The School of Public Health, Anhui Medical University, Hefei, China.
Wuxi Maternity and Child Health Care Hospital, Wuxi, China.
Mol Med. 2024 Sep 4;30(1):138. doi: 10.1186/s10020-024-00908-6.
Premature rupture of the membranes (PROM) is a key cause of preterm birth and represents a major cause of neonatal mortality and morbidity. Natural products N-acetyl-d-galactosamine (GalNAc), which are basic building blocks of important polysaccharides in biological cells or tissues, such as chitin, glycoproteins, and glycolipids, may improve possible effects of wound healing.
An in vitro inflammation and oxidative stress model was constructed using tumor necrosis-α (TNF-α) and lipopolysaccharide (LPS) action on WISH cells. Human amniotic epithelial cells (hAECs) were primarily cultured by digestion to construct a wound model. The effects of GalNAc on anti-inflammatory and anti-oxidative stress, migration and proliferation, epithelial-mesenchymal transition (EMT), glycosaminoglycan (GAG)/hyaluronic acid (HA) production, and protein kinase B (Akt) pathway in hAECs and WISH cells were analyzed using the DCFH-DA fluorescent probe, ELISA, CCK-8, scratch, transwell migration, and western blot to determine the mechanism by which GalNAc promotes amniotic wound healing.
GalNAc decreased IL-6 expression in TNF-α-stimulated WISH cells and ROS expression in LPS-stimulated WISH cells (P < 0.05). GalNAc promoted the expression of Gal-1 and Gal-3 with anti-inflammatory and anti-oxidative stress effects. GalNAc promoted the migration of hAECs (50% vs. 80%) and WISH cells through the Akt signaling pathway, EMT reached the point of promoting fetal membrane healing, and GalNAc did not affect the activity of hAECs and WISH cells (P > 0.05). GalNAc upregulated the expression of sGAG in WISH cells (P < 0.05) but did not affect HA levels (P > 0.05).
GalNAc might be a potential target for the prevention and treatment of PROM through the galectin pathway, including (i) inflammation; (ii) epithelial-mesenchymal transition; (iii) proliferation and migration; and (iv) regression, remodeling, and healing.
胎膜早破（PROM）是早产的主要原因，也是新生儿死亡和发病的主要原因。天然产物 N-乙酰-D-氨基半乳糖（GalNAc）是生物细胞或组织中重要多糖的基本组成部分，如几丁质、糖蛋白和糖脂，可能改善伤口愈合的效果。
采用肿瘤坏死-α（TNF-α）和脂多糖（LPS）作用于 WISH 细胞构建体外炎症和氧化应激模型。通过消化原代培养人羊膜上皮细胞（hAECs）构建伤口模型。采用 DCFH-DA 荧光探针、ELISA、CCK-8、划痕、Transwell 迁移和 Western blot 分析 GalNAc 对 hAECs 和 WISH 细胞的抗炎、抗氧化应激、迁移和增殖、上皮-间充质转化（EMT）、糖胺聚糖（GAG）/透明质酸（HA）产生及蛋白激酶 B（Akt）通路的影响，探讨 GalNAc 促进羊膜伤口愈合的机制。
GalNAc 降低了 TNF-α刺激的 WISH 细胞中 IL-6 的表达和 LPS 刺激的 WISH 细胞中 ROS 的表达（P<0.05）。GalNAc 促进了具有抗炎和抗氧化应激作用的 Gal-1 和 Gal-3 的表达。GalNAc 通过 Akt 信号通路促进 hAECs（50%比 80%）和 WISH 细胞的迁移，达到促进胎膜愈合的 EMT 点，且不影响 hAECs 和 WISH 细胞的活性（P>0.05）。GalNAc 上调了 WISH 细胞中 sGAG 的表达（P<0.05），但不影响 HA 水平（P>0.05）。
GalNAc 可能通过半乳糖凝集素途径成为预防和治疗 PROM 的潜在靶点，包括（i）炎症；（ii）上皮-间充质转化；（iii）增殖和迁移；和（iv）退行、重塑和愈合。
