基于Hspa5/PrPc信号研究健脑解郁方对脑小血管病并发抑郁症大鼠海马和皮质星形胶质细胞铜稳态的影响OA

Objective:To investigate the interventional mechanisms of Jiannao Jieyu Formula(JNJYF)on the copper homeostasis of astrocytes(AS)in rats with cerebral small vessel disease related with with depression(CSVDD).Methods:A total of 40 spontaneously hypertensive(SHR)rats were randomly divided into 5 groups:the model group,the positive control group,and the low,medium,and high dose groups of JNJYF,8 rats in each group,and 8 WKY rats were taken as the control group.D-galactose was injected subcutaneously into the back of rats for 8 weeks,together with bilateral common carotid artery stenosis for 4 weeks and chronic unpredicted mild stress for 4 weeks.The CSVDD model was established.The depression-like behavior of rats was evaluated by using the open field experiment and novel feeding experiment;LFB staining was used to observe the pathological changes in the corpus callosum;content of copper ions in hippocampus and cortex were detected by ELISA and electron microprobe for metal ions,immunohistochemical techniques were used to detect the total protrusion length and number of glial fibrillary acidic protein(GFAP)-positive cells in the hippocampal and cortex.Immunofluorescence were used to detect the expression of copper ion transporter protein 1(Ctrl),Western Blot and RT-qPCR techniques were used to detect the expression of heat shock protein family A member 5(Hspa5),prion protein(PrPc)in hippocampus and cortex.Results:Compared with the control group,rats in the model group exhibited reduced spontaneous activity,myelin damage in the corpus callosum,a significant increase in AS copper ion content(P＜0.01),a significant increase in Ctr1 expression(P＜0.01),and a significant decrease in Hspa5,PrPcprotein,and mRNA expression(P＜0.01).Compared with the model group,rats in the high dose group of JNJYF exhibited significantly increased spontaneous activity(P＜0.01)and significantly reduced feeding latency(P＜0.01),corpus callosum vacuolization improved,total dendritic length in AS significantly increased(P＜0.01,P＜0.05),the number of GFAP-positive cells was significantly reduced(P＜0.01),and copper ion level in the hippocampus and cortex was significantly decreased(P＜0.01),in the Ctrl expression in the hippocampus and cortex was significantly reduced(P＜0.01),and the protein and mRNA expression levels of Hspa5,PrPc in the hippocampus and cortex were significantly increased(P＜0.01,P＜0.05).Conclusion:JNJYF can effectively ameliorate depressive-like behaviors in CSVDD rats,the underlying mechanism may be closely associated with the regulation of Hspa5/PrPc signal,maitaining copper homeostasis and suppressing reactive AS.