针刺调控脑缺血再灌注损伤大鼠缺血侧海马差异miRNA信号归属通路及mir-34c-3p表达的研究

关键词：针刺  脑缺血再灌注损伤  信号通路  mir-34c-3p  大鼠

Keywords:acupuncture  cerebral ischemia reperfusion injury  signaling pathway  mir-34c-3p  rats

摘要：

      摘要 目的：观察针刺调控脑缺血再灌注损伤大鼠缺血侧海马组织差异miRNA信号归属通路及mir-34c-3p表达的作用，探讨mir-34c-3p在脑缺血再灌注损伤中的作用机制。 方法：通过线栓法制作局灶性脑缺血再灌注模型，将60只健康SD大鼠随机分为空白组、假手术组、模型组、针刺组，每组15只，共4组。造模成功约180min后开始针刺，每12h一次，连续进行6次。干预结束后，用Zea Longa评分观察大鼠神经功能缺损状况，测定脑梗死面积比，采用miRNA微阵列芯片技术筛选出缺血侧海马组织组间差异基因并归纳信号传导通路，RT-PCR法检测缺血侧海马组织mir-34c-3p的表达水平。 结果：造模后，与空白组、假手术组比较，模型组神经功能缺失评分升高，梗死面积比均明显增加（P<0.01）；与模型组比较，针刺组神经功能缺失评分降低，梗死面积进一步缩小（P<0.05）。与空白组、假手术组比较，模型组差异基因数分别为4、5个，主要归属MAPK、P53、Calcium、Jak-STAT以及Neurotrophin信号通路；与模型组比较，针刺组差异基因数为16个，主要归属MAPK、P53、Calcium、Jak-STAT以及Neurotrophin信号通路，且每条信号通路差异表达的miRNA不完全相同。与空白组和假手术组比较，模型组大鼠脑缺血侧海马mir-34c-3p表达量明显降低（P<0.05）；与模型组比较，针刺组mir-34c-3p的表达趋势上升明显（P<0.01）。 结论：针刺可降低脑缺血再灌注损伤大鼠神经功能缺失评分，减少脑梗死面积，其机制可能与针刺可激活多种miRNA的表达，多途径、多网络调控脑缺血再灌注及上调mir-34c-3p的表达量有关。

Abstract:

      Abstract Objective：To observe the effects of acupuncture on the differential miRNA signaling pathways and mir-34c-3p expression in ischemic hippocampus of cerebral ischemia reperfusion injury rats， and to explore the mechanism of mir-34c-3p in cerebral ischemia reperfusion injury. Method：A focal cerebral ischemia reperfusion model was established by suture method， and 60 healthy SD rats were randomly divided into blank group， sham operation group， model group and acupuncture group， with 15 rats in each group， 4 groups in total. After successful modeling for about 180min， acupuncture was started every 12h for 6 consecutive times. After the intervention， Zea Longa score was used to observe the neurological defects in rats and measure the cerebral infarction area ratio.MiRNA microarray chip technology was used to screen out differentially expressed genes and analyze signaling pathways.RT-PCR was used to detect the expression level of mir-34c-3p in the hippocampal tissue of ischemic side. Result：After modeling，compared with the blank group and the sham operation group， neurological function deficit score and cerebral infarct area ratio in the model group were significantly increased(P<0.01)；Compared with the model group，the neurological function deficit score in acupuncture group decreased， and the cerebral infarct area further decreased(P<0.05).Compared with the blank group and the sham group， the number of different genes in the model group was 4 and 5， respectively， mainly attributed to MAPK， P53， Calcium， Jak-STAT and Neurotrophin signaling pathways.Compared with the model group， the number of differentially expressed genes in the acupuncture group was 16， mainly attributed to MAPK， P53， Calcium， Jak-STAT and Neurotrophin signaling pathways， and the miRNA differentially expressed in each signaling pathway was not identical.Compared with the blank group and the sham group， the expression level of mir-34c-3p in the model group was significantly reduced (P<0.05). Compared with the model group， the expression of mir-34c-3p increased significantly in the acupuncture group (P<0.01). Conclusion：Acupuncture can reduce the neurological deficit score and cerebral infarction area of CIRI rats. The mechanism may be related to the fact that acupuncture can activate the expression of various miRNAs， regulate cerebral ischemia reperfusion through multiple channels and networks， and up-regulate the expression of mir-34c-3p.