Modified Xiaoyaosan Alleviates Depression-like Behaviors by Regulating Activation of Hippocampal Microglia Cells in Rat Model of Juvenile Depression / 中国实验方剂学杂志

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.

Overexpression of NRF1 alleviates mitochondrial and cognitive dysfunction in mice models of Alzheimer's disease / 安徽医科大学学报

Objective To investigate the effects of nuclear respiratory factor 1(NRF1)on mitochondrial and cog-nitive dysfunction in Alzheimer's disease(AD)model mice.Methods The 5 × FAD mice were utilized as a mod-el for Alzheimer's disease,and the sparsely labeled AAV virus overexpressing NRF1(AAV-NRF1)was adminis-tered via stereotaxic injection into the brain.The expression of NRF1 in hippocampus was determined by Western blot,the morphology of mitochondria in hippocampus was observed by transmission electron microscope,the den-dritic spines of sparsely labeled neurons in the CA1 region were visualized and quantified using confocal laser mi-croscopy,cognitive and memory functions of mice were evaluated using the Morris water maze test,while electro-physiological methods were employed to detect long-term potentiation(LTP)of synaptic efficacy.Results The ex-pression of NRF1 in the hippocampus was significantly upregulated following stereotactic injection of AAV-NRF1(P＜0.001).This intervention led to notable improvements in mitochondrial morphology within hippocampal neurons,as well as enhanced cognitive and memory functions in mice(P＜0.01).Moreover,there was a significant in-crease in dendritic spine density among neurons located in the CA1 region of the hippocampus(P＜0.001),ac-companied by long-lasting and stable long-term potentiation(LTP)and a substantial elevation in fEPSP slope(P＜0.01).Conclusion The overexpression of NRF1 in a 5 × FAD mouse model of Alzheimer's disease(AD)initia-ted the restoration of mitochondrial dysfunction and enhanced synaptic plasticity,indicating that these alterations may contribute to the therapeutic efficacy of NRF1 overexpression in ameliorating cognitive dysfunction associated with AD.

Effects of five-element acupuncture on clinical symptoms, brain metabolism and immunoglobulin levels in patients with chronic fatigue syndrome / 国际中医中药杂志

Objective:To evaluate the effect of five-element acupuncture on clinical symptoms, brain metabolism and immunoglobulin level in patients with chronic fatigue syndrome.Methods:It was a randomized controlled trial. A total of 94 patients with chronic fatigue syndrome in our hospital from January 2021 to January 2022 were selected and divided into two groups according to the random number table method, with 47 in each group. The conventional western medicine group was treated with conventional western medicine, and the five-element acupuncture group was treated with five-element acupuncture on the basis of the conventional western medicine group. Both groups were treated for 4 weeks. Before and after treatment, the serum levels of interferon-γ (IFN-γ), corticosterone (CORT), IL-2 and 5-hydroxytryptamine (5-HT) were detected by ELISA; the levels of natural killer (NK) cells, CD4 +, CD8 +, IgG and IgM were detected by flow cytometry; the whole body superconducting MRI scanner was used to scan T2 Flair, T2WI and TlWI sequences of the hippocampus, and the spectral curves and the areas under the peak of N-acetylaspartic acid (NAA), creatine (Cr) and choline (Cho) were obtained, and the ratios of Cho/Cr and NAA/Cr were calculated. the fatigue Scale-14 (FS-14) and Fatigue Severity Scale (FSS) were used to evaluate the fatigue state of the patients, and the Depression-Anxiety-Stress Scale-21 (DASS-21) and Beck Anxiety Inventory (BAI) were used to evaluate the anxiety state of the patients. Symptom Checklist 90 (SCL-90) and Somatic and Mental Health Report Score (SPHERE) were used to evaluate the quality of life of patients. And the clinical efficacy was evaluated. Results:After treatment, the levels of IgG, CD4 + and NK in the five-element acupuncture group were significantly higher than those in the conventional western medicine group ( t values were 4.76, 3.65, 6.42, respectively, P<0.01), and the level of IgM, CD8 + was significantly lower than that in the conventional western medicine group ( t values were 7.30, 4.79, P<0.01); the levels of IFN-γ, IL-2 and 5-HT in the observation group were significantly higher than those in the conventional western medicine group ( t values were 7.60, 4.05, 2.79, respectively, respectively, P<0.01), and the level of CORT was significantly lower than that in the conventional western medicine group ( t=6.72, P<0.01); the NAA/Cr levels in the left [(1.10±0.04) vs. (1.05±0.03), t=6.86] and right [(1.18±0.02) vs. (1.21±0.03), t=8.23] hippocampus of the experimental group were significantly higher than those in the conventional western medicine group ( P<0.01), and the Cho/Cr levels in the left [(1.08±0.04) vs. (1.03±0.03), t=5.70] and right [(1.17±0.02) vs. (1.20±0.03), t=5.71] hippocampus of the experimental group were significantly lower than those of the conventional western medicine group ( P<0.01). After treatment, the scores of physical fatigue, mental fatigue and FSS in the five-element acupuncture group were significantly lower than those in the conventional western medicine group ( t values were 8.08, 9.08 and 7.07, respectively, P<0.01). The scores of DASS-21, BAI, SCL-90 and SPHERE in the conventional western medicine group were significantly lower than those in the conventional western medicine group ( t values were 3.63, 5.77, 8.74, 5.92, respectively, P<0.01).The total effective rate was 95.74% (45/47) in the five-element acupuncture group and 82.98% (39/47) in the conventional western medicine group, and there was no significant difference between the two groups ( χ2=2.80, P=0.094). Conclusion:Five-elements acupuncture can improve the expression of T lymphocytes, increase the levels of immunoglobulin and NK, reduce the level of CORT, regulate the brain metabolism of NAA in the left and right hippocampus, improve the clinical symptoms and negative emotions, and improve the clinical efficacy and quality of life in patients with chronic fatigue syndrome.

The effects of minocycline on neuroinflammation in rats with post-traumatic stress disorder / 神经解剖学杂志

Objective:To investigate the effect of minocycline on neuroinflammation of rats with post-traumatic stress disorder(PTSD).Methods:The rat model of PTSD was prepared by a single prolonged stress(SPS)method,and the rats were treated with minocycline(PTSD+Mino group)or normal saline(PTSD group)by gavage.The behavioral changes of rats were detected by light-dark box test.The expression of ionized calcium-binding adapter molecule 1(Iba-1)in hippocampus was detected by immunohistochemical staining.The contents of IL-1β and TNF-α in hippocampus were detected by ELISA,and the expression levels of IL-1β and TNF-α mRNA in hippocampus were detected by real-time RT-PCR(qRT-PCR).Results:After 3 days of SPS stimulation,the anxiety-like behavior of rats was obvious,the expression of Iba-1 in hippocampus was increased,and the contents of IL-1β and TNF-α in hippocampus were in-creased.Minocycline treatment significantly reduced anxiety-like behavior and decreased the expression of Iba-1 in the hippocampus of PTSD rats.Meanwhile,minocycline treatment also decreased the levels of IL-1β and TNF-α mRNA and protein in the hippocampus.Conclusion:Minocycline can improve the anxiety-like behavior of PTSD rats by inhibiting the activation of microglia.

Shuxuening injection protects rats from cerebral ischemia through the Wnt/β-catenin signaling pathway / 神经解剖学杂志

Objective:To study the effects of Shuxuening injection(SXN)on Wnt/β-catenin signaling pathway in hippocampus of rats with cerebral ischemia.Methods:SD rats were divided into three groups:Sham operation group(Sham),middle cerebral artery occlusion group(MCAO)and MCAO+SXN treatment group(MCAO+SXN).The model of cerebral ischemia in rats was prepared by MCAO.The rats with cerebral ischemia were treated with SXN by caudal vein injection.Zea-Longa scoring criteria and balance beam test were employed to evaluated neurological function of rats.HE staining were used to observe the changes of inflammatory cells infiltration the hippocampal CAI region.The expression of β-catenin in hippocampal CA1 region was observed by immunofluorescence staining.The mRNA and pro-tein expressions of caspase-3,cyclooxygenase-2(COX-2)and endothelial nitric oxide synthetase(eNOS)in hippocam-pus were detected by real time RT-PCR and Western Blot,respectively.Results:SXN can SXN can improve the neuro-logical dysfunction of cerebral ischemia rats.The inflammatory cells infiltration in hippocampal CAI region was decreased,and the expression of β-catenin,caspase-3 and COX-2 was decreased,while the expression of eNOS was upregulated.Conclusion:SXN protects against cerebral ischemia by inhibiting Wnt/β-catenin signaling pathway against inflammation response,oxidative stress and apoptosis of nerve cells in rats.

To Explore the Effect and Mechanism of Shenghui Granule on EC and CA1 Regions of AD Model Rats Based on p38 MAPK Signal Pathway / 世界科学技术-中医药现代化

Objective To observe the effect of Shenghui Granule on EC and CA1 regions of scopolamine-induced dementia rats and explore its mechanism based on p38 MAPK signal pathway.Methods 40 SD rats were randomly divided into four groups(blank group,model group,Shenghui granule group,donepezil group),and were treated with scopolamine.Morris water maze and open field test were used to evaluate the cognition and anxiety behavior of rats.The nerve injury of EC and CA1 was observed by HE staining.The activity of neurons in EC and CA1 regions was observed by c-Fos immunofluorescence staining.Western blot was used to detect p38 MAPK pathway related proteins.Results The behavioral experiment found that Shenghui Granule could improve the cognitive impairment and anxiety-like behavior of AD model rats.The results of HE staining showed that Shenghui granules had protective effects on EC and CA1 regions.The results of c-Fos immunofluorescence staining showed that Shenghui granules could increase the activity of neurons in EC and CA1 regions.Western blot results showed that Shenghui Granule could down-regulate the expression of Bax,reduce the levels of phosphorylated p38 and Tau,and increase the expression of Bcl-2.Conclusion Shenghui granule has protective effect on EC and CA1 regions of AD model rats,and may play a therapeutic role through p38 MAPK signal pathway.

circ_HIPK3 regulates function and morphology of Aβ induced hippocampal neurons by targeting miR-381-3p/ZNF217 axis / 天津医药

Objective To analyze the influence of cyclic RNA homologous domain interacting protein kinase 3(circ_HIPK3)on function and morphology of myloid β-protein(Aβ)induced hippocampal neurons by targeting miR-381-3p/zinc finger protein 217(ZNF217)axis.Methods Hippocampal neurons of neonatal rats were prepared and divided into the control group,the Aβ group,the si NC1 group,the si HIPK3 group,the si HIPK3+inhibitor NC group,the si HIPK3+miR-381-3p inhibitor group,the si HIPK3+miR-381-3p inhibitor+si NC2 group and the si HIPK3+miR-381-3p inhibitor+si ZNF217 group.Except the control group,all the other groups were modeled by 40 μmol/L Aβ1～42.qRT-PCR was used to determine the circ of hippocampal neurons circ_HIPK3,miR-381-3p and ZNF217 mRNA levels.Cell morphology was observed by transmission electron microscope,and the survival rate of hippocampal neurons was measured by CCK-8 method.Hochesst 33342 method was used to measure apoptosis of hippocampal neurons.The intracellular Ca2+ fluorescence intensity of hippocampal neurons was detected by flow cytometry.The expression levels of P-Tau,B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),Caspase-3 and ZNF217 proteins in hippocampal neurons were measured by Western blot assay.Double luciferase reporter genes were used to analyze the targeting relationship between miR-381-3p and circ_HIPK3,ZNF217.Results In the control group,the structure of hippocampal neurons was normal,the morphology of nucleus was normal,and there were no pathological changes in mitochondria and endoplasmic reticulum.In the Aβ group,hippocampal neurons showed degenerative changes,abnormal nuclear morphology,membrane invagination,a large number of mitochondria swelling and a large number of lipid droplets vacuoles in cytoplasm.Compared with the Aβ group,the hippocampal neuronal structure was partially restored in the si HIPK3 group.Compared with the si HIPK3 group,the hippocampal neuronal structure was severely damaged in the si HIPK3+miR-381-3p inhibitor group.Compared with the si HIPK3+miR-381-3p inhibitor group,the damage of hippocampal neurons in the si HIPK3+miR-381-3p inhibitor+si ZNF217 group was reduced.Compared with the control group,the circ_HIPK3,ZNF217 mRNA and ZNF217 protein levels,apoptosis rate,Ca2+ fluorescence intensity,P-Tau,Bax,Caspase-3 protein expression of hippocampal neurons were increased in the Aβ group,and the miR-381-3p level,survival rate and Bcl-2 protein expression decreased(P＜0.05).Compared with the Aβ group,the circ_HIPK3,ZNF217 mRNA and ZNF217 protein levels,apoptosis rate,Ca2+ fluorescence intensity,P-Tau,Bax and Caspase-3 protein expression of hippocampal neurons were decreased in the si HIPK3 group,and miR-381-3p level,survival rate and Bcl-2 protein expression increased(P＜0.05).Compared with the si HIPK3 group,the circ_HIPK3,ZNF217 mRNA and ZNF217 protein levels,apoptosis rate,Ca2+ fluorescence intensity,P-Tau,Bax and Caspase-3 protein expression of hippocampal neurons in the si HIPK3+miR-381-3p inhibitor group were increased,and the miR-381-3p level,survival rate and Bcl-2 protein expression decreased(P＜0.05).Compared with the si HIPK3+miR-381-3p inhibitor group,the ZNF217 mRNA and ZNF217 protein levels,apoptosis rate,Ca2+ fluorescence intensity,P-Tau,Bax and Caspase-3 protein expression of hippocampal neurons in the si HIPK3+miR-381-3p inhibitor+si ZNF217 group were decreased,and the survival rate and Bcl-2 protein expression increased(P＜0.05).miR-381-3p targeted and combined with HIPK3 and ZNF217.Conclusion circ_HIPK3 silencing may ameliorate Aβ-induced damage of hippocampal neuronal structure and function by regulating miR-381-3p/ZNF217 axis.

Effects of Electroacupuncture on the Hippocampus Glutamate System of Acute Myocardial Ischemia Rats / 中国中医药信息杂志

Objective To observe the effects of electroacupuncture(EA)on glutamate(Glu),metabotropic glutamate receptor 2/3(mGluR2/3)and apoptosis related proteins expression in hippocampus in rats with acute myocardial ischemia(AMI);To explore the mechanism of EA against AMI.Methods Totally 50 SD rats were randomly divided into sham-operation group,model group,EA group and inhibitor group,with 10 rats in each group.Except for the sham-operation group,the rats were treated with ligation at the left anterior descending coronary artery to establish AMI model.The rats in the EA group was treated with EA at"Shenmen"and"Tongli",30 minutes each time,once a day for 3 consecutive days.The rats in the inhibitor group were treated with injection of LY341459 via the lateral ventricle 30 min after modeling.HE staining was used to observe myocardial tissue morphology,and ELISA was used to detect Caspase-3 activity in myocardial tissue and Glu content in hippocampal tissue,immunofluorescence staining was used to detect mGluR2/3 expression in hippocampal tissue,TUNEL staining was used to detect apoptosis in hippocampal tissue cells,Western blot was used to detect the expressions of PI3K,Akt,and Caspase-3 protein in hippocampal tissue.Results Compared with the sham-operation group,the myocardial cells of the model group rats showed sparse and swelling with severe infiltration of inflammatory cells;the activity of Caspase-3 in myocardial tissue significantly increased,and the Glu content,positive expression of mGluR2/3,number of apoptotic cells in hippocampal tissue significantly increased(P<0.01),and the expressions of PI3K and Akt proteins in hippocampal tissue were significantly decreased,while the expression of Caspase-3 protein significantly increased(P<0.01).Compared with the model group,myocardial cell edema and inflammatory cell infiltration were reduced in the EA group and inhibitor group,the activity of Caspase-3 in myocardial tissue was significantly decreased,the Glu content,positive expression of mGluR2/3,and number of apoptotic cells in hippocampal tissue were significantly reduced(P<0.01),the expressions of PI3K and Akt proteins in hippocampal tissue significantly increased,while the expression of Caspase-3 protein significantly decreased(P<0.01).Conclusion EA can improve myocardial injury in AMI rats,and its mechanism may be related to activation of PI3K/Akt signaling pathway,inhibition of hippocampal mGluR2/3 overexpression,reduction of Glu accumulation,inhibition of apoptosis of hippocampal neurons and reduction of neurotoxicity.

Effects of Regulation of SHH/Gli1 Signaling Pathway by Baishile Capsules on Hippocampal Neurogenesis Depression Model Rats / 中国中医药信息杂志

Objective To explore the effects and mechanism of Baishile Capsules regulating SHH/Gli1 signaling pathway on hippocampal neurogenesis of depression model rats.Methods Totally 32 SD rats were randomly divided into control group,model group,fluoxetine(5.4 mg/kg)group and Baishile Capsules(2.88 g/kg)group,with 8 rats in each group.A depression rat model was established using chronic unpredictable mild stress and single cage feeding method.The model was established and administered simultaneously for 21 consecutive days.Depression-like behavior in rats were evaluated by sucrose preference experiment and open field experiment,ELISA was used to detect brain derived neurotrophic factor(BDNF)contents in rat serum and hippocampal tissue,the number of BrdU,BrdU/DCX,BrdU/NeuN positive cells in dentate gyrus of the hippocampus was observed by immunofluorescence,immunofluorescence and Western blot were used to detect the fluorescence intensity and protein expression of SHH,Gli1,Smo,Ptch in hippocampal tissue.Results Compared with the control group,the degree of sucrose preference significantly decreased in the model group(P<0.01),the number of horizontal and vertical movements significantly decreased(P<0.01),the contents of BDNF in serum and hippocampal tissue significantly decreased(P<0.05),the number of BrdU,BrdU/DCX,BrdU/NeuN positive cells in dentate gyrus of the hippocampus significantly decreased(P<0.01),and the fluorescence intensity and protein expression of SHH,Gli1,Smo,Ptch in hippocampal tissue significantly decreased(P<0.01,P<0.05).Compared with the model group,the degree of sucrose preference and the number of horizontal and vertical movements in fluoxetine group and Baishile Capsule group increased significantly(P<0.05,P<0.01),the contents of BDNF in serum and hippocampal tissue significantly increased(P<0.05,P<0.01),and the number of BrdU,BrdU/DCX,BrdU/NeuN positive cells in dentate gyrus of the hippocampus significantly increased(P<0.01,P<0.05),the fluorescence intensity and protein expressions of SHH,Gli1,Smo,Ptch in hippocampal tissue significantly increased(P<0.01,P<0.05).Conclusion Baishile Capsule can promote the hippocampus neurogenesis in depression model rats by regulating SHH/Gli1 signaling pathway,and play an antidepressant role.

Application of optimized hippocampus-avoidance prophylactic cranial irradiation in limited-stage small cell lung cancer / 中华放射肿瘤学杂志

Objective:To analyze the treatment efficacy, safety and dose parameters of optimized hippocampus-avoidance prophylactic cranial irradiation (HA-PCI) in limited-stage small cell lung cancer (LS-SCLC) and explore the corresponding dosimetric parameters under the condition of narrowing the hippocampus avoidance region as hippocampus region plus 2 mm in three dimensions.Methods:Clinical data of patients with LS-SCLC receiving HA-PCI (hippocampus avoidance region defined as hippocampus region plus 2 mm in three dimensions) in Cancer Hospital Chinese Academy of Medical Sciences from August 2014 to June 2020 were retrospectively analyzed. Dose parameters of HA-PCI and adverse events were analyzed using descriptive statistics analysis. Changes of neurocognitive function, such as mini-mental state examination (MMSE) and Hopkins verbal learning test-revised (HVLT-R) scores, were evaluated by analysis of variance and Kruskal-Wallis H test. Overall survival (OS), progression-free survival (PFS) and intracranial PFS (iPFS) were calculated using Kaplan-Meier method. The cumulative incidence of local-regional recurrence (LRR), extracranial distant metastases (EDM), and locoregional recurrence (LR) were investigated under competing risk analysis. Results:A total of 112 patients were included, the median follow-up time was 50 months (95% CI: 45.61-54.38). The median volume of hippocampus was 4.85 ml (range: 2.65-8.34 ml), with the average dose ≤9 Gy in 106 patients (94.6%), ≤8 Gy in 92 patients (82.1%). The median volume of hippocampus avoidance area was 15.00 ml (range: 8.61-28.06 ml), with the average dose ≤12 Gy in 109 patients (97.3%), ≤10 Gy in 101 patients (90.2%). The 2-year cumulative LRR, EDM, LR rates were 16.9%, 23.2% and 28.5%, respectively. The 5-year cumulative LRR, EDM, LR rates were 23.2%, 26.9% and 33.3%, respectively. The 2-year iPFS, PFS and OS rates were 66.1% (95% CI: 57.9%-75.4%), 53.6% (95% CI: 45.1%-63.7%) and 80.4% (95% CI: 73.3%-88.1%), respectively. The most common grade I-Ⅱ adverse events were nausea (33.9%) and dizziness (31.3%), and only 1 patient developed grade Ⅲ nausea and dizziness. MMSE ( n=57) and HVLT-R tests ( n=56) showed no significant decline. Conclusions:Optimized HA-PCI can achieve similar dose limitation with favorable efficacy and light toxicity. No significant decline is observed in short-term neurocognitive function in evaluable patients.

Effects of Ziziphi Spinosae Semen-Albiziae Flos on the IRE1α/ASK1/JNK Pathway of Endoplasmic Reticulum Stress in Hippocampus of Depression Model Rats / 中药新药与临床药理

Objective Based on the endoplasmic reticulum stress(ERS)inositol-requiring enzyme-1α(IRE1α)/apoptosis signal-regulated kinase 1(ASK1)/c-Jun N-terminal kinase(JNK)pathway to investigate the intervention effect of Ziziphi Spinosae Semen(ZSS)-Albiziae Flos(AF)on the depression model of rats,which were prepared by solitary rearing combined with chronic unpredictable mild stress(CUMS).Methods A total of 144 rats were randomly divided into normal group,model group,high-,medium-and low-dose groups of ZSS-AF,and Venlafaxine group.In addition to the normal group,the rats in other groups were isolated for 21 days combined with CUMS to prepare the depression model.The behavioral changes of rats were observed by open field test and Morris water maze test.The ultrastructural changes of hippocampal neurons were observed by transmission electron microscope.TUNEL staining was used to observe the apoptosis of nerve cells in hippocampus.The protein expression levels of IRE1α,phosphorylated(P)-IRE1α,ASK1,P-ASK1,JNK,P-JNK,B cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax)and cysteme aspartate specific protease-3(Caspase-3)in hippocampus were detected by Western Blot.The mRNA expression levels of IRE1α,ASK1,JNK,Bax,Bcl-2 and Caspase-3 in hippocampus were detected by Real-Time PCR.Results Compared with the normal group,the scores of horizontal movement and vertical movement in the open field test of rats in the model group were decreased(P＜0.01).In the water maze test,the escape latency was increased and the number of crossing the original platform was decreased(P＜0.01).The number of hippocampal apoptosis was increased(P＜0.01).The protein expression levels of P-IRE1 α/IRE1 α,P-ASK1/ASK1,P-JNK/JNK,Bax,Caspase-3 and mRNA expressions of IRE1α,ASK1,JNK,Bax,Caspase-3 in hippocampus were increased,while the protein and mRNA expression levels of Bcl-2 were decreased(P＜0.01).Compared with model group,the scores of horizontal movement and vertical movement in the open field test of rats in each dose ZSS-AF groups and Venlafaxine group were increased(P＜0.01).In the water maze test,the escape latency was decreased and the times of crossing the original platform was increased(P＜0.01).The number of hippocampal apoptosis was decreased(P＜0.01).The mRNA expression levels of P-IRE1α/IRE1α,P-ASK1/ASK1,P-JNK/JNK,Bax,Caspase-3 protein and IRE1α,ASK1,JNK,Bax,Caspase-3 in hippocampus were decreased,while the protein and mRNA expression levels of Bcl-2 were increased(P＜0.05,P＜0.01).The effect of medium-dose ZSS-AF group was better than that of high-and low-dose groups.Conclusion ZSS-AF may play an antidepressant role by regulating IRE1α/ASK1/JNK pathway of endoplasmic reticulum stress.

Expression profile analysis of long non-coding RNAs in hippocampus of alcohol-dependent mice induced by double-bottle selective drinking / 中国生物制品学杂志

@#Objective To analyze the expression profiles of long non-coding RNAs(lncRNA)in hippocampus of alcoholdependent mice induced by double-bottle selective drinking.Methods The alcohol-dependent mouse model was established by double-bottle selective drinking method,and the control group was set up(drinking water). Three male mice with alcohol preference more than 60% and alcohol consumption more than 10 g/(kg·24 h)in alcohol group and random three male mice in control group were selected,of which bilateral hippocampal brain tissues were isolated and stored in liquid nitrogen. LncRNA and mRNA of mouse hippocampal brain tissue RNA samples were sequenced by using Agilent-084388 microarray,and the differential expression of lncRNA in samples was detected by using ncRNA microarray. The biological processes and signaling pathways involved in differential expression of lncRNA were clustered and enriched by Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis. Pearson correlation analysis was used to predict the coding genes co-expressed by each differentially expressed lncRNA. Hypergeometric distribution test was used to calculate the significance of differential gene enrichment in each corresponding transcription factor item,and Cytoscape software was used to draw a visual network diagram.Results Compared with the control group,totally 855 lncRNAs(FC ≥ 2. 0,P < 0. 05)were differentially expressed in the hippocampus of mice in alcohol group,of which 337 lncRNAs were up-regulated significantly,with NONMMUT025786.2 and NONMMUT072246.2 being the most up-regulated,and 518 significant downward adjustments were observed,with the largest downward adjustments being NONMMUT113098.1 and NONMMUT076455.1. There were 361 mRNAs differentially expressed in the two groups(FC ≥ 2. 0,P < 0. 05)with 271 mRNAs up-regulated significantly and 90 significant downward adjustments,among which,the most obvious up-regulated were Upf3b and Zfp943,and Adamts 13 and Ift 27 showed the largest downward adjustments. The differential expression of lncRNAs was most obvious in the positive regulation of cell surface,GTPase activity and cell vesicle transport;The main signaling pathways involved were propanoate metabolism,taurine metabolism,extracellular matrix receptor interaction and AMPK signaling pathway. The most abundant transcription factors were FOXL1 and LHX3,with 25 and 21 corresponding co-expressed genes,respectively.Conclusion Through high-throughput gene expression profile microarray analysis,the possible key regulatory sites of lncRNAs and mRNAs were obtained,which provided experimental basis for research of the molecular mechanism of alcohol dependence in the hippocampus.

Intervention effect of HSP60 on learning and memory impairment induced by combined exposure to lead and hypertension in mice / 中国职业医学

ObjectiveTo investigate the intervention effect of heat shock protein 60 (HSP60) on learning and memory impairment induced by combined exposure to lead and hypertension in mice, and the relative mechanism of triggering receptor expressed on myeloid cells 2 (TREM2). Methods Specific pathogen-free C57BL/6J male mice were randomly divided into control group, hypertension group, lead-exposed group and lead-exposed + hypertension group, or into control group, heat shock protein 60 (HSP60) control group, lead-exposed + hypertension group and HSP60 intervention group, with 10 mice in each group. Mice of hypertension group and lead-exposed + hypertension group were intraperitoneally injected with angiotensin Ⅱ at a dose of 0.5 mg/(kg·d) for seven consecutive days to induce hypertension model. Mice of the lead-exposed group, lead-exposed + hypertension group, and HSP60 intervention group were given lead acetate drinking water with a mass concentration of 250.0 mg/L, while mice in the control group, hypertension group, and HSP60 control group were given purified water for 12 weeks. Mice of the HSP60 control group and HSP60 intervention group were intraperitoneally injected with a solution of HSP60 at a dose of 4 mg/kg body weight, every other day for a total of three times at the 12th week. The learning and memory ability of mice was detected using the Morris water maze test. The enzyme-linked immunosorbent assay was used to detect the levels of interleukin (IL)-1β, IL-6 and tumor necrosis factor-α (TNF-α) in the hippocampal tissues of the mice. The relative expression of ionized calcium binding adaptor molecule-1 (IBA1) and TREM2 protein in the hippocampus of mice was detected using Western blot. Results i) The number of platform crossings of the mice in the hypertension group and the lead-exposed group was lower than that in the control group (both P<0.05). The escape latency of the mice on the third day was longer and the number of platform crossings was lower in the lead-exposed + hypertension group compared with the control group, hypertension group and lead-exposed group (all P<0.05). The levels of IL-1β, IL-6, and TNF-α in the hippocampus of the other three groups increased compared with the control group (all P<0.05). The relative expression of IBA1 protein in the hippocampus of lead-exposed group and lead-exposed + hypertension group increased (all P<0.05), while the relative protein expression of TREM2 decreased compared with the control group (all P<0.05). The levels of IL-1β, IL-6, TNF-α, and the relative protein expression of IBA1 protein in the hippocampus of the lead-exposed+hypertension group were higher (all P<0.05), and relative expression of TREM2 protein was lower (P<0.05) than those in the hypertension group. The level of TNF-α and the relative expression of IBA1 protein in the hippocampus of lead-exposed+hypertension group were higher than those in lead-exposed group (all P<0.05). ii) The escape latency of mice in the lead-exposed + hypertension group was longer than that in the control group (P<0.05), and the number of platform crossings was fewer than that in the control group (P<0.05). The escape latency of mice in the HSP60 intervention group was shortened (P<0.05), the number of platform crossings increased (P<0.05), and the levels of IL-1β, IL-6, TNF-α and relative expression of IBA1 protein decreased in the hippocampus (all P<0.05), while the relative expression of TREM2 protein increased (P<0.05) compared with the lead-exposed+hypertension group. Conclusion Combined exposure of lead and hypertension has a synergistic effect on learning and memory impairment in mice. The mechanism may be related to the inhibition of TREM2 expression by lead in the hippocampus of hypertensive mice and aggravating the neuroinflammatory response. Intervention with TREM2 receptor agonist HSP60 can alleviate learning and memory impairment in mice exposed to lead and hypertension by up-regulating TREM2 expression in the hippocampus.

The Mechanism of miR-124 in Depression / 生物化学与生物物理进展

Depression is a prevalent mental illness worldwide, its multifaceted pathogenesis is still in the exploratory stage. MicroRNA (miRNA), as a crucial epigenetic regulator, plays an important role in depression. miR-124 is one of the most abundant miRNAs in the central nervous system including neurons and microglia, and involved in various biological events like neuron development and differentiation, synaptic and axonal growth, neural plasticity, inflammation and autophagy. Recent studies have reported abnormal expression of miR-124 in both depression patients and animal models. Most of the studies showed that miR-124 is upregulated in the hippocampus or prefrontal cortex in stress-induced rodent depression animal models such as CUMS, CSDS, CORT, CRS and LH but some evidence for divergence. Upregulation of miR-124 expression may be involved in depression-like behavior via CREB/BDNF/TrkB pathway, GR pathway, SIRT1 pathway, apoptosis and autophagy pathways by directly targeting these genes including Creb, Bdnf, Sirt1, Nr3c1, Ezh2 and Stat3. The downregulation of miR-124 expression in neurons is mainly involved in the neurogenesis and neuroplasticity impairments in depression by targeting the Notch signaling pathway and DDIT4/TSC1/2/mTORC1 pathway. The downregulation of miR-124 expression also was found in the activated microglia in the stress-induced models, and resulted in neuroinflammation. In summary, the abnormal expression of miR-124 in the brain of depression-related models and its related mechanisms are complex and even contradictory, and still need further research. This review provides a summary of the research progress of miR-124 in depression.

Effect and Mechanism of Epigallocatechin-3-gallate on Learning and Memory Abilities of Epileptic Rats / 中国实验方剂学杂志

ObjectiveTo investigate the effects of epigallocatechin-3-gallate （EGCG） on learning and memory abilities of amygdala electrical kindling-induced epilepsy in rats and its mechanism. MethodMale SD rats were randomly divided into the normal group， model group， intervention group （model+25 mg·kg-1 EGCG）， and EGCG group （25 mg·kg-1 EGCG）. Rats in the EGCG group were only given EGCG intraperitoneal injection， those in the normal group were only given electrode implantation， and those in the other experimental groups were given amygdala electrical kindling stimulation to establish a chronic kindling epilepsy model. EGCG was injected intraperitoneally daily before electrical stimulation. Twenty-four hours after the last electrical stimulation， the escape latency and percentage of target quadrant were recorded by the Morris water maze. Twenty-four hours after the behavioral test， rats in each group were sacrificed by decapitation. The number of hippocampal neurons was observed by Nissl staining. The thickness of postsynaptic density in the hippocampus， synaptic cleft， length of active zone and the curvature of synaptic interface were observed by transmission electron microscopy （TEM）. The expressions of synapse-related proteins synaptotagmin （Syt）， postsynaptic density-95 （PSD-95） and Kalirin-7 in the hippocampus were examined by Western blot. ResultCompared with those in the normal group， the escape latency was significantly prolonged （P<0.05， P<0.01） and the target quadrant ratio was significantly decreased in the model group （P<0.05）. The number of hippocampus neurons decreased significantly （P<0.01）. The synaptic cleft of the hippocampus was widened significantly， and the length of active zone and the thickness of postsynaptic density were significantly decreased （P<0.05， P<0.01）. The expressions of synapse-related proteins Syt， PSD-95 and Kalirin-7 in the hippocampus were significantly decreased （P<0.05，P<0.01）. Compared with those in the model group， the escape latency was significantly shortened and the percentage of target quadrant was significantly increased in the intervention group （P<0.05， P<0，01）. The number of hippocampal neurons significantly increased （P<0.01）. The synaptic cleft of the hippocampus was significantly shortened， and the length of active zone and postsynaptic density were significantly increased （P<0.05， P<0.01）. The expressions of synaptic related proteins Syt， PSD-95 and Kalirin-7 were significantly increased （P<0.05， P<0.01）. ConclusionEGCG can effectively improve cognitive dysfunction after epilepsy. Its protective effect may be achieved by protecting the ultrastructure of hippocampal synapses and regulating the expressions of synapse-related proteins Syt， PSD-95 and Kalirin-7.

Effects of Lactobacillus Rhamnosus GG on Activated Microglia and Tau Phosphorylation in the Hippocampus of Aged Mice Induced by Anesthesia and Surgery / 中山大学学报(医学科学版)

ObjectiveTo investigate the effects of Lactobacillus rhamnosus GG （LGG）on microglia and Tau phosphorylation in the hippocampus of aged mice induced by anesthesia and surgery. MethodsA total of thirty 18-month-old C57BL/6J mice were randomly divided into three groups： control group， anesthesia surgery group， and anesthesia surgery + LGG group （10 mice/group）. The aged mice were oral administered by NS or LGG 109 CFU 150 μL once a day for 20 days. Then anesthesia surgery group and anesthesia surgery +LGG group received anesthesia with isoflurane and exploratory laparotomy. The activation status of microglia in the hippocampus was detected by immunofluorescence staining 12 hours after surgery. IL-6 concentration changes was detected by ELISA. The expression changes of Tau protein phosphorylation site （Tau-pS202/pT205） and total Tau protein was detected by western blot. ResultsThe microglia in the hippocampus of the control group were in a resting state， and the concentration of inflammatory factor IL-6 was （82.08 ± 12.07） pg/mL in control group. Compared to the control group， the anesthesia surgery group showed microglial cell Microglia were activated， the concentration of inflammatory factors IL-6 increased significantly to （123.7±5.72） pg/mL （P=0.000）， and the expression of phosphorylated Tau-pS202/pT205 increased the hippocampus （P=0.002）. Compared to the anesthesia surgery group， the activated microglia were inhibited， the concentration of IL-6 decreased to （96.68±9.59） pg/mL （P=0.008）， and the expression of phosphorylated Tau-pS202/pT205 reduced significantly in the AS+LGG group （P=0.002）. While there were no significant changes in total Tau protein among 3 groups. ConclusionPreoperative administration of probiotic LGG can alleviate the activation of microglia， increased secretion of inflammatory factors， and increased Tau protein phosphorylation levels in the hippocampus of elderly mice caused by anesthesia surgery.

Wenyang Jieyu Prescription Regulates Hippocampal Neuron Apoptosis and Improves Synaptic Plasticity in Depressed Mice via BDNF/Akt/mTOR Pathway / 中国实验方剂学杂志

ObjectiveTo explore the mechanism of Wenyang Jieyu prescription in regulating hippocampal neuron apoptosis and improving synaptic plasticity in the mouse model of depression induced by maternal separation combined with restraint stress. MethodThe mice on postnatal day 0 （PD0） were randomly assigned into a control group （n=10） and a modeling group （n=50）. Maternal separation combined with restraint stress was adopted to establish the mouse model of depression， and the modeled mice were randomized into model， Wenyang prescription， Jieyu prescription， Wenyang Jieyu prescription， and fluoxetine groups （n=10） on the weaning day （PD21）. From PD21 to PD111， the mice were fed with the diets mixed with corresponding medicines. The sucrose preference test， open field test， O-maze test， and novel object recognition test were then conducted to evaluate the depression， memory， and learning abilities of mice. Immunohistochemistry （IHC） was employed to measure the atomic absorbance （AA） of postsynaptic density protein 95 （PSD95） in the hippocampus. Terminal-deoxynucleoitidyl transferase-mediated nick-end labeling （TUNEL） was employed to detect the apoptosis of hippocampal neurons. Western blot was employed to determine the protein levels of brain-derived neurotrophic factor （BDNF）， phosphorylated tyrosine kinase receptor B/tyrosine kinase receptor B （p-TrkB/TrkB）， phosphorylated protein kinase B/protein kinase B （p-Akt/Akt）， phosphorylated mammalian target of rapamycin/mammalian target of rapamycin （p-mTOR/mTOR）， B-cell lymphoma-2 （Bcl-2）， Bcl-2-associated X （Bax）， cysteinyl aspartate-specific proteinase-3 （Caspase-3）， synaptophysin （Syn）， and PSD95. ResultCompared with the control group， the modeling decreased the sucrose preference rate， time spent in central zone within 5 min， total movement distance， time spent in the open arm， and cognition index （P<0.01）. Furthermore， it decreased the expression of PSD95， increased the neuron apoptosis in the hippocampus （P<0.01）， down-regulated the protein levels of BDNF， p-TrkB/TrkB， p-Akt/Akt， p-mTOR/mTOR， Bcl-2， PSD95， and Syn （P<0.01）， and up-regulated the protein levels of Bax and Caspase-3 （P<0.05） in the hippocampus. Compared with the model group， Wenyang Jieyu prescription and fluoxetine increased the sucrose preference rate， time spent in central zone within 5 min， total movement distance， time spent in the open arm， and cognition index （P<0.05， P<0.01）. Moreover， the drugs increased the expression of PSD95， reduced the neuron apoptosis （P<0.01）， up-regulated the protein levels of BDNF， p-TrkB/TrkB， p-Akt/Akt， p-mTOR/mTOR， Bcl-2， PSD95， and Syn （P<0.01）， and down-regulated the protein levels of Bax and Caspase-3 （P<0.01）. ConclusionWenyang Jieyu prescription outperformed Wenyang prescription and Jieyu prescription in the treatment of the depressive behavior induced by maternal separation combined with restraint stress in mice. It exerted the therapeutic effect by reducing the hippocampal neuron apoptosis and improving the synaptic plasticity via the BDNF/Akt/mTOR pathway.

Role of 5-hydroxytryptamine type 3 receptors in the regulation of anxiety reactions / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

5-Hydroxytryptamine (5-HT) type 3 receptor (5-HT3R) is the only type of ligand-gated ion channel in the 5-HT receptor family. Through the high permeability of Na+, K+, and Ca2+ and activation of subsequent voltage-gated calcium channels (VGCCs), 5-HT3R induces a rapid increase of neuronal excitability or the release of neurotransmitters from axon terminals in the central nervous system (CNS). 5-HT3Rs are widely expressed in the medial prefrontal cortex (mPFC), amygdala (AMYG), hippocampus (HIP), periaqueductal gray (PAG), and other brain regions closely associated with anxiety reactions. They have a bidirectional regulatory effect on anxiety reactions by acting on different types of cells in different brain regions. 5-HT3Rs mediate the activation of the cholecystokinin (CCK) system in the AMYG, and the γ‍-aminobutyric acid (GABA) "disinhibition" mechanism in the prelimbic area of the mPFC promotes anxiety by the activation of GABAergic intermediate inhibitory neurons (IINs). In contrast, a 5-HT3R-induced GABA "disinhibition" mechanism in the infralimbic area of the mPFC and the ventral HIP produces anxiolytic effects. 5-HT2R-mediated regulation of anxiety reactions are also activated by 5-HT3R-activated 5-HT release in the HIP and PAG. This provides a theoretical basis for the treatment of anxiety disorders or the production of anxiolytic drugs by targeting 5-HT3Rs. However, given the circuit specific modulation of 5-HT3Rs on emotion, systemic use of 5-HT3R agonism or antagonism alone seems unlikely to remedy anxiety, which deeply hinders the current clinical application of 5-HT3R drugs. Therefore, the exploitation of circuit targeting methods or a combined drug strategy might be a useful developmental approach in the future.

Theta Oscillations Support Prefrontal-hippocampal Interactions in Sequential Working Memory / 神经科学通报·英文版

The prefrontal cortex and hippocampus may support sequential working memory beyond episodic memory and spatial navigation. This stereoelectroencephalography (SEEG) study investigated how the dorsolateral prefrontal cortex (DLPFC) interacts with the hippocampus in the online processing of sequential information. Twenty patients with epilepsy (eight women, age 27.6 ± 8.2 years) completed a line ordering task with SEEG recordings over the DLPFC and the hippocampus. Participants showed longer thinking times and more recall errors when asked to arrange random lines clockwise (random trials) than to maintain ordered lines (ordered trials) before recalling the orientation of a particular line. First, the ordering-related increase in thinking time and recall error was associated with a transient theta power increase in the hippocampus and a sustained theta power increase in the DLPFC (3-10 Hz). In particular, the hippocampal theta power increase correlated with the memory precision of line orientation. Second, theta phase coherences between the DLPFC and hippocampus were enhanced for ordering, especially for more precisely memorized lines. Third, the theta band DLPFC → hippocampus influence was selectively enhanced for ordering, especially for more precisely memorized lines. This study suggests that theta oscillations may support DLPFC-hippocampal interactions in the online processing of sequential information.

Clinical evaluation of brain structural changes in patients with alcohol dependence complicated by major depressive disorder using functional magnetic resonance imaging / 中国基层医药

Objective:Utilizing functional magnetic resonance imaging (fMRI) to investigate changes in brain structure and function in patients with alcohol dependence (AD) complicated by major depressive disorder (MDD), and assessing the clinical significance of fMRI in diagnosing alcohol dependence complicated by MDD.Methods:From August 2019 to October 2022, 90 patients with AD complicated by MDD and 90 healthy subjects who concurrently received physical examination in our hospital were included in the study. All participants underwent magnetic resonance imaging (MRI) and fMRI to observe the brain tissue structure of patients with AD complicated by MDD and to assess differences in N-acetylaspartic acid/creatine (NAA/Cr) and factional anisotropy (FA) values across different brain tissue regions.Results:The widths of the left and right ventricular temporal angles in the AD complicated by MDD group [(2.67 ± 0.24) mm, (2.63 ± 0.25) mm] were significantly higher than those observed in the healthy control group [(2.29 ± 0.21) mm, (2.31 ± 0.23) mm, t = 22.48, 20.64, both P < 0.001]. Additionally, the volumes of the left and right hippocampus and nucleus accumbens in the AD complicated by MDD group [(2 673.46 ± 155.74) mm 3, (2 692.29 ± 154.61) mm 3, (682.04 ± 65.37) mm 3, (729.65 ± 68.49) mm 3] were significantly lower compared with those in the healthy control group [(2 826.53 ± 158.95) mm 3, (2 849.17 ± 157.23) mm 3, (766.28 ± 69.51) mm 3, and (805.43 ± 71.36) mm 3, t = -9.53, -8.44, -15.62, -13.92, all P < 0.001]. Moreover, the NAA/Cr values in the left and right frontal lobes, temporal lobes, hippocampi, and nucleus accumbens in the AD complicated by MDD group were significantly lower than those in the healthy control group ( t = -11.36, -7.19, -9.96, -7.84, -14.59, -8.25, -7.64, -6.84, all P < 0.001). Similarly, the FA values of the left and right frontal lobes, temporal lobes, hippocampi, and right nucleus accumbens in the AD complicated by MDD group were significantly lower compared with those in the healthy control group ( t = -9.48, -11.74, -9.22, -10.36, -16.85, -14.67, -5.28, all P < 0.001). Conclusion:Patients with AD accompanied by MDD exhibit alterations in brain tissue structure, neuronal metabolic function, and the integrity of white matter nerve fibers. fMRI is effective in identifying changes in brain neuron metabolism and the integrity of white matter nerve fibers, making it invaluable for the diagnosis and assessment of AD accompanied by MDD.