参芪补气颗粒对慢性心衰大鼠血浆TMAO相关代谢物及肠道菌群的影响

Effect of Shenqi Buqi Granule on TMAO-related Metabolites in Plasma and Gut Microbiota in Rats with Chronic Heart Failure

目的探讨参芪补气颗粒(Shenqi Buqi granule,SQBQ)对慢性心衰大鼠血浆氧化三甲胺(trimetlylamine oxide,TMAO)相关代谢物及肠道菌群的影响。方法SPF级雄性SD大鼠随机分为假手术组、模型组、参芪补气颗粒组(3.33 g/kg)和福辛普利钠组(1.05 mg/kg),每组8只。采用大鼠心梗后心衰模型,连续灌胃给药28 d,应用心脏超声检测大鼠心脏功能,HE染色观察心肌组织病理变化,液相色谱质谱联用(liquid chromatography-tandem mass spectrometry,LC-MS/MS)技术测定血浆TMAO相关代谢物含量,16S rRNA高通量测序观察肠道菌群组成和丰度,并分析TMAO相关代谢物与差异菌群的相关性。结果与假手术组比较,模型组大鼠心脏结构明显改变,心功能显著下降(P<0.01),心肌细胞水肿变性坏死,炎细胞大片浸润,纤维组织增生;血浆TMAO、三甲胺(trimethylamine,TMA)和γ-丁基甜菜碱(γ-butyrobetaine,BBet)水平显著升高(P<0.05),氯化胆碱(choline chloride,Cho)和甜菜碱(betaine,Bet)显著降低(P<0.01);肠道菌群多样性明显改变,拟杆菌门、放线菌门、软壁菌门、拟杆菌属、乳酸杆菌属、异杆菌属、梭菌属等菌群比例显著降低(P<0.05),变形菌门、颤螺菌属、螺杆菌属等菌群比例显著升高(P<0.05),厚壁菌门与拟杆菌门(F/B)比值也显著升高(P<0.05)。与模型组比较,SQBQ能够显著改善大鼠心脏结构和功能(P<0.05),减轻心肌组织损伤;使血浆TMAO及其前体物质Cho、Bet和BBet含量均发生逆向调节(P<0.05);能够增加肠道菌群多样性,使差异菌群发生显著性的逆向调节(P<0.05)。与TMAO代谢具有较强相关性的菌群有拟杆菌门、螺杆菌属、F/B比值、罗氏菌属、拟杆菌属、异杆菌属及脱硫弧菌属等。结论SQBQ能够改善心衰大鼠TMAO及其相关代谢物的血浆水平,调节与TMAO代谢密切相关的菌群丰度,同时促进肠道微生物内环境稳态,改善心脏功能,减轻心肌损伤,从而发挥抗心力衰竭的作用。

Objective This study aims to investigate the impact of Shenqi Buqi granule(SQBQ)on plasma trimethylamine oxide(TMAO)and its associated metabolites,as well as intestinal flora in rats with chronic heart failure.Methods Male SPF SD rats were randomly assigned to the sham group,model group,SQBQ group(3.33 g/kg),and Fosinopril sodium group(1.05 mg/kg).The rat model of heart failure following myocardial infarction was utilized,with the drugs administered via gavage for a continuous 28-day period.The study utilized echocardiography to assess the cardiac structure and function of rats,HE staining to evaluate pathological changes in the myocardium,liquid chromatography-tandem mass spectrometry(LC-MS/MS)to measure plasma TMAO and its related metabolites,and 16S rRNA high-throughput sequencing to observe the intestinal microbial composition and abundance in rats.These analyses were conducted to investigate the correlation between plasma levels of TMAO-related metabolites and differential flora.Results In the model group,significant changes were observed in the cardiac structure,with a notable decrease in cardiac function(P<0.01).Myocardial cells exhibited edema,degeneration,and necrosis,accompanied by infiltration of inflammatory cells in large areas and proliferation of fibrous tissue.Plasma levels of TMAO,trimethylamine(TMA),and γ-butyrobetaine(BBet)were significantly increased(P<0.05),while choline chloride(Cho)and betaine(Bet)were significantly decreased(P<0.01).Additionally,the composition of intestinal microbiota exhibited significant changes,with notable decreases in the relative abundances of Bacteroidetes,Actinobacteria,Tenericutes,Bacteroides,Lactobacillus,Allobaculum,and Clostridium(P<0.05),alongside significant increases in the relative abundances of Proteobacteria,Oscillospira,and Helicobacter(P<0.05).Additionally,the Firmicutes to Bacteroidetes ratio(F/B Ratio)showed a significant increase(P<0.05).The SQBQ group demonstrated significant improvements in cardiac structure and function in rats(P<0.05)and mitigated myocardial injury.Plasma levels of TMAO,Cho,Bet,and BBet were found to be inversely regulated(P<0.05).SQBQ treatment resulted in a significant reversal of the aforementioned changes in gut microbiota composition(P<0.05)and ameliorated the dysbiosis of intestinal flora.Bacteria such as Bacteroidetes,Helicobacter,F/B ratio,Roseburia,Bacteroides,Allobaculum,and Desulfovibrio were identified as having a strong correlation with the plasma levels of TMAO-related metabolites.Conclusion SQBQ could improve plasma levels of TMAO and its associated metabolites in rats with heart failure,and modulated the composition of gut microbiota involved in TMAO metabolism,contributed to the maintenance of gut microbiota homeostasis,ultimately resulting in improved cardiac function and decreased myocardial damage.