[Wenyang Huazhuo Tongluo Formula promotes angiogenesis in systemic sclerosis dermal microvascular endothelial cells...

To investigate the effect of Wenyang Huazhuo Tongluo Formula (WHT) for promoting angiogenesis in systemic sclerosis (SSc) dermal microvascular endothelial cells and its possible mechanism.

Wistar rats were gavaged with 15, 30, or 60 g/kg WHT and normal saline for 7 consecutive days to prepare low-, medium-, or high-concentration WHT-medicated sera and blank serum, respectively. Human dermal microvascular endothelial cells (HDMECs) in routine culture were treated with the sera from SSc patients to establish an SSc cell model. The cells were treated with the blank serum, low-, medium-, or high-concentration WHT-medicated sera, or blank serum combined with EGCG (a Sema3A inhibitor). The changes in cell proliferation, migration and angiogenesis were assessed using CCK-8 assay, wound-healing assay and Matrigel tube formation assay, and the mRNA and protein expressions of Sema3A, Nrp1, VEGFA, CD31 and α-SMA were analyzed using qRT-PCR and Western blotting.

HDMECs treated with the serum from SSc patients exhibited significantly reduced cell proliferation and migration rates, increased α-SMA, Sema3A and VEGFA protein and mRNA expression levels, decreased CD31 and Nrp1 protein and mRNA expression levels, and suppressed angiogenesis. In SSc serum-induced cells, treatments with low-, medium-, or high-concentration WHT-medicated sera or EGCG all significantly improved cell survival and migration rates, reduced α-SMA, Sema3A and VEGFA protein and mRNA expression levels, enhanced CD31 and Nrp1 protein and mRNA expressions, and promoted angiogenesis of the cells.

WHT promotes angiogenesis of SSc sera-induced HDMECs by modulating the Sema3A/Nrp1 signaling pathway.