Department of Neurosurgery, The People’s Hospital of Chizhou, Chizhou 247000, Anhui, China.
Department of Anesthesiology, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230001, Anhui, China.
Aging (Albany NY). 2020 Nov 16;13(1):877-893. doi: 10.18632/aging.202194.
Ischemic stroke is a devastating disease that causes long-term disability. However, its pathogenesis is unclear, and treatments for ischemic stroke are limited. Recent studies indicate that oxidative stress is involved in the pathological progression of ischemic stroke and that angiogenesis participates in recovery from ischemic stroke. Furthermore, previous studies have shown that Coicis Semen has antioxidative and anti-inflammatory effects in a variety of diseases. In the present study, we investigated whether Coicis Semen has a protective effect against ischemic stroke and the mechanism of this protective effect.
Coicis Semen administration significantly decreased the infarct volume and mortality and alleviated neurological deficits at 3, 7 and 14 days after MCAO. In addition, cerebral edema at 3 days poststroke was ameliorated by Coicis Semen treatment. DHE staining showed that ROS levels in the vehicle group were increased at 3 days after reperfusion and then gradually declined, but Coicis Semen treatment reduced ROS levels. The levels of GSH and SOD in the brain were increased by Coicis Semen treatment, while MDA levels were reduced. Furthermore, Coicis Semen treatment decreased the extravasation of EB dye in MCAO mouse brains and elevated expression of the tight junction proteins ZO-1 and Occludin. Double immunofluorescence staining and western blot analysis showed that the expression of angiogenesis markers and TGFβ pathway-related proteins was increased by Coicis Semen administration. Consistent with the , cytotoxicity assays showed that Coicis Semen substantially promoted HUVEC survival following OGD/RX . Additionally, though LY2109761 inhibited the activation of TGFβ signaling in OGD/RX model animals, Coicis Semen cotreatment markedly reversed the downregulation of TGFβ pathway-related proteins and increased VEGF levels.
Adult male wild-type C57BL/6J mice were used to develop a middle cerebral artery occlusion (MCAO) stroke model. Infarct size, neurological deficits and behavior were evaluated on days 3, 7 and 14 after staining. In addition, changes in superoxide dismutase (SOD), GSH and malondialdehyde (MDA) levels were detected with a commercial kit. Blood-brain barrier (BBB) permeability was assessed with Evans blue (EB) dye. Western blotting was also performed to measure the levels of tight junction proteins of the BBB. Additionally, ELISA was performed to measure the level of VEGF in the brain. The colocalization of CD31, angiogenesis markers, and Smad1/5 was assessed by double immunofluorescent staining. TGFβ pathway-related proteins were measured by western blotting. Furthermore, the cell viability of human umbilical vein endothelial cells (HUVECs) following oxygen-glucose deprivation/reoxygenation (OGD/RX) was measured by Cell Counting Kit (CCK)-8 assay.
Coicis Semen treatment alleviates brain damage induced by ischemic stroke through inhibiting oxidative stress and promoting angiogenesis by activating the TGFβ/ALK1 signaling pathway.
缺血性中风是一种导致长期残疾的破坏性疾病。然而，其发病机制尚不清楚，缺血性中风的治疗方法也有限。最近的研究表明，氧化应激参与了缺血性中风的病理进展，血管生成参与了缺血性中风的恢复。此外，先前的研究表明，薏苡仁具有抗氧化和抗炎作用，可用于多种疾病。在本研究中，我们探讨了薏苡仁是否对缺血性中风具有保护作用，以及这种保护作用的机制。
薏苡仁给药可显著降低 MCAO 后 3、7 和 14 天的梗死体积和死亡率，并减轻神经功能缺损。此外，薏苡仁治疗可改善中风后 3 天的脑水肿。DHE 染色显示，再灌注后 3 天，载体组 ROS 水平升高，然后逐渐下降，但薏苡仁治疗降低了 ROS 水平。薏苡仁治疗可增加脑内 GSH 和 SOD 水平，降低 MDA 水平。此外，薏苡仁治疗可减少 MCAO 小鼠脑内 EB 染料的外渗，并增加紧密连接蛋白 ZO-1 和 Occludin 的表达。双免疫荧光染色和 Western blot 分析显示，薏苡仁给药可增加血管生成标志物和 TGFβ 通路相关蛋白的表达。与 MCAO 模型动物相一致，细胞毒性测定表明，薏苡仁可显著促进 HUVEC 在 OGD/RX 后的存活。此外，尽管 LY2109761 抑制了 OGD/RX 模型动物中 TGFβ 信号通路的激活，但薏苡仁共处理明显逆转了 TGFβ 通路相关蛋白的下调和 VEGF 水平的增加。
使用成年雄性野生型 C57BL/6J 小鼠建立大脑中动脉闭塞（MCAO）中风模型。在 MCAO 后第 3、7 和 14 天通过染色评估梗死面积、神经功能缺损和行为。此外，通过商业试剂盒检测超氧化物歧化酶（SOD）、GSH 和丙二醛（MDA）水平的变化。用 Evans 蓝（EB）染料评估血脑屏障（BBB）通透性。还进行了 Western blot 以测量 BBB 紧密连接蛋白的水平。此外，通过 ELISA 测量脑内 VEGF 水平。通过双免疫荧光染色评估 CD31、血管生成标志物和 Smad1/5 的共定位。通过 Western blot 测量 TGFβ 通路相关蛋白。此外，通过 Cell Counting Kit (CCK)-8 测定人脐静脉内皮细胞（HUVEC）在氧葡萄糖剥夺/再氧合（OGD/RX）后的细胞活力。
薏苡仁通过抑制氧化应激和通过激活 TGFβ/ALK1 信号通路促进血管生成来减轻缺血性中风引起的脑损伤。
