Yen Ting-Lin, Chen Ray-Jade, Jayakumar Thanasekaran, Lu Wan-Jung, Hsieh Cheng-Ying, Hsu Ming-Jen, Yang Chih-Hao, Chang Chao-Chien, Lin Yen-Kuang, Lin Kuan-Hung, Sheu Joen-Rong
Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei, Taiwan; Department of Pharmacology, College of Medicine, Taipei Medical University, Taipei, Taiwan.
School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan; Division of General Surgery, Department of Surgery, Taipei Medical University Hospital, Taipei, Taiwan; Department of Medical Research, Taipei Medical University Hospital, Taipei, Taiwan.
Transl Res. 2016 Apr;170:57-72. doi: 10.1016/j.trsl.2015.12.002. Epub 2015 Dec 17.
Stroke pathogenesis involves complex oxidative stress-related pathways. The nuclear factor erythroid-2-related factor 2 (Nrf2) and heme oxygenase 1 (HO-1) pathways have been considered molecular targets in pharmacologic intervention for ischemic diseases. Andrographolide, a labdane diterpene, has received increasing attention in recent years because of its various pharmacologic activities. We determined that andrographolide modulates the mitogen-activated protein kinase (MAPK)-Nrf2-HO-1 signaling cascade in primary cerebral endothelial cells (CECs) to provide positive protection against middle cerebral artery occlusion (MCAO)-induced ischemic stroke in rats. In the present study, andrographolide (10 μM) increased HO-1 protein and messenger RNA expressions, Nrf2 phosphorylation, and nuclear translocation in CECs, and these activities were disrupted by a p38 MAPK inhibitor, SB203580, but not by the extracellular signal-regulated kinase inhibitor PD98059 or c-Jun amino-terminal kinase inhibitor SP600125. Similar results were observed in confocal microscopy analysis. Moreover, andrographolide-induced Nrf2 and HO-1 protein expressions were significantly inhibited by Nrf2 small interfering RNA. Moreover, HO-1 knockdown attenuated the protective effect of andrographolide against oxygen-glucose deprivation-induced CEC death. Andrographolide (0.1 mg/kg) significantly suppressed free radical formation, blood-brain barrier disruption, and brain infarction in MCAO-insulted rats, and these effects were reversed by the HO-1 inhibitor zinc protoporphyrin IX. The mechanism is attributable to HO-1 activation, as directly evidenced by andrographolide-induced pronounced HO-1 expression in brain tissues, which was highly localized in the cerebral capillary. In conclusion, andrographolide increased Nrf2-HO-1 expression through p38 MAPK regulation, confirming that it provides protection against MCAO-induced brain injury. These findings provide strong evidence that andrographolide could be a therapeutic agent for treating ischemic stroke or neurodegenerative diseases.
中风发病机制涉及复杂的氧化应激相关途径。核因子红细胞2相关因子2（Nrf2）和血红素加氧酶1（HO-1）途径被认为是缺血性疾病药物干预的分子靶点。穿心莲内酯，一种半日花烷二萜，因其多种药理活性近年来受到越来越多的关注。我们确定穿心莲内酯可调节原代脑内皮细胞（CECs）中的丝裂原活化蛋白激酶（MAPK）-Nrf2-HO-1信号级联反应，为大鼠大脑中动脉闭塞（MCAO）诱导的缺血性中风提供积极保护。在本研究中，穿心莲内酯（10μM）增加了CECs中HO-1蛋白和信使RNA表达、Nrf2磷酸化及核转位，这些活性被p38 MAPK抑制剂SB203580破坏，但未被细胞外信号调节激酶抑制剂PD98059或c-Jun氨基末端激酶抑制剂SP600125破坏。共聚焦显微镜分析也观察到类似结果。此外，Nrf2小干扰RNA显著抑制了穿心莲内酯诱导的Nrf2和HO-1蛋白表达。此外，HO-1基因敲低减弱了穿心莲内酯对氧-葡萄糖剥夺诱导的CEC死亡的保护作用。穿心莲内酯（0.1mg/kg）显著抑制了MCAO损伤大鼠的自由基形成、血脑屏障破坏和脑梗死，而HO-1抑制剂锌原卟啉IX可逆转这些作用。该机制归因于HO-1激活，穿心莲内酯诱导脑组织中明显的HO-1表达可直接证明这一点，其高度定位于脑毛细血管中。总之，穿心莲内酯通过p38 MAPK调节增加了Nrf2-HO-1表达，证实其可预防MCAO诱导的脑损伤。这些发现提供了有力证据，表明穿心莲内酯可能是治疗缺血性中风或神经退行性疾病的治疗药物。
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