调任通督针法通过调控Notch1/Hes1信号轴改善卒中后抑郁大鼠海马自噬稳态及神经修复OA

Objective To evaluate the effects of Tiao Ren Tong Du(TRTD)electroacupuncture on hippocampal neural repair in rats modeled with post-stroke depression(PSD),focusing on regulation of hippocampal autophagy homeostasis via the Notch1/Hes1 signaling axis,and to compare its efficacy with fluoxetine.Methods A PSD model was established in adult male Sprague-Dawley rats using middle cerebral artery occlusion(MCAO)followed by chronic unpredictable mild stress for 21 days.Rats were divided randomly into sham,MCAO,PSD,fluoxetine,and TRTD electroacupuncture groups.Rats in the treatment groups received either TRTD electroacupuncture or fluoxetine by gavage for 14 days.Brain injury and neurological recovery were assessed using 2,3,5-triphenyltetrazolium chloride staining and modified neurological severity scoring.Depressive-like behaviors were evaluated using the open-field test(OFT)and sucrose preference test.Glial fibrillary acidic protein(GFAP)-positive signals in the hippocampal CA1 region were detected using immunofluorescence.Hippocampal protein expression levels of p62,C3,LAMP2,LC3-Ⅱ,Beclin-1,GFAP,and neuronal nuclei(NeuN)in the hippocampus were detected using Western blot,and mRNA expression levels of Notch1 and Hes1 were determined using reverse transcription-quantitative PCR(RT-qPCR).Results Compared with the PSD group,TRTD electroacupuncture treatment significantly improved neurological deficits(P＜0.05)and depressive-like behaviors,as shown by increased sucrose preference(P＜0.05)and increased total distance traveled in the OFT(P＜0.001)in PSD rats.The TRTD group exhibited greater improvement in the OFT than the fluoxetine group.At the mRNA level,the TRTD electroacupuncture group showed significantly downregulated hippocampal Notch1 expression compared with the MCAO group(P＜0.05)and Hes1 expression compared with the PSD group(P＜0.000 1),indicating inhibition of Notch1/Hes1 signaling overactivation as a core mechanism.At the protein level,immunofluorescence showed that TRTD electroacupuncture significantly reduced the number of GFAP-positive cells in the hippocampal CA1 region(P＜0.01).Western blot showed that GFAP,C3,Beclin-1 expression exhibited a decreasing trend in the TRTD group compared with that in the PSD group,and an increasing trend in LAMP2,but the difference was not statistically significant(P＞0.05).Similarly,the TRTD group exhibited significantly decreased LC3-Ⅱ expression levels compared with the PSD group(P＜0.05)and p62 compared with the MCAO group(P＜0.05).Suggesting that treatment may have restored autophagic homeostasis.Collectively,these molecular and cellular changes ultimately promoted the recovery of NeuN expression.Conclusions TRTD electroacupuncture may exert antidepressant and neuroprotective effects by inhibiting overactivation of the Notch1/Hes1 signaling axis,thereby attenuating reactive astrogliosis and restoring hippocampal autophagy homeostasis.The multitargeted regulation of TRTD may underlie its therapeutic advantages over fluoxetine.