Mechanism of Zuogui Jiangtang Jieyu Formula in treating diabetes-related depression by regulating GluR2-mediated mitophagy
10.3969/j.issn.1006-2157.2025.09.009
- VernacularTitle:左归降糖解郁方调控GluR2介导的线粒体自噬治疗糖尿病并发抑郁症的机制研究
- Author:
Jian LIU
1
;
Shuxia YAO
;
Hui YANG
;
Wei LI
;
Yuhong WANG
;
Hu TAN
Author Information
1. 湖南中医药大学第一附属医院 长沙 410007
- Publication Type:Journal Article
- Keywords:
Zuogui Jiangtang Jieyu Formula;
diabetes-related depression;
glutamate receptor 2;
mitophagy;
hippocampal neuron
- From:
Journal of Beijing University of Traditional Chinese Medicine
2025;48(9):1242-1256
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effect and mechanism of Zuogui Jiangtang Jieyu Formula(ZGJTJYF)in treating diabetes-related depression by regulating glutamate receptor 2(GluR2).Methods The primary isolated and cultured hippocampal neurons of SD rats were used.The experiment consisted of normal,model,blank serum(10％blank serum),positive drug(10％[metformin+fluoxetine]drug-containing serum),20％ZGJTJYF group,10％ZGJTJYF group,10％ZGJTJYF+GluR2 knockdown group,and 10％ZGJTJYF+GluR2 overexpression group(with corresponding volume fractions of ZGJTJYF drug-containing serum added).The ZGJTJYF+GluR2 knockdown and overexpression groups,were transfected with lentivirus to obtain hippocampal neurons with either GluR2 overexpression or knockdown.The glucose(150 mmol/L)and corticosterone(200 μmol/L)were used for 18 h to establish an in vitro cell model of hippocampal neurons in diabetes-related depression.After 24 h of successful modeling,the corresponding serum was added to each group for intervention.After 24 h of intervention,the morphological structure of hippocampal neurons was observed using an optical microscope.Biochemical methods were used to determine the glucose and insulin content in cell supernatant.An enzyme-linked immunosorbent assay was used to detect 5-hydroxytryptamine(5-HT)and dopamine(DA)levels in the cell supernatant,and the microtubule-associated protein 1A/1B light chain 3 autophagy double-labeled adenovirus(mRFP-GFP-LC3)autophagy fluorescence double labeling method was used to detect the average fluorescence intensity of LC3 protein in hippocampal neurons.Nissl staining was used to observe synaptic damage in hippocampal neurons,and an immunofluorescence method was used to detect the protein expression of Parkin,phosphatase and tensin homolog-induced putative kinase 1(PINK1),regulating synaptic membrane exocytosis 3(RIMS3),synapsin 1(SYN1),postsynaptic density-95(PSD-95),synapse-associated protein 102(SAP 102),and GluR2 in hippocampal neurons.Realtime fluorescence PCR was used to detect GluR2 mRNA expression in hippocampal neurons,while Western blotting was employed to assess the expression of mitophagy proteins Parkin and PINK1 in these neurons.Results Compared to the normal group,the model group and blank serum group showed structural damage to hippocampal neurons,increased glucose content in cell supernatant,decreased insulin,5-HT,and DA content,increased average fluorescence intensity of LC3,Parkin,and PINK1,decreased average fluorescence intensity of RIMS3,SYN1,PSD-95,SAP 102,and GluR2,decreased GluR2 mRNA expression,increased protein expression of Parkin and PINK1(P＜0.05),and decreased Nissl bodies.Compared to the model group and blank serum group,the above indicators in each administration group were improved to varying degrees(P＜0.05).Compared to the positive drug group,the average fluorescence intensity of LC3,Parkin,and PINK1 decreased,Parkin and PINK1 protein expression decreased,and the average fluorescence intensity of GluR2,SYN1,and PSD-95 increased in 10％ZGJTJYF,20％ZGJTJYF group,and 10％ZGJTJYF+GluR2 overexpression group(P＜0.05).Compared to 10％and 20％ZGJTJYF groups,10％ZGJTJYF+GluR2 knockdown group showed a decrease in 5-HT content,an increase in average fluorescence intensity of LC3 and Parkin,a decrease in average fluorescence intensity of SYN1,PSD-95,and GluR2,a decreased in GluR2 mRNA expression,and an increase of Parkin and PINK1 protein expression(P＜0.05).In contrast,the above indicators were improved to varying degrees in 10％ZGJTJYF+GluR2 overexpression group(P＜0.05).Compared to 10％ZGJTJYF+GluR2 knockdown group,the above abnormal indicators in 10％ZGJTJYF+GluR2 overexpression group were reversed to varying degrees(P＜0.05).Conclusion ZGJTJYF has a protective effect on synaptic damage of hippocampal neurons in diabetes-related depression,and its mechanism may be related to the upregulation of GluR2 and the inhibition of mitophagy over activation.
