隔药饼灸调节大鼠免疫抑制机制的转录组测序分析  

Mechanism of immunosuppression in rats by herb-partitioned moxibustion based on transcriptome sequencing technology

作　　者：田岳凤 熊罗节 王慧芳 翟春涛 李玮 Tian Yuefeng;Xiong Luojie;Wang Huifang;Zhai Chuntao;Li Wei(The Second Clinical College of Shanxi University of Chinese Medicine,Jinzhong 030619,Shanxi Province,China;College of Acupuncture,Massage and Rehabilitation,Hunan University of Chinese Medicine,Changsha 410208,Hunan Province,China)

机构地区：[1]山西中医药大学第二临床学院,山西省晋中市030619 [2]湖南中医药大学针灸推拿与康复学院,湖南省长沙市410208

出　　处：《中国组织工程研究》2025年第5期978-988,共11页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金资助项目(82174504,81674062),项目负责人:田岳凤;山西中医药大学基础研究项目(ZJJC2022005),项目负责人:田岳凤。

摘　　要：背景:免疫抑制会导致机体免疫功能受损,加重病情。隔药饼灸能有效调节机体免疫功能,提高机体免疫力,但其调节机制目前仍不清楚。目的:基于转录组学的生物信息学技术对隔药饼灸干预的免疫抑制模型大鼠进行测序,探究隔药饼灸调控免疫的机制。方法:将24只SD大鼠随机分为空白组、模型组和隔药饼灸组,每组8只,模型组与隔药饼灸组采用腹腔注射环磷酰胺制备免疫抑制模型,35 mg/kg,连续3 d,造模结束后空白组与模型组不做干预,隔药饼灸组在大鼠中脘、神阙、关元及足三里穴位进行艾炷与药饼结合的隔药饼灸干预,连续10 d,1次/d,干预结束后次日取材。取各组大鼠外周血进行白细胞数量检测;采用Illumina测序平台进行RNA-seq,筛选差异基因,结合GO与KEGG数据库,对差异基因进行生物信息学相关分析。结果与结论:①与空白组比较,模型组白细胞数量显著减少(P<0.001)。②RNA-seq共筛选出模型组与空白组间差异基因3026个,其中1565个上调、1461个下调,隔药饼灸组与模型组间差异基因535个,其中280个上调、255个下调。③韦恩图分析获得模型组与空白组159个下调基因经隔药饼灸干预后上调,蛋白互作网络分析获得Oasl,Oas2,Isg15,Herc6,Mx2,Helz2,Mx1,Syk,Hspa1a及Ret等10个核心靶点;④GO与KEGG分析隔药饼灸调节机体的机制有免疫应答途径、病毒相关、血管新生和自身免疫系统等通路。⑤上述结果证实,实验筛选出隔药饼灸干预免疫抑制与Oasl,Oas2,Isg15,Herc6,Mx2,Helz2及Mx1靶点有明显关联,并且在免疫应答、病毒相关及血管新生等通路中发挥调控作用。BACKGROUND:Immunosuppression leads to impaired body immune function and aggravates the disease.Herb-partitioned moxibustion can effectively regulate immune function and improve immunity in the body,but its regulatory mechanism has not been elucidated.OBJECTIVE:To sequence immunosuppressed model rats treated with herb-partitioned moxibustion using bioinformatics techniques based on transcriptomics and to explore the mechanisms by which it regulates immunity.METHODS:Twenty-four Sprague-Dawley rats were randomly assigned to three groups:control,model,and herb-partitioned moxibustion groups,with eight rats in each group.The model and herb-partitioned moxibustion groups were subjected to establishment of an immune suppression model by intraperitoneal injection of cyclophosphamide at a dose of 35 mg/kg for 3 consecutive days.No interventions were administered to the control and model groups after modeling.In contrast,the herb-partitioned moxibustion group received moxibustion treatment at Zhongwan,Shenque,Guanyuan,and Zusanli acupoints using a combination of moxa and herbal cakes,once a day,for 10 consecutive days,with samples being collected the day after the end of the intervention.Peripheral blood was collected from all groups of rats to measure their white blood cell count.RNA-seq was performed on the Illumina sequencing platform,and differentially expressed genes were selected for bioinformatics analysis using the GO and KEGG databases.RESULTS AND CONCLUSION:Compared with the control group,the model group exhibited a significant decrease in white blood cell count(P<0.001).RNAseq analysis identified 3026 differentially expressed genes between the model and control groups,with 1565 upregulated and 1461 downregulated.There were 535 differentially expressed genes identified between the herb-partitioned moxibustion group and the model group,with 280 upregulated and 255 downregulated.The Venn diagram analysis revealed that 159 genes were downregulated in the model group compared with the control group.However,after moxi

关 键 词：免疫抑制 隔药饼灸 转录组测序 寡腺苷酸合成酶样蛋白 寡腺苷酸合成酶2 干扰素诱导基因15 E3泛素蛋白连接酶家族成员6 抗黏病毒蛋白2 锌指解旋酶2 抗黏病毒蛋白1 

分 类 号：R459.9[医药卫生—治疗学] R363[医药卫生—临床医学] R364