加味逍遥散通过外泌体miRNA途径发挥抗肝癌作用  

Jiawei Xiaoyao San exerts anti-liver cancer effects via exosomal miRNA pathway

作　　者：刘晓明 程金来 黎如霜 李妞妞 秦秋云 夏猛[1] 姚春[2] Liu Xiaoming;Cheng Jinlai;Li Rushuang;Li Niuniu;Qin Qiuyun;Xia Meng;Yao Chun(Guangxi University of Chinese Medicine,Nanning 530200,Guangxi Zhuang Autonomous Region,China;First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530200,Guangxi Zhuang Autonomous Region,China)

机构地区：[1]广西中医药大学,广西壮族自治区南宁市530200 [2]广西中医药大学第一附属医院,广西壮族自治区南宁市530200

出　　处：《中国组织工程研究》2025年第19期4052-4062,共11页Chinese Journal of Tissue Engineering Research

基　　金：广西自然科学基金(2021GXNSFAA220128),项目负责人:夏猛;广西重点研发项目(桂科AB23026137),项目负责人:夏猛。

摘　　要：背景:在课题组先前研究中发现加味逍遥散具有明显的抗肝癌作用,但具体作用机制不明。目的:基于高通量测序结合生物信息学,探讨加味逍遥散对二乙基亚硝胺慢性诱导的原发性肝癌模型大鼠血浆外泌体miRNA水平的调节作用。方法:将SD大鼠随机分为空白对照组、肝癌模型组、加味逍遥散组。以二乙基亚硝胺持续给药12周诱导肝癌模型,从第17周开始,加味逍遥散组大鼠给予加味逍遥散灌服,每日1次,直至第20周末停药,空白对照组和肝癌模型组大鼠给予等量生理盐水灌胃,通过检测大鼠肝组织的形态结构、肝癌标志物Glypican-3蛋白和血清甲胎蛋白表达验证抗肝癌效应。使用超速离心法分离提取各组大鼠血浆外泌体,利用高通量测序技术筛选出大鼠血浆外泌体中差异表达的miRNA,生物信息学预测加味逍遥散通过肝癌血浆来源外泌体miRNA发挥抗肝癌作用的潜在生物标志物,并进行功能分析。结果与结论:①加味逍遥散对肝癌模型大鼠肝组织的形态结构有显著改善作用,与肝癌模型组相比,加味逍遥散组肝癌标志物Glypican-3蛋白和血清甲胎蛋白的表达均显著降低;②生物信息学分析显示,加味逍遥散组对肝癌模型组上调的miR-223-3p与基因E2F1、NCOA1存在靶向结合位点,并且与肝癌生存率及预后密切相关。由此可见,加味逍遥散对肝癌有治疗作用,可能是通过肝癌血浆来源外泌体miR-223-3p靶向负调控NCOA1/E2F1进而发挥抗肝癌效应。BACKGROUND:Previous studies by our research group discovered that Jiawei Xiaoyao San has a significant anti-liver cancer effect,but the specific mechanism of action was unclear.OBJECTIVE:To investigate the regulatory effects of the traditional Chinese medicine formula Jiawei Xiaoyao San on the levels of miRNAs in plasma exosomes of rats with diethylnitrosamine chronically induced primary liver cancer,based on high-throughput sequencing combined with bioinformatics.METHODS:SD rats were randomly divided into a blank control group,a liver cancer model group,and a Jiawei Xiaoyao San treatment group.Liver cancer models were induced by continuous administration of diethylnitrosamine for 12 weeks.Starting from the 17th week,rats in the Jiawei Xiaoyao San treatment group were administered Jiawei Xiaoyao San once daily until the end of the 20th week,while rats in the blank control and liver cancer model groups were given an equivalent volume of saline.Anti-hepatocellular carcinoma effects were validated by assessing the morphological structure of rat liver tissues,along with the expression of the hepatocellular carcinoma markers,Glypican-3 protein and serum alpha-fetoprotein.Plasma exosomes from each group of rats were isolated using ultracentrifugation.High-throughput sequencing technology was used to screen for differentially expressed miRNAs in rat plasma exosomes.Bioinformatics was used to predict the potential biomarkers through which Jiawei Xiaoyao San exerts its anti-liver cancer effects via liver cancer-derived exosomal miRNAs,followed by functional analysis.RESULTS AND CONCLUSION:(1)Jiawei Xiaoyao San significantly improved the morphological structure of liver tissues in a rat model of liver cancer.Compared with the liver cancer model group,the expression of liver cancer markers Glypican-3 protein and serum alpha-fetoprotein was significantly reduced in the Jiawei Xiaoyao San treatment group.(2)Bioinformatics analysis showed that in the Jiawei Xiaoyao San group,upregulated miR-223-3p in the liver cancer model gro

关 键 词：肝癌 加味逍遥散 外泌体 MIRNA 生物信息学 miR-223-3p 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R285.5