Background <p>The SPHKS/S1P pathway has a complex yet close association with the progression of diabetic retinopathy (DR). Oxymatrine (OMT), is believed to potentially regulate this pathway; however, systematic clinical evidence remains insufficient.</p> Methods <p>A total of 84 DR cases was finally included in the observation cohort, among which 41 cases were assigned to the OMT group and 43 cases to the non-OMT group. Additionally, 20 non-DR cases were included as blank controls. The levels of 4 target proteins, including S1P, VEGF, SPHK1, and SPHK2, were detected in the peripheral blood of cases in each group. The temporal changes in these protein levels were compared between the OMT group and the non-OMT group. Furthermore, the progression of DR between the two groups was statistically analyzed before and after treatment.</p> Results <p>Compared with non-DR cases, the levels of 4 target proteins of DR cases were significantly increased. No statistically significant differences in the levels of these proteins were observed between the OMT group and the non-OMT group before treatment; the temporal change trends in the expression levels of these proteins in the OMT group were significantly lower than those in the non-OMT group after treatment. However, no statistically significant differences were found between the OMT group and the non-OMT group in terms of DR progression.</p> Conclusion <p>OMT could inhibit the activity of the SPHKS/S1P pathway and downregulate the expression of key proteins; however, the clinical efficacy of OMT in relieving the progression of DR remained unclear in this study.</p>