Table2_Buyang Huanwu Decoction alleviates cerebral ischemic injury through modulating caveolin-1-mediated mitochondrial quality control.XLSX

Introduction: Mitochondrial quality control (MQC) is an important mechanism of neural repair after cerebral ischemia (CI). Recent studies have shown that caveolin-1 (Cav-1) is an important signaling molecule in the process of CI injury, but its mechanism of regulating MQC after CI is still unclear. Buyang Huanwu Decoction (BHD) is a classic traditional Chinese medicine formula that is often used to treat CI. Unfortunately, its mechanism of action is still obscure.
Methods: In this study, we tested the hypothesis that BHD can regulate MQC through Cav-1 and exert an anti-cerebral ischemia injury effect. We used Cav-1 knockout mice and their homologous wild-type mice, replicated middle cerebral artery occlusion (MCAO) model and BHD intervention. Neurobehavioral scores and pathological detection were used to evaluate neurological function and neuron damage, transmission electron microscopy and enzymology detection of mitochondrial damage. Finally, western blot and RT-qPCR expression of MQC-related molecules were tested.
Results: After CI, mice showed neurologic impairment, neuronal damage, and significant destruction of mitochondrial morphology and function, and MQC was imbalanced. Cav-1 deletion aggravated the damage to neurological function, neurons, mitochondrial morphology and mitochondrial function after CI, aggravated the imbalance of mitochondrial dynamics, and inhibited mitophagy and biosynthesis. BHD can maintain MQC homeostasis after CI through Cav-1 and improve CI injury.
Discussion: Cav-1 can affect CI injury by regulating MQC, and this mechanism may be another target of BHD for anti-cerebral ischemia injury.

引言：线粒体质量控制（mitochondrial quality control, MQC）是脑缺血（cerebral ischemia, CI）后神经修复的重要机制。近期研究表明，小窝蛋白1（caveolin-1, Cav-1）是脑缺血损伤过程中的关键信号分子，但其调控脑缺血后线粒体质量控制的具体机制仍未明确。补阳还五汤（Buyang Huanwu Decoction, BHD）是临床常用于治疗脑缺血的经典中药方剂，但其作用机制尚不清楚。
方法：本研究验证了"补阳还五汤可通过小窝蛋白1调控线粒体质量控制，进而发挥抗脑缺血损伤作用"的假说。实验采用小窝蛋白1基因敲除小鼠及其同窝野生型小鼠，构建大脑中动脉闭塞（middle cerebral artery occlusion, MCAO）模型并给予补阳还五汤干预。通过神经行为学评分与病理检测评估神经功能与神经元损伤；借助透射电镜与酶学检测分析线粒体损伤情况；最后采用蛋白质印迹法与实时定量聚合酶链反应（RT-qPCR）检测线粒体质量控制相关分子的表达水平。
结果：脑缺血造模后，小鼠出现神经功能缺损、神经元损伤，线粒体形态与功能显著破坏，且线粒体质量控制失衡。敲除小窝蛋白1可加重脑缺血后小鼠的神经功能损伤、神经元损伤、线粒体形态与功能异常，加剧线粒体动力学失衡，并抑制线粒体自噬与生物合成过程。补阳还五汤可通过小窝蛋白1维持脑缺血后线粒体质量控制稳态，从而改善脑缺血损伤。
讨论：小窝蛋白1可通过调控线粒体质量控制影响脑缺血损伤，这一机制或为补阳还五汤发挥抗脑缺血损伤作用的另一潜在靶点。