目的 研究龙琥醒脑颗粒对脑缺血性再灌注损伤（cerebral ischemia reperfusion injury，CIRI）大鼠受损脑细胞线粒体动力学的影响及其机制。方法 大鼠制备CIRI模型，将造模成功的40只大鼠随机分为模型组及龙琥醒脑颗粒低、中、高剂量（75、150、300 mg/kg）组和金纳多（150 mg/kg）组，另设置假手术组8只，给予相应药物干预4 d后，TTC染色法检测大鼠脑组织梗死体积；透射电镜（TEM）观察梗塞侧脑皮层组织超微结构的变化；Western blotting和qRT-PCR法检测各组大鼠线粒体分裂蛋白1（mitochondrial fission protein 1，Fis1）、线粒体融合蛋白2（mitochondrial fusion protein 2，Mfn2）、视神经萎缩蛋白1（optic atrophy 1，Opa1）、动力相关蛋白1（dynamin related protein 1，Drp1）、核受体相关因子1（nuclear receptor-related factor 1，Nurr1）、Yes相关蛋白1（Yes-associated protein 1，YAP1）和反式形成蛋白2（inverted formin 2，INF2）蛋白及mRNA表达情况。结果 与模型组相比，各给药组大鼠神经功能评分和脑组织梗死面积均显著下降（P＜0.05、0.01）；大鼠大脑皮层神经元病理损伤显著减轻；受损脑组织Fis1、INF2和Drp1 mRNA表达水平显著下调（P＜0.01、0.001），Mfn2、Nurr1、Opa1和YAP1 mRNA表达水平显著上调（P＜0.05、0.01、0.001）；受损脑组织Drp1和Fis1蛋白表达水平显著下调（P＜0.05、0.01、0.001），Mfn2、Opa1、Nurr1和YAP1蛋白表达水平均显著上调（P＜0.05、0.01、0.001）。结论 龙琥醒脑颗粒可以改善CIRI大鼠脑梗死面积，减轻缺血病理损伤，延缓CIRI进展，其机制可能为通过Nurr1调控YAP-INF2信号通路，纠正线粒体动力学失衡，从而发挥神经保护作用。

Objective To study the effect and mechanism of Longhu Xingnao Granules (龙琥醒脑颗粒) on mitochondrial dynamics of damaged brain cells in rats with cerebral ischemia reperfusion injury (CIRI). Methods CIRI rats model was established, and 40 successful rats were randomly divided into model group, Longhu Xingnao Granules low-, medium-and high-dose (75, 150, 300 mg/kg) groups and Ginadol (150 mg/kg) groups, and 8 rats were set as sham group. After 4 d of corresponding drug intervention, infarct volume of rats brain tissue was detected by TTC staining; Ultrastructure of infarcted cerebral cortex was observed by transmission electron microscope (TEM); Western blotting and qRT-PCR were used to detect protein and mRNA expressions of mitochondrial fission protein 1 (Fis1), mitochondrial fusion protein 2 (Mfn2), optic atrophy 1 (Opa1), dynamin related protein 1 (Drp1), nuclear receptor-related factor 1 (Nurr1), Yes-associated protein 1 (YAP1) and inverted formin 2 (INF2). Results Compared with model group, neurological function score and brain infarction area of rats in each administration group were significantly decreased (P < 0.05, 0.01); Pathological damage of cerebral cortex neurons of rats was significantly alleviated; Fis1, INF2 and Drp1 mRNA expressions in damaged brain tissue were significantly down-regulated (P < 0.01, 0.001), Mfn2, Nurr1, Opa1 and YAP1 mRNA expressions were significantly up-regulated (P < 0.05, 0.01, 0.001); Drp1 and Fis1 protein expressions in damaged brain tissue were significantly down-regulated (P < 0.05, 0.01, 0.001), Mfn2, Opa1, Nurr1 and YAP1 protein expressions were significantly up-regulated (P < 0.05, 0.01, 0.001). Conclusion Longhu Xingnao Granules can improve the cerebral infarct size in CIRI rats, reduce ischemic pathological damage, and delay the progression of CIRI. Its mechanism may be that Nurr1 regulates YAP-INF2 signaling pathway and corrects the imbalance of mitochondrial dynamics, thereby exerting neuroprotective effects.

R285.5

湖南省中医药科研计划项目(202015);湖南省卫健委科研计划项目(20200308);湖南省自然科学基金面上项目(2021JJ30529,2021JJ30499);湖南中医药大学中西医结合一流学科重点项目(2021ZXYJH01)