北京中医药大学学报2025，Vol.48Issue(3)：354-369,16.DOI:10.3969/j.issn.1006-2157.2025.03.009
芪参益气滴丸调节肠道菌群和ROS/TXNIP/NLRP3信号通路改善慢性心力衰竭大鼠的作用机制
Mechanism of Qishen Yiqi Dropping Pills in regulating gut microbiota and ROS/TXNIP/NLRP3 signaling pathway to improve chronic heart failure in rats
摘要
Abstract
Objective This study explored the regulatory effects of QiShen Yiqi Dropping Pills(QSYQ)on chronic heart failure(CHF)in rats and their related mechanisms based on the gut microbiota and reactive oxygen species(ROS)/thioredoxin interacting protein(TXNIP)/NOD-like receptor protein 3(NLRP3)signaling pathway.Methods Sixty-five SPF-grade male SD rats were used to establish a CHF model through subcutaneous multiple injections of isoproterenol(ISO)combined with exhaustion and food control methods.The modeled rats were randomly divided into model,captopril(5.30 mg/kg),and QSYQ low-,medium-,and high-dose groups(0.08,0.16,and 0.32 g/kg,respectively),with 11 rats per group,plus a blank group of seven rats.The medication groups were given corresponding drugs by gavage,whereas the blank and model groups were administered an equivalent volume of purified water continuously for four weeks.Rat heart function was assessed via transthoracic echocardiography,and myocardial tissue pathology changes were observed through hematoxylin and eosin staining.Serum levels of brain natriuretic peptide(BNP),lipopolysaccharide(LPS),interleukin-18(IL-18),and interleukin-1β(IL-1β)were measured using an enzyme-linked immunosorbent assay.Automated biochemical analyzers were used to determine creatine kinase(CK),lactate dehydrogenase(LDH),and MB isoenzyme of creatine kinase(CK-MB)content.Myocardial ROS levels were examined using flow cytometry;myocardial TXNIP and NLRP3 expression were detected using immunohistochemistry.Real-time qPCR and Western blotting were used to examine myocardial mRNA and protein expression of TXNIP,NLRP3,apoptosis-related spot-like protein(ASC),caspase-1,and IL-1β,as well as myocardial thioredoxin(Trx)and colonic tight junction proteins(zonula occludens-1,ZO-1),occludin,and claudin-5.Differences in the gut microbiota of the blank,model,and QSYQ high-dose groups were determined using high-throughput 16S rDNA sequencing.Results Compared to the blank group,the model group exhibited significantly reduced left ventricular ejection fraction(LVEF)and left ventricular fraction shortening(LVFS)(P<0.01);increased serum BNP,LPS,IL-18,and IL-1β(P<0.01)levels;increased CK,LDH,and CK-MB(P<0.01)contents;visible myocardial tissue fibrous edema,wavy appearance,cytoplasmic loosening,round vacuolar degeneration,local tissue fibrous dissolution replaced by proliferative connective tissue,accompanied by inflammatory cell infiltration;significantly increased myocardial ROS levels(P<0.01);and significantly increased myocardial TXNIP and NLRP3 expression(P<0.01).TXNIP,NLRP3,ASC,caspase-1,and IL-1β mRNA and protein expression were significantly increased(P<0.05,P<0.01,respectively),whereas Trx,ZO-1,occludin,and claudin-5 expression was significantly decreased(P<0.01).Compared to the model group,the QSYQ high-dose group showed the most significant changes(P<0.05,P<0.01),with significant increases in LVEF and LVFS(P<0.01);significant decreases in serum BNP,LPS,IL-18,and IL-1β levels(P<0.01);significant reductions in CK,LDH,and CK-MB content(P<0.01);improved myocardial tissue damage;significantly decreased myocardial ROS levels(P<0.01);and significantly reduced myocardial TXNIP and NLRP3 expression(P<0.01).TXNIP,NLRP3,ASC,caspase-1,and IL-1β mRNA and protein expression were significantly decreased(P<0.05,P<0.01),whereas Trx,ZO-1,occludin,and claudin-5 expression was significantly increased(P<0.01).16S rDNA sequencing results confirmed that the gut microbiota of rats changed after modeling and drug intervention,with significant differences in both α-and β-diversity.Compared to the blank group,at the family level,the abundance of Oscillospiraceae decreased(P<0.05),whereas the abundance of Lactobacillaceae increased.At the species level,the abundance of Segatella copri and Treponema succinifaciens increased,whereas the abundance of Kineothrix alysoides(P<0.05),Ruminococcus callidus,and Prevotellamassilia timonensis decreased.Compared to the model group,at the family level,the abundance of Oscillospiraceae increased(P<0.05)in the QSYQ high-dose group,whereas the abundance of Lactobacillaceae decreased.At the species level,the abundance of Segatella copri and Treponema succinifaciens decreased,whereas the abundance of Kineothrix alysoides increased(P<0.05).Conclusion QSYQ can regulate the relative abundance of symbiotic bacteria Kineothrix alysoides in the intestines,reduce serum LPS levels,inhibit the ROS/TXNIP/NLRP3 signaling pathway,and improve inflammatory responses,thereby exerting therapeutic effects on CHF.关键词
芪参益气滴丸/肠道菌群/活性氧/硫氧还蛋白相互作用蛋白/核苷酸结合结构域富含亮氨酸重复序列和含热蛋白结构域受体3信号通路/脂多糖/慢性心力衰竭/炎症反应/Kineothrix alysoides/大鼠Key words
Qishen Yiqi Dropping Pills/gut microbiota/reactive oxygen species/thioredoxin interacting protein/NOD-like receptor protein 3 signaling pathway/lipopolysaccharide/chronic heart failure/inflammatory response/Kineothrix alysoides/rats分类
中医学引用本文复制引用
吕李飞,朱婷婷,丁帆,路迎冬,崔向宁..芪参益气滴丸调节肠道菌群和ROS/TXNIP/NLRP3信号通路改善慢性心力衰竭大鼠的作用机制[J].北京中医药大学学报,2025,48(3):354-369,16.基金项目
国家自然科学基金项目(No.81973842,No.82305016) National Natural Science Foundation of China(Nos.81973842 and 82305016) （No.81973842,No.82305016）
