Objective To analyze the relationship between serum myocardial enzyme expression and disease occurrence and progression in neonatal hyperbilirubinemia children. Methods 94 neonates with hyperbilirubinemia treated in our hospital from January 2022 to July 2024 were included in the hyperbilirubinemia group. Another 94 healthy newborns born in our hospital during the same period were included in the health group. Children were divided into mild group, moderate group and severe group according to different disease progression. Baseline data were collected and compared between the two groups. The levels of myocardial enzymes [aspartate aminotransferase (AST), creatine kinase (CK), creatine kinase isoenzyme (CK-MB), lactate dehydrogenase (LDH)] in the two groups were analyzed and compared. The serum myocardial enzyme expression levels of neonatal hyperbilirubinemia with different disease progression were analyzed and compared. Spearman correlation analysis was used to analyze the correlation between myocardial enzymes and hyperbilirubinemia. Results There were no significant differences in gender, gestational age, birth weight, delivery mode, feeding regimen and age between hyperbilirubinemia group and healthy group (P>0.05). Serum AST, CK, CK-MB and LDH levels in hyperbilirubinemia group were significantly higher than those in healthy group (P<0.05). According to different disease progression, the children were divided into mild group (n=50), moderate group (n=23) and severe group (n=21). With the severity of disease, serum AST, CK, CK-MB and LDH levels showed an increasing trend, and there were significant differences between groups (P<0.05). Spearman correlation analysis showed that AST, CK, CK-MB, LDH levels were positively correlated with the progression of hyperbilirubinemia (P<0.05). Conclusion The expression levels of AST, CK, CK-MB, LDH and other myocardial enzymes in neonatal hyperbilirubinemia are increased, which is related to the progression of the disease.