淫羊藿素调控巨噬细胞极化治疗肝内胆管癌的药效作用及机制初探
Preliminary exploration of the pharmacological effects and mechanisms of icaritin in regulating macrophage polarization for the treatment of intrahepatic cholangiocarcinoma
王婧雯 1李珍 2黄秀勤 2徐子婧 2耿嘉豪 2许燕瑜 2梁天祎 2湛小燕 3康利平 4王伽伯 5宋鑫华2
作者信息
- 1. 南方医科大学中医药学院,广东广州 510000;首都医科大学中医药学院,北京 100071
- 2. 首都医科大学中医药学院,北京 100071
- 3. 解放军总医院第五医学中心,北京 100071
- 4. 道地药材品质保障与资源持续利用全国重点实验室,北京 100070
- 5. 南方医科大学中医药学院,广东广州 510000;首都医科大学中医药学院,北京 100071;道地药材品质保障与资源持续利用全国重点实验室,北京 100070
- 折叠
摘要
肝内胆管癌(intrahepatic cholangiocarcinoma,ICC)发病率持续攀升,目前尚无有效治疗药物.免疫微环境在ICC发展中扮演重要角色,是当前研究热点.淫羊藿素(icaritin,ICA)是治疗晚期肝细胞癌的创新中药,被认为有潜在的免疫调控抗肿瘤作用,与中医"扶正"治疗肿瘤的认识潜在相合.然而,ICA是否可用于ICC治疗尚未见研究报道.为此,本研究选取免疫系统完整的原位ICC小鼠模型sgp19/kRas,首次在体内评估了 ICA治疗ICC的药效,并通过流式细胞术分析ICA对ICC小鼠肿瘤免疫微环境的影响[本实验所有动物实验均获得首都医科大学动物伦理委员会(中国北京)批准,批准号:AEEI-2023-138].本研究应用100 mg·kg-1 ICA灌胃治疗sgp19/kRas ICC原位小鼠模型,结果显示,ICA治疗3周后能够显著抑制sgp19/kRas ICC小鼠模型的肿瘤生长及肿瘤细胞增殖.通过流式细胞术分析ICA对ICC小鼠肿瘤免疫微环境的影响,结果表明,ICA治疗后能够显著降低M2型巨噬细胞占比,并且显著增加CD3+CD4+T细胞的数量.体外实验阐明ICA能促进巨噬细胞向M1型巨噬细胞极化,抑制其向M2型巨噬细胞极化.此外,转录组学分析结果提示,ICA能够增强Toll样受体9(TLR9)的表达,影响巨噬细胞一氧化氮代谢合成途径及膜受体相关活性,发挥其巨噬细胞极化调节功能.综上,本研究发现ICA治疗能显著延缓sgp19/kRas ICC小鼠肿瘤进展.机制上,ICA可能通过上调TLR9受体表达水平,促进巨噬细胞向M1细胞的极化,改变肿瘤免疫微环境,进而达到抗ICC的作用.
Abstract
The incidence of intrahepatic cholangiocarcinoma(ICC)continues to rise,and there are no effective drugs to treat it.The immune microenvironment plays an important role in the development of ICC and is currently a research hotspot.Icaritin(ICA)is an innovative traditional Chinese medicine for the treatment of advanced hepatocellular carcinoma.It is considered to have potential immunoregulatory and anti-tumor effects,which is potentially consistent with the understanding of"Fuzheng"in the treatment of tumor in traditional Chinese medicine.However,whether ICA can be used to treat ICC has not been reported.Therefore,in this study,sgpl9/kRas,an in situ ICC mouse model with intact immune system,was selected to evaluate the efficacy of ICA in the treatment of ICC in vivo for the first time,and the effects of ICA on the tumor immune microenvironment of ICC mice were analyzed by flow cytometry.This experiment was approved by the Experimental Animal Ethics Committee of Capital Medical University(approval number:AEEI-2023-138).In this study,sgp19/kRas ICC mouse model was treated with oral gavage of 100 mg·kg-1 ICA.We found that ICA significantly inhibited tumor growth and tumor cell proliferation in the sgp19/kRas ICC mouse model after 3 weeks of treatment.Flow cytometry analysis indicated that ICA treatment markedly reduced the proportion of M2 macrophages and increased the number of CD3+CD4+T cells.In vitro experiments demonstrated that ICA promoted macrophage polarization towards the Ml phenotype while inhibiting polarization towards the M2 phenotype.Furthermore,transcriptomic analysis suggested that ICA enhanced Toll-like receptor 9(TLR9)expression,influencing macrophage nitric oxide metabolism synthesis pathways and receptor-related activities,thereby regulating macrophage polarization.In summary,this study demonstrates that ICA treatment significantly delays tumor progression in sgp19/kRas ICC mouse models.Mechanistically,ICA may achieve its anti-ICC effects by upregulating TLR9 receptor expression levels,promoting macrophage polarization towards the M1 phenotype,and altering the tumor immune microenvironment.
关键词
肝内胆管癌/淫羊藿素/免疫微环境/巨噬细胞极化/Toll样受体9Key words
intrahepatic cholangiocarcinoma/icaritin/immune microenvironment/macrophage polarization/Toll-like receptor 9引用本文复制引用
基金项目
国家自然科学基金资助项目(82204680)
中央本级重大增减支项目名贵中药资源可持续利用能力建设项目(2060302-2303-19)
国家中医药管理局高水平中医药重点学科建设项目(ZYYZDXK-2023004)
国家中医药管理局中医药创新团队及人才支持计划项目(ZYYCXTD-C-202005)
出版年
2024
NETL
NSTL
万方数据