背景:脑缺血再灌注损伤(CIRI)是缺血性卒中的一种破坏性不良反应,导致高残疾和死亡率。丹参。(丹参,DS)用猪心血(PCB-DS)处理,一种独特的加工产品,具有有希望的抗缺血作用。然而,PCB-DS抗CIRI的潜在机制尚不清楚.
目的:铁凋亡被证明与CIRI有关。本研究的目的是探讨PCB-DS抑制GLRX5介导的铁凋亡减轻CIRI的分子机制。这与DS不同。
方法:采用UPLC对PCB-DS和DS进行质量评价。使用神经行为评分比较PCB-DS和DS对CIRI的药理活性,梗死体积,促炎因子,和病理检查。采用蛋白质组学方法探索PCB-DS抗CIRI的潜在特异性机制,这与DS不同。基于差异蛋白GLRX5,铁凋亡相关铁,GSH,MDA,SOD,ROS,liperfluo,和线粒体形态学分析。然后,分析了GLRX5介导的铁饥饿反应和SLC7A11/GPX4蛋白。最后,构建了GLRX5沉默后OGD/R诱导的SH-SY5Y细胞,以证明PCB-DS通过GLRX5介导的铁凋亡改善了CIRI。
结果:PCB-DS通过降低神经评分更好地缓解CIRI,减少梗死体积,和抑制炎症细胞因子的释放比DS。蛋白质组学提示PCB-DS可能通过抑制GLRX5介导的铁凋亡来改善CIRI,这与DS不同。PCB-DS逆转了异常的线粒体形态,铁,GSH,MDA,SOD,ROS,和liperfluo在体外和体内抑制铁凋亡。PCB-DS直接激活GLRX5抑制铁饥饿反应并下调SLC7A11/GPX4信号通路以抑制铁凋亡。最后,GLRX5沉默后,SH-SY5Y细胞和PCB-DS未改善的OGD/R损伤的铁饥饿反应激活。
结论:与DS不同,PCB-DS通过抑制GLRX5介导的铁饥饿反应抑制铁死亡以减轻CIRI。这些发现全面了解了PCB-DS对CIRI影响的分子机制,并为临床研究中评估该产品提供了证据。
BACKGROUND: Cerebral ischemia-reperfusion injury (CIRI) is a destructive adverse reaction of ischemic stroke, leading to high disability and mortality rates. Salvia miltiorrhiza Bge. (Danshen, DS) processed with porcine cardiac blood (PCB-DS), a characteristic processed product, has promising anti-ischemic effects. However, the underlying mechanism of PCB-DS against CIRI remains unclear.
OBJECTIVE: Ferroptosis is demonstrated to be involved in CIRI. The aim of this study was to explore the molecular mechanism underlying PCB-DS inhibited GLRX5-mediated ferroptosis alleviating CIRI, which was different from DS.
METHODS: Quality evaluation of PCB-DS and DS was conducted by UPLC. Pharmacological activities of PCB-DS and DS against CIRI were compared using neurobehavioral scores, infarct volume, proinflammatory factors, and pathological examinations. Proteomics was employed to explore the potential specific mechanism of PCB-DS against CIRI, which was different from DS. Based on the differential protein GLRX5, ferroptosis-related iron, GSH, MDA, SOD, ROS, liperfluo, and mitochondrial morphology were analyzed. Then, the proteins of GLRX5-mediated iron-starvation response and SLC7A11/GPX4 were analyzed. Finally, OGD/R-induced SH-SY5Y cells upon GLRX5 silencing were constructed to demonstrate that PCB-DS improved CIRI by GLRX5-mediated ferroptosis.
RESULTS: PCB-DS better alleviated CIRI through decreasing neurological score, reducing the infarct volume, and suppressing the release of inflammatory cytokines than DS. Proteomics suggested that PCB-DS may ameliorate CIRI by inhibiting GLRX5-mediated ferroptosis, which was different from DS. PCB-DS reversed the abnormal mitochondrial morphology, iron, GSH, MDA, SOD, ROS, and liperfluo to inhibit ferroptosis in vitro and in vivo. PCB-DS directly activated GLRX5 suppressing the iron-starvation response and downregulated the SLC7A11/GPX4 signaling pathway to inhibit ferroptosis. Finally, silencing GLRX5 activated the iron-starvation response in SH-SY5Y cells and PCB-DS unimproved OGD/R injury upon GLRX5 silencing.
CONCLUSIONS: Different from DS, PCB-DS suppressed ferroptosis to alleviate CIRI through inhibiting GLRX5-mediated iron-starvation response. These findings give a comprehensive understanding of the molecular mechanism underlying the effect of PCB-DS against CIRI and provide evidence to assess the product in clinical studies.