<p>Myopia is the most widespread refractive error caused by an increase in the axial length (AL) of the eyeball, and is also a major risk factor for other blinding eye diseases, seriously endangering human health and quality of life. Investigating practical methods to control the progression of myopia is therefore crucial. The only medication that has been shown to effectively delay the progression of myopia over an extended period of time is atropine. Although they have more adverse effects, atropine eye drops with a high concentration work best to correct myopia. Atropine at low concentrations can slow the progression of myopia, however the results might not be very noticeable. It is of great significance to explore the application of drugs to suppress myopia in combination with low-concentration atropine in the treatment of myopia. This study investigated the role and mechanism of 0.01% atropine combined with carteolol hydrochloride in the treatment of myopia. By establishing a model of form-deprivation myopia in guinea pigs, we examined the protective effect of 0.01% atropine combined with carteolol hydrochloride on myopia in guinea pigs and further explored the mechanism of scleral remodeling mediated by mitochondrial dysfunction. We found that 0.01% atropine combined with carteolol hydrochloride refined mitochondrial dysfunction-induced extracellular matrix degradation by activating the PGC-1α/NRF2/HO-1 signaling pathway, thereby suppressing scleral remodeling and the progression of form-deprivation myopia in guinea pigs. In conclusion, 0.01% atropine combined with carteolol hydrochloride may be an effective strategy for the treatment of myopia.</p>