化浊胶囊对肥胖大鼠肠道菌群的影响

Effect of Huazhuo Capsule(化浊胶囊)on Intestinal Flora in Obese Rat

目的:观察化浊胶囊对高脂饮食诱导的肥胖大鼠肠道菌群多样性、种群结构的影响及血脂、炎症指标的变化。方法:选取Wistar雄性大鼠随机分正常对照组(KB)及造模组,高脂饲料造模,造模成功后随机分为高脂模型组(MX)、二甲双胍组(EJ)、化浊胶囊高剂量组(HG)和化浊胶囊低剂量组(HD),各组饲料不变,在药物干预0、14、28 d时分别收集大鼠粪便,检测粪便细菌DNA并扩增其16S rRNA V1-V3区,采用高通量测序分析肠道菌群多样性和结构变化。并于实验结束后检测血脂4项、C-反应蛋白(CRP)、白介素-6(IL-6)、肿瘤坏死因子(TNF-α)。结果:(1)与KB组比较,MX组厚壁菌门丰度上升,拟杆菌门丰度下降;与MX组相比,EJ、HG、HD组厚壁菌门丰度下降;拟杆菌门丰度上升,其中以HG组较明显。与KB组比较,MX组中拟杆菌属、毛螺菌属、乳酸杆菌属、罗斯拜瑞氏菌属、瘤胃球菌属丰度有下降,拟普雷沃菌属丰度有上升,螺杆菌、大肠杆菌丰度增加;与MX组相比,EJ、HG、HD组中拟杆菌属、毛螺菌属、乳酸杆菌属、罗斯拜瑞氏菌属、瘤胃球菌属丰度上升,HG组丁酸弧菌属、Akkermansia丰度上升,拟普雷沃菌属在HG组下降。(2)与MX组比较,HG组TG、TC、LDL-C有所降低,差异有统计学意义(P<0.05);EJ组TG、LDL-C、HDL-C效果明显,具有统计学差异差异(P<0.05)。(3)与MX组比较,HG组TNF-α、IL-6、CRP有所降低,但略高于KB组,差异有统计学意义(P<0.05,P<0.01);EJ组TNF-α、IL-6、CRP有较好的治疗效果,具有显著性差异(P<0.05,P<0.01)。结论:(1)化浊胶囊可改善肥胖大鼠肠道菌群,表现为厚壁菌门丰度下降,拟杆菌门丰度上升,其中以HG组较明显;拟杆菌属、毛螺菌属、乳酸杆菌属、罗斯拜瑞氏菌属、瘤胃球菌属、丁酸弧菌属、Akkermansia等有益微生物丰度上升,条件致病菌拟普雷沃菌属丰度下降。(2)化浊胶囊调节血脂、减轻炎症反应,其机制可能与改善肥胖大鼠肠道菌群的结构及多样性有关。

Objective:To observe the effects of Huazhuo Capsule on intestinal microflora diversity and population structure in obese rats induced by high-fat diet,and to explore the relationship between Huazhuo Capsule and the changes of intestinal microflora in reducing blood lipid,inflammatory index and intestinal microflora in obese rats induced by high-fat diet.Methods:SPF-grade Wistar male rats were fed adaptively for 1 week and randomly divided into normal control group(KB)and model group.After the model group was successfully made,it was randomly divided into high fat model group(MX),metformin group(EJ),Huazhuo Capsule high dose group(HG)and Huazhuo Capsule low dose group(HD),and the feed remained unchanged in each group,and the model group was divided into two groups:high dose group(HG)and low dose group(HD),The feces of rats were collected at 0,14 and 28 days after drug intervention.The bacterial DNA in feces was detected and its 16s rRNA V1-V3 region was amplified.The diversity and structural changes of intestinal flora were analyzed by high-throughput sequencing.And in the test knot serum lipids,C-reactive protein(CRP),interleukin-6(IL-6)and tumor necrosis factor(TNF-α)were measured.Results:(1)Compared with KB group,the abundance of thick-walled bacteria increased and the abundance of Bacteroides decreased in MX group,and abundance of thick-walled bacteria was decreased in EJ,HG,HD groups,the abundance of Bacteroides increased in EJ,HG,HD group,especially in HG group.Compared with KB group,in MX group,the abundance of Bacteroides,Spirulina,Lactobacillus,Rothbergii and Rumen cocci decreased,while the abundance of preprevous bacteria increased,and the abundance of Helicobacter and Escherichia coli increased.In comparison with MX group,the abundance of Bacteroides,Spirulina,Lactobacillus,Rossellae and Rumen cocci in EJ,HG,HD group was higher than that in control group.The abundance of Akkermansia increased in HG group and decreased in HG group.(2)Compared with MX group,TG,TC and LDL-C in HG group decreased significantly(P<0.05);TG,LDL-C,HDL-C improvement in EJ group were better had statistical difference(P<0.05).(3)Compared with MX group,TNF-αand IL-6,CRP in HG group were lower than those in KB group,but slightly higher than those in KB group(P<0.05,P<0.01),TNF-αand IL-6,CRP had better therapeutic effect in EJ group(P<0.05,P<0.01).Conclusion:(1)Huazhuo Capsule can improve intestinal microflora of obese rats,which shows that the abundance of thick-walled bacteria decreases and the abundance of mycorrhizal bacteria increases,especially in HG group.The abundance of beneficial microorganisms,such as Bacteroides,Spirulina,Lactobacillus,Rosbereli-a,Rumen cocci,Vibrio butyrate,Akkermansia,and so on,increased.(2)Huazhuo Capsule can regulate disorder of blood lipid and alleviate inflammation reaction,that is related to improving the structure and diversity of intestinal flora.