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.

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 investigate the effect of Zuogui Jiangtang Jieyu Formula(左归降糖解郁方,ZJJF)on hippocampal neuron apoptosis in diabetic rats with depression and to ascertain whether its mechanism involves the regulation of JNK signaling pathway. Methods(i)A total of 72 specific pathogen-free(SPF)grade male Sprague Dawley(SD)rats were randomly divided into six groups,with 12 rats in each group:control,model,metformin(Met,0.18 g/kg)+fluoxetine(Flu,1.8 mg/kg),and the high-,medium-,and low-ZJJF dosages(ZJJF-H,20.52 g/kg;ZJJF-M,10.26 g/kg;ZJJF-L,5.13 g/kg)groups.All groups except control group were injected once via the tail vein with streptozotocin(STZ,38 mg/kg)combined with 28 d of chronic unpredictable mild stress(CUMS)to establish diabetic rat models with de-pression.During the CUMS modeling period,treatments were administered via gavage,with control and model groups receiving an equivalent volume of distilled water for 28 d.The effi-cacy of ZJJF in reducing blood sugar and alleviating depression was evaluated by measuring fasting blood glucose,insulin,and glycated hemoglobin levels,along with behavioral assess-ments,including the open field test(OFT),forced swim test(FST),and sucrose preference test(SPT).Hippocampal tissue damage and neuronal apoptosis were evaluated using hema-toxylin-eosin(HE)staining and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL)staining.Apoptosis-related proteins Bax,Bcl-2,caspase-3,and the ex-pression levels of JNK/Elk-1/c-fos signaling pathway were detected using Western blot and real-time quantitative polymerase chain reaction(RT-qPCR).(ii)To further elucidate the role of JNK signaling pathway in hippocampal neuronal apoptosis and the pharmacological ef-fects of ZJJF,an additional 50 SPF grade male SD rats were randomly divided into five groups,with 10 rats in each group:control,model,SP600125(SP6,a JNK antagonist,10 mg/kg),ZJJF(20.52 g/kg),and ZJJF(20.52 g/kg)+Anisomycin(Aniso,a JNK agonist,15 mg/kg)groups.Ex-cept for control group,all groups were established as diabetic rat models with depression,and treatments were administered via gavage for ZJJF and intraperitoneal injection for SP6 and Aniso for 28 d during the CUMS modeling period.Behavioral changes in rats were evaluated through the OFT,FST,and SPT,and hippocampal neuron damage and apoptosis were ob-served using HE staining,Nissl staining,TUNEL staining,and transmission electron mi-croscopy(TEM).Changes in apoptosis-related proteins and JNK signaling pathway in the hippocampal tissues of rats were also analyzed. Results(i)ZJJF significantly reduced the high blood glucose,insulin,and glycated he-moglobin levels in model rats(P<0.01).It increased autonomous activity and decreased de-spair-like behaviors(P<0.01),improved the pathological damage of hippocampal neurons,increased the number of neuronal nuclei(P<0.01),and reduced the number of mechanocytes,vacuolar cells,and apoptotic neurons(P<0.05,P<0.01,and P<0.01,respec-tively).ZJJF down-regulated the expression levels of pro-apoptotic proteins Bax and caspase-3(P<0.01),up-regulated the anti-apoptotic protein Bcl-2(P<0.01),and significantly inhibit-ed the overexpression of phosphorylated JNK(p-JNK),Elk-1,and c-fos(P<0.01).(ii)SP6 in-creased autonomous activity and reduced despair time in model rats(P<0.05),although it had no significant effects on sucrose preference(P>0.05).It increased the number of Nissl bodies in hippocampal neurons(P<0.01),reduced the protein expression levels of Bax(P<0.01)and caspase-3(P<0.05),and decreased the number of apoptotic neurons(P<0.05).SP6 also increased the expression level of Bcl-2(P<0.01),and inhibited the high expression levels of p-JNK,Elk-1,and c-fos(P<0.01,P<0.01,and P<0.05,respectively),suggesting that hip-pocampal neuronal apoptosis in diabetic rats with depression is associated with abnormal ac-tivation of JNK signaling pathway.Compared with ZJJF group,ZJJF+Aniso group showed a decrease in sucrose preference(P<0.05)and an increase in despair time(P<0.01)with more notable hippocampal neuronal damage.This group also exhibited a decrease in expression level(P<0.01)Bcl-2 and an increase in expression levels of Bax,caspase-3,p-JNK,Elk-1,and c-fos(P<0.01,P<0.05,P<0.05,P<0.01,and P<0.05,respectively),indicating that the antidepressant effects of ZJJF,its improvement of neuronal apoptosis,and regulation of JNK signaling molecules could all be reversed by a specific JNK agonist. Conclusion ZJJF exerts a significant hypoglycemic effect and ameliorates the apoptosis of hippocampal neurons by inhibiting the activation of JNK signaling pathway,which is a promising formula for the treatment of diabetic depression in clinical settings.

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.