安寐丹调控BDNF通路改善睡眠剥夺斑马鱼模型睡眠的作用机制研究OA

Objective To explore the effects and potential mechanisms of Anmei Dan(安寐丹,AMD)in improving sleep disorders and brain damage induced by sleep deprivation(SD)in zebrafish models through regulating the BDNF signaling pathway.Methods A total of 240 4-month-old wild-type AB strain zebrafish were randomly divided into a blank group,a model group,low/medium/high-dose AMD groups(0.009,0.027 and 0.081 mg·mL⁻¹·d⁻¹),and a melatonin group(0.4 mg·mL⁻¹·d⁻¹).The zebrafish SD model was estab‑lished via the LED light induction method.The resting time was detected by 24 h locomotor behavior assay;hematoxylin-eosin(HE)staining was used to observe the morphology of brain neurons,and transmission electron microscopy(TEM)was adopted to examine the ultrastructure of neurons.Real-time fluorescent quantitative polymerase chain reaction(RT-qPCR)was applied to detect the mRNA expression levels of inflammatory factors TNF-α,IL-1β and IL-6 in brain tissue;enzyme-linked immunosorbent assay(ELISA)was used to measure the levels of antioxidant indices(MDA,SOD)and neurotransmitters(5-HT,DA,NE);immunofluorescence(IF)was employed to detect the protein expression levels of related molecules in the BDNF signaling pathway.Results Compared with the blank group,the resting time of zebrafish in the model group was significantly reduced(P<0.01);the number of brain neurons decreased with abnormal morphology and obvious ultrastructural damage;the expression of inflammatory factors was elevated,while the levels of anti‑oxidant indices and neurotransmitters were decreased(P<0.001);the protein expressions of BDNF,p-TRKB,p-ERKB and p-CREB were significantly down-regulated(P<0.05 or P<0.01).Compared with the model group,the above behavioral,pathological and bio‑chemical indices were significantly improved in all AMD groups and the melatonin group(P<0.05 or P<0.01)in a dose-dependent manner,with the high-dose AMD group exerting the optimal ameliorative effect.Conclusion AMD can improve sleep in SD-induced zebrafish by activating the BDNF signaling pathway,which further inhibits cerebral inflammatory response,alleviates oxidative stress damage and up-regulates the levels of neurotransmitters.