火针疗法对大脑中动脉栓塞大鼠神经功能及肠道菌群的影响

Effects of fire-acupuncture therapy on neurological function and intestinal flora in MCAO rats

关键词:  缺血性脑卒中  火针疗法  炎症因子  肠道菌群  大脑中动脉栓塞大鼠

Key words:  ischaemic stroke  fire needle therapy  inflammatory factors  intestinal flora  middle cerebral artery occlusion rat

摘要:

[目的] 观察火针对大脑中动脉栓塞（MCAO）模型大鼠神经功能及肠道菌群的影响，探讨火针治疗缺血性脑卒中的作用机制。[方法] SPF级健康SD大鼠分为假手术组、MCAO组、火针组，其中MCAO组和火针组建立大脑中动脉阻塞模型，假手术组仅进行颈总动脉、颈内动脉与颈外动脉的分离，不进行造模干预；造模结束后第2天火针组进行火针干预，应用北京贺氏单头细火针予针刺中脘、关元、大椎、命门，快速点刺不留针，隔日干预1次，7天内共干预4次。第7天干预结束后采用Zausinger六分法来评估大鼠的神经功能恢复情况，采用酶联免疫吸附试验（ELISA）法检测大鼠肠黏膜白细胞介素-17（IL-17）和肿瘤坏死因子-α（TNF-α）表达情况，采用苏木精-伊红（HE）染色法检测大鼠肠黏膜整体结构、16SrRNA基因测序技术检测肠道微生物分布情况；采用ELISA法检测外周血和梗死侧脑组织中TNF-α、IL-17水平。[结果] 与假手术组比较，模型组的Zausinger评分降低，肠黏膜中IL-17和TNF-α水平升高，肠道黏膜整体结构异常，肠道微生物丰度降低，致病菌增多，外周血和梗死侧脑组织中TNF-α、IL-17水平升高（均P＜0.05），火针疗法显著提高了MCAO大鼠Zausinger评分，降低肠黏膜中IL-17和TNF-α水平，提高肠道微生物多样性并减少致病菌相对丰度，降低外周血和梗死侧脑组织中TNF-α、IL-17水平（均P＜0.05）。[结论] 火针疗法可促进MACO大鼠神经功能的恢复，其机制可能与调节大鼠肠道菌群，降低外周和脑组织的炎症反应相关。

Abstract:

[Objective] To observe the effect of fire needle on the neurological function and intestinal flora of rats with middle cerebral artery occlusion（MCAO） model，and to explore the mechanism of action of fire needle in the treatment of ischemic stroke. [Methods] SPF-grade healthy SD rats were divided into the sham operation group，MCAO group and fire-acupuncture group，in which the middle cerebral artery occlusion model was established in the MCAO group and fire-acupuncture group，and the sham operation group only carried out the separation of the common carotid artery，the internal carotid artery and the external carotid artery，and did not carry out the intervention of modelling；the fire-acupuncture group was subjected to the intervention of fire-acupuncture on the 2nd day after the completion of modelling，and the single-head fine fire-acupuncture of He was applied to the Zhongwan，Guan Yuan，Dazhui，and Mingmen. Rapid puncture was applied without needle retention，and the intervention was carried out once every other day，with a total of 4 interventions in 7 days. At the end of the intervention on the 7th day，Zausinger’s six-point method was used to assess the neurological recovery of the rats，ELISA was used to detect the expression of IL-17 and TNF-α in the intestinal mucosa of the rats，HE staining was used to detect the overall structure of the intestinal mucosa of the rats，and the distribution of intestinal microorganisms was detected by 16SrRNA sequencing technology；ELISA was used to detect TNF-α in the peripheral blood and brain tissues of infarcted sides. TNF-α，IL-17 levels in brain tissue. [Results] Compared with the sham-operated group，the model group showed lower Zausinger score，higher levels of IL-17 and TNF-α in the intestinal mucosa，abnormal overall structure of the intestinal mucosa，lower abundance of intestinal microorganisms，more pathogenic bacteria，and higher levels of TNF-α and IL-17 in the peripheral blood and infarcted-side brain tissues（all P<0.05），and the fire-acupuncture therapy significantly increased the MCAO rats’ Zausinger score，decreased IL-17 and TNF-α levels in the intestinal mucosa，increased intestinal microbial diversity and decreased the relative abundance of pathogenic bacteria，and decreased TNF-α and IL-17 levels in peripheral blood and infarcted-side brain tissue（all P<0.05）. [Conclusion] Fire-needle therapy can promote the recovery of neurological function in MACO rats，and its mechanism may be related to the regulation of intestinal flora and the reduction of inflammatory response in peripheral and brain tissues.