基于“肠-脑”相通理论的针刺对癫痫模型大鼠肠道菌群及血清炎症因子的影响

Effect of Acupuncture Based on Intestine-Brain Communicating Theory on Intestinal Flora and Serum Inflammatory Factors in Epilepsy Model Rats

目的:探讨针刺对癫痫大鼠的肠道菌群、炎症水平的可能作用机制。方法:根据实验要求,将SD大鼠分为对照组、模型组和针刺组,每组各10只。采用戊四氮(PTZ)腹腔注射的方法建立癫痫模型,针刺组针刺“百会”“大椎”穴,每次20 min,对照组和模型组每天固定束缚20 min,Morris水迷宫观察大鼠行为学;海马组织病理变化采用HE染色观察;ELISA检测大鼠血清IL-6、IL-10和肿瘤坏死因子-α(TNF-α)含量;大鼠肠道菌群变化采用16S rDNA测序技术。结果:与对照组比较,模型组大鼠穿越平台次数减少,差异具有统计学意义(P<0.05),逃避潜伏期明显延长,差异具有统计学意义(P<0.05);模型组IL-6和TNF-α表达明显降低,IL-10表达明显升高;肠道有害菌相对丰度明显增加,差异具有统计学意义(P<0.05)。大鼠经针刺治疗后,穿越平台次数显著增加,逃避潜伏期明显缩短,差异具有统计学意义(P<0.05);IL-6和TNF-α表达明显升高,IL-10表达明显降低;与对照组比较,模型组大鼠肠道菌群中Lactobacillus_reuteri、Prevotella_9、Lactobacillus_murinus、Ruminococcaceae_UCG_014、Lactobacillus_intestinalis、Dubosiella、Ruminococcus_2、Prevotellaceae_NK3B31_group及Akkermansia相对丰度显著升高,差异具有统计学意义(P<0.05),Oribacterium、Prevotellaceae_UCG_003、Holdemanella及Lachnospiraceae_UCG-008相对丰度显著降低,差异具有统计学意义(P<0.05);与模型组比较,针刺组肠道菌群g_Rikenellaceae_RC9_gut_group、g_Romboutsia、g_Lachnospiraceae_ND3007_group、g_Quinella及g_Allobaculum相对丰度显著升高,差异具有统计学意义(P<0.05)。结论:戊四氮可能破坏了肠道菌群的稳态,并影响相关代谢物及炎性因子的产生,针刺百会、大椎穴可减少癫痫的发作,其机制可能与针刺调节肠道菌群结构及抑制炎性反应有关。

Objective:To explore the possible mechanism of acupuncture on intestinal flora and inflammation level in epilepsy model rats.Methods:SD rats were randomly divided into the control group,the model group and the acupuncture group,with 10 rats in each group.Epilepsy model was established by intraperitoneal injection of Pentatetrazene(PTZ).The rats in the acupuncture group were treated with needling Baihui(DU20)and Dazhui(DU14),20 min each time;the rats in the control group and the model group were fixed for 20 min each day.The behavior of rats was observed by Morris water maze.The pathological changes of hippocampus were observed by HE staining.The serum levels of IL-6,IL-10 and TNF-αwere detected by ELISA.The changes in intestinal flora were observed by 16S rDNA sequencing technique.Results:Compared with those in the control group,the number of rats crossing the platform was decreased(P<0.05),and the escape latency was significantly extended(P<0.05),the expressions of IL-6 and TNF-αwere decreased significantly(P<0.05),the expression of IL-10 was increased significantly(P<0.05),and the relative abundance of intestinal harmfulness was significantly increased in the mode group(P<0.05).After acupuncture treatment,the number of crossing the platform was significantly increased,and the escape latency was significantly shortened(P<0.05).The expressions of IL-6 and TNF-αwere significantly increased,whereas the expression of IL-10 was significantly decreased.Compared to those in the control group,the relative abundance of intestinal flora of Lactobacillus_reuteri,Prevotella_9,Lactobacillus_murinus,Ruminococcaceae_UCG_014,Lactobacillus_intestinalis,Dubosiella,Ruminococcus_2,Prevotellaceae_NK3B31_group and Akkermansia were significantly increased(P<0.05);the relative abundance of intestinal flora of Oribacterium,Prevotellaceae_UCG_003,Holdemanella and Lachnospiraceae_UCG-008 were significantly decreased in the model group(P<0.05).Compared to those in the model group,the relative abundance of g_Rikenellaceae_RC9_gut_group,g_Romboutsia,g_Lachnospiraceae_ND3007_group and g_Quinella and g_Allobaculum were significantly increased in the acupuncture group(P<0.05).Conclusion:Pentenetetrazon may destroy the homeostasis of intestinal flora,and affect the production of related metabolites and inflammatory factors.Needling DU20 and DU14 can reduce the onset of epilepsy,and the mechanism may be related to the regulation of intestinal flora structure and inhibition of inflammatory response.