An Integrated Approach Combining Chemical Profiling, Network Pharmacology, and Experimental Validation Is Used to Clarify the Pharmacological Basis of the Yiqi‐Tongluo‐Huoxue‐Mingmu Formula in Diabetic Retinopathy

Background According to traditional Chinese medical (TCM) principles diabetic retinopathy (DR) is categorized as a type of “Wasting‐Thirst Eye Disease.” According to TCM, the pathophysiology of DR is primarily attributed to blood stasis, obstructed collaterals, and concurrent Qi and Yin deficiencies. Yiqi‐Tongluo‐Huoxue‐Mingmu Formula (YQMM), a TCM commonly prescribed for DR, has yet to have its active components and underlying molecular mechanisms fully elucidated. Methods The therapeutic efficacy of YQMM was assessed in mice with diabetes induced by streptozotocin (STZ). The chemical profiles of YQMM and its absorbed prototype components in vivo were characterized using LC‐Q‐TOF/MS. After predicting potential targets and pathways using network pharmacology methods and molecular docking, this study validated these computational results through RT‐qPCR experiments. To further evaluate the antiproliferative and antiangiogenic effects of YQMM and its active components, HUVECs cultured under high‐glucose conditions were used. Results The retinal protective effects of YQMM were observed without significant alterations in systemic blood glucose levels. Chemical analysis identified 142 compounds, with five prototype constituents (caffeic acid, isoferulic acid, daidzin, daidzein, and mirificin) detected in plasma. Network pharmacology indicated 118 shared targets between YQMM and DR, with key nodes including TP53, ESR1, JUN, STAT3, and MAPK1, primarily linked to the AGE‐RAGE, PI3K‐Akt, and HIF‐1 signaling pathways. Molecular docking predictions revealed strong binding affinities between the targets and the compounds, particularly for daidzin. Both in vivo and in vitro experiments confirmed that YQMM and its active components downregulated the expression of core targets and inhibited high glucose‐induced endothelial cell proliferation and tube formation. Conclusion YQMM confers its protective benefits against DR via a complex interplay of multiple active ingredients acting on diverse molecular targets and signaling networks. These findings elucidate the chemical and mechanistic basis for the clinical efficacy of YQMM, supporting its further development as a promising therapeutic agent for DR.