Departments of Ophthalmology and Anatomy and Cell Biology, Kresge Eye Institute, Wayne State University School of Medicine, Detroit, Michigan, United States.
Invest Ophthalmol Vis Sci. 2024 Mar 5;65(3):35. doi: 10.1167/iovs.65.3.35.
To investigate the mechanisms underlying the differential roles of TGFβ1 and TGFβ3 in accelerating corneal epithelial wound healing (CEWH) in diabetic (DM) corneas, with normoglycemia (NL) corneas as the control.
Two types of diabetic mice, human corneal organ cultures, mouse corneal epithelial progenitor cell lines, and bone marrow-derived macrophages (BMDMs) were employed to assess the effects of TGFβ1 and TGFβ3 on CEWH, utilizing quantitative PCR, western blotting, ELISA, and whole-mount confocal microscopy.
Epithelial debridement led to an increased expression of TGFβ1 and TGFβ3 in cultured human NL corneas, but only TGFβ1 in DM corneas. TGFβ1 and TGFβ3 inhibition was significantly impeded, but exogenous TGFβ1 and, more potently, TGFβ3 promoted CEWH in cultured TKE2 cells and in NL and DM C57BL6 mouse corneas. Wounding induced similar levels of p-SMAD2/SMAD3 in NL and DM corneas but weaker ERK1/2, Akt, and EGFR phosphorylation in DM corneas compared to NL corneas. Whereas TGFβ1 augmented SMAD2/SMAD3 phosphorylation, TGFβ3 preferentially activated ERK, PI3K, and EGFR in healing DM corneas. Furthermore, TGFβ1 and TGFβ3 differentially regulated the expression of S100a9, PAI-1, uPA/tPA, and CCL3 in healing NL and DM corneas. Finally, TGFβ1 induced the expression of M1 macrophage markers iNOS, CD86, and CTGF, whereas TGFβ3 promoted the expression of M2 markers CD206 and NGF in BMDMs from db/db or db/+ mice.
Hyperglycemia disrupts the balanced expression of TGFβ3/TGFβ1, resulting in delayed CEWH, including impaired sensory nerve regeneration in the cornea. Supplementing TGFβ3 in DM wounds may hold therapeutic potential for accelerating delayed wound healing in diabetic patients.
研究转化生长因子β1（TGFβ1）和转化生长因子β3（TGFβ3）在加速糖尿病（DM）角膜上皮伤口愈合（CEWH）中的差异作用机制，以正常血糖（NL）角膜作为对照。
使用两种类型的糖尿病小鼠、人角膜器官培养物、小鼠角膜上皮祖细胞系和骨髓来源的巨噬细胞（BMDMs），通过定量 PCR、western blot、ELISA 和全层共聚焦显微镜评估 TGFβ1 和 TGFβ3 对 CEWH 的影响。
上皮清创导致培养的 NL 角膜中 TGFβ1 和 TGFβ3 的表达增加，但 DM 角膜中仅 TGFβ1 增加。TGFβ1 和 TGFβ3 抑制明显受阻，但外源性 TGFβ1 更能促进培养的 TKE2 细胞以及 NL 和 DM C57BL6 小鼠角膜的 CEWH。与 NL 角膜相比，伤口诱导导致 NL 和 DM 角膜中 p-SMAD2/SMAD3 水平相似，但 DM 角膜中 ERK1/2、Akt 和 EGFR 磷酸化较弱。虽然 TGFβ1 增强了 SMAD2/SMAD3 磷酸化，但 TGFβ3 优先在愈合的 DM 角膜中激活 ERK、PI3K 和 EGFR。此外，TGFβ1 和 TGFβ3 以不同的方式调节愈合的 NL 和 DM 角膜中 S100a9、PAI-1、uPA/tPA 和 CCL3 的表达。最后，TGFβ1 诱导 db/db 或 db/+ 小鼠 BMDMs 中 M1 巨噬细胞标志物 iNOS、CD86 和 CTGF 的表达，而 TGFβ3 促进 M2 标志物 CD206 和 NGF 的表达。
高血糖破坏了 TGFβ3/TGFβ1 的平衡表达，导致 CEWH 延迟，包括角膜中感觉神经再生受损。在 DM 伤口中补充 TGFβ3 可能为加速糖尿病患者的延迟伤口愈合提供治疗潜力。
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