电针及电针后粪菌液灌肠对肥胖大鼠肠道菌群的影响

Effect of Electroacupuncture and Fecal Microbiota Enema after Electroacupuncture on the Intestinal Flora of Obese Rats

目的探究电针和电针后粪菌液灌肠对肥胖的影响及可能作用机制。方法120只Wistar大鼠随机抽取8只作为正常组,其余大鼠采用高脂饲料诱导建立肥胖大鼠模型。造模成功的78只大鼠中选取40只随机分为模型组、电针组、非经非穴组、粪菌移植组、模型对照组,每组8只。1)正常组和模型组大鼠不进行干预,电针组造模同时进行电针“中脘”“关元”、双侧“足三里”和“丰隆”;非经非穴组大鼠取电针组穴位旁约5 mm处皮下浅刺并连接电针夹,不予通电,两组均每次10 min,每周3次,干预8周。2)另在剩余的38只造模成功的大鼠中随机选出5只作为粪菌采集组,采取和电针组大鼠相同的干预方式干预8周后,继续电针干预3周,并采集此3周每周最后一次干预后的新鲜粪便制备成粪菌液备用。模型对照组和粪菌移植组大鼠造模8周后进行灌肠,模型对照组大鼠用2 ml生理盐水灌肠,粪菌移植组大鼠用粪菌采集组大鼠新鲜粪便制成的2 ml粪菌液灌肠,每周1次,共3周,两组同时继续高脂饲料喂养。正常组、模型组、电针组和非经非穴组于干预8周后记录大鼠体重并计算Lee’s指数,并取新鲜粪便分析肠道菌群丰度、多样性和代谢功能。模型对照组和粪菌移植组于灌肠3周后取材,检测指标和方法同前。结果1)与正常组比较,模型组大鼠体重、Lee’s指数升高,肠道拟杆菌属、罗氏菌属和普雷沃氏菌属等丰度降低,乳杆菌属、肠球菌丰度升高,肠道COG功能在一般功能预测、氨基酸转运与代谢、细胞修复、能量转化和脂代谢方面降低(P<0.05或P<0.01)。与模型组比较,电针组大鼠体重和Lee’s指数降低,拟杆菌属、罗氏菌属丰度升高,乳杆菌属和肠球菌属丰度降低,肠道COG功能在一般功能预测、氨基酸转运与代谢、能量转化、脂代谢方面均升高(P<0.05或P<0.01);非经非穴组大鼠体重、Lee’s指数、各菌属差异均无统计学意义(P>0.05)。2)与模型对照组比较,粪菌移植组大鼠Lee’s指数降低,肠道乳杆菌属和布劳特氏菌属丰度升高,拟杆菌属、罗姆布茨菌属和柯林斯菌属丰度降低,肠道COG功能在碳水化合物运输和代谢、氨基酸转运与代谢、信号转导、能量转化、脂代谢方面明显升高,而胞膜形成和胞内运输功能降低(P<0.05或P<0.01)。结论电针可降低肥胖大鼠体重,其机制可能与提高肠道优势菌有关,从而改善肠道菌群在能量转化与脂代谢方面的功能,并且通过电针后粪菌液灌肠的方式可继续发挥电针的良性效应。

Objective To explore the effect of electroacupuncture and fecal microflora transplantation after electroacupuncture on obesity,as well as the possible mechanism.Methods Eight out of 120 Wistar rats were randomly selected as the normal group,and the remaining rats were fed with high-fat diet to establish the obesity rat model.Forty out of the 78 successfully modeled rats were randomly divided into model group,electroacupuncture(EA)group,sham EA group(acupuncture inserted not on the channels or the acupoints),fecal microbiota transplantation(FMT)group and model control group,with eight rats in each group.The rats in the normal group and the model group received no interventions.The rats in the EA group received EA inserted on“Zhongwan”(RN 12),“Guanyuan”(RN 4),bilateral“Zusanli”(ST 36),and bilateral“Fenglong”(ST 40),while those in sham EA group were administered with shallow needles on the points,5 mm away from the acupoints used in EA group and with no electricity,both for 10 minutes each time,three times a week for eight weeks.In addition,five rats were randomly selected from the remaining 38 successfully modeled rats as the fecal microbiota collection group.The five rats were given the same8-week intervention with the EA group for an additional three weeks,and the fresh feces were collected after the weekly last treatment to prepare the spare fecal mibrobiota liquid.Rats in model control group and FMT group were given2 ml normal saline and 2 ml fecal mibrobiota liquid from the fecal microbiota collection group by enema,respectively,at eight weeks after modelling,once weekly for three weeks;simultaneously,both groups continued to be fed with high-fat diet.The changes of body weight and Lee’s index were assessed,and the abundance,diversity and metabolic function of intestinal flora were analyzed after eight-week intervention in the normal group,model group,EA group and sham EA group.The same outcomes were assessed in the model control group and FMP group at three weeks after enema.Results creased;the abundance of Enterobacteriaceae,Roche and Prevotella decreased,while the abundance of Lactobacillus and Enterococcus increased;the intestinal COG function was reduced in terms of general function prediction,amino acid transport and metabolism,cell repair,energy conversion and lipid metabolism(P<0.05 or P<0.01).Compared to those in the model group,the body weight and Lee’s index decreased;the abundance of Bacteroides and Roche increased,while the abundance of Lactobacillus and Enterococcus decreased,and the intestinal COG function in terms of general function prediction,amino acid transport and metabolism,energy conversion and lipid metabolism increased in the EA group(P<0.05 or P<0.01).There were no significantly statistical differences between model group and sham EA group in body weight,Lee’s index,and the abundance of varied bacteria genus(P>0.05).Compared to those in the model control group,the Lee’s index of rats in FMT group was reduced;the abundance of Lactobacillus enterica and Brucella increased,while the abundance of Bacteroides,Rhombus sp.and Collins were reduced;the intestinal COG function in carbohydrate transport and metabolism,amino acid transport and metabolism,energy conversion,and lipid metabolism increased,while membrane formation and intracellular transport function decreased(P<0.05 or P<0.01).Conclusion EA can lower the body weight of obesity rats,which may be related to the increase of intestinal dominant bacteria,thereby improving the function of intestinal flora in energy conversion and lipid metabolism.Moreover,fecal microbiota enema after EA helps maintain the benigh effect of EA.