Table_1_Dl-3-n-Butylphthalide Alleviates Hippocampal Neuron Damage in Chronic Cerebral Hypoperfusion via Regulation of the CNTF/CNTFRα/JAK2/STAT3 Signaling Pathways.docx

Chronic cerebral hypoperfusion (CCH) contributes to cognitive impairments, and hippocampal neuronal death is one of the key factors involved in this process. Dl-3-n-butylphthalide (D3NB) is a synthetic compound originally isolated from the seeds of Apium graveolens, which exhibits neuroprotective effects against some neurological diseases. However, the protective mechanisms of D3NB in a CCH model mimicking vascular cognitive impairment remains to be explored. We induced CCH in rats by a bilateral common carotid artery occlusion (BCCAO) operation. Animals were randomly divided into a sham-operated group, CCH 4-week group, CCH 8-week group, and the corresponding D3NB-treatment groups. Cultured primary hippocampal neurons were exposed to oxygen-glucose deprivation/reperfusion (OGD/R) to mimic CCH in vitro. We aimed to explore the effects of D3NB treatment on hippocampal neuronal death after CCH as well as its underlying molecular mechanism. We observed memory impairment and increased hippocampal neuronal apoptosis in the CCH groups, combined with inhibition of CNTF/CNTFRα/JAK2/STAT3 signaling, as compared with that of sham control rats. D3NB significantly attenuated cognitive impairment in CCH rats and decreased hippocampal neuronal apoptosis after BCCAO in vivo or OGD/R in vitro. More importantly, D3NB reversed the inhibition of CNTF/CNTFRα expression and activated the JAK2/STAT3 pathway. Additionally, JAK2/STAT3 pathway inhibitor AG490 counteracted the protective effects of D3NB in vitro. Our results suggest that D3NB could improve cognitive function after CCH and that this neuroprotective effect may be associated with reduced hippocampal neuronal apoptosis via modulation of CNTF/CNTFRα/JAK2/STAT3 signaling pathways. D3NB may be a promising therapeutic strategy for vascular cognitive impairment induced by CCH.

慢性脑灌注不足（CCH）是导致认知障碍的重要因素，其中海马体神经元的死亡是这一过程中涉及的关键因素之一。Dl-3-正丁基邻苯二甲酸酯（D3NB）是一种从芹菜（Apium graveolens）种子中分离出的合成化合物，其对某些神经系统疾病具有神经保护作用。然而，D3NB在模拟血管性认知障碍的CCH模型中的保护机制尚待研究。本研究通过双侧颈动脉结扎术（BCCAO）诱导大鼠CCH，将动物随机分为假手术组、CCH 4周组、CCH 8周组和相应的D3NB治疗组。培养的原代海马神经元暴露于缺氧/复氧（OGD/R）以模拟体外CCH。本研究旨在探讨D3NB治疗对CCH后海马神经元死亡的影响及其潜在的分子机制。与假手术组相比，CCH组观察到记忆力障碍和海马神经元凋亡增加，伴随CNTF/CNTFRα/JAK2/STAT3信号通路的抑制。D3NB显著减轻了CCH大鼠的认知障碍，并在BCCAO体内或OGD/R体外降低了海马神经元凋亡。更重要的是，D3NB逆转了CNTF/CNTFRα表达的抑制并激活了JAK2/STAT3通路。此外，JAK2/STAT3通路抑制剂AG490在体外抵消了D3NB的保护作用。我们的结果表明，D3NB可能改善CCH后的认知功能，其神经保护作用可能与通过调节CNTF/CNTFRα/JAK2/STAT3信号通路减少海马神经元凋亡有关。D3NB可能成为治疗由CCH引起的血管性认知障碍的有望策略。