六君子汤加味灌肠对肥胖大鼠肠道菌群影响实验研究

Experimental Study on the Effect of Enema of Modified Liujunzi Decoction(六君子汤加味)on Intestinal Microflora in Obese Rats

目的探讨六君子汤加味灌肠法对肥胖大鼠肠道菌群的影响。方法将40只雄性SD大鼠按体质量随机分为4组,分别为正常组、模型组、菌液灌肠组、六君子汤加味灌肠组,每组10只。除正常组外,其他组采用高脂饲料喂养复制肥胖大鼠模型,造模成功后,给予菌液灌肠组粪菌液2 mL灌肠,六君子汤加味灌肠组给予六君子汤加味药液2 mL灌肠,正常组与模型组给予0.9%氯化钠溶液2 mL灌肠。灌肠操作1周3次,隔天1次,连续3周。实验结束后,称重检测大鼠体质量,生物化学法检测大鼠血清中甘油三酯含量,并收集各组大鼠粪便进行16SrRNA高通量测序。结果与正常组比较,模型组大鼠体质量显著升高(P<0.01);与模型组比较,菌液灌肠组与六君子汤加味灌肠组体质量均有明显下降(P<0.01)。与正常组相比,模型组和六君子汤加味灌肠组大鼠血清甘油三酯水平升高(P<0.01);与模型组比较,菌液灌肠组大鼠血清甘油三酯水平下降(P<0.05);六君子汤加味灌肠组较菌液灌肠组大鼠血清甘油三酯水平升高(P<0.05)。高脂饮食诱导的肥胖大鼠存在肠道菌群失调,其肠道菌群物种丰富度与多样性均出现上升。六君子汤加味灌肠可上调肥胖大鼠的迷踪菌门、迷踪菌纲、氨基酸球菌科、产碱杆菌科、红螺菌科、肠杆菌科、迷踪菌科、琥珀酸弧菌科、未分类的脱硫弧菌科、蛭弧菌科、毛螺菌科、脱硫弧菌属、粪芽胞菌属、变形杆菌属、考拉杆菌属、寡源杆菌属、未分类的红螺菌属、镰刀菌属、丁酸球菌属的丰度;下调肥胖大鼠的ε变形菌纲、巴斯德氏菌科、消化链球菌科、螺杆菌科、未分类的根瘤菌科、梭菌科1、链球菌科、罗姆布茨菌属、幽门螺杆菌属、狭义梭菌属、罗氏菌属、链球菌属、苏黎世杆菌属、韦荣氏球菌属的丰度。结论菌液灌肠与六君子汤加味灌肠均能在一定程度上减缓高脂饮食诱导的肥胖大鼠体质量增加。肥胖大鼠出现肠道菌群失调,六君子汤加味灌肠可帮助肥胖大鼠重构体内肠道菌群组成,上调有益菌,下调致病菌丰度以达到新的平衡从而减缓肥胖发展进程。提示六君子汤加味灌肠可以调节肥胖大鼠肠道菌群紊乱。

Objective To investigate the effect of Modified Liujunzi Decoction(六君子汤加味)enema on intestinal flora in obese rats.Methods Forty male SD rats were randomly divided into four groups according to their body mass:normal group,model group,bacterial liquid enema group and Modified Liujunzi Decoction enema group,with 10 rats in each group.In addition to the normal group,other groups were fed with high-fat feed to replicate the obese rat model.After the model was successfully established,the bacteria liquid enema group was given 2 mL of fecal bacteria liquid enema,the Modified Liujunzi Decoction enema group was given 2 mL of Modified Liujunzi Decoction enema,and the normal group and the model group were given 2 mL of normal saline enema.The enema was performed three times a week and once every other day for three consecutive weeks.After the experiment,the body weight of rats was measured by weighing;The content of triglyceride in rat serum was detected by biochemical method;The feces of rats in each group were collected for high-throughput 16SrRNA sequencing.Results Compared with the normal group,the body mass of rats in the model group increased significantly(P<0.01);Compared with the model group,the body mass of the bacteria liquid enema group and the Modified Liujunzi Decoction enema group decreased significantly(P<0.01).Compared with the normal group,the serum triglyceride level of rats in the model group and Modified Liujunzi Decoction enema group increased(P<0.01);Compared with the model group,the serum triglyceride level in the bacterial liquid enema group decreased(P<0.05);The serum triglyceride level in the Modified Liujunzi Decoction enema group was higher than that in the bacterial liquid enema group(P<0.05).Obese rats induced by high-fat diet had intestinal flora imbalance,and the species richness and diversity of intestinal flora increased.The Modified Liujunzi Decoction enema can increase the abundance of intestinal flora in obese rats,such as Elusimicrobia,Elusimicrobia,Acidaminococcaceae,Alcaligenaceae,Rhodospirillaceae,Enterobacteriaceae,Elusimicrobiaceae,Succinivibrionaceae,unclassified_Desulfovibrionales,Bdellovibrionaceae,Lachnospiraceae,Desulfovibrio,Coprobacillus,Proteus,Phascolarctobacterium,Oligella,unclassified_Rhodospirillaceae,Fusicatenibacter,Butyricicoccus;Down-regulate the Abundance of Epsilonproteobacteria,Pasteurellaceae,Peptostreptococcaceae,Helicobacteraceae,unclassified_Rhizobiales,Clostridiaceae_1,Streptococcaceae,Romboutsia,Helicobacter,Clostridium_sensu_stricto,Rothia,Streptococcus,Turicibacter,Veillonella.Conclusion Both bacterial liquid enema and Modified Liujunzi Decoction enema can slow down the increase of body mass of obese rats induced by high-fat diet to a certain extent.Obese rats have intestinal flora imbalance.The Modified Liujunzi Decoction enema can help obese rats reconstruct the composition of intestinal flora in their body,upregulate beneficial bacteria,and down-regulate the abundance of pathogenic bacteria to achieve a new balance and slow down the development of obesity.It is suggested that Modified Liujunzi Decoction enema can regulate intestinal flora disorder in obese rats.