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郑慧娥,何灏龙,陈芯仪,田浩梅,陈楚淘.针刺对CIRI大鼠缺血侧海马区miRNA表达及miR-20a-5p和miR-22-5p的影响[J].湖南中医药大学学报,2020,40(10):1226-1231[点击复制] |
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| 针刺对CIRI大鼠缺血侧海马区miRNA表达及miR-20a-5p和miR-22-5p的影响 |
| 郑慧娥,何灏龙,陈芯仪,田浩梅,陈楚淘 |
| (湖南中医药大学针灸推拿学院, 湖南 长沙 410208) |
| 摘要: |
| 目的 观察针刺对脑缺血再灌注损伤（cerebral ischemia reperfusion injury，CIRI）大鼠缺血侧海马区miRNA表达及miR-20a-5p和miR-22-5p的影响，以探究针刺修复CIRI的可能机制。方法 60只大鼠采用随机数字表法分成正常组、假手术组（Sham组）、CIRI组及CIRI+针刺组，每组15只。CIRI组及CIRI+针刺组参照优化版Zea Longa线栓法复制大脑中动脉缺血再灌注模型；Sham组只插入线栓不阻断中动脉。Sham组及CIRI组造模后不予针刺，CIRI+针刺组造模后于人中、百会、大椎穴予针刺干预，正常组不予任何处理。分别于治疗前后行神经功能症状缺损评分观察大鼠神经症状变化；治疗后，采用TTC染色检测脑梗死面积比；基因芯片技术检测CIRI大鼠缺血侧海马区miRNA表达；qPCR检测CIRI大鼠缺血侧miR-20a-5p、miR-22-5p的表达量。结果 （1）与治疗前比较，治疗后CIRI+针刺组的神经功能症状缺损评分显著下降（P<0.01），其余各组差异均无统计学意义（P>0.05）。治疗后，与正常组及Sham组比较，CIRI组及CIRI+针刺组神经功能症状缺损评分及脑梗死面积比均显著升高（P<0.01）；与CIRI组比较，CIRI+针刺组神经功能症状缺损评分及脑梗死面积比明显下降（P<0.05）。（2）基因芯片检测结果示，与正常组比较，CIRI组差异表达4个miRNA，且其中2个参与调控细胞增殖、3个参与调控细胞凋亡；与Sham组比较，CIRI组差异表达5个miRNA，且其中3个参与调控细胞增殖、1个参与调控细胞凋亡；与CIRI组比较，CIRI+针刺组差异表达16个miRNA，且其中12个参与调控细胞增殖、10个参与调控细胞凋亡。CIRI+针刺组与CIRI组改变了相同功用的miRNA，其中包括miR-20a-5p和miR-22-5p。（3）qPCR结果示，与正常组比较，CIRI组miR-20a-5p表达增加（P<0.01）、miR-22-5p表达降低（P<0.01）；与Sham组比较，CIRI组miR-20a-5p表达增加（P<0.01）、miR-22-5p表达降低（P<0.05）；与CIRI组比较，CIRI+针刺组miR-20a-5p表达降低（P<0.01）、miR-22-5p表达增加（P<0.05）。结论 针刺能明显改善CIRI大鼠神经功能症状缺损评分及脑梗死面积比，促进脑缺血再灌注后修复，其机制可能与激发多种类、多功能miRNA的改变，特别是与下调miR-20a-5p、上调miR-22-5p的表达量，促进细胞增殖、抑制细胞凋亡功能相关。 |
| 关键词: 脑缺血再灌注损伤 针刺 miRNA miR-20a-5p miR-22-5p |
| DOI：10.3969/j.issn.1674-070X.2020.10.010 |
| 投稿时间：2020-04-03 |
| 基金项目:国家自然科学基金项目（81874508，81303051）；湖南省自然科学基金项目（2016JJ3101）；湖南省大学生研究性学习和创新性实验计划（2017-287）；湖南省中医药管理局课题（201961）。 |
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| Effects of Acupuncture on Expression of miRNA, miR-20a-5p and miR-22-5p in the Ischemic Hippocampus of Rats with CIRI |
| ZHENG Huie,HE Haolong,CHEN Xinyi,TIAN Haomei,CHEN Chutao |
| (College of Acupuncture-Moxbustion and Tuina, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China) |
| Abstract: |
| Objective To observe the effects of acupuncture on expression of miRNA, miR-20a-5p and miR-22-5p in the ischemic hippocampus of rats with cerebral ischemia reperfusion injury (CIRI), and to explore the possible mechanism of acupuncture in repairing CIRI. Methods A total of 60 rats were divided into a normal group, a sham operation group (sham group), a CIRI group and a CIRI+acupuncture group by random number table, with 15 rats in each group. The CIRI group and the CIRI+acupuncture group established CIRI model by referring to reformed Zea Longa suture-occluded method; and line was inserted without blocking the middle artery in the sham group. The sham group and the CIRI group received no acupuncture treatment after modeling. The CIRI+acupuncture group received acupuncture at Renzhong (GV26), Baihui (GV20) and Dazhui (GV14). The normal group did not receive any treatment. The neurological deficit scores were performed before and after treatment to observe the changes in neurological symptoms in rats; after treatment, TTC staining was used to detect the ratio of cerebral infarction area; gene chip technology was used to detect miRNA expression in the hippocampus of the ischemic side of CIRI rats; qPCR was used to detect the expression of miR-20a-5p and miR-22-5p on the ischemic side of CIRI rats. Results (1) Compared with before treatment, the neurological deficit score of the CIRI+acupuncture group decreased significantly after treatment (P<0.01), and the difference in the other groups was not statistically significant (P>0.05). After treatment, compared with the normal group and the sham group, the neurological deficit score and the cerebral infarction area ratio in the CIRI group and the CIRI+acupuncture group were significantly increased (P<0.01); compared with the CIRI group, the neurological symptoms defect score and cerebral infarction area ratio of the CIRI+acupuncture group decreased significantly (P<0.05). (2) The gene chip test results showed that compared with the normal group, the CIRI group differentially expressed 4 miRNAs. 2 were involved in regulating cell proliferation, and 3 were involved in regulating cell apoptosis; compared with the sham group, the CIRI group differentially expressed 5 miRNAs. 3 were involved in regulating cell proliferation, 2 were involved in regulating inflammation and 1 was involved in regulating cell apoptosis, gene expression, axon extension and neuron differentiation; Compared with CIRI group, the CIRI+acupuncture group has differential expression of 16 miRNAs. 12 were involved in regulating cell proliferation, 10 were involved in regulating cell apoptosis. The CIRI+ acupuncture group and the CIRI group changed miRNAs with the same function, including miR-20a-5p and miR-22-5p. (3) qPCR results showed that the expression of miR-20a-5p in the CIRI group increased compared with the normal group (P<0.01), and the expression of miR-22-5p decreased (P<0.01); compared with the sham group, the expression of miR-20a-5p increased (P<0.01), and the expression of miR-22-5p decreased (P<0.05) in the CIRI group; Compared with the CIRI group, the expression of miR-20a-5p in the CIRI + acupuncture group decreased (P<0.01), and the expression of miR-22-5p increased (P<0.05). Conclusion Acupuncture can significantly improve the neurological deficit score and cerebral infarction area ratio in CIRI rats, and promote the repair of cerebral ischemia-reperfusion. The mechanism may be related to stimulating the changes of various and multifunctional miRNAs, especially the down-regulating miR-20a -5p, up-regulating the expression of miR-22-5p is related to promoting cell proliferation and inhibiting apoptosis. |
| Key words: cerebral ischemia reperfusion injury acupuncture miRNA miR-20a-5p miR-22-5p |
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