摘要
目的基于脑源性神经营养因子(BDNF)/酪氨酸激酶受体B(TrkB)通路探讨镇静安神颗粒对乳腺癌相关性失眠小鼠的作用机制。方法采用鼠源4T1细胞制备乳腺癌荷瘤小鼠模型,并采用足底电刺激为主的慢性不可预知刺激复制乳腺癌相关性失眠小鼠模型。将乳腺癌相关性失眠小鼠随机分为模型组、艾司唑仑组、镇静安神低剂量组、镇静安神高剂量组、镇静安神高剂量+K-252a(BDNF/TrkB通路抑制剂)组,每组12只,另外选取12只乳腺癌荷瘤小鼠作为对照组。艾司唑仑组灌胃给予0.15 mg·kg^(-1)艾司唑仑片;镇静安神低、高剂量组分别灌胃0.54、1.08 g·kg^(-1)镇静安神颗粒;镇静安神高剂量+K-252a组在灌胃1.08 g·kg^(-1)镇静安神颗粒同时,腹腔注射25 mg·kg^(-1)K-252a;每日1次,药物干预持续7 d。观察小鼠一般活动状态;采用戊巴比妥钠翻正实验测定小鼠睡眠质量;旷场实验、高架十字迷宫实验评价小鼠失眠样行为;ELISA法检测小鼠血清5-羟色胺(5-HT)、多巴胺(DA)、γ-氨基丁酸(GABA)含量;RT-qPCR、Western Blot法检测小鼠脑组织中BDNF、TrkB mRNA及蛋白表达水平。结果(1)与对照组比较,模型组小鼠暴躁易激惹,毛发竖起且粗糙、无光泽,昼夜节律消失,体形消瘦,体质量显著降低(P<0.01);睡眠潜伏期显著延长(P<0.01),睡眠时长显著缩短(P<0.01);运动总距离、中心区停留时间、开臂停留时间百分比及进入开臂次数百分比均显著缩短或降低(P<0.01);血清5-HT、GABA含量显著降低(P<0.01),DA含量显著升高(P<0.01);脑组织BDNF、TrkB mRNA及蛋白表达水平显著降低(P<0.01)。(2)与模型组比较,艾司唑仑组及镇静安神低、高剂量组小鼠的一般状态明显改善,体质量显著升高(P<0.01);睡眠潜伏期显著缩短(P<0.01),睡眠时长显著延长(P<0.01);运动总距离、中心区停留时间、开臂停留时间百分比及进入开臂次数百分比均显著延长或提高(P<0.01);血清5-HT、GABA含量显著升高(P<0.01),DA含量显著降低(P<0.01);脑组织BDNF、TrkB mRNA及蛋白表达水平显著升高(P<0.01)。(3)与镇静安神高剂量组比较,镇静安神高剂量+K-252a组小鼠的一般状态变差,体质量显著降低(P<0.01);睡眠潜伏期显著延长(P<0.01),睡眠时长显著缩短(P<0.01);运动总距离、中心区停留时间、开臂停留时间百分比及进入开臂次数百分比均显著缩短或降低(P<0.01);血清5-HT、GABA含量显著降低(P<0.01),DA含量显著升高(P<0.01);脑组织BDNF、TrkB mRNA及蛋白表达水平显著降低(P<0.01)。结论镇静安神颗粒可能通过激活BDNF/TrkB信号通路,调节神经递质水平,改善失眠样行为,提高乳腺癌相关性失眠小鼠的睡眠质量。
Objective To investigate the mechanism of Zhenjing Anshen Granules(ZASG)in treating breast cancerrelated insomnia(BCRI)in mice based on the brain-derived neurotrophic factor(BDNF)/tropomyosin receptor kinase B(TrkB)pathway.Methods A mouse model bearing breast cancer was established using murine 4T1 cells,followed by chronic unpredictable stimulation primarily consisting of plantar electrical stimulation to replicate the BCRI mouse model.The BCRI mice were randomly divided into five groups(n=12 per group):model group,estazolam group,lowdose ZASG group,high-dose ZASG group,and high-dose ZASG+K-252a(a BDNF/TrkB pathway inhibitor)group.An additional 12 tumor-bearing mice served as the control group.The estazolam group received intragastric administration of 0.15 mg·kg^(-1) estazolam tablets;the low-and high-dose ZASG groups received 0.54 g·kg^(-1) and 1.08 g·kg^(-1) ZASG,respectively;the high-dose ZASG+K-252a group received 1.08 g·kg^(-1) ZASG intragastrically along with intraperitoneal injection of 25μg·kg^(-1) K-252a.All treatments were administered once daily for 7 consecutive days.The general activity and state of the mice were observed;sleep quality was assessed using the pentobarbital sodium-induced sleep-wake test;insomnia-like behaviors were evaluated via the open field test and elevated plus maze test;serum levels of serotonin(5-HT),dopamine(DA),andγ-aminobutyric acid(GABA)were measured by ELISA;and the mRNA and protein expression levels of BDNF and TrkB in brain tissue were detected by RT-qPCR and Western Blot,respectively.Results(1)Compared with the control group,mice in the model group exhibited irritability,erected and rough fur,loss of circadian rhythm,emaciation,and a significant decrease in body mass(P<0.01);sleep latency was significantly prolonged(P<0.01),while total sleep duration was significantly shortened(P<0.01);the total movement distance,time spent in the central zone,percentage of time spent in the open arms,and percentage of entries into the open arms were all significantly reduced(P<0.01);serum levels of 5-HT and GABA were significantly decreased(P<0.01),while DA levels were significantly increased(P<0.01);the mRNA and protein expression levels of BDNF and TrkB in brain tissue were significantly downregulated(P<0.01).(2)Compared with the model group,mice in the estazolam group and the low-and high-dose ZASG groups showed significant improvement in general state and a significant increase in body mass(P<0.01);sleep latency was significantly shortened(P<0.01),while total sleep duration was significantly prolonged(P<0.01);the total movement distance,time spent in the central zone,percentage of time spent in the open arms,and percentage of entries into the open arms were all significantly increased(P<0.01);serum levels of 5-HT and GABA were significantly elevated(P<0.01),while DA levels were significantly reduced(P<0.01);the mRNA and protein expression levels of BDNF and TrkB in brain tissue were significantly upregulated(P<0.01).(3)Compared with the high-dose ZASG group,mice in the high-dose ZASG+K-252a group exhibited a worsened general state and a significant decrease in body mass(P<0.01);sleep latency was significantly prolonged(P<0.01),while total sleep duration was significantly shortened(P<0.01);the total movement distance,time spent in the central zone,percentage of time spent in the open arms,and percentage of entries into the open arms were all significantly reduced(P<0.01);serum levels of 5-HT and GABA were significantly decreased(P<0.01),while DA levels were significantly increased(P<0.01);the mRNA and protein expression levels of BDNF and TrkB in brain tissue were significantly downregulated(P<0.01).Conclusion Zhenjing Anshen Granules may improve insomnia-like behaviors and enhance sleep quality in mice with breast cancer-related insomnia by activating the BDNF/TrkB signaling pathway and modulating neurotransmitter levels.
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(The Fifth Ward,Department of Encephalopathy,The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450099 Henan,China;Moscow State Sechenov First Medical University,Moscow 101135,Russia;School of Medicine,Henan University of Chinese Medicine,Zhengzhou 450046,China;Department of Hematology and Oncology,The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450099 Henan,China;Traditional Chinese Medicine Hall,The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450099 Henan,China)
出处
《中药新药与临床药理》
北大核心
2026年第1期22-30,共9页
Traditional Chinese Drug Research and Clinical Pharmacology
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)。
