Modified Ziyun Ointment protects against high glucose-induced endothelial injury through miR-29c-3p-dependent modulation of VEGFA and AGE-RAGE signaling.

BACKGROUND: Diabetic ulceration is a major diabetes-related complication characterized by impaired angiogenesis, persistent inflammation, oxidative stress, and endothelial dysfunction. Modified Ziyun Ointment (MZYO), derived from the traditional Zi-Yun ointment, has shown clinical benefits in wound repair, but its molecular mechanisms remain unclear. METHODS: A high glucose-induced human umbilical vein endothelial cell (HUVEC) injury model was established. miR-29c-3p expression was measured by quantitative real-time PCR. Cell viability, apoptosis, and migration were evaluated using CCK-8, flow cytometry, wound-healing, and Transwell assays. VEGFA targeting by miR-29c-3p was examined using dual-luciferase reporter assays. VEGFA, AGE, and RAGE protein levels were assessed by Western blotting. The protective effects of MZYO-containing serum (MZYO-S) were investigated under high glucose conditions. RESULTS: High glucose increased miR-29c-3p expression and induced endothelial injury, reflected by reduced viability, enhanced apoptosis, and impaired migration. miR-29c-3p overexpression worsened these effects, whereas inhibition attenuated them. VEGFA was confirmed as a direct target of miR-29c-3p, and miR-29c-3p upregulation suppressed VEGFA while activating the AGE-RAGE pathway. MZYO-S significantly improved cell viability and migration, reduced apoptosis, increased VEGFA expression, and decreased AGE/RAGE levels in high glucose-treated HUVECs. Rescue experiments showed that miR-29c-3p inhibition strengthened the protective effects of MZYO-S, while miR-29c-3p overexpression partially reversed them. CONCLUSION: MZYO-S protects endothelial cells from high glucose-induced injury through modulation of the miR-29c-3p/VEGFA/AGE-RAGE signaling axis, supporting its potential as a candidate multi-target strategy for diabetic ulcer repair. Because the present design did not resolve the direction of regulation between VEGFA and the AGE-RAGE pathway, this axis is described as a coordinated regulatory network rather than a strictly linear cascade, and these in vitro findings require confirmation in in vivo models.