ObjectiveTo investigate the mechanism by which 2，3，5，4'-tetrahydroxyldiphenylethylene-2-O-glucoside （THSG） mitigates cerebral ischemia/reperfusion （CI/R） injury by regulating mitochondrial calcium overload and promoting mitophagy. MethodsSixty male SD rats were randomized into sham， model， SAS （40 mg·kg^-1）， and low-， medium- and high-dose （10， 20， 40 mg·kg^-1， respectively） THSG groups， with 10 rats in each group. The middle cerebral artery occlusion/reperfusion （MCAO/R） model was established by the modified Longa suture method. An oxygen-glucose deprivation/reoxygenation （OGD/R） model was constructed in PC12 cells. Neurological deficits were assessed via Zea Longa scoring， and cerebral infarct volume was measured by 2，3，5-triphenyltetrazolium chloride （TTC） staining. Structural and functional changes of cortical neurons in MCAO/R rats were assessed by hematoxylin-eosin and Nissl staining. PC12 cell viability was detected by cell counting kit-8 （CCK-8） assay， and mitochondrial calcium levels were quantified by Rhod-2 AM. Immunofluorescence was used to detect co-localization of PTEN-induced kinase 1 （PINK1） and leucine zipper/EF-hand-containing transmembrane protein 1 （LETM1） in neurons. Transmission electron microscopy （TEM） was employed to observe mitochondrial morphology in neurons. Western blot was employed to analyze the expression of translocase of outer mitochondrial membrane 20 （TOMM20）， autophagy-associated protein p62， microtubule-associated protein light chain 3 （LC3）， cysteinyl aspartate-specific proteinase-9 （Caspase-9）， B-cell lymphoma 2-associated protein X （Bax）， and cytochrome C （Cyt C）. ResultsCompared with the sham group， the model group exhibited increased infarct volume （P<0.01） and neurological deficit scores （P<0.01）， neuronal structure was disrupted with reduced Nissl bodies. （P<0.01）， mitochondrial swelling/fragmentation， decreased PINK1/LETM1 co-localization （P<0.01）， upregulated protein levels of LC3Ⅱ/LC3Ⅰ， TOMM20， Caspase-9， Bax， and Cyt C （P<0.01）， downregulated protein level of p62 （P<0.05）， weakened PC12 viability （P<0.01）， and elevated mitochondrial calcium level （P<0.01）. Compared with the model group， THSG and SAS groups showed reduced infarct volumes （P<0.05，P<0.01） and neurological deficit scores （P<0.05，P<0.01）， mitigated mitochondrial damage， and increased PINK1/LETM1 co-localization （P<0.01）. Medium/high-dose THSG and SAS alleviated the neurological damage， increased Nissl bodies （P<0.05，P<0.01）， downregulated the protein levels of p62， TOMM20， Caspase-9， Bax， and Cyt C （P<0.05，P<0.01）， and elevated the LC3Ⅱ/LC3Ⅰ level （P<0.05，P<0.01）. High-dose THSG enhanced PC12 cell viability （P<0.01）， increased PINK1/LETM1 co-localization （P<0.01）， and reduced mitochondrial calcium （P<0.01）. ConclusionTHSG may exert the neuroprotective effect on CI/R injury by activating the PINK1-LETM1 signaling pathway， reducing the mitochondrial calcium overload， and promoting mitophagy.

ObjectiveTo investigate the mechanism by which 2，3，5，4'-tetrahydroxyldiphenylethylene-2-O-glucoside （THSG） mitigates cerebral ischemia/reperfusion （CI/R） injury by regulating mitochondrial calcium overload and promoting mitophagy. MethodsSixty male SD rats were randomized into sham， model， SAS （40 mg·kg^-1）， and low-， medium- and high-dose （10， 20， 40 mg·kg^-1， respectively） THSG groups， with 10 rats in each group. The middle cerebral artery occlusion/reperfusion （MCAO/R） model was established by the modified Longa suture method. An oxygen-glucose deprivation/reoxygenation （OGD/R） model was constructed in PC12 cells. Neurological deficits were assessed via Zea Longa scoring， and cerebral infarct volume was measured by 2，3，5-triphenyltetrazolium chloride （TTC） staining. Structural and functional changes of cortical neurons in MCAO/R rats were assessed by hematoxylin-eosin and Nissl staining. PC12 cell viability was detected by cell counting kit-8 （CCK-8） assay， and mitochondrial calcium levels were quantified by Rhod-2 AM. Immunofluorescence was used to detect co-localization of PTEN-induced kinase 1 （PINK1） and leucine zipper/EF-hand-containing transmembrane protein 1 （LETM1） in neurons. Transmission electron microscopy （TEM） was employed to observe mitochondrial morphology in neurons. Western blot was employed to analyze the expression of translocase of outer mitochondrial membrane 20 （TOMM20）， autophagy-associated protein p62， microtubule-associated protein light chain 3 （LC3）， cysteinyl aspartate-specific proteinase-9 （Caspase-9）， B-cell lymphoma 2-associated protein X （Bax）， and cytochrome C （Cyt C）. ResultsCompared with the sham group， the model group exhibited increased infarct volume （P<0.01） and neurological deficit scores （P<0.01）， neuronal structure was disrupted with reduced Nissl bodies. （P<0.01）， mitochondrial swelling/fragmentation， decreased PINK1/LETM1 co-localization （P<0.01）， upregulated protein levels of LC3Ⅱ/LC3Ⅰ， TOMM20， Caspase-9， Bax， and Cyt C （P<0.01）， downregulated protein level of p62 （P<0.05）， weakened PC12 viability （P<0.01）， and elevated mitochondrial calcium level （P<0.01）. Compared with the model group， THSG and SAS groups showed reduced infarct volumes （P<0.05，P<0.01） and neurological deficit scores （P<0.05，P<0.01）， mitigated mitochondrial damage， and increased PINK1/LETM1 co-localization （P<0.01）. Medium/high-dose THSG and SAS alleviated the neurological damage， increased Nissl bodies （P<0.05，P<0.01）， downregulated the protein levels of p62， TOMM20， Caspase-9， Bax， and Cyt C （P<0.05，P<0.01）， and elevated the LC3Ⅱ/LC3Ⅰ level （P<0.05，P<0.01）. High-dose THSG enhanced PC12 cell viability （P<0.01）， increased PINK1/LETM1 co-localization （P<0.01）， and reduced mitochondrial calcium （P<0.01）. ConclusionTHSG may exert the neuroprotective effect on CI/R injury by activating the PINK1-LETM1 signaling pathway， reducing the mitochondrial calcium overload， and promoting mitophagy.

ObjectiveTo investigate the mechanism of Baihuan Xiaoyao Decoction （Xiaoyaosan added with Lilii Bulbus and Albiziae Cortex） in alleviating depression-like behaviors of juvenile rats by regulating the polarization of microglia. MethodSixty juvenile SD rats were randomized into normal control， model， fluoxetine， and low-， medium-， and high-dose （5.36， 10.71， 21.42 g·kg^-1， respectively） Baihuan Xiaoyao decoction groups. The rat model of juvenile depression was established by chronic unpredictable mild stress （CUMS）. The sucrose preference test （SPT） was carried out to examine the sucrose preference of rats. Forced swimming test （FST） was carried out to measure the immobility time of rats. The open field test （OFT） was conducted to measure the total distance， the central distance， the number of horizontal crossings， and the frequency of rearing. Morris water maze （MWM） was used to measure the escape latency and the number of crossing the platform. The immunofluorescence assay was employed to detect the expression of inducible nitric oxide synthase （iNOS， the polarization marker of M1 microglia） and CD206 （the polarization marker of M2 microglia）. Real-time polymerase chain reaction was employed to determine the mRNA levels of iNOS， CD206， pro-inflammatory cytokines ［tumor necrosis factor （TNF）-α， interleukin （IL）-1β， and IL-6］ and anti-inflammatory cytokines （IL-4 and IL-10） in the hippocampus. Western blotting was employed to determine the protein levels of iNOS and CD206 in the hippocampus. The levels of IL-4 and IL-6 in the hippocampus were detected by enzyme-linked immunosorbent assay. ResultCompared with the normal control group， the model rats showed a reduction in sucrose preference （P<0.05）， an increase in immobility time （P<0.05）， decreased motor and exploratory behaviors （P<0.05）， and weakened learning and spatial memory （P<0.05）. In addition， the model rats showed up-regulated mRNA and protein levels of iNOS and mRNA levels of IL-1β， IL-6， and TNF-α （P<0.05）. Compared with the model group， Baihuan Xiaoyao decoction increased the sucrose preference value （P<0.05）， shortened the immobility time （P<0.01）， increased the motor and exploratory behaviors （P<0.05）， and improved the learning and spatial memory （P<0.01）. Furthermore， the decoction down-regulated the positive expression and protein level of iNOS， lowered the levels of TNF-α， IL-1β， and IL-6 （P<0.01）， promoted the positive expression of CD206， and elevated the levels of IL-4 and IL-10 （P<0.01） in the hippocampus of the high dose group. Moreover， the high-dose Baihuan Xiaoyao decoction group had higher sucrose preference value （P<0.01）， shorter immobility time （P<0.01）， longer central distance （P<0.01）， stronger learning and spatial memory （P<0.01）， higher positive expression and protein level of iNOS （P<0.01）， lower levels of TNF-α， IL-1β， and IL-6 （P<0.05， P<0.01）， lower positive expression and mRNA level of iNOS （P<0.05）， and higher levels of IL-4 and IL-10 （P<0.05， P<0.01） than the fluoxetine group. ConclusionBaihuan Xiaoyao decoction can improve the depression-like behavior of juvenile rats by inhibiting the M1 polarization and promoting the M2 polarization of microglia in the hippocampus.

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.

Objective To explore the protective mechanism of Zuogui Jiangtang Jieyu Formula(ZGF)on synaptic damage of hippocampal neurons based on leukocyte mono-immunoglobulin-like receptor 3(CD300f)/glucose transporter 1(GLUT1)signal-mediated microglial glucose metabolism in rats with diabetes-related depression.Methods Eighty male SD rats were randomly selected using random number table method,with 10 rats serving as the normal group.The remaining 70 rats were fed a high-fat diet for 4 weeks and then injected once with 38 mg/kg of streptozotocin via the tail vein to replicate the diabetes rat model.Sixty rats were screened and successfully modeled,which were randomly divided into the model,CD300f blocker,CD300f agonist,metformin+fluoxetine(metformin 0.18 g/kg+fluoxetine 1.8 mg/kg),and ZGF high-and low-dose(20.52 and 10.26 g/kg,respectively)groups using random number table method.In addition to the normal group,the rats in the other groups underwent chronic unpredictable mild stress combined with isolation feeding for 28 days to replicate the diabetes-related depression rat model.The metformin+fluoxetine and ZGF high-and low-dose groups were subjected to continuous intragastrial administration for 14 days after the second week of modeling.The normal and model groups were administered an equal amount of distilled water by gavage.The CD300f blocker group and agonist group received microinjection into the hippocampus,with injection of myeloid cell trigger receptor inhibitory factor(CLM1,2 μg/kg)and immunoglobulin Fc surface protein(Fcγ,5 μg/kg)once a week,respectively.Depression-like behavior in rats was evaluated using open-field and forced swimming tests after the intervention.Biochemical analysis was used to detect the glucose,lactic acid,and adenosine diphosphate(ADP)/adenosine triphosphate(ATP)ratio contents.The insulin,5-hydroxytryptamine(5-HT),and dopamine(DA)levels in the hippocampus were detected using an enzyme-linked immunosorbent assay.Immunofluorescence was used to detect the average fluorescence intensity of CD300f,GLUT1,regulating synaptic membrane wxocytosis 3(RIMS3),and synapse-associated protein 102(SAP102)in hippocampal tissue.Western blotting was used to detect the CD300f,GLUT1,RIMS3,and SAP102 protein expression levels in the hippocampus.The synaptic damage of hippocampal neurons was observed using Nissl staining and transmission electron microscope.Results Compared with the normal group,the model group showed a decrease in the total active distance in the open-field test and an increase in forced swimming immobility time,with an increase in glucose and lactic acid contents and ADP/ATP ratio,whereas a decrease in insulin,5-HT,and DA levels was observed in the hippocampus.The average fluorescence intensity and relative protein expression levels of CD300f,GLUT1,RIMS3,and SAP102 in hippocampal tissue decreased(P<0.05),and the synaptic ultrastructure of hippocampal neurons was damaged.Compared with the model group,depression-like behavioral changes,glucose metabolism,and monoamine neurotransmitter imbalance were alleviated in the CD300f agonist group and ZGF high-and low-dose group(P<0.05).The average fluorescence intensity and relative protein expression levels of CD300f,GLUT1,RIMS3,and SAP102 in the hippocampus of the CD300f agonist group and the ZGF high-dose group were all increased(P<0.05),and synaptic damage was alleviated.The abnormal levels of glucose,lactate,ADP/ATP,5-HT,and CD300f protein expression were aggravated in the CD300f blocker group(P<0.05),and synaptic damage was aggravated.Conclusion ZGF can alleviate glucose metabolism disorders in hippocampal microglia and synaptic damage in hippocampal neurons in rats with diabetes-related depression.Its mechanism may be related to regulating the CD300f/GLUT1 signaling pathway.

Objective To explore the protective mechanism of Zuogui Jiangtang Jieyu Formula(ZGF)on synaptic damage of hippocampal neurons based on leukocyte mono-immunoglobulin-like receptor 3(CD300f)/glucose transporter 1(GLUT1)signal-mediated microglial glucose metabolism in rats with diabetes-related depression.Methods Eighty male SD rats were randomly selected using random number table method,with 10 rats serving as the normal group.The remaining 70 rats were fed a high-fat diet for 4 weeks and then injected once with 38 mg/kg of streptozotocin via the tail vein to replicate the diabetes rat model.Sixty rats were screened and successfully modeled,which were randomly divided into the model,CD300f blocker,CD300f agonist,metformin+fluoxetine(metformin 0.18 g/kg+fluoxetine 1.8 mg/kg),and ZGF high-and low-dose(20.52 and 10.26 g/kg,respectively)groups using random number table method.In addition to the normal group,the rats in the other groups underwent chronic unpredictable mild stress combined with isolation feeding for 28 days to replicate the diabetes-related depression rat model.The metformin+fluoxetine and ZGF high-and low-dose groups were subjected to continuous intragastrial administration for 14 days after the second week of modeling.The normal and model groups were administered an equal amount of distilled water by gavage.The CD300f blocker group and agonist group received microinjection into the hippocampus,with injection of myeloid cell trigger receptor inhibitory factor(CLM1,2 μg/kg)and immunoglobulin Fc surface protein(Fcγ,5 μg/kg)once a week,respectively.Depression-like behavior in rats was evaluated using open-field and forced swimming tests after the intervention.Biochemical analysis was used to detect the glucose,lactic acid,and adenosine diphosphate(ADP)/adenosine triphosphate(ATP)ratio contents.The insulin,5-hydroxytryptamine(5-HT),and dopamine(DA)levels in the hippocampus were detected using an enzyme-linked immunosorbent assay.Immunofluorescence was used to detect the average fluorescence intensity of CD300f,GLUT1,regulating synaptic membrane wxocytosis 3(RIMS3),and synapse-associated protein 102(SAP102)in hippocampal tissue.Western blotting was used to detect the CD300f,GLUT1,RIMS3,and SAP102 protein expression levels in the hippocampus.The synaptic damage of hippocampal neurons was observed using Nissl staining and transmission electron microscope.Results Compared with the normal group,the model group showed a decrease in the total active distance in the open-field test and an increase in forced swimming immobility time,with an increase in glucose and lactic acid contents and ADP/ATP ratio,whereas a decrease in insulin,5-HT,and DA levels was observed in the hippocampus.The average fluorescence intensity and relative protein expression levels of CD300f,GLUT1,RIMS3,and SAP102 in hippocampal tissue decreased(P<0.05),and the synaptic ultrastructure of hippocampal neurons was damaged.Compared with the model group,depression-like behavioral changes,glucose metabolism,and monoamine neurotransmitter imbalance were alleviated in the CD300f agonist group and ZGF high-and low-dose group(P<0.05).The average fluorescence intensity and relative protein expression levels of CD300f,GLUT1,RIMS3,and SAP102 in the hippocampus of the CD300f agonist group and the ZGF high-dose group were all increased(P<0.05),and synaptic damage was alleviated.The abnormal levels of glucose,lactate,ADP/ATP,5-HT,and CD300f protein expression were aggravated in the CD300f blocker group(P<0.05),and synaptic damage was aggravated.Conclusion ZGF can alleviate glucose metabolism disorders in hippocampal microglia and synaptic damage in hippocampal neurons in rats with diabetes-related depression.Its mechanism may be related to regulating the CD300f/GLUT1 signaling pathway.

Objective To explore the protective mechanism of Zuogui Jiangtang Jieyu Formula(ZGF)on synaptic damage of hippocampal neurons based on leukocyte mono-immunoglobulin-like receptor 3(CD300f)/glucose transporter 1(GLUT1)signal-mediated microglial glucose metabolism in rats with diabetes-related depression.Methods Eighty male SD rats were randomly selected using random number table method,with 10 rats serving as the normal group.The remaining 70 rats were fed a high-fat diet for 4 weeks and then injected once with 38 mg/kg of streptozotocin via the tail vein to replicate the diabetes rat model.Sixty rats were screened and successfully modeled,which were randomly divided into the model,CD300f blocker,CD300f agonist,metformin+fluoxetine(metformin 0.18 g/kg+fluoxetine 1.8 mg/kg),and ZGF high-and low-dose(20.52 and 10.26 g/kg,respectively)groups using random number table method.In addition to the normal group,the rats in the other groups underwent chronic unpredictable mild stress combined with isolation feeding for 28 days to replicate the diabetes-related depression rat model.The metformin+fluoxetine and ZGF high-and low-dose groups were subjected to continuous intragastrial administration for 14 days after the second week of modeling.The normal and model groups were administered an equal amount of distilled water by gavage.The CD300f blocker group and agonist group received microinjection into the hippocampus,with injection of myeloid cell trigger receptor inhibitory factor(CLM1,2 μg/kg)and immunoglobulin Fc surface protein(Fcγ,5 μg/kg)once a week,respectively.Depression-like behavior in rats was evaluated using open-field and forced swimming tests after the intervention.Biochemical analysis was used to detect the glucose,lactic acid,and adenosine diphosphate(ADP)/adenosine triphosphate(ATP)ratio contents.The insulin,5-hydroxytryptamine(5-HT),and dopamine(DA)levels in the hippocampus were detected using an enzyme-linked immunosorbent assay.Immunofluorescence was used to detect the average fluorescence intensity of CD300f,GLUT1,regulating synaptic membrane wxocytosis 3(RIMS3),and synapse-associated protein 102(SAP102)in hippocampal tissue.Western blotting was used to detect the CD300f,GLUT1,RIMS3,and SAP102 protein expression levels in the hippocampus.The synaptic damage of hippocampal neurons was observed using Nissl staining and transmission electron microscope.Results Compared with the normal group,the model group showed a decrease in the total active distance in the open-field test and an increase in forced swimming immobility time,with an increase in glucose and lactic acid contents and ADP/ATP ratio,whereas a decrease in insulin,5-HT,and DA levels was observed in the hippocampus.The average fluorescence intensity and relative protein expression levels of CD300f,GLUT1,RIMS3,and SAP102 in hippocampal tissue decreased(P<0.05),and the synaptic ultrastructure of hippocampal neurons was damaged.Compared with the model group,depression-like behavioral changes,glucose metabolism,and monoamine neurotransmitter imbalance were alleviated in the CD300f agonist group and ZGF high-and low-dose group(P<0.05).The average fluorescence intensity and relative protein expression levels of CD300f,GLUT1,RIMS3,and SAP102 in the hippocampus of the CD300f agonist group and the ZGF high-dose group were all increased(P<0.05),and synaptic damage was alleviated.The abnormal levels of glucose,lactate,ADP/ATP,5-HT,and CD300f protein expression were aggravated in the CD300f blocker group(P<0.05),and synaptic damage was aggravated.Conclusion ZGF can alleviate glucose metabolism disorders in hippocampal microglia and synaptic damage in hippocampal neurons in rats with diabetes-related depression.Its mechanism may be related to regulating the CD300f/GLUT1 signaling pathway.

Objective To explore the protective mechanism of Zuogui Jiangtang Jieyu Formula(ZGF)on synaptic damage of hippocampal neurons based on leukocyte mono-immunoglobulin-like receptor 3(CD300f)/glucose transporter 1(GLUT1)signal-mediated microglial glucose metabolism in rats with diabetes-related depression.Methods Eighty male SD rats were randomly selected using random number table method,with 10 rats serving as the normal group.The remaining 70 rats were fed a high-fat diet for 4 weeks and then injected once with 38 mg/kg of streptozotocin via the tail vein to replicate the diabetes rat model.Sixty rats were screened and successfully modeled,which were randomly divided into the model,CD300f blocker,CD300f agonist,metformin+fluoxetine(metformin 0.18 g/kg+fluoxetine 1.8 mg/kg),and ZGF high-and low-dose(20.52 and 10.26 g/kg,respectively)groups using random number table method.In addition to the normal group,the rats in the other groups underwent chronic unpredictable mild stress combined with isolation feeding for 28 days to replicate the diabetes-related depression rat model.The metformin+fluoxetine and ZGF high-and low-dose groups were subjected to continuous intragastrial administration for 14 days after the second week of modeling.The normal and model groups were administered an equal amount of distilled water by gavage.The CD300f blocker group and agonist group received microinjection into the hippocampus,with injection of myeloid cell trigger receptor inhibitory factor(CLM1,2 μg/kg)and immunoglobulin Fc surface protein(Fcγ,5 μg/kg)once a week,respectively.Depression-like behavior in rats was evaluated using open-field and forced swimming tests after the intervention.Biochemical analysis was used to detect the glucose,lactic acid,and adenosine diphosphate(ADP)/adenosine triphosphate(ATP)ratio contents.The insulin,5-hydroxytryptamine(5-HT),and dopamine(DA)levels in the hippocampus were detected using an enzyme-linked immunosorbent assay.Immunofluorescence was used to detect the average fluorescence intensity of CD300f,GLUT1,regulating synaptic membrane wxocytosis 3(RIMS3),and synapse-associated protein 102(SAP102)in hippocampal tissue.Western blotting was used to detect the CD300f,GLUT1,RIMS3,and SAP102 protein expression levels in the hippocampus.The synaptic damage of hippocampal neurons was observed using Nissl staining and transmission electron microscope.Results Compared with the normal group,the model group showed a decrease in the total active distance in the open-field test and an increase in forced swimming immobility time,with an increase in glucose and lactic acid contents and ADP/ATP ratio,whereas a decrease in insulin,5-HT,and DA levels was observed in the hippocampus.The average fluorescence intensity and relative protein expression levels of CD300f,GLUT1,RIMS3,and SAP102 in hippocampal tissue decreased(P<0.05),and the synaptic ultrastructure of hippocampal neurons was damaged.Compared with the model group,depression-like behavioral changes,glucose metabolism,and monoamine neurotransmitter imbalance were alleviated in the CD300f agonist group and ZGF high-and low-dose group(P<0.05).The average fluorescence intensity and relative protein expression levels of CD300f,GLUT1,RIMS3,and SAP102 in the hippocampus of the CD300f agonist group and the ZGF high-dose group were all increased(P<0.05),and synaptic damage was alleviated.The abnormal levels of glucose,lactate,ADP/ATP,5-HT,and CD300f protein expression were aggravated in the CD300f blocker group(P<0.05),and synaptic damage was aggravated.Conclusion ZGF can alleviate glucose metabolism disorders in hippocampal microglia and synaptic damage in hippocampal neurons in rats with diabetes-related depression.Its mechanism may be related to regulating the CD300f/GLUT1 signaling pathway.

Objective To explore the protective mechanism of Zuogui Jiangtang Jieyu Formula(ZGF)on synaptic damage of hippocampal neurons based on leukocyte mono-immunoglobulin-like receptor 3(CD300f)/glucose transporter 1(GLUT1)signal-mediated microglial glucose metabolism in rats with diabetes-related depression.Methods Eighty male SD rats were randomly selected using random number table method,with 10 rats serving as the normal group.The remaining 70 rats were fed a high-fat diet for 4 weeks and then injected once with 38 mg/kg of streptozotocin via the tail vein to replicate the diabetes rat model.Sixty rats were screened and successfully modeled,which were randomly divided into the model,CD300f blocker,CD300f agonist,metformin+fluoxetine(metformin 0.18 g/kg+fluoxetine 1.8 mg/kg),and ZGF high-and low-dose(20.52 and 10.26 g/kg,respectively)groups using random number table method.In addition to the normal group,the rats in the other groups underwent chronic unpredictable mild stress combined with isolation feeding for 28 days to replicate the diabetes-related depression rat model.The metformin+fluoxetine and ZGF high-and low-dose groups were subjected to continuous intragastrial administration for 14 days after the second week of modeling.The normal and model groups were administered an equal amount of distilled water by gavage.The CD300f blocker group and agonist group received microinjection into the hippocampus,with injection of myeloid cell trigger receptor inhibitory factor(CLM1,2 μg/kg)and immunoglobulin Fc surface protein(Fcγ,5 μg/kg)once a week,respectively.Depression-like behavior in rats was evaluated using open-field and forced swimming tests after the intervention.Biochemical analysis was used to detect the glucose,lactic acid,and adenosine diphosphate(ADP)/adenosine triphosphate(ATP)ratio contents.The insulin,5-hydroxytryptamine(5-HT),and dopamine(DA)levels in the hippocampus were detected using an enzyme-linked immunosorbent assay.Immunofluorescence was used to detect the average fluorescence intensity of CD300f,GLUT1,regulating synaptic membrane wxocytosis 3(RIMS3),and synapse-associated protein 102(SAP102)in hippocampal tissue.Western blotting was used to detect the CD300f,GLUT1,RIMS3,and SAP102 protein expression levels in the hippocampus.The synaptic damage of hippocampal neurons was observed using Nissl staining and transmission electron microscope.Results Compared with the normal group,the model group showed a decrease in the total active distance in the open-field test and an increase in forced swimming immobility time,with an increase in glucose and lactic acid contents and ADP/ATP ratio,whereas a decrease in insulin,5-HT,and DA levels was observed in the hippocampus.The average fluorescence intensity and relative protein expression levels of CD300f,GLUT1,RIMS3,and SAP102 in hippocampal tissue decreased(P<0.05),and the synaptic ultrastructure of hippocampal neurons was damaged.Compared with the model group,depression-like behavioral changes,glucose metabolism,and monoamine neurotransmitter imbalance were alleviated in the CD300f agonist group and ZGF high-and low-dose group(P<0.05).The average fluorescence intensity and relative protein expression levels of CD300f,GLUT1,RIMS3,and SAP102 in the hippocampus of the CD300f agonist group and the ZGF high-dose group were all increased(P<0.05),and synaptic damage was alleviated.The abnormal levels of glucose,lactate,ADP/ATP,5-HT,and CD300f protein expression were aggravated in the CD300f blocker group(P<0.05),and synaptic damage was aggravated.Conclusion ZGF can alleviate glucose metabolism disorders in hippocampal microglia and synaptic damage in hippocampal neurons in rats with diabetes-related depression.Its mechanism may be related to regulating the CD300f/GLUT1 signaling pathway.

Objective To explore the protective mechanism of Zuogui Jiangtang Jieyu Formula(ZGF)on synaptic damage of hippocampal neurons based on leukocyte mono-immunoglobulin-like receptor 3(CD300f)/glucose transporter 1(GLUT1)signal-mediated microglial glucose metabolism in rats with diabetes-related depression.Methods Eighty male SD rats were randomly selected using random number table method,with 10 rats serving as the normal group.The remaining 70 rats were fed a high-fat diet for 4 weeks and then injected once with 38 mg/kg of streptozotocin via the tail vein to replicate the diabetes rat model.Sixty rats were screened and successfully modeled,which were randomly divided into the model,CD300f blocker,CD300f agonist,metformin+fluoxetine(metformin 0.18 g/kg+fluoxetine 1.8 mg/kg),and ZGF high-and low-dose(20.52 and 10.26 g/kg,respectively)groups using random number table method.In addition to the normal group,the rats in the other groups underwent chronic unpredictable mild stress combined with isolation feeding for 28 days to replicate the diabetes-related depression rat model.The metformin+fluoxetine and ZGF high-and low-dose groups were subjected to continuous intragastrial administration for 14 days after the second week of modeling.The normal and model groups were administered an equal amount of distilled water by gavage.The CD300f blocker group and agonist group received microinjection into the hippocampus,with injection of myeloid cell trigger receptor inhibitory factor(CLM1,2 μg/kg)and immunoglobulin Fc surface protein(Fcγ,5 μg/kg)once a week,respectively.Depression-like behavior in rats was evaluated using open-field and forced swimming tests after the intervention.Biochemical analysis was used to detect the glucose,lactic acid,and adenosine diphosphate(ADP)/adenosine triphosphate(ATP)ratio contents.The insulin,5-hydroxytryptamine(5-HT),and dopamine(DA)levels in the hippocampus were detected using an enzyme-linked immunosorbent assay.Immunofluorescence was used to detect the average fluorescence intensity of CD300f,GLUT1,regulating synaptic membrane wxocytosis 3(RIMS3),and synapse-associated protein 102(SAP102)in hippocampal tissue.Western blotting was used to detect the CD300f,GLUT1,RIMS3,and SAP102 protein expression levels in the hippocampus.The synaptic damage of hippocampal neurons was observed using Nissl staining and transmission electron microscope.Results Compared with the normal group,the model group showed a decrease in the total active distance in the open-field test and an increase in forced swimming immobility time,with an increase in glucose and lactic acid contents and ADP/ATP ratio,whereas a decrease in insulin,5-HT,and DA levels was observed in the hippocampus.The average fluorescence intensity and relative protein expression levels of CD300f,GLUT1,RIMS3,and SAP102 in hippocampal tissue decreased(P<0.05),and the synaptic ultrastructure of hippocampal neurons was damaged.Compared with the model group,depression-like behavioral changes,glucose metabolism,and monoamine neurotransmitter imbalance were alleviated in the CD300f agonist group and ZGF high-and low-dose group(P<0.05).The average fluorescence intensity and relative protein expression levels of CD300f,GLUT1,RIMS3,and SAP102 in the hippocampus of the CD300f agonist group and the ZGF high-dose group were all increased(P<0.05),and synaptic damage was alleviated.The abnormal levels of glucose,lactate,ADP/ATP,5-HT,and CD300f protein expression were aggravated in the CD300f blocker group(P<0.05),and synaptic damage was aggravated.Conclusion ZGF can alleviate glucose metabolism disorders in hippocampal microglia and synaptic damage in hippocampal neurons in rats with diabetes-related depression.Its mechanism may be related to regulating the CD300f/GLUT1 signaling pathway.