基于1H-NMR和16S rDNA测序技术探讨针与灸不同刺激方法对大鼠结肠代谢物和肠道菌群的影响  

Effects of different stimulation methods,acupuncture and moxibustion,on colonic metabolites and intestinal flora in rats analyzed by 1H-NMR and 16S rDNA sequencing

作　　者：曹思慧 陈琳 何灏龙 李祖强 刘琼[1] 刘密[1] CAO Sihui;CHEN Lin;HE Haolong;LI Zuqiang;LIU Qiong;LIU Mi(College of Acupuncture and Moxibustion,Massage and Rehabilitation,Hunan University of Chinese Medicine,Changsha 410208,China)

机构地区：[1]湖南中医药大学针灸推拿与康复学院,湖南长沙410208

出　　处：《中国病理生理杂志》2023年第9期1620-1633,共14页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81774438);湖南省教育厅资助科研项目(No.21A0235);湖南省中医药科研计划项目(No.C2022027);湖南省自然科学基金资助项目(No.2023JJ30457);长沙市自然科学基金资助项目(No.kq2208183)。

摘　　要：目的:观察针刺与艾灸两种不同干预方法对健康SD大鼠结肠代谢物和肠道菌群调节作用的差异。方法:将33只健康SD大鼠随机分为空白组、针刺组和艾灸组,每组各11只。空白组大鼠仰卧固定在治疗台上,不予任何干预,连续7 d;针刺组以不锈钢针灸针针刺大鼠双侧上巨虚穴和天枢穴,留针15 min,连续干预7 d;艾灸组采用温和灸,将艾条置于大鼠双侧上巨虚穴和天枢穴上方3~5 cm处,使皮肤温度维持在(45±5)℃之间,每次15 min,连续干预7 d。干预结束后,取各组大鼠结肠组织和粪便样品,运用质子核磁共振(proton nuclear magnetic resonance,1H-NMR)技术检测各组大鼠结肠组织代谢物,分析并筛选大鼠结肠组织差异代谢物;16S核糖体DNA(ri-bosomal DNA,rDNA)测序技术检测各组大鼠肠道菌群丰度和多样性的变化情况,并比较差异菌群。结果:代谢组学检测结果显示,与空白组相比,针刺组大鼠结肠组织中代谢物组氨酸、缬氨酸和丁酸含量显著上升(P<0.05),艾灸组大鼠结肠组织中代谢物胆碱和肌醇含量显著上升(P<0.05或P<0.01);与针刺组相比,艾灸组大鼠结肠组织中代谢物甲酸和乙酸含量显著上升(P<0.05)。菌群多样性分析显示,与空白组相比,针刺组和艾灸组大鼠肠道菌群的Chao1指数和基于丰度的覆盖估计值(abundance-based coverage estimator,ACE)均显著上升(P<0.05);在菌群门和属水平上,针、灸干预后部分有益菌丰度上升,部分条件致病菌丰度下降。结论:针刺与艾灸两种不同干预方法对健康SD大鼠结肠代谢物和肠道菌群的调节存在差异性:针刺更擅长调节酸类代谢失衡所导致的胃肠道疾病,艾灸更适用于脂质代谢异常引发的代谢类疾病;针刺与艾灸对肠内微生态环境具有良性调节作用;针刺能通过影响肠道菌群的结构和丰度缓解胃肠道疾病。AIM:To observe the differences in the regulatory effects of acupuncture and moxibustion on colonic metabolites and gut microbiota in healthy SD rats.METHODS:A total of 33 healthy SD rats were randomly divided into 3 groups:blank group,acupuncture group,and moxibustion group,with 11 rats in each group.The rats in blank group was placed in a supine position on the treatment table without interventions for 7 consecutive days.In acupuncture group,stainless steel acupuncture needles were used to puncture bilateral Shangjuxu and Tianshu points,which was retained for 15 min,and the intervention was performed for 7 consecutive days.In moxibustion group,mild moxibustion was applied by placing moxa sticks on both sides of Shangjuxu and Tianshu points,approximately 3~5 cm above the skin.The skin temperature was maintained at(45±5)℃for 15 min during each session,and the intervention continued for 7 days.After collecting colon tissues and fecal samples from the rats in each group,proton nuclear magnetic resonance(1H-NMR)tech-nology was utilized to detect the metabolites in colon tissues,and the differential metabolites were screened and identified.Additionally,the abundance and diversity characteristics of intestinal flora,including differential flora,were examined by 16S ribosomal DNA(rDNA)sequencing.RESULTS:The results of metabolomics detection revealed that the levels of histidine,valine and butyric acid in the colons of rats in acupuncture group were significantly higher than those in blank group(P<0.05).Additionally,the levels of choline and inositol in the colons of rats in moxibustion group showed signifi-cant increases(P<0.01 or P<0.05).Furthermore,there was a significant increase in the levels of formic acid and acetic acid in the colons of rats in moxibustion group compared with acupuncture group(P<0.05).The analysis of the diversity of intestinal flora showed that the Chao1 index and abundance-based coverage estimator(ACE)of the intestinal flora in acupuncture group and moxibustion group were significantly incre

关 键 词：针灸 代谢组学 肠道菌群 质子核磁共振 16S核糖体DNA测序 

分 类 号：R245[医药卫生—针灸推拿学] R363.2[医药卫生—中医临床基础] R333[医药卫生—中医学]