基于BDNF/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OA

Objective To investigate the mechanism of Zhenjing Anshen Granules(ZASG)in treating breast cancer-related 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,low-dose 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 25 μg·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),and γ-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.