摘要
目的基于网络药理学和动物实验探讨酸枣仁-远志药对治疗乳腺癌相关性失眠(breast cancer related insomnia,BCRI)的作用机制。方法通过TCMSP数据库、HERB数据库、DisGeNET数据库筛选酸枣仁-远志药对治疗BCRI的核心靶点;STRING数据库构建酸枣仁-远志药对治疗BCRI的核心靶点蛋白互作网络图;利用Metascape平台对核心靶点进行GO及KEGG富集分析。60只小鼠分为对照组、BCRI组、酸枣仁-远志药对组、地西泮组、酸枣仁-远志药对+LY294002(PI3K抑制剂)组,每组12只。对照组小鼠仅构建乳腺癌模型,不进行慢性不可预知刺激及腹腔注射环磷酰胺处理;其他组小鼠均通过乳腺癌模型+慢性不可预知刺激+腹腔注射环磷酰胺的方法构建BCRI模型,建模成功后给药,1次/d,持续7 d。检测小鼠睡眠潜伏期、睡眠持续时长;HE染色检测下丘脑病理;ELISA检测下丘脑中4-氨基丁酸(4-aminobutyric acid,GABA)、5-羟色胺(5-hydroxytryptamine,5-HT)水平;qRT-PCR检测下丘脑中脑和肌肉芳香烃受体核转运蛋白样蛋白-1(brain and muscle arnt-like 1,BMAL1)、昼夜运动输出周期蛋白(circadian locomotor output cycles kaput,CLOCK)mRNA表达;检测下丘脑中磷酸化磷脂酰肌醇3-激酶(phosphorylated phosphatidylinositol 3-kinase,p-PI3K)、磷酸化蛋白激酶B(phosphorylated protein kinase B,p-AKT)蛋白。结果酸枣仁-远志药对治疗BCRI的核心靶点分别为AKT1、TP53、TNF、ALB、HIF1A、STAT3、ESR1、BCL2、HSP90AA1、CASP3、PPARG、HSP90AB1、MAPK3、TGFB1、MMP9、MTOR、CCND1。对上述17个核心靶点进行GO富集分析得到酸枣仁-远志药对治疗BCRI的细胞组分(cellular component,CC)共23条,分子功能(molecular function,MF)共47条,生物过程(biological process,BP)共211条;KEGG富集分析得到116条通路,预测PI3K/AKT通路可能是酸枣仁-远志药对治疗BCRI的的重要机制。动物实验表明,与对照组相比,BCRI组小鼠下丘脑病理损伤严重,睡眠潜伏期延长,睡眠持续时长减少,下丘脑中GABA、5-HT水平,BMAL1、CLOCK mRNA表达及p-PI3K、p-AKT蛋白降低(P<0.05);与BCRI组相比,酸枣仁-远志药对组、地西泮组小鼠下丘脑病理损伤减轻,睡眠潜伏期缩短,睡眠持续时长增加,下丘脑中GABA、5-HT水平,BMAL1、CLOCK mRNA表达及p-PI3K、p-AKT蛋白升高(P<0.05);LY294002逆转了酸枣仁-远志药对对BCRI小鼠下丘脑病理、神经递质及昼夜节律的影响。结论酸枣仁-远志药对能改善BCRI小鼠下丘脑病理,促进神经递质平衡,恢复昼夜节律,进而改善睡眠,且其作用机制可能与调控P13K/AKT通路有关。
Objective To explore the mechanism of Suanzaoren-Yuanzhi drug pair in the treatment of breast cancer related insomnia(BCRI)based on network pharmacology and animal experiments.Methods The core targets of Suanzaoren-Yuanzhi drug pair for treating BCRI were screened through TCMSP database,HERB database,and DisGeNET database.The STRING database was applied to construct an interaction network diagram of the core target proteins for the treatment of BCRI with Suanzaoren-Yuanzhi drug pair.The core targets were subjected to GO and KEGG enrichment analysis through the Metascape platform.Sixty mice were divided into control group,BCRI group,Suanzaoren-Yuanzhi drug pair group,diazepam group,Suanzaoren-Yuanzhi drug pair+LY294002(PI3K inhibitor)group,with 12 mice in each group.The control group mice were only used to establish a breast cancer model,without undergoing chronic unpredictable stress or intraperitoneal injection of cyclophosphamide.In other groups,the BCRI models were established using the method of breast cancer modeling+chronic unpredictable stress+intraperitoneal injection of cyclophosphamide.After successful modeling,administration was performed once daily for 7 consecutive days.The latency and duration of sleep in mice were detected.HE staining was used to detect hypothalamic pathology.ELISA was used to detect the levels of 4-aminobutyric acid(GABA)and 5-hydroxytryptamine(5-HT)in the hypothalamus.QRT-PCR was used to detect the mRNA expressions of brain and muscle arnt-like 1(BMAL1)and circadian locomotor output cycles kaput(CLOCK)in the hypothalamus.The phosphorylated phosphatidylinositol 3-kinase(p-PI3K)and phosphorylated protein kinase B(p-AKT)proteins in the hypothalamus were detected.Results The core targets for the treatment of BCRI using Suanzaoren-Yuanzhi drug pair were AKT1,TP53,TNF,ALB,HIF1A,STAT3,ESR1,BCL2,HSP90AA1,CASP3,PPARG,HSP90AB1,MAPK3,TGFB1,MMP9,MTOR,and CCND1.GO enrichment analysis for the aforementioned 17 core targets revealed that the cellular component(CC)of Suanzaoren-Yuanzhi drug pair for treating BCRI comprised 23 entries,the molecular function(MF)comprised 47 entries,and the biological process(BP)comprised 211 entries.KEGG enrichment analysis identified 116 pathways,predicting that the PI3K/AKT signaling pathway might be a crucial mechanism for Suanzaoren-Yuanzhi drug pair in the treatment of BCRI.Animal experiments indicated that,compared with the control group,the pathological damage to the hypothalamus of mice in the BCRI group was severe,the sleep latency extended,the duration of sleep decreased,the levels of GABA and 5-HT decreased,the expressions of BMAL1 and CLOCK mRNA,and p-PI3K and p-AKT proteins in the hypothalamus were reduced(all P<0.05).Compared with the BCRI group,the pathological damage to the hypothalamus of mice in the Suanzaoren-Yuanzhi drug pair group and diazepam group was reduced,the sleep latency shortened,the duration of sleep increased,the levels of GABA and 5-HT incresed,the expressions of BMAL1 and CLOCK mRNA,and p-PI3K and p-AKT proteins in the hypothalamus were elevated(all P<0.05).LY294002 reversed the effects of Suanzaoren-Yuanzhi drug pair on hypothalamus pathology,neurotransmitter and circadian rhythm in BCRI mice.Conclusion Suanzaoren-Yuanzhi drug pair can improve the hypothalamic pathology of BCRI mice,promote neurotransmitter balance,restore circadian rhythm,and improve sleep.Its mechanism of action may be related to the regulation of the PI3K/AKT pathway.
作者
古春青
郭睿思
周勤勤
刘恒辉
巴婉玉
孙士玲
王冰
郑玉玲
吴宿慧
GU Chunqing;GUO Ruisi;ZHOU Qinqin;LIU Henghui;BA Wanyu;SUN Shiling;WANG Bing;ZHENG Yuling;WU Suhui(First Clinical Medical College,Henan University of Chinese Medicine,Zhengzhou 450046,Henan,China;Encephalopathy Hospital,The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450099,Henan,China;I.M.Sechenov First Moscow State Medical University,Moscow 101135,Russia;Department of Hematology and Oncology,The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450099,Henan,China;School of Medicine,Henan University of Chinese Medicine,Zhengzhou 450046,Henan,China)
出处
《山东大学学报(医学版)》
北大核心
2026年第1期99-108,130,共11页
Journal of Shandong University(Health Sciences)
基金
河南省中医药科学研究专项课题(20-21ZY2106)
河南省卫生健康委国家中医临床研究基地科研专项(2021JDZY077)
全国名老中医药专家传承工作室建设项目(豫财社〔2021〕177号)。
关键词
酸枣仁-远志药对
乳腺癌相关性失眠
神经递质
昼夜节律
Suanzaoren-Yuanzhi drug pair
Breast cancer related insomnia
Neurotransmitters
Circadian rhythm
