目的 探讨丁香酚调控硫氧还蛋白互作蛋白/硫氧还蛋白/谷胱甘肽过氧化物酶4（TXNIP/TRX-1/GPX4）通路介导的铁死亡对血管性痴呆（VD）大鼠术后认知功能障碍的影响。 方法 将大鼠随机分为模型组、丁香酚低剂量组、丁香酚高剂量组、丁香酚高剂量+OE-NC组、丁香酚高剂量+OE-TXNIP组，每组12只。除对照组外，其余组别均构建VD模型。水迷宫实验检测大鼠认知功能；ELISA检测海马组织丙二醛（MDA）和还原型谷胱甘肽（GSH）含量；尼氏染色法及HE染色观察海马CA1区神经元形态；普鲁士蓝染色观察海马组织铁沉积；免疫组化检测铁死亡蛋白GPX4、溶质载体家族7成员11（SLC7A11）表达；Western blot检测海马组织TXNIP/TRX-1/GPX4通路相关蛋白表达。 结果 模型组较对照组海马组织病理损伤严重，CA1区神经元细胞形态不规则，胞体水肿、胞核固缩现象明显，细胞间隙增大、边界模糊，神经元排列稀疏、紊乱，丢失明显，尼氏体明显减少，铁颗粒沉积增加，大鼠平均逃避潜伏期增加，穿越平台次数减少，MDA含量及TXNIP表达升高，GSH含量及SLC7A11、TRX-1、GPX4表达降低（P<0.05）；丁香酚低剂量组、丁香酚高剂量组较模型组海马组织及CA1区神经元损伤明显改善，铁颗粒沉积减少，平均逃避潜伏期减短，穿越平台次数增多，MDA含量及TXNIP表达降低，GSH含量及SLC7A11、TRX-1、GPX4表达升高（P<0.05）；丁香酚高剂量+OE-TXNIP组部分逆转了丁香酚对海马组织的保护作用。 结论 丁香酚可改善血管性痴呆大鼠术后认知功能障碍，与抑制TXNIP/TRX-1/GPX4通路介导的铁死亡相关。
Abstract
Objective To investigate the effect of eugenol on postoperative cognitive dysfunction in vascular dementia (VD) rats via modulation of thioredoxin-interacting protein/thioredoxin-1/glutathione peroxidase 4 (TXNIP/TRX-1/GPX4) pathway-mediated ferroptosis. Methods The rats were randomly divided into control group, model group, low-dose eugenol group, high-dose eugenol group, high-dose eugenol+OE-NC group, and high-dose eugenol+OE-TXNIP group, with 12 rats in each group. Except for control group, VD models were constructed in all other groups. The water maze experiment was used to exam the cognitive function of rats. ELISA was used to detect the contents of malondialdehyde (MDA) and reduced glutathione (GSH) in hippocampal tissue. Nissl staining and HE staining were used to observe the morphology of neurons in hippocampal CA1 area. Prussian blue staining was used to observe iron deposition in hippocampal tissue. Immunohistochemistry was used to detect the expressions of ferroptosis protein GPX4 and solute carrier family 7 member 11 (SLC7A11). Western blot was used to detect the expression of proteins related to TXNIP/TRX-1/GPX4 pathway in hippocampal tissues. Results Compared with control group, the pathological damage of hippocampal tissue in model group was severe. The morphology of neuronal cells in hippocampal CA1 area was irregular, with obvious phenomena of cell edema and nuclear condensation, enlarged cell gaps, blurred boundaries, disordered arrangement, obvious neuronal loss, significant reduction of Nissl body, and increased deposition of iron particles. The rats of model group had increased average escape latency, and decreased number of crossing platforms (P<0.05). The content of MDA and the expression of TXNIP were increased, while the content of GSH and the expressions of SLC7A11, TRX-1, and GPX4 were decreased in model group (P<0.05). Compared with model group, hippocampal tissue and neuronal damage in CA1 area in low-dose eugenol group and high-dose eugenol group were significantly improved, iron particle deposition was reduced, the average escape latency was shortened, the number of crossing platforms increased. Meanwhile, MDA content and TXNIP expression were decreased. The content of GSH and the expressions of SLC7A11, TRX-1, and GPX4 were increased in two group (P<0.05). In addition, high-dose eugenol+OE-TXNIP group partially reversed the protective effect of eugenol on the hippocampal tissue. Conclusions Eugenol can improve postoperative cognitive dysfunction in VD rats and is associated with the inhibition of ferroptosis mediated by TXNIP/TRX-1/GPX4 pathway.
关键词
丁香酚 /
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TXNIP/TRX-1/GPX4通路 /
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铁死亡 /
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血管性痴呆 /
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术后认知功能障碍
Key words
Eugenol /
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TXNIP/TRX-1/GPX4 pathway /
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Ferroptosis /
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Vascular dementia /
Postoperative cognitive dysfunction
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基金
黑龙江省自然科学基金项目（LH2023H052）
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