Treatment with pyridostigmine attenuated cardiac hypertrophy and improved cardiac function, which was related to improvement of cholinergic transmission efficiency via inhibition of AChE, suppression of Orai1/STIM1 complex formation and inhibition of the downstream CaN/NFAT3/p‐GATA4 signalling pathway in the pathogenesis of cardiac conditions in the presence of stress. This evidence concerns the gene ACHE and cardiac hypertrophy.