NMPA Key Laboratory for Research of Traditional Chinese Medicine Syndrome, School of Pharmaceutics, Guangzhou University of Chinese Medicine, Guangzhou, 51006, Guangdong, China; Institute of Formula and Syndrome, Guangzhou University of Chinese Medicine, Guangzhou, 51006, Guangdong, China.
NMPA Key Laboratory for Research of Traditional Chinese Medicine Syndrome, School of Pharmaceutics, Guangzhou University of Chinese Medicine, Guangzhou, 51006, Guangdong, China.
J Ethnopharmacol. 2024 Mar 25;322:117567. doi: 10.1016/j.jep.2023.117567. Epub 2023 Dec 18.
Patients with ischemic stroke (IS) often continue to exhibit cerebral microcirculatory dysfunction even after receiving thrombolytic therapy. Enhancing the function of cerebral microvascular endothelia represents a pivotal advancement in the therapeutic strategy for ischemic microcirculatory disturbances. A traditional Chinese medicinal formulation named Shexiang Tongxin Dropping Pills (STDP), has been clinically employed to ameliorate microcirculatory abnormalities. Existing literature attests to the beneficial role of STDP on endothelial cells (ECs). Nevertheless, specific impacts and underlying mechanisms of STDP in rectifying IS-induced cerebral microvascular dysfunction warrant further exploration.
This investigation seeks to delineate the effects of STDP on cerebral microvascular endothelial damage induced by ischemic stroke and to elucidate the underlying mechanism involved.
Middle cerebral artery occlusion and reperfusion (MCAO/R) technique was employed to established ischemic stroke model in mice. The therapeutic efficacy of STDP on cerebral microvascular function was assessed through laser speckle contrast imaging, behavioral assays, and histological evaluations. Biochemical markers in the brain tissue, including GSH, SOD, MDA, and ROS, were quantified using specific assay kits. In vitro study, oxygen-glucose deprivation and reperfusion (OGD/R) was performed in bEnd.3 cells. The cytoprotective potential of STDP was then evaluated by measuring cell viability, LDH activity, endothelial permeability, and oxidative stress parameters. Important targets in critical pathway were verified by immunoblotting and immunofluorescence both in mice brain slices and bEnd.3 cells.
STDP decrease brain infarct size, repaired microvascular cerebral blood flow and attenuated neurological deficiency in MCAO/R mice. Moreover, STDP abolished MCAO/R-induced oxidative stress which was reflected by rescuing GSH content, restoration of SOD activity and T-AOC, reduction of MDA and ROS. Ex vivo, STDP increased cerebral microvascular endothelial cells viability, abolished oxidative stress and decreased their permeability after ODG/R. Mechanistically, STDP significantly suppressed endothelial ROS-TXNIP mediated the activation of NLRP3 inflammasome in vivo and in vitro.
STDP improves ischemic stroke-induced cerebral microcirculatory deficits by regulating cerebral microvascular endothelial ROS/TXNIP/NLRP3 signaling pathway.
ETHNOPHARMACOLOGICAL 相关性：缺血性中风（IS）患者即使接受溶栓治疗后，仍常存在脑微循环功能障碍。增强脑微血管内皮细胞的功能是治疗缺血性微循环障碍的关键策略。一种名为麝香通心滴丸（STDP）的中药制剂已在临床上用于改善微循环异常。现有文献证明 STDP 对内皮细胞（ECs）有益。然而，STDP 纠正 IS 引起的脑微血管功能障碍的具体影响和潜在机制仍需进一步探讨。
本研究旨在探讨 STDP 对缺血性中风引起的脑微血管内皮损伤的影响，并阐明其潜在机制。
采用大脑中动脉闭塞再灌注（MCAO/R）技术建立小鼠缺血性中风模型。通过激光散斑对比成像、行为学评估和组织学评估评估 STDP 对脑微血管功能的治疗效果。使用特定的测定试剂盒测定脑组织中的生化标志物，包括 GSH、SOD、MDA 和 ROS。在体外，对 bEnd.3 细胞进行氧葡萄糖剥夺和再灌注（OGD/R）。然后通过测量细胞活力、LDH 活性、内皮通透性和氧化应激参数来评估 STDP 的细胞保护潜力。通过免疫印迹和免疫荧光在小鼠脑切片和 bEnd.3 细胞中验证关键途径中的重要靶点。
STDP 减少脑梗死面积，修复脑微血管血流，减轻 MCAO/R 小鼠的神经功能缺损。此外，STDP 消除了 MCAO/R 引起的氧化应激，这反映在恢复 GSH 含量、恢复 SOD 活性和 T-AOC、减少 MDA 和 ROS 上。在离体条件下，STDP 增加了脑微血管内皮细胞的活力，消除了 ODG/R 后的氧化应激并降低了其通透性。在体内和体外，STDP 均显著抑制内皮 ROS-TXNIP 介导的 NLRP3 炎性体的激活。
STDP 通过调节脑微血管内皮细胞 ROS/TXNIP/NLRP3 信号通路改善缺血性中风引起的脑微循环缺陷。
