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Background The incidence of non-alcoholic fatty liver disease (NAFLD) is increasing by years and has become a major public health problem. Serum osteocalcin (OCN), as a marker of bone formation, may increase the risk of NAFLD. Objective To conduct a Meta-analysis to quantitatively evaluate the association between imaging or biopsy-confirmed NAFLD and serum OCN levels. Methods English databases including Medline, Embase, Web of Science, Cochrane, and PROSPERO, as well as Chinese databases including CNKI, Wanfang Data, and VIP were searched for case-control studies, cross-sectional studies, and cohort studies on serum OCN and NAFLD. Two investigators screened the literature, extracted data and performed quality evaluation. Meta-analysis was performed using Stata 16.0 software. Results A total of 13 papers involving 11 772 participants were included. Meta-analysis showed that serum OCN levels were lower in patients with NAFLD than in those without NAFLD (SMD=-0.73, 95%CI=-1.20 to -0.27, P