accompanied by a decrease in endogenous hydrogen sulphide (H2S) synthesis and the development of mitochondrial dysfunction. The aim of our work was to study the possible participation of exercise training‐induced regulation of endogenous H2S production in the restoration of mitochondrial function in old rats. Materials and Methods Male rats were divided into three groups: adult, old and exercise‐trained old. Exercise training of old rats was performed for 4 weeks. The mRNA expression cystathionine‐γ‐lyase (CSE) and 3‐mercaptopyruvate sulfurtransferase (3‐MST) were determined using reverse transcription and real‐time polymerase chain reaction analysis. Mitochondrial dysfunction was determined by mPTP opening, which was investigated by spectrophotometric registration of the swelling of mitochondria isolated from the rat heart. We also studied the effect of exercise on H2S content, oxidative stress and mtNOS activity. Results Exercise training in old animals significantly increased the expression of H2S‐synthesizing enzymes CSE and 3‐MST and restored endogenous H2S production in cardiac tissue and cardiac mitochondria to levels of adult animals. In addition, the training significantly reduced oxidative stress in old rats, in particular the rate of formation of •O2− and H2O2, diene conjugates and malondialdehyde levels in the mitochondria of the heart. Simultaneously, in the hearts of these animals, resistance of mPTP to the inducer of its opening of calcium ions was increased. Conclusions Thus, exercise training restores endogenous H2S production, and significantly reduces oxidative stress in cardiac mitochondria of old rats that are associated with the inhibition of calcium‐induced mPTP opening as an indicator of mitochondrial dysfunction.

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