解毒化瘀颗粒通过库普弗细胞外泌体拮抗ACLF的效用机制研究  

Mechanism of Jiedu Huayu Granules in Antagonizing ACLF through Kupffer Exosome miRNA

作　　者：林镛 颜耿杰 张衎[2] 黄小桃 刘美燕 毛德文[2] 曹音 龙富立[2] LIN Yong;YAN Gengjie;ZHANG Kan;HUANG Xiaotao;LIU Meiyan;MAO Dewen;CAO Yin;LONG Fuli(Graduate School of Guangxi University of Chinese Medicine,Nanning 530000,China;Department of Hepatology,The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,China;Guangxi School of Traditional Chinese Medicine,Nanning 530022,China)

机构地区：[1]广西中医药大学研究生院,南宁530000 [2]广西中医药大学第一附属医院肝病科,南宁530023 [3]广西中医学校,南宁530022

出　　处：《世界中医药》2024年第24期3791-3801,共11页World Chinese Medicine

基　　金：国家自然科学基金项目(82260907,82260899,82274434);广西中医药大学“桂派中医药传承创新团队”资助项目(2022A001);国家青年岐黄学者培养项目(国中医药人教函〔2022〕256号)。

摘　　要：目的:基于库普弗细胞(Kuffer)外泌体微RNA(miRNA)的角度,观察解毒化瘀颗粒(JDHY)对慢加急性肝功能衰竭(ACLF)大鼠的干预作用,并探讨其可能的效应机制。方法:将70只大鼠随机分为正常组(10只)、模型组(30只)以及中药组(30只)。采用四氯化碳(CCl_(4))与脂多糖(LPS)和D-氨基半乳糖(D-GalN)结合构建ACLF大鼠模型,JDHY于造模前48 h灌胃,正常组和模型组给予等量蒸馏水代替。在中药干预及造模完成后,收集各组大鼠肝组织、血液等。提取每组大鼠肝组织库普弗细胞外泌体,并阐明JHDY和库普弗细胞外泌体之间的关系。使用高通量miRNA芯片技术检测各组大鼠库普弗细胞外泌体miRNA表达变化,并确定ACLF组与中药组之间的差异miRNA。最后,通过对库普弗细胞外泌体miRNA进行生物信息学分析,并使用信号相关通路阻断剂和miRNA干扰技术进行反向验证,确定了JDHY发挥药理作用的可能途径。结果:体内研究表明,JDHY颗粒能有效保护ACLF大鼠的肝功能,减轻肝组织病理损伤;体外研究表明,JDHY颗粒可以通过上调库普弗细胞外泌体中miR-9a-5p的表达,减轻ACLF大鼠肝细胞的损伤,提高肝细胞的存活率,这可能是抑制了低氧诱导因子(HIF)信号通路的过度激活,该结果与生物信息学分析一致。结论:JDHY通过上调库普弗细胞外泌体中miR-9a-5p的表达并抑制HIF信号通路的过度激活,在体外和体内保护ACLF大鼠的肝脏损伤,表明其在治疗肝脏疾病中的潜在作用。Objective:To investigate the intervention effect of Jiedu Huayu Granules(JDHY Granules)on acute-on-chronic liver failure(ACLF)in rats from the perspective of Kupffer cell exosome microRNA(miRNA)and explore its potential mechanism of action.Methods:Seventy rats were randomly divided into a normal group(n=10),a model group(n=30),and a Chinese medicine group(n=30).The ACLF rat model was established by administering a combination of carbon tetrachloride(CCl_(4)),lipopolysaccharide(LPS),and D-galactosamine(D-GalN).JDHY Granules were administered orally 48 hours before model induction,and the normal and model groups were given an equal amount of distilled water.After the intervention and model completion,liver tissues and blood samples were collected from the rats.Kupffer cell exosomes were extracted from the liver tissues of each group,and the relationship between JDHY Granules and Kupffer cell exosomes was explored.High-throughput miRNA microarray technology was used to detect changes in miRNA expression in Kupffer cell exosomes from each group,and the differentially expressed miRNAs between the model and Chinese medicine groups were identified.Bioinformatics analysis of miRNA from Kupffer cell exosomes was performed,and pathway inhibitors and miRNA interference techniques were used for reverse validation to identify the potential pathways through which JDHY Granules exerted its pharmacological effects.Results:In vivo studies showed that JDHY Granules significantly protected liver function in ACLF rats and alleviated liver tissue pathological damage.In vitro studies indicated that JDHY Granules increased the expression of miR-9a-5p in Kupffer cell exosomes,which reduced liver cell damage and improved liver cell survival.This effect was likely due to the inhibition of excessive activation of the hypoxia-inducible factor(HIF)signaling pathway,a result consistent with bioinformatics analysis.Conclusion:JDHY Granules protect liver damage in ACLF rats both in vivo and in vitro by upregulating miR-9a-5p expression in Kupff

关 键 词：解毒化瘀颗粒 慢加急性肝衰竭 库普弗细胞 外泌体 miRNA 低氧诱导因子-1信号 中医药 炎症反应 

分 类 号：R285.5[医药卫生—中药学]