龙柴降血方对原发性血小板增多症转录组关键节点的调控  

Longchai Jiangxue Formula Regulates Key Nodes of the Transcriptome in Treating Essential Thrombocythemia

作　　者：赵霈 杨二鹏 孙妍 李雨蒙 郑茂凤 陈卓 王明镜[2] 吕妍[2] 胡晓梅[2] ZHAO Pei;YANG Er-peng;SUN Yan;LI Yu-meng;ZHENG Mao-feng;CHEN Zhuo;WANG Ming-jing;LYU Yan;HU Xiao-mei(Graduate School,Beijing University of Chinese Medicine,Beijing 100029;Department of Hematology,Xiyuan Hospi-tal,China Academy of Chinese Medical Sciences,Beijing 100091;Graduate School,China Academy of Chinese Medical Sciences,Beijing 100700)

机构地区：[1]北京中医药大学研究生院,北京100029 [2]中国中医科学院西苑医院血液科,北京100091 [3]中国中医科学院研究生院,北京100700

出　　处：《世界中西医结合杂志》2025年第1期126-135,共10页World Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金面上项目(82174360);中国中医科学院科技创新工程重大攻关项目(CI2021A01702,CI2021A01708)。

摘　　要：目的基于转录组学,构建原发性血小板增多症(Essential thrombocythemia,ET)相关lncRNA-miR⁃NA-mRNA调控网络,并寻找龙柴降血方发挥治疗作用的靶点。方法纳入28例ET患者和14例健康人。其中10例患者治疗前的标本和5例健康人的标本用于lncRNA、miRNA、mRNA高通量测序;剩余9例健康人和18例ET患者治疗前的标本用做荧光定量PCR(Quantitative realtime PCR,qPCR)检测验证高通量测序的结果;最后将上述患者按1∶1纳入观察组(龙柴降血方+基础治疗)和对照组(基础治疗),用qPCR检测他们在治疗4个月后标本中的关键差异基因表达水平。结果5个lncRNA、12个miRNA和19个mRNA被纳入lncRNA-miRNA-mRNA网络,共包含7对miRNA-lncRNA、35对miRNA-mRNA关系。差异mRNA的GO和KEGG富集分析表明,血小板活化信号通路和PI3K-AKT信号通路在ET中显著富集。两条通路涉及的关键靶点有:F2R、ITGA2B、ITGB1、ITGB3、PTGS1、GP1BB、THBS1和YWHAH,这些与血栓和增殖相关。利用qPCR验证上述网络节点,与健康组相比,18例ET患者GP1BB、ITGB1、THBS1、F2R、ITGA2B、ITGB3和YWHAH的表达水平显著上调,差异有统计学意义(P<0.05),与高通量测序结果一致,PTGS1无显著差异(P>0.05)。使用qPCR检测上述患者治疗后样本中除PTGS1外的关键节点。与对照组比较,观察组治疗后THBS1、F2R和ITGB3的表达水平显著降低,差异有统计学意义(P<0.05)。结论龙柴降血方可能通过下调THBS1、F2R和ITGB3的表达水平,调控ET相关lncRNA-miRNA-mRNA网络中的血小板活化信号通路和PI3K-AKT信号通路。Objective To find the targets of Longchai Jiangxue Formula by building the essential thrombocythe⁃mia(ET)-related lncRNA-miRNA-mRNA regulatory network based on transcriptomics.Methods This study enrolled 28 patients with ET and 14 healthy individuals.Samples from 10 patients before treatment and 5 healthy individuals were used for high-throughput sequencing of lncRNAs,miRNAs,and mRNAs.Samples from the remaining 9 healthy individu⁃als and 18 patients with ET before treatment were used for qPCR to evaluate the results of high-throughput sequencing.Finally,the patients were assigned into an observation group(Longchai Jiangxue Formula+basic treatment)and a control group(basic treatment)based on a ratio of 1∶1.After 4 months of treatment,qPCR was employed to determine the expres⁃sion levels of key differentially expressed genes.Results A total of 5 lncRNAs,12 miRNAs,and 19 mRNAs were included in the lncRNA-miRNA-mRNA network,which included seven miRNA-lncRNA pairs and 35 miRNA-mRNA pairs.GO and KEGG enrichment analyses of differential mRNAs showed that platelet activation signaling pathway and PI3K-AKT signaling pathway were significantly enriched in ET.The key targets involved in the two pathways were F2R,ITGA2B,ITGB1,ITGB3,PTGS1,GP1BB,THBS1,and YWHAH,which were related to thrombosis and tumor proliferation.Then,qPCR was employed to verify the above targets.Compared with healthy individuals,the 18 patients with ET showed up-regulated expression levels of F2R,ITGA2B,ITGB1,ITGB3,GP1BB,THBS1,and YWHAH(P<0.05),which were consistent with the high-throughput sequencing results,and there was no significant difference in the expression level of PTGS1(P>0.05).Subsequently,qPCR was employed to determine the expression levels of the above key targets except PTGS1.Com⁃pared with the control group,the observation group showed down-regulated expression levels of THBS1,F2R,and ITGB3(P<0.05).Conclusion Longchai Jiangxue Formula may regulate the platelet activation and PI3K-Akt signaling path⁃ways in the ET-related lncRNA

关 键 词：龙柴降血方 原发性血小板增多症 lncRNA MIRNA MRNA 调控网络 转录组学 

分 类 号：R558[医药卫生—血液循环系统疾病]