基于肠道菌群探讨心脉隆注射液干预慢性心衰大鼠的作用机制

Exploring the mechanism of action of cardioplegia injection in rats with

关键词:  心脉隆注射液  慢性心衰  肠道菌群  阿霉素  作用机制

Key words:  xinmailong injection  chronic heart failure  intestinal flora  doxorubicin  mechanism

摘要:

目的:基于肠道菌群研究心脉隆注射液对阿霉素诱导的慢性心力衰竭大鼠的作用及机制。 方法:将48只SD大鼠随机分为正常组、模型组、地高辛组(0.025 mg·kg-1)及心脉隆注射液低、中、高剂量组(60、120、240 mg·kg-1),每组8只。除正常组外,其余各组大鼠采用2 mg/kg阿霉素腹腔注射造模,2次/周,持续6周,再进行药物干预14d。对各组大鼠进行心脏超声检测,测定心功能指标左心室射血分数(LVEF)、缩短分数(FS)；腹主动脉取血并收集大鼠血清,测定血清生化指标二胺氧化酶(DAO)、D-乳酸(D-LA)的含量；收集心脏样本并对其进行苏木素-伊红(HE)染色,观察各组大鼠心肌组织病理变化；收集大鼠粪便并采用16S rDNA测序技术对大鼠菌群的物种差异性进行分析。 结果:与正常组比较,模型组LVEF和FS显著下降(P ＜ 0.01)；与模型组比较,心脉隆注射液低、中、高剂量组和地高辛组均显著升高(P ＜ 0.01)。与正常组比较,模型组DAO和D-LA含量显著升高(P ＜ 0.01)；与模型组比较,心脉隆注射液低、中、高剂量组和地高辛组均显著降低(P ＜ 0.01)。与正常组比较,模型组心肌细胞出现水肿、变性、坏死,排列紊乱等；与模型组比较,心脉隆注射液高剂量组和地高辛组心肌细胞形态基本正常,未见明显病理损伤改变,心脉隆注射液中、低剂量组可见心肌细胞形态改变,低剂量组较为明显。肠道菌群结果表明,与正常组比较,模型组肠道菌群Alpha多样性降低,肠道菌群结构、优势菌及各菌群丰度产生变化,在门水平上,螺旋体门(Spirochaetes)丰度明显增加,拟杆菌门(Bacteroidota)丰度降低。属水平上,模型组的异杆菌属(Allobaculum),密螺旋体属(Treponema)等菌群丰富增加；双歧杆菌属(Bifidobacterium)、普雷沃氏菌属Prevotellaceae_Prevotella和瘤胃球菌属(Ruminococcaceae_Ruminococcus)等菌群丰度显著降低。与模型组比较,心脉隆注射液组能够降低潜在致病菌而增加有益菌占比,调节肠道菌群。 结论:心脉隆注射液能够改善阿霉素诱导的慢性心衰,其作用机制可能与调节肠屏障功能,改善肠道菌群紊乱有关。

Abstract:

Objective:Study of the effects and mechanisms of cardioplegia injection on doxorubicin induced chronic heart failure in rats based on intestinal flora.Methods:Forty-eight SD rats were randomly divided into control group,model group,digoxin group (0.025 mg·kg-1),and the low,medium,and high dose groups of xinmailong injection (60,120,and 240 mg·kg-1),8 rats in each group. The rats in all groups except the control group were modeled by intraperitoneal injection of 2 mg/kg doxorubicin twice/week for 6 weeks,followed by pharmacological intervention for 14 d. Ultrasonic cardiogram was performed on each group of rats to determine the left ventricular ejection fraction (LVEF) and shortening fraction (FS),which are indicators of cardiac function；blood was taken from the abdominal aorta and serum was collected from the rats to determine the levels of serum biochemical indicators of diamine oxidase (DAO) and D-lactic acid (D-LA)；heart samples were collected and stained with Hematoxylin-Eosin (HE) to observe the myocardial histopathological changes in the rats in each group；and feces were collected from the rats and analyzed for the species variability of the rat flora by 16S rDNA sequencing technology. Results:Compared with the control group, LVEF and FS were significantly decreased in the model group (P < 0.01)； compared with the model group,they were significantly elevated in the low,medium,and high dose groups of xinmailong injection and digoxin group (P < 0.01). Compared with the control group, DAO and D-LA levels were significantly increased in the model group (P < 0.01)；compared with the model group,they were significantly decreased in the low,medium,and high dose groups of xinmailong injection and digoxin group (P < 0.01). Compared with the control group,cardiomyocytes in the model group showed edema,degeneration,necrosis,and disordered arrangement,etc.Compared with the model group,the morphology of cardiomyocytes in the high dose group of xinmailong injection and Digoxin group was basically normal,and no obvious pathological damage changes were seen,while morphological changes of cardiomyocytes could be seen in the medium and low-dose groups of xinmailong injection and were more obvious in the low-dose group. The results of the intestinal flora showed that the Alpha diversity of the intestinal flora in the model group decreased compared with the control group,and the structure of the intestinal flora,the dominant bacteria,and the abundance of each bacterial gr-oup producedchanges,and at the level of phylum,the abundance of the Spirochaetes increas-ed significantly,and the abundance of the Bacteroidota decreased. At the genus level,the model group showed an increase in the abundance of Allobaculum and Treponema,and the a-bundance of Bifido-bacterium,Prevotellaceae_Prevotella and Ruminococcaceae_Ruminococcus and other flora abun-dance decreased significantly. Compared with the model group,the xinmailong injection group was able to reduce potential pathogenic bacteria and increase the percentage of beneficial bacteria,regulating the intestinal flora.Conclusion:xinmailong injection can improve adriamycin-induced chronic heart failure,and its mechanism of action may be related to the regulation of intestinal barrier function and improvement of intestinal flora disorders.