Single-cell analysis reveals IGF-1 potentiation of inhibition of the TGF-β/Smad pathway of fibrosis in human keratocytes in vitro.

摘要: Corneal wound healing is often affected by TGF-β–mediated fibrosis and scar formation. Guided with IGF-1 antifibrotic substances might maintain corneal transparency. Primary human keratocytes under serum-free conditions were used as a model of stromal wounding, markers opacity studied TGF-β2 stimulation. Single-cell imaging flow cytometry was to determine nuclearization Smad3 intracellular fluorescence intensity Smad7 the crystallin aldehyde dehydrogenase 3A1. Extracellular matrix proteoglycans keratocan biglycan quantified using ELISAs. On background, treated suberoylanilidehydroxamic acid (SAHA) or halofuginone ± IGF-1. alone decreased increased 3A1 expression, favorable extracellular proteoglycan composition. SAHA induced higher levels inhibited translocation nucleus, also when combined IGF-1. Immunofluorescence showed that myofibroblast transdifferentiation attenuated appearance fibroblasts favored in combination substances. The TGF-β/Smad pathway further particular halofuginone. thus valid aid treatment, potential for effective transparent healing.