Cai Gui-Xian, Guo Kai-Kai
Department of Anesthesiology, The PLA Rocket Force Characteristic Medical Center, Beijing 100088, China.
Department of pain medicine, The First Medical Center, Chinese PLA General Hospital, Beijing, 100853, China.
J Stroke Cerebrovasc Dis. 2025 Nov;34(11):108447. doi: 10.1016/j.jstrokecerebrovasdis.2025.108447. Epub 2025 Sep 5.
Ischemic stroke is a leading cause of neurological disability. Current therapies fail to address its multifactorial pathologies. Miltirone, a bioactive compound from Salvia miltiorrhiza, has shown antioxidative and anti-inflammatory potential. However, its neuroprotective mechanisms in stroke remain unexplored.
Using young/aged dMCAO models and OGD/R-treated BV2 microglia, we evaluated Miltirone's effects on infarct volume, neurological function, microglial polarization, lipid metabolism. Cerebral infarct volume was quantified by TTC staining. Neurological deficits were assessed via mNSS, rotarod, and adhesive removal tests. Cell viability was determined by CCK-8 assay. Pro-/anti-inflammatory cytokines, SOD activity and MDA content were measured by ELISA. Microglial polarization was analyzed via immunofluorescence and RT-qPCR. TLR4/MyD88/NF-κB pathway proteins and PLN2 were analyzed by Western blot. Lipid metabolism was evaluated by BODIPY staining. ROS was measured by flow cytometry RESULTS: Miltirone reduced cerebral infarct volume, attenuated brain edema, and improved neurological/motor recovery in dMCAO mice. It shifted microglial polarization toward the anti-inflammatory M2 phenotype by suppressing M1 markers and enhancing M2 markers. Miltirone downregulated pro-inflammatory cytokines while elevating anti-inflammatory cytokines. Miltirone restored lipid homeostasis by inhibiting lipid synthesis genes and activating lipolysis genes. This reduced lipid accumulation. Mechanistically, Miltirone suppressed LBP expression and TLR4/MyD88/NF-κB pathway. Moreover, Miltirone mitigated oxidative stress by lowering ROS, restoring SOD activity, and reducing lipid peroxidation.
Miltirone confers neuroprotection through multi-target actions. It simultaneously provides neuroinflammation, regulates lipid metabolism, and counters oxidative stress. This occurs via LBP/TLR4/NF-κB axis modulation. Its multitarget action addresses the complexity of ischemic stroke pathophysiology, positioning it as a promising therapeutic candidate for clinical translation.
缺血性中风是导致神经功能障碍的主要原因。目前的治疗方法无法解决其多因素病理问题。丹参酮，一种来自丹参的生物活性化合物，已显示出抗氧化和抗炎潜力。然而，其在中风中的神经保护机制仍未被探索。
使用年轻/老年大脑中动脉闭塞（dMCAO）模型和氧糖剥夺/再灌注（OGD/R）处理的BV2小胶质细胞，我们评估了丹参酮对梗死体积、神经功能、小胶质细胞极化和脂质代谢的影响。通过TTC染色定量脑梗死体积。通过mNSS、转棒试验和黏附去除试验评估神经功能缺损。通过CCK-8法测定细胞活力。通过ELISA测量促炎/抗炎细胞因子、超氧化物歧化酶（SOD）活性和丙二醛（MDA）含量。通过免疫荧光和RT-qPCR分析小胶质细胞极化。通过蛋白质印迹法分析Toll样受体4（TLR4）/髓样分化因子88（MyD88）/核因子κB（NF-κB）信号通路蛋白和PLIN2。通过BODIPY染色评估脂质代谢。通过流式细胞术测量活性氧（ROS）。
丹参酮减少了dMCAO小鼠的脑梗死体积，减轻了脑水肿，并改善了神经/运动恢复。它通过抑制M1标志物并增强M2标志物，使小胶质细胞极化转向抗炎M2表型。丹参酮下调促炎细胞因子，同时上调抗炎细胞因子。丹参酮通过抑制脂质合成基因并激活脂肪分解基因来恢复脂质稳态。这减少了脂质积累。机制上，丹参酮抑制脂多糖结合蛋白（LBP）表达和TLR4/MyD88/NF-κB信号通路。此外，丹参酮通过降低ROS、恢复SOD活性和减少脂质过氧化来减轻氧化应激。
丹参酮通过多靶点作用赋予神经保护。它同时减轻神经炎症、调节脂质代谢并对抗氧化应激。这是通过LBP/TLR4/NF-κB轴调节实现的。其多靶点作用解决了缺血性中风病理生理学的复杂性，使其成为临床转化中有前景的治疗候选药物。
