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颜思阳,杨仁义,刘利娟,马强,李俊熙,周德生.基于“STAT3/miR-17”反馈环探讨活血荣络方对脑缺血再灌注损伤模型大鼠的保护作用机制[J].湖南中医药大学学报英文版,2021,41(4):489-497.[Click to copy
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| 基于“STAT3/miR-17”反馈环探讨活血荣络方对脑缺血再灌注损伤模型大鼠的保护作用机制 |
| 颜思阳,杨仁义,刘利娟,马强,李俊熙,周德生 |
| (湖南中医药大学, 湖南 长沙 410007;湖南中医药大学第一附属医院, 湖南 长沙 410007) |
| 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 反馈环 线粒体自噬 |
| DOI：10.3969/j.issn.1674-070X.2021.04.001 |
| Received:December 13, 2020 |
| 基金项目:国家自然科学基金项目（81874463）；湖南省科技厅项目（2017SK4005）；湖南省财政中医药项目名院工程院内优势病种项目（rsk-010-013/006-09）；湖南省中医药管理局资助项目（201824，202046，202038）；湖南省教育厅一般项目（20C1405）；湖南中医药大学中西医结合一流学科开放基金项目（2019ZXYJH08，2018ZXYJH10）；湖南省研究生创新项目（CX20190587）；湖南中医药大学研究生培养质量工程专项（2020CX62，2018CX07）。 |
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| Explore the Protective Mechanism of Huoxue Rongluo Decoction on Cerebral Ischemia Reperfusion Injury Model Rats Based on the Feedback Loop of “STAT3/miR-17” |
| YAN Siyang,YANG Renyi,LIU Lijuan,MA Qiang,LI Junxi,ZHOU Desheng |
| (Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;The First Affiliated Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China) |
| 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. |
| Key words: cerebral infarction Huoxue Rongluo Decoction STAT3 miR-17 feedback loop mitochondrial autophagy |
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