摘要
目的:利用转录组测序技术(RNA-Seq)揭示原发性肝癌不同中医证型外周免疫环境的微观差异。方法:纳入原发性肝癌患者81例及健康人50例,分别为原发性肝癌组和健康对照组,其中原发性肝癌组包括脾虚湿困证17例、湿热蕴毒证20例、气虚血瘀证26例及肝肾阴虚证18例,提取受试者外周血单个核细胞(PBMC)用于RNA-Seq测序,利用生物信息学分析技术对两组外周28种免疫细胞浸润丰度进行分析。结果:健康对照组与原发性肝癌组的循环免疫细胞群存在广泛差异,其中原发性肝癌组效应CD8^(+)T细胞、效应记忆CD8^(+)T细胞、滤泡辅助T细胞、自然杀伤样T细胞、中央记忆CD4^(+)T细胞、效应CD4^(+)T细胞、Th1辅助细胞、肥大细胞、幼稚及活化B淋巴细胞丰度均低于健康对照组,而中性粒细胞、巨噬细胞、CD56^(+)自然杀伤细胞、活化树突状细胞、幼稚树突状细胞、单核细胞及髓源性抑制细胞(MDSC)丰度均高于健康对照组(P<0.05)。相关性分析显示,28种循环免疫细胞之间存在广泛的交互作用。效应CD8^(+)T细胞、效应记忆CD8^(+)T细胞、效应CD4^(+)T细胞、γ-δ-T细胞、巨噬细胞及Th17辅助细胞在原发性肝癌不同中医证型中存在差异(P<0.05)。其中气虚血瘀证及肝肾阴虚证效应CD8^(+)T细胞表达水平低于脾虚湿困证及湿热蕴毒证(P<0.05);效应记忆CD8^(+)T细胞在肝肾阴虚证患者中丰度最低,与脾虚湿困证、湿热蕴毒证比较,差异均有统计学意义(P<0.05)。效应CD4^(+)T细胞在气虚血瘀证中浸润丰度最低,与脾虚湿困证、湿热蕴毒证比较,差异有统计学意义(P<0.05)。γ-δ-T细胞在脾虚湿困证水平最低,Th17辅助细胞和巨噬细胞均在气虚血瘀证丰度最低,与肝肾阴虚证比较,差异均有统计学意义(P<0.05)。结论:不同中医证型原发性肝癌循环免疫细胞丰度存在差异,可能成为中医药宏观论治指导下的微观干预靶标。
Objective:To reveal the microscopic differences of peripheral immune environment in different traditional Chinese medicine(TCM)syndromes of primary liver cancer by using Transcriptome sequencing technology(RNA-Seq).Methods:A total of 81 patients with primary liver cancer(PLC)and 50 healthy ordinary people were included as the primary liver cancer group and the healthy control group.The primary liver cancer group included 17 cases of spleen deficiency and dampness syndrome,20 cases of damp-heat accumulation and toxicity syndrome,26 cases of qi deficiency and blood stasis syndrome and 18 cases of liver and kidney yin deficiency syndrome.Peripheral blood mononuclear cells(PBMC)of examinee were extracted for RNA-Seq sequencing.The infiltration abundance of 28 kinds of peripheral immune cells in two groups was analyzed by bioinformatics.Results:There were extensive differences in the circulating immune cell populations between the healthy control group and the primary liver cancer group.Among them,the primary liver cancer group has lower abundance of effector CD8+T cells,effector memory CD8+T cells,follicular helper T cells,natural killer like T cells,central memory CD4+T cells,effector CD4+T cells,Th1 co helper cells,mast cells,immature and activated B lymphocytes compared to the healthy control group,while neutrophils,macrophages,CD56+natural killer cells,activated dendritic cells,immature dendritic cells,monocytes,and myeloid suppressor cells(MDSC)were significantly higher than those of the healthy control group(P<0.05).Correlation analysis showed that there were extensive interactions between 28 circulating immune cells.There were differences in the abundance of effector CD8+T cells,effector memory CD8+T cells,effector CD4+T cells,γ-delta-T cells,macrophages and Th17 helper cells in different TCM syndrome types of PLC(P<0.05).The expression level of CD8+T cells in qi deficiency and blood stasis syndrome and liver-kidney yin deficiency syndrome was lower than that in spleen deficiency and dampness syndrome and dam
作者
胡锐
孙嘉玲
钟欣
李静
张卫
孙新锋
冯文杏
周小舟
HU Rui;SUN Jialing;ZHONG Xin;LI Jing;ZHANG Wei;SUN Xinfeng;FENG Wenxing;ZHOU Xiaozhou(Guangzhou University of Traditional Chinese Medicine,Guangzhou 510006,China;Shenzhen Hospital of Traditional Chinese Medicine,Shenzhen 518033,China;Macao University of Science and Technology,Macao 999078,China)
出处
《山东中医杂志》
2023年第9期955-962,共8页
Shandong Journal of Traditional Chinese Medicine
基金
国家自然科学基金青年科学基金项目(编号:82205209)
深圳市科技计划项目(编号:JCYJ20210324120405015)
广东省中医药局项目(编号:20231302)
广州中医药大学研究生科研创新项目(编号:A1-2606-21-429-001Z38)。
关键词
原发性肝癌
脾虚湿困证
湿热蕴毒证
气虚血瘀证
肝肾阴虚证
循环免疫细胞
转录组测序技术
primary liver cancer
spleen deficiency and dampness syndrome
dampness-heat accumulated toxicity syndrome
qi deficiency and blood stasis syndrome
liver-kidney yin deficiency syndrome
circulating immune cells
RNA sequencing
