目的 探讨当归补血汤（DBT）能否通过抑制内质网应激、增强内皮祖细胞（EPCs）动员及自噬活性，促进 急性脑缺血再灌注（I/R）小鼠脑血管生成与功能恢复，为急性缺血性脑卒中（AIS）的防治提供新的机制依据和治 疗策略。方法 采用 C57BL/6 雄性小鼠建立大脑中动脉栓塞（MCAO）模型。动物随机分为假手术组、模型组和 当归补血汤干预组。当归补血汤经灌胃给药，给药起始时间为术后 2 h，每日 1 次，连续 7 d。通过 TTC 染色 评估脑梗死体积；尼氏染色观察神经元形态；免疫荧光检测血管性血友病因子（vWF）、血管内皮细胞标志物 （CD31）以及包括造血干细胞标志物 CD34、造血干/祖细胞标志物 CD133 和血管内皮生长因子受体 2（VEGFR2） 在内的内皮祖细胞的表达，用以评价血管修复与内皮祖细胞归巢情况；Western Blot 检测蛋白激酶样内质网激 酶（PERK）、肌醇需求酶 1α（IRE1α）、真核翻译起始因子 2α（eIF2α）、转录因子 C/EBP 同源蛋白（CHOP）、微 管相关蛋白 1 轻链 3-Ⅱ（LC3-Ⅱ）、自噬相关蛋白 SQSTM1 以及凋亡相关蛋白 Bcl-2、Bax，探讨当归补血汤对 内质网应激与自噬的影响。神经功能缺损采用 Bederson 评分进行行为学评价。结果 与模型组比较，当归补 血汤干预组小鼠脑梗死体积明显减小（P＜0.01），尼氏染色提示存活神经元数量明显增加。免疫荧光结果显示， 当归补血汤组脑组织中微血管密度（vWF、CD31 阳性）明显升高（P＜0.05），内皮祖细胞归巢能力增强，CD34/ CD133/VEGFR2 三重阳性细胞数量明显增加（P＜0.01）。Western Blot 结果表明，当归补血汤组 PERK、IRE1α、 eIF2α 及 CHOP 蛋白表达水平均较模型组明显下降（P＜0.01），而 LC3-Ⅱ表达上调、SQSTM1 表达下调（P＜ 0.01），提示当归补血汤可能通过抑制内质网应激并促进自噬通量发挥神经保护作用。行为学方面，当归补血 汤组在术后第 3 与第 5 天的贝德森神经功能缺损评分（Bederson）较模型组明显降低（P＜0.05，P＜0.01）。 结论 当归补血汤能明显缩小 MCAO 模型小鼠的脑梗死体积，减轻神经功能缺损，改善组织病理形态，并促 进神经元存活、微血管再生及内皮祖细胞归巢。其机制可能与抑制内质网应激、促进自噬通量、抑制细胞凋亡 及增强血管修复能力有关。

Objective To investigate whether Danggui Buxue Decoction （DBT） can promote cerebral angiogenesis and functional recovery in mice with acute cerebral ischemia-reperfusion （I/R） injury by inhibiting endoplasmic reticulum stress，enhancing the mobilization of endothelial progenitor cells （EPCs），and regulating autophagy activity，thereby providing new mechanistic insights and therapeutic strategies for the prevention and treatment of acute ischemic stroke （AIS）. Methods A middle cerebral artery occlusion （MCAO） model was established using male C57BL/6 mice. The animals were randomly divided into sham-operated， model， and DBT-treated groups. DBT was administered via intragastric gavage starting at 2 hours post-surgery， once daily for 7 consecutive days. Cerebral infarct volume was assessed by TTC staining；neuronal morphology was observed via Nissl staining；immunofluorescence was employed to detect the expression of von Willebrand factor （vWF），the vascular endothelial cell marker CD31，and EPC markers （including the hematopoietic stem cell marker CD34， the hematopoietic stem/progenitor cell marker CD133， and vascular endothelial growth factor receptor 2（VEGFR2） to evaluate vascular repair and EPC homing. Western Blot analysis was performed to examine the protein expression levels of protein kinase R-like endoplasmic reticulum kinase （PERK），inositol-requiring enzyme 1α（IRE1α），eukaryotic translation initiation factor 2α（eIF2α），the transcription factor C/EBP homologous protein （CHOP），microtubule-associated protein 1 light chain 3-Ⅱ（LC3-Ⅱ），and the autophagy-related protein SQSTM1 and apoptosis-related proteins Bcl-2 and Bax， to explore the effects of DBT on endoplasmic reticulum stress and autophagy. Neurological deficits were evaluated using the Bederson score. Results Compared with the model group，mice in the DBT-treated group exhibited a significant reduction in cerebral infarct volume （P＜0.01）. Nissl staining indicated a marked increase in the number of surviving neurons. Immunofluorescence results showed that the DBT group had significantly increased microvessel density （positive for vWF and CD31） in brain tissue （P＜0.05），enhanced EPC homing ability，and a significantly greater number of CD34/CD133/VEGFR2 triplepositive cells （P＜0.01）. Western Blot analysis revealed that the expression levels of PERK， IRE1α， eIF2α， and CHOP proteins were significantly lower in the DBT group compared with the model group （P＜0.01），while LC3-Ⅱ expression was upregulated and SQSTM1 expression was downregulated （P＜0.01），suggesting that DBT might exert neuroprotective effects by inhibiting endoplasmic reticulum stress and promoting autophagic flux. Regarding behavioral assessment，the Bederson neurological deficit scores on post-operative days 3 and 5 were significantly lower in the DBT group than in the model group （P＜0.05，P＜0.01）. Conclusion DBT significantly reduces cerebral infarct volume， alleviates neurological deficits，improves histopathological morphology，and promotes neuronal survival，microvascular regeneration， and endothelial progenitor cell homing in MCAO model mice. The underlying mechanisms may be associated with the inhibition of endoplasmic reticulum stress，promotion of autophagic flux，suppression of apoptosis， and enhancement of vascular repair capacity.

R285.5

广东省基础与应用基础研究基金企业联合基金项目 （2022A1515220121）。