颜思阳 杨仁义 刘利娟 马强 李俊熙 周德生

〔摘要〕 目的 探讨活血荣络方对脑缺血再灌注损伤大鼠的保护作用及对线粒体自噬的影响。方法 采用生物信息学方法挖掘脑梗死差异表达miRNAs，并预测活血荣络方可能通过调节“STAT3-miR-17”反馈环路发挥作用。将SD雄性大鼠随机分为假手术组、模型组、活血荣络方组，运用线栓法建立脑缺血再灌注模型。造模前36、12 h及术后12 h给药。术后24 h行神经功能评分，并行TTC染色评估脑梗死体积;透射电镜观察自噬小体;Western blot检测线粒体及自噬相关蛋白，评估线粒体自噬情况;Real-time PCR检测“STAT3-miR-17”反馈环中miRNA、mRNA的表达。结果 与模型组相比，活血荣络方可明显改善模型大鼠神经功能损伤，减少脑梗死体积（P<0.01），下调p62、TOMM20的表达（P<0.05），上调LC3B的表达（P<0.01），增加损伤侧脑组织自噬小体;下调miR-17 miRNA的表达（P<0.01），上调STAT3 mRNA的表达（P<0.01）。结论 活血荣络方可通过上调线粒体自噬发挥神经保护作用，其机制可能与调控“STAT3-miR-17”反馈环路相关。

〔关键词〕 脑梗死;活血荣络方;STAT3;miR-17;反饋环;线粒体自噬

〔中图分类号〕R285.5       〔文献标志码〕A       〔文章编号〕doi：10.3969/j.issn.1674-070X.2021.04.001

〔Abstract〕 Objective To investigate the protective effect of Huoxue Rongluo Decoction on ischemia/reperfusion injury rats and its influence on mitochondrial autophagy. Methods Bioinformatics method was used to mine the differentlyexpressed miRNAs in cerebral infarction， and it was predicted that Huoxue Rongluo Decoction might play a role by regulating the "STAT3-miR-17" feedback loop. SD male rats were randomly divided into sham operation group， model group， and Huoxue Rongluo Decoction group， and the cerebral ischemia reperfusion model was established by thread plug method. Drugs were administered at 36 and 12 hours before modeling and 12 hours after surgery. Neural function score was performed 24 hours after operation， and cerebral infarction volume was assessed by TTC staining; autophagosomes were observed by transmission electron microscope; mitochondria and autophagy-related proteins were detected by Western blot to assess mitochondrial autophagy; real-time PCR was used to detect the expression of miRNA and mRNA in the feedback loop of "STAT3-miR-17". Results Compared with the model group， Huoxue Rongluo Decoction can significantly improve the neurological damage， reduce cerebral infarction volume of rats （P<0.01）， down-regulate the expressions of p62 and TOMM20 （P<0.05）， up-regulate the expression of LC3II （P<0.01）， and increase autophagosomes in the injured brain tissue; down-regulate the expression of miR-17 miRNA （P<0.01）， up-regulate the expression of STAT3 mRNA （P<0.01）. Conclusion Huoxue Rongluo Decoction exerts neuroprotective effect by moderately up-regulating mitochondrial autophagy， and its mechanism may be related to the regulation of "STAT3-miR-17" feedback loop.

〔Keywords〕 cerebral infarction; Huoxue Rongluo Decoction; STAT3; miR-17; feedback loop; mitochondrial autophagy

脑血管病是全球第二大死亡原因，也是全世界致残的第三常见原因，具有高死亡率、致残率和复发率的特点[1-2]。同时，研究显示我国脑血管病疾病负担在脑出血呈缓慢下降趋势时，脑梗死上升趋势较明显[3]。因此，脑梗死的防治具有重要的社会意义。线粒体自噬作为一种特殊的自噬类型，可以选择性地清除受损线粒体，其在应激状态中（如缺氧、钙超载等）起着至关重要的作用。脑缺血再灌注引发的级联反应可激活线粒体自噬这一过程[4]，但目前对于线粒体自噬在脑缺血再灌注损伤（ischemia/reperfusion injury， I/R）的作用仍存在争议。

脑梗死属中医学“缺血性中风”范畴。荣气，为一切精微物质的总称[5]。因其具备精微物质属性，又具有气化运动的功能，而荣气虚与荣气滞互为因果、消长共存，导致气化失司、内邪交互、神机失用，发为脑梗死[6]。……