Objective Age-related cataract (ARC) is an eye disease characterized by lens opacity. This study analyzed the correlation between 25 hydroxyvitamin D [25 (OH) D] levels and Interleukin 6 (IL-6) levels in aqueous humor from the patients with ARC, as well as between 25 (OH) D levels and lens opacity, and explored the effect of 25 (OH) D on the pathogenesis of ARC. Methods A retrospective analysis was performed, which included the clinical data from 120 ARC patients and 40 healthy individuals. They were divided into three groups based on the objective scattering index (OSI): 40 healthy individuals (control group), 84 patients with the early cataract (EC group), and 36 patients with the mature cataract (MC group). An electrochemical luminescence method was used to detect the levels of 25 (OH) D and IL-6 in aqueous humor, the OSI of the eye was measured using a visual quality analysis system. The correlation between the changes in the above indicators and ARC was analyzed. Results The OSI and IL-6 levels in the control group, EC group, and MC group increased sequentially (all p < .001), while the levels of 25 (OH) D showed an opposite trend (all p < .001). Conclusion The decrease in 25 (OH) D levels is related to the occurrence and development of ARC, and the relationship between 25 (OH) D deficiency and the pathological mechanism of ARC needs further research.