低氧诱导因子-1α对牙周炎氧化应激与铁死亡影响的初步研究

A preliminary investigation into the role of hypoxia-inducible factor-1α in oxidative stress and ferroptosis associated with periodontitis

摘要:

目的 探讨低氧诱导因子-1α（HIF-1α）在牙周组织中的表达及对氧化应激与铁死亡的影响。方法 将40只6～8周龄SD雄性大鼠分为常氧对照组（A组），常氧牙周炎组（B组），低氧对照组（C组），低氧牙周炎组（D组），每组10只，分别复制牙周炎模型和低氧模型。模型复制1、3、6周时取牙龈组织，采用苏木精-伊红（HE）染色检测软组织炎症状态，采用免疫组织化学染色检测HIF-1α、铁重链蛋白1（FTH1）和谷胱甘肽过氧化物酶（GPX4）蛋白阳性表达，采用酶联免疫吸附试验（ELISA）检测血清HIF-1α、谷胱甘肽（GSH）及活性氧（ROS）表达。结果 HE染色结果显示：与A组比较，B组炎症随时间延长逐渐加重，C组与A组相似，D组前、中期炎症细胞浸润明显，后期炎症减轻。免疫组织化学染色结果显示：与A组比较，B、C、D组HIF-1α、FTH1表达均升高（P <0.05），D组前、中期HIF-1α、FTH1表达高于B组（P <0.05），后期GPX4表达高于B组（P <0.05）。ELISA结果显示，D组前期HIF-1α、ROS表达升高（P <0.05），中期C组和D组GSH表达高于B组（P <0.05），后期C组和D组ROS表达低于A组和B组（P <0.05）。结论 低氧环境激活的HIF-1α与牙周组织炎症的发生、发展相关，且HIF-1α可能通过影响ROS生成与铁代谢参与牙周组织氧化应激与铁死亡。

Abstract:

Objective To investigate the expression level of hypoxia-inducible factor-1α (HIF-1α) in periodontal tissues and its effects on oxidative stress and ferroptosis, while exploring their interrelationships.Methods Forty 6-8-week-old male Sprague-Dawley (SD) rats were randomly divided into four groups: normoxia control (Group A), normoxia periodontitis (Group B), hypoxia control (Group C), and hypoxia periodontitis (Group D). Periodontitis and hypoxia models were established accordingly. At 1, 3, and 6 weeks post-modeling, gingival tissues were collected for hematoxylin-eosin (HE) staining to evaluate inflammatory status. Immunohistochemistry (IHC) was performed to detect protein expression levels of HIF-1α, ferritin heavy chain 1 (FTH1), and glutathione peroxidase 4 (GPX4). Serum levels of reactive oxygen species (ROS), GPX4, and HIF-1α were measured by enzyme-linked immunosorbent assay (ELISA).Results HE staining: Compared with Group A, Group B showed progressively aggravated inflammation, while Group D exhibited significant inflammatory cell infiltration during early and middle stages with attenuated inflammation at the later stage. IHC staining: HIF-1α and FTH1 expression levels were significantly elevated in Groups B, C and D compared with Group A (P < 0.05). During early and middle stages, Group D demonstrated higher HIF-1α and FTH1 expression than Group B, while at the later stage Group D showed increased GPX4 expression compared to Group B. ELISA: Group D displayed significantly elevated HIF-1α and ROS levels at early stage (P < 0.05). During middle stage, the hypoxia groups exhibited higher GSH expression than Group B (P < 0.05), while at the later stage the Group C and Group D showed lower ROS levels than Group A and Group B (P < 0.05).Conclusion The HIF-1α activated under hypoxic conditions, is associated with the initiation and progression of periodontal inflammation. Furthermore, HIF-1α may contribute to oxidative stress and ferroptosis in periodontal tissues by regulating reactive oxygen species production and iron metabolism.