基于“肠-脑”相关理论的针刺对偏头痛模型大鼠肠道菌群及中枢炎症的影响

Effects of Acupuncture on Gut Microbiota and Central Iinflammation in Rat Model of Migraine based on“Gut-Brain”Theory

目的基于“肠-脑”相关理论观察针刺对偏头痛模型大鼠肠道菌群和中枢炎症的影响,以阐明针刺治疗偏头痛的“肠-脑”作用机制。方法采用硝酸甘油皮下注射法制作偏头痛大鼠模型,并随机分为模型组和针刺组,每组6只,另设空白对照组6只进行常规捆绑固定。在造模前及开始造模后第1、5、9天各组均采用VonFrey测定大鼠足底机械痛阈。实验结束后应用ELISA检测各组大鼠中枢三叉神经脊束核炎性因子白介素-6(Interleukin-6,IL-6)、肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)的表达含量。采用三代Pacbio全长微生物多样性测序对各组粪便样本进行16S全长rDNA测序,检测各组样本间操作分类单元(Operational taxonomic units,OTU)聚类及其丰度、Alpha多样性指数、Beta多样性指数、物种丰度等方面的差异。结果偏头痛模型大鼠存在足底机械痛阈显著降低(P<0.01)、中枢IL-6、TNF-α的含量显著升高(P<0.01)、肠道菌群的结构和丰度发生异常改变(P<0.01);持续针刺治疗可显著升高偏头痛大鼠的足底机械痛阈(P<0.01),调节偏头痛大鼠肠道菌群多样性,并升高鼠乳杆菌、降低肠乳杆菌菌群丰度(P<0.05),降低偏头痛模型大鼠中枢IL-6、TNF-α的含量(P<0.01)。结论针刺可能通过调节偏头痛大鼠肠道菌群结构和中枢IL-6、TNF-α炎症因子的表达,发挥针刺治疗偏头痛的“肠-脑”抗炎镇痛作用。

Objective Based on the theory of"gut-brain",this study explored the effect of acupuncture on the gut microbiota and central inflammation in migraine model rats,in order to explore the mechanism of acupuncture in the treatment of migraine from the perspective of"gut-brain".Methods The migraine rat model was established by subcutaneous injection of nitroglycerin.They were randomly divided into a model group and an acupuncture group,with 6 rats in each group,and a control group with 6 rats for conventional binding and fixation.Before modeling and on the 1st,5th,and 9th days after modeling,each group used electronic VonFrey to measure the plantar mechanical pain threshold of rats.After the experiment,Elisa was used to detect the expression levels of inflammatory factors IL-6 and TNF-αin the central trigeminal spinal nucleus of the rats in each group.Three-generation Pacbio full-length microbial diversity sequencing was used to perform 16S full-length rDNA sequencing on each group of fecal samples to detect the operational taxonomic unit(OTU)clustering and its abundance,Alpha diversity index,Beta diversity index,species among the samples in each group.differences in abundance.Results In migraine model rats,plantar mechanical pain threshold was significantly decreased(P<0.01),central IL-6 and TNF-αcontents were significantly increased(P<0.01),and the structure and abundance of gut microbiota were abnormal.change(P<0.01).Continuous acupuncture treatment can significantly increase the plantar mechanical pain threshold in migraine rats(P<0.01),regulate the diversity of gut microbiota in migraine rats,increase Lactobacillus murine,and reduce the abundance of Lactobacillus enterobacteriaceae.degree(P<0.05),and decreased the levels of IL-6 and TNF-αin the central nervous system of migraine model rats(P<0.01).Conclusion Acupuncture can exert the"gut-brain"anti-inflammatory and analgesic effect of acupuncture in the treatment of migraine by regulating the gut microbiota structure and the expression of central IL-6 and TNF-αinflammatory factors in migraine model rats.