| 李明哲,张英杰,单春雷.电针结合经颅交流电刺激对脑缺血大鼠神经炎症和凋亡相关基因表达的影响[J].中国康复医学杂志,2024,(2):163~169 |
| 电针结合经颅交流电刺激对脑缺血大鼠神经炎症和凋亡相关基因表达的影响 点此下载全文 |
| 李明哲 张英杰 单春雷 |
| 上海市针灸经络研究所，上海市，200030 |
| 基金项目：上海市卫生健康委中西医结合康复诊疗提升项目（ZY（2018-2020-FWTX-8002））；上海市卫生健康委加快中医药事业发展三年行动计划项目（ZY（2018-2020-CCCX-2004-05））；上海领军人才项目 |
| DOI：10.3969/j.issn.1001-1242.2024.02.003 |
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| 摘要： |
| 摘要
目的：观察电针（electroacupuncture， EA）结合经颅交流电刺激（transcranial alternating current stimulation， tACS）对大脑中动脉闭塞（middle cerebral artery occlusion， MCAO）模型大鼠神经功能、脑血流、炎症-细胞凋亡基因表达的影响，探讨脑缺血再灌注后脑功能康复的神经调控机制。
方法：将40只SD大鼠随机分为假手术组（S组）、模型组（M组）、电针组（EA组）、经颅交流电刺激组（T组）及电针结合经颅交流电组（EA+T组），每组8只。缺血再灌注后2h，EA组选取双侧曲池、足三里进行电针干预；T组选取右侧M1区（motor cortex M1，运动皮质M1区）进行tACS干预；EA+T组选取电针结合tACS干预；S组与M组均进行气麻30min/次，连续7天。记录大鼠造模前（B）—造模后7天（D7）的神经缺损评分；B—D7右侧大脑中动脉供血区的血流值，记录血流量；D7时取脑RT-PCR方法检测缺血侧的炎症-细胞凋亡基因的表达。
结果：神经缺损评分（neurological deficit scores， NDS）：2h、D1时，M组、EA组、T组、EA+T组与S组相比显著增加（P<0.05）；D3、D5、D7时，S组与其他各组相比显著降低、M组与其他各组相比显著增加（P<0.05）。EA组、T组、EA+T组各时间均有显著性差异（P<0.05）；M组2h、D1、D3、D5、D7与B相比显著上升；D1、D3、D5、D7与2h相比显著下降（P<0.05）。血流量：EA+T组与S组均在2h时下降、D1时上升、D3时下降；EA组则从D3时上升；而M组在D1、D3、D5时与其他各相比均显著下降（P<0.05）。RT-PCR：运动皮质ΔCt法分析（参照基因）：EA+T组的天冬氨酸蛋白水解酶12（cysteinyl aspartate specific proteinase12， Caspase 12）表达显著低于T组（P<0.05）；EA组、EA+T组的细胞因子白介素-1β（interleukin-1β， IL-1β）、核苷酸结合寡聚化结构域样受体1（nucleotide-binding oligomerization domain-like receptor pyrin domain containing 1， NLRP1a）均显著低于S组（P<0.05）。缺血区2-ΔΔCt分析：M组的内质网应激相关蛋白激活转录因子4（activating transcription factor 4， ATF4）表达显著高于其他各组（P<0.05）；M组的B淋巴细胞瘤-2基因（B-cell lymphoma， l-2）显著高于S组、EA组、EA+T组（P<0.01）；M组的Bcl-2相关X蛋白（Bcl-2 associated X potein，Bax）、Caspase 12、细胞癌基因fos（cellular oncogene fos， fos）均显著高于S组（P<0.05）。
结论：电针结合tACS对缺血性脑卒中的干预可能通过调控ATF4、Bcl-2、Bax、Caspase 12、C-fos的表达发挥调节作用，联合应用在减轻炎症反应、抑制细胞凋亡方面，可能优于单独电针或tACS干预，可为缺血性脑卒中的治疗提供新的策略。 |
| 关键词：电针 颅交流电刺激 大脑中动脉闭塞模型 脑血流 炎症-细胞凋亡基因表达 |
| Effects of electroacupuncture combined with transcranial alternating current stimulation on inflammatory-apoptotic genes expression of MCAO rats Download Fulltext |
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| Shanghai Research Institute of Acupuncture and Meridian， Shanghai， 200030 |
| Fund Project: |
| Abstract: |
| Abstract
Objective： To establish right middle cerebral artery occlusion（MCAO） model in rats and to investigate the mechanism underlying motor function regulation by transcranial alternating current stimulation（tACS） intervention. Furthermore， to dynamically observe the effects of electroacupuncture combined with tACS on neurological deficit scores(NDS)， cerebral blood flow， inflammatory-cell apoptosis gene of MCAO model rats and to explore the mechanism of cerebral and neural regulation on motor function rehabilitation after cerebral ischemia reperfusion.
Method： Forty SD rats were randomly divided into sham-operation group （S group）， model group （M group）， electroacupuncture group （EA group）， transcranial alternating current stimulation group （T group） and electroacupuncture combined with transcranial alternating current stimulation group （EA+T group）. After 2 h ischemia-reperfusion， EA group was given bilateral Qu chi （LI 11） and Zu san li （ST 36）electroacupuncture under anesthesia. Right M1 was selected for tACS in T group. EA+T group was treated with EA and tACS together. S and M group were treated with anesthesia for 30min per time， for 7 days. The data from before modling（B） to after modling（D7） were recorded, including neurological deficit score and blood flow of the right middle cerebral artery by laser doppler flowmetry. RT-PCR was used to analyze inflammation-cell apoptosis gene expression at D7.
Result：Neurological deficit score: at 2h， D1， M group， EA group， T group and EA+T group increased significantly compared with other time（P<0.05）. At D3， D5， D7， S group decreased significantly compared with other time（P<0.05）, while M group increased significantly compared with other group（P<0.05）. EA group， T group and EA+T group were significantly different in all times（P<0.05）. At 2h， D1， D3， D5， D7, M group increased significantly compared with B. At D1， D3， D5， D7, NDS decreased significantly compared with that at 2h（P<0.05）. Blood flow： EA+T group and S group decreased at 2h， increased at D1 and decreased at D3. EA group increased at D3. However, M group decreased significantly compared with other group at D1， D3 and D5（P<0.05）. RT-PCR： motor cortex ΔCt analysis： Caspase 12 in EA+T group decreased significantly compared with T group（P<0.05）， IL-1β and NLRP1a in EA group and T group decreased significantly compared with those in S group（P<0.05）. 2-ΔΔCt analysis in ischemic region： ATF4 in M group increased significantly compared with that in other groups （P<0.05）； Bcl 2 in M group increased significantly compared with that in S group， EA group and EA+T group（P<0.05）； Bax， Caspase 12， C-fos in M group increased significantly compared with those in S Group（P<0.05）.
Conclusion： Electroacupuncture combined with tACS can modulate the inflammatory response and inhibit cell apoptosis through regulating ATF4、Bcl-2、Bax、Caspase 12、C-fos and can be a new strategy for the treatment of ischemic stroke. |
| Keywords:electroacupuncture transcranial alternating current stimulation middle cerebral artery occlusion model blood flow inflammatory-apoptotic genes expression |
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