Hydrogen sulfide promotes proliferation and regeneration of human cerebral microvascular endothelial cells via the sonic hedgehog signaling pathway.

We investigated the effects of hydrogen sulfide (H(2)S) and sonic hedgehog (SHH) on the proliferation, autophagy, and apoptosis of human microvascular endothelial cells (HCMEC/D3). We also explored the regulatory relationship between cystathionine-β-synthase (CBS) and the SHH pathway. Human microglia cells (HMC3) were stimulated under hypoxia to secrete H(2)S and SHH proteins, which were then co-cultured with HCMEC/D3 cells. The relationship between H(2)S and SHH was investigated by inhibiting the CBS or SHH pathways. Vascular endothelial growth factor (VEGF) and H(2)S levels were detected using ELISA. The mRNA and Protein levels of VEGF, Beclin-1, light chain 3 (LC3), Cysteine aspartic acid protease-3(caspase-3), CBS, SHH, extracellular regulated kinase 1/2 (ERK1/2) and phospho-ERK1/2 (P-ERK1/2) were determined by RT-PCR and western blot. The results indicated that H(2)S secretion by HMC3 increased during hypoxia, with both CBS and SHH proteins being up-regulated. The inhibition of CBS resulted in decreased levels of H(2)S and SHH in HMC3. When the SHH pathway is inhibited, H(2)S secretion levels remain unaffected. H(2)S and SHH proteins increased VEGF, P-ERK1/2, Beclin-1, and LC3 expression while reducing caspase-3 expression in HCMEC/D3 cells. H(2)S secretion by HMC3 promotes the proliferation and regeneration of HCMEC/D3 by regulating SHH protein and alleviating hypoxic injury.