注射用益气复脉(冻干)对慢性心力衰竭大鼠肠道菌群的影响

Effect of Yiqi Fumai Lyophilized Injection on gut microbiota in rats with chronic heart failure

目的探讨注射用益气复脉(冻干)(YQFM)对慢性心力衰竭(CHF)大鼠的肠道菌群的影响。方法采用腹主动脉缩窄法制备CHF大鼠模型,选取造模成功的大鼠随机分为3组:模型组和YQFM低、高剂量(464.3、928.6 mg·kg^(-1),464.3 mg·kg^(-1)为临床等效剂量)组;假手术组大鼠进行切口和缝合,但不进行结扎缩窄。每天尾iv给药1次,连续给药14 d,假手术组和模型组尾iv给予0.9%氯化钠注射液。给药前后采用超声诊断仪检测大鼠心功能;给药14 d后取血,ELISA法检测大鼠血清心钠肽(ANP)、脑钠肽(BNP)、肌酸激酶同工酶(CK-MB)、内皮素(ET)、丙二醛(MDA)水平;取出盲肠位置粪便,进行高通量16S rRNA测序分析。结果与模型组比较,YQFM组大鼠的左室短轴缩短率(LVFS)和左室射血分数(LVEF)均显著升高(P<0.01、0.001),ANP、BNP、CK-MB、ET、MDA水平均显著下降(P<0.001)。Venn图结果显示,CHF模型可以使大鼠肠道菌群数量下降;α多样性分析结果显示,与模型组比较,YQFM组的Chao1、Shannon、Simpson指数均显著上升(P<0.05、0.01);在非度量多维尺度(NMDS)和主坐标分析(PCoA)中,YQFM组与假手术组的差异比模型组与假手术组的差异小;在门水平下,与模型组相比,YQFM组的厚壁菌门相对丰度升高,高剂量组差异显著(P<0.01),YQFM组的拟杆菌门相对丰度有降低趋势,YQFM低、高剂量组的厚壁菌门/拟杆菌门(F/B)呈升高趋势。在丰度等级曲线中,模型组与假手术组的差距大,YQFM组更接近假手术组,肠道菌群的均匀度明显回升。结论YQFM改善CHF大鼠的心功能,缓解炎症反应,改善肠道菌群紊乱。

Objective To investigate the effect of Yiqi Fumai Lyophilized Injection on intestinal microbiota in rats with chronic heart failure(CHF).Method The rat model of CHF was established by constriction of abdominal aorta.The rats that successfully established the model were randomly divided into three groups:model group and YQFM low and high dose(464.3,928.6 mg·kg^(-1),464.3 mg·kg^(-1) as clinical equivalent dose)groups.The rats in the sham-operated group group were cut and sutured,but not ligated or narrowed.The drug was administered iv once a day for 14 consecutive days,and 0.9%sodium chloride injection was administered intravenously in the sham-operated group and the model group.Ultrasound diagnostic equipment was used to detect the cardiac function of rats before and after administration;Blood samples were taken 14 days after administration,and the serum levels of cardiac natriuretic peptide(ANP),brain natriuretic peptide(BNP),creatine kinase isozyme(CK-MB),endothelin(ET),and malondialdehyde(MDA)were detected by ELISA.Remove feces from the cecal site and perform high-throughput 16Sr RNA sequencing analysis.Result Compared with the model group,the left ventricular short axis shortening rate(LVFS)and left ventricular ejection fraction(LVEF)of YQFM group rats were significantly increased(P<0.01,0.001),and the levels of ANP,BNP,CK-MB,ET,MDA were significantly decreased(P<0.001).Venn diagram results showed that CHF model could reduce the number of gut microbiota in rats.The diversity analysis results showed that compared with the model group,the Chao1,Shannon,and Simpson indices of the YQFM group significantly increased(P<0.05,0.01).In non metric multidimensional scale(NMDS)and principal coordinate analysis(PCoA),the difference between YQFM group and sham-operated group was smaller than that between model group and sham-operated group.At the phyla level,compared with the model group,the relative abundance of Bacillota in the YQFM group increased,with a significant difference in the high-dose group(P<0.01).The relative abundance of Bacteroidota in the YQFM group decreased,and the relative abundance of Bacillota/Bacteroidota(F/B)in the low-dose and high-dose YQFM groups increased.In the abundance grade curve,there was a big gap between the model group and the sham-operated group.The YQFM group was closer to the sham-operated group,and the evenness of gut microbiota obviously rised.Conclusion YQFM can improve the heart function of CHF rats,alleviate inflammatory reaction,and improve gut microbiota disorder.