中国病理生理杂志2024，Vol.40Issue(6)：997-1007,11.DOI:10.3969/j.issn.1000-4718.2024.06.005
基于胶质细胞GR/CX3CR1双信号探讨柴金解郁安神片含药血清减轻体外抑郁模型大鼠ACC神经元突触损伤的机制
Chaijin-Jieyu-Anshen tablet-medicated serum protects rat ACC neurons from synaptic damage via GR/CX3CR1 double signaling in an in vitro de-pression model
摘要
Abstract
AIM:To explore the mechanism by which Chaijin-Jieyu-Anshen tablet(CJJY)-medicated serum prevents synaptic injury in rat anterior cingulate cortex(ACC)neurons using an in vitro depression model.METHODS:Cells(astrocytes,microglia and neurons)were isolated from the ACC of SD rats.The isolated cells were characterized by immunofluorescence staining.An in vitro depression model was developed using 1 mg/L lipopolysaccharide(LPS)com-bined with 200 μmol/L corticosterone(CORT).These cells were divided into control group,model group(CORT+LPS),glucocorticoid receptor(GR)blocker(GR-)group(CORT+LPS+RU486),GR agonist(GR+)group(CORT+LPS+dexa-methasone),CX3C chemokine receptor 1(CX3CR1)blocker(CX3-)group(CORT+LPS+AZD8797),CX3CR1 agonist(CX3+)group(CORT+LPS+fractalkine),CJJY group(CORT+LPS+CJJY-medicated serum),CJJY/GR+group(CORT+LPS+CJJY-medicated serum+dexamethasone),and CJJY/CX3+group(CORT+LPS+CJJY-mediated serum+fractalkine).The morphological characteristics of all ACC cells were observed by high-content analysis.The levels of neuroendocrine-related factors,adrenocorticotropic hormone(ACTH),corticotropin-releasing hormone(CRH)and CORT,and neuroin-flammatory mediators,tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),IL-6 and glutamate(Glu),in the cell supernatants were quantified by ELISA.Immunofluorescence staining was used to analyze the protein expression of GR and vesicular glutamate transporter 1(VGluT1)in astrocytes,as well as CX3CR1 and adenosine A2A receptor(A2AR)in microglia.Immunofluorescence staining with Nissl and β-tubulin was performed to evaluate synaptic damage in ACC neurons.RESULTS:In an in vitro model of depression,CJJY-medicated serum prevented morphological damage to ACC neurons,microglia and astrocytes.Moreover,CJJY-medicated serum reversed abnormal increases in the levels of ACTH,CRH,CORT,TNF-α,IL-1β,IL-6 and Glu in cell supernatants(P<0.05 or P<0.01).It was also found that CJJY-medi-cated serum reduced abnormal expression of GR,VGluT1,CX3CR1 and A2AR(P<0.05 or P<0.01),alleviating damage to the neuronal dendrites and dendritic spines of ACC neurons.CONCLUSION:The CJJY-medicated serum regulates GR/CX3CR1 double signaling in glia,and attenuates rat ACC neuronal synaptic damage in an in vitro depression model,indicating that CJJY-medicated serum controls depression by affecting the GR/CX3CR1 double signaling.关键词
抑郁症/柴金解郁安神片/前扣带皮层/神经元/突触损伤/糖皮质激素受体/CX3C趋化因子受体1Key words
depression/Chaijin-Jieyu-Anshen tablet/anterior cingulate cortex/neurons/synaptic damage/glucocorticoid receptor/CX3C chemokine receptor 1分类
医药卫生引用本文复制引用
刘检,杨蕙,赵洪庆,孟盼,王宇红..基于胶质细胞GR/CX3CR1双信号探讨柴金解郁安神片含药血清减轻体外抑郁模型大鼠ACC神经元突触损伤的机制[J].中国病理生理杂志,2024,40(6):997-1007,11.基金项目
国家重大新药创制科技重大专项(No.2017ZX09309026) （No.2017ZX09309026）
国家自然科学基金资助项目(No.82104793) （No.82104793）
湖南省自然科学基金资助项目(No.2022JJ30451) （No.2022JJ30451）
湖南省卫健委科研课题(No.W20243072) （No.W20243072）
