Mouse Model of IVS

All animal care and experiments were performed after ethical approval from the Animal Care Committee of Nanjing University of Chinese Medicine (approval numbers ACU210305). Male C57BL/6 mice aged 8 weeks were divided into six groups, namely, sham/normal saline (NS; n = 3), sham/diosmin A (n = 3), sham/diosmin B (n = 3), IVS/NS (n = 6), IVS/diosmin A (n = 7), and IVS/diosmin B (n = 7). The doses of diosmin A (Chia Tai Tianqing Pharmaceutical, Nanjing, China) and B (Servier Tianjin Pharmaceutical Co., Ltd. Tianjin, China) were 40 mg/kg. Mice in all groups were administered the treatments intragastrically once a day.

An experimental model of IVS was created based on the model described previously (12). Under narcosis with isoflurane in a supine position, mice underwent stenosis of the left external iliac vein. A groin incision was made, and muscle tissues were separated to expose the vein. The left external iliac vein was separated from the corresponding artery while avoiding damage to the vein and artery. The left external iliac vein was ligated over a 30-G needle with a 7–0 non-absorbable suture. Subsequently, the needle was removed, and the skin was closed by 5–0 non-absorbable sutures.