Antidepressant effects of Yuanzhi (Polygalae Radix) extract on chronic unpredictable mild stress-induced depression in rats: modulation of the NLRP3 inflammasome and NF-κB pathway

Objective To investigate the antidepressant effects of Yuanzhi(Polygalae Radix;PR)aqueous extract on chronic unpredictable mild stress(CUMS)-induced depression rat models and the underlying mechanisms.Methods A total of 40 male Sprague Dawley(SD)rats were randomly divided into control;model;low dose of PR(PR-L;0.5 g/kg);high dose of PR(PR-H;1 g/kg);and fluoxetine(10 mg/kg)groups;with 8 rats in each group.Except for the rats in control group;those in the other four groups underwent CUMS-induced depression modeling.PR and fluoxetine were administered intragastrically once daily;30 min prior to the CUMS procedure;for 14 consecu-tive days until the behavioral tests were performed.After CUMS modeling;the sucrose prefer-ence test(SPT);open field test(OFT);novelty-suppressed feeding test(NSFT);forced swim test(FST);and tail suspension test(TST)were employed to assess the pharmacological ef-fects of PR on the mitigation of depressive-like behaviors in rat models.Additionally;the en-zyme-linked immunosorbent assay(ELISA)was utilized to quantify the serum levels of tumor necrosis factor(TNF)-α;interleukin(IL)-6;and IL-1βin the rats.Western blot analysis was al-so conducted to evaluate the protein expression levels of nuclear factor kappa-B(NF-κB);in-ducible nitric oxide synthase(iNOS);cyclooxygenase-2(COX-2);nucleotide-binding oligomerization domain(NOD)-like receptor family pyrin domain containing 3(NLRP3);apoptosis-associated speck-like protein containing caspase recruitment domain(ASC);and caspase-1 in the hippocampal tissues of the rats.Immunofluorescence staining was per-formed to observe the morphological changes in ionized calcium-binding adapter molecule 1 positive(Iba-1+)cells in the dentate gyrus(DG)of rats with CUMS-induced depression.Results(i)Treatment with PR-H and fluoxetine resulted in significant enhancements in both the total distance and time the rats moved during tests(P<0.01 and P<0.05;respectively).Post-administration of PR-H and fluoxetine also led to statistically significant increase in su-crose preference among rats(P<0.05).Besides;PR-L;PR-H;and fluoxetine treatment markedly decreased the latency of ingestion(P<0.05;P<0.05;and P<0.01;respectively).As observed from the FST;PR-L;PR-H;and fluoxetine presented antidepressant effects on rats with CUMS-induced depression;leading to the reduction in time of their immobility(P<0.05;P<0.01;and P<0.01;respectively).The results of TST indicated reduced immobility time in rats receiving PR-H and fluoxetine treatment as well(P<0.01).(ii)Rats in model group showed an increase in the levels of Iba-1+microglia in their left and right brains in compari-son with control group(P<0.01).However;such increase was negated post PR treatment(P<0.01).Treatment with PR-L;PR-H;and fluoxetine considerably reduced the levels of inflam-matory factors(TNF-α;IL-1β;and IL-6;P<0.01).In addition;treatment of PR-L and PR-H ef-fectively counteracted the elevated levels of NLRP3;ASC;and caspase-1;and markedly down-regulated the expression levels of phosphorylated p65(p-p65);COX-2;and iNOS in rats’hip-pocampus(P<0.01).Conclusion Collectively;these findings indicate that PR exerts an antidepressant effect on rats with CUMS-induced depression partially through the modulation of the NLRP3 and NF-κB signaling pathways.

Shuganheweitang Ameliorates Chronic Unpredictable Mild Stress-Induced Depression-Like Behaviors in Rats through the PI3K/AKT/mTOR Pathway: Involvement of Amino Acids, Glycerophospholipids, and Energy Metabolism

Background: Depression is a typical psychosomatic disease. Shuganheweitang (SGHWT) is a clinical formula that effectively treats depression. However, the potential mechanism used by SGHWT to ameliorate depression-like behaviors is still unclear. This study investigated the effects of SGHWT on metabolic change in the liver and hypothalamus with signaling pathways involved in chronic unpredictable mild stress (CUMS)-induced depression in rats to explore the mechanism of the anti-depressive effect. Methods: A total of 52 rats were used to create a model of depression by CUMS combined with solitary rearing for 6 weeks. Open field test (OFT), sucrose preference test (SPT), forced swim test (FST), and body weight (BW) were performed to analyze the pharmacodynamic effects of SGHWT. H&E staining, Nissl staining, immunofluorescence, immunohistochemistry, and western blot were used to evaluate the mechanism of action. Untargeted metabolomics techniques by ultra-performance liquid chromatography-quantitative time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS) were used to analyze all the metabolic differences in the liver and hypothalamus. Results: SGHWT improved CUMS-induced depression-like behaviors in vivo. SGHWT reduced hepatic c-Fos protein expression and increased hypothalamic c-Fos protein expression. Moreover, p-PI3K, p-AKT473, p-AKT308, and p-mTOR protein expressions were significantly downregulated in the liver and hypothalamus of CUMS rats. Notably, these alterations were reversed by the SGHWT administration. Furthermore, the metabolomic analysis identified 15 and 5 key differential SPT-associated metabolites in the liver and hypothalamus, respectively. Conclusion: This study suggests that SGHWT ameliorates chronic unpredictable mild stress-induced depression-like behaviors, by the involvement of amino acids, glycerophospholipids, energy metabolism, and the PI3K/AKT/mTOR pathway. Highlights: 1) Shuganheweitang was derived from the TCM herbal formula Sinisan. 2) SGHWT treatment reverses depression-like behaviors in CUMS-induced rats. 3) The mechanism of SGHWT on depression by the liver and hypothalamus metabolomics. 4) SGHWT regulates amino acids, glycerophospholipids, and energy metabolism. 5) SGHWT exerts antidepressant effects through the PI3K/AKT/mTOR pathway.

Dynamic changes of behaviors, dentate gyrus neurogenesis and hippocampal miR-124 expression in rats with depression induced by chronic unpredictable mild stress

The depression-like behavior phenotype,neurogenesis in the dentate gyrus and miR-124 expression in the hippocampus are the focus of current research on the pathogenesis of depression and antidepressant therapy.The present study aimed to clarify the dynamic changes of depression-like behavior,dentate gyrus neurogenesis and hippocampal miR-124 expression during depression induced by chronic stress to reveal pathological features at different stages of depression and to further provide insight into depression treatment.Chronic unpredictable mild stress depression models were established by exposing Sprague-Dawley rats to various mild stressors,including white noise,thermal swimming,stroboscopic illumination,soiled cages,pairing with three other stressed animals,cold swimming,tail pinch,restraint and water and food deprivation.Chronic unpredictable mild stress model rats underwent dynamic observation from 1 to 8 weeks and were compared with a control group(normal feeding without any stressors).To observe changes in the depression-like behavior phenotype during chronic unpredictable mild stress-induced depression,a sucrose preference test was used to evaluate the degree of anhedonia.An open-field test was used to evaluate locomotor activity and anxiety status.Compared with the control group,chronic unpredictable mild stress rats lost weight but did not have a depression-like behavioral phenotype at 1-4 weeks.Chronic unpredictable mild stress rats presented decreased sucrose preference and locomotor activity at 5-8 weeks.In addition,chronic unpredictable mild stress rats did not have significant anxiety-like behavior during 1-8 weeks of modeling.To observe neurogenesis dysfunctions and changes in neuronal number in the dentate gyrus during chronic unpredictable mild stress-induced depression,markers(DCX and DCX/BrdU)of neural proliferation and differentiation and the neuronal marker NeuN were assessed by immunofluorescence.Compared with the control group,neurogenesis and the neuronal number in the dentate gyrus did not change from 2 to 6 weeks;however,neural proliferation and differentiation in the dentate gyrus decreased,and the number of neurons decreased until the eighth week in the chronic unpredictable mild stress group.Real-time quantitative reverse transcription polymerase chain reaction assays and fluorescence in situ hybridization were used to measure the expression of hippocampal miR-124 during chronic unpredictable mild stress-induced depression.The results showed that the expression of hippocampal miR-124 was unchanged during the first 4 weeks but increased from 5 to 6 weeks and decreased from 7 to 8 weeks compared with the control group.These findings indicate that during chronic unpredictable mild stress-induced depression,the behavioral phenotype,miR-124 expression in the hippocampus,neurogenesis in the dentate gyrus and neuronal numbers showed dynamic changes,which suggested that various pathological changes occur at different stages of depression.All experimental procedures and protocols were approved by the Experimental Animal Ethics Committee of Guangzhou University of Chinese Medicine of China in March 2015.

Brain region-specific roles of brain-derived neurotrophic factor in social stress-induced depressive-like behavior

Brain-derived neurotrophic factor is a key factor in stress adaptation and avoidance of a social stress behavioral response.Recent studies have shown that brain-derived neurotrophic factor expression in stressed mice is brain region–specific,particularly involving the corticolimbic system,including the ventral tegmental area,nucleus accumbens,prefrontal cortex,amygdala,and hippocampus.Determining how brain-derived neurotrophic factor participates in stress processing in different brain regions will deepen our understanding of social stress psychopathology.In this review,we discuss the expression and regulation of brain-derived neurotrophic factor in stress-sensitive brain regions closely related to the pathophysiology of depression.We focused on associated molecular pathways and neural circuits,with special attention to the brain-derived neurotrophic factor–tropomyosin receptor kinase B signaling pathway and the ventral tegmental area–nucleus accumbens dopamine circuit.We determined that stress-induced alterations in brain-derived neurotrophic factor levels are likely related to the nature,severity,and duration of stress,especially in the above-mentioned brain regions of the corticolimbic system.Therefore,BDNF might be a biological indicator regulating stress-related processes in various brain regions.

Quercetin regulates depression-like behavior in CUMS rat models via TLR4/NF-κB signaling

Background:Depression is becoming increasingly prevalent around the world,imposing a substantial burden on individuals,families,as well as society.Quercetin is known to be highly effective in treating depression.However,additional research is needed to dissect the mechanisms of its anti-depressive effects.Methods:For this study,Sprague-Dawley(SD)rats were randomized into the control,model,quercetin,or fluoxetine group.The latter three groups were exposed to chronic unpredictable mild stress(CUMS)for 42 d.The first two groups received saline solution daily via oral gavage.Meanwhile,the quercetin group was orally administered a quercetin suspension(52.08 mg/kg)every day,while the fluoxetine group was orally administered a fluoxetine solution(2.08 mg/kg).Here,fluoxetine served as the positive control drug to compare the therapeutic effects of quercetin.The experimental period was 6 weeks.Depressive behaviors in rats were assessed through various physiological and behavioral measures.Additionally,pathological changes in hippocampal tissues were examined using Nissl staining.Serum cytokines were detected using an enzymelinked immunosorbent assay(ELISA),and immunohistochemistry was employed to quantify the levels and integral optical density(IOD)values of ionized calcium binding adaptor molecule-1(Iba-1)expression in the brain.Real-time fluorescence quantitative PCR(RT-qPCR)was utilized to evaluate the mRNA levels of inflammatory indicators as well as toll-like receptor 4(TLR4),and nuclear factor-κappa B P65(NF-κB P65)in hippocampus.Western blot(WB)technique was employed to observe the protein levels of TLR4,NF-κB P65,and phospho-NF-κB P65(p-NF-κB P65).Results:After 42 d of exposure to CUMS,rats exhibited a slow increase in body weight,a reduction in food intake,an abnormal preference for sugar water,and aberrant open-field behaviors.Pathological analysis revealed the disintegration,rupture,interruption,and disorganization of hippocampal neuronal cells after CUMS exposure,along with a decrease in Nissl bodies in the CA1 region.This was accompanied by the elevated expression of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),and interleukin-6(IL-6)in the serum and the upregulation of IL-1β,IL-6,and TNF-αmRNA expression in the hippocampus.Increases in Iba-1-positive cells and the IOD values of Iba-1 were detected in hippocampal microglia.Furthermore,TLR4 and NF-κB P65 mRNA and protein levels were upregulated in hippocampal tissues.Quercetin,an antidepressant,could alleviate depression-like symptoms in rats and downregulate inflammatory factors associated with the TLR4/NF-κB signaling pathway in hippocampal microglia,and its therapeutic effect was comparable to fluoxetine.Conclusion:In rat models of CUMS,quercetin may act as an antidepressant by inhibiting inflammation in hippocampal microglia via TLR4/NF-κB signaling pathway.These results offer experimental and theoretical support for applying quercetin in the clinical management of depression.

Effect and mechanism of LW-AFC on chronic unpredictable mild stress-induced mood and cognition impairment of mice

OBJECTIVE To explore the effect and mechanisms of LW-AFC,a new formula derived fromLiuwei Dihuang decoction,on chronic unpredictable mild stress(CUMS)-induced mood and cogni.tion impairment in mice.METHODS C57 BL/6 J mice were randomly placed into seven groups(n=10):normal control group,CUMS group,Fluoxetine(10 mg·kg^(-1),once per day) group,Liuwei Dihuang decoction group(LW,10 g·kg^(-1),once per day),and LW-AFC(0.8 g·kg^(-1),1.6 g·kg^(-1),3.2 g·kg^(-1),once per day) group.The stressed group was given CUMS for 4 weeks to set up a chronic multiple-stressed model.LW and LW-AFC was oral administered a week prior to CUMS and until the end of the study(a total of 35 d),while fluoxetine was administrated orally for 4 weeks.The anxiety behavior was analyzed using the open field test(OFT) and elevated plus maze test(EPM).The depression behavior was ana.lyzed using the sucrose preference test(SPT) and forced swimming test(FST).Spatial cognition was evaluated using Morris water maze(MWM) test and working memory was evaluated using new object recognition test(NORT).RESULTS CUMS for 28 d increased depressive-and anxiety-like behaviors.LW-AFC(1.6 g·kg^(-1)) significantly increased the numbers of entries into the open arm and time in the open arm of CUMS mice(P<0.05).LW-AFC(3.2 g·kg^(-1)) increased sucrose consumption and de.creased the immobility time of FST(P<0.01) of CUMS mice.The MWM test showed that spatial learning andmemory in CUMS mice were remarkably affected relative to controls,whereas LW-AFC(3.2 g·kg^(-1)) im.proves cognitive functions(P<0.05).CONCLUSION The mood and theability of learning and memory of thestressed group can be affected after exposure to CUS.Oral administration of LW-AFC significant.ly improved CUMS-induced impairments of mood and cognition in mice.

Pathogenesis of chronic social defeat stress model induced depressive-like mouse model according to LC-MS/MS-based metabolomics

OBJECTIVE To explore the pathogenesis of depression according to the LC-MS/MS-based metabolomics in the mouse model which exhibits social avoidance state induced by the chronic social defeat stress model(CSDS).METHODS Twenty male C57BL/6N mice were randomly divided into control group and model group suffering CSDS,and the ICR retired breeder mice were used to attack the model group for 14 d of chronic social defeated stress.The open field test and source preference test were both used to observe depression-like behavior.Besides,the social interaction test is used to observe the social interaction state,especially.After the stress,the serum samples of mice were collected,and the changes of endogenous metabolites were analyzed by LC-MS metabolomics technology,and the pathway analysis of the differential metabolites was performed to explore the pathogenesis of the CSDS induced depressive-like mouse model.RESULTS After the stress of CSDS was completed,the mice in the model group showed a significant slowdown in body weight growth,a reduction in the source preference rate,and a significant reduction in the total distance and the number of rearing in the open field test.Distinctively,the social interaction rate is remarkably decreasing.There are 24 differential metabolites found in the serum of CSDS model mice.CONCLUSION The mouse who suffered CSDS stress would show depressive-like behavior.Based on the LC-MS/MS metabolomics,24 differential metabolites were found in the serum of CSDS model mice.The amino acid metabolism might be significant to the pathogenesis of the CSDS induced depressive-like mouse model.

Antidepressant Effect of Shenwei Ningyu Tablets on Rat Chronic Stress Model and Mouse Tail Suspension Model

[Objectives]To explore the antidepressant effect of Shenwei Ningyu Tablet,a new antidepressant traditional Chinese medicine,on rat chronic stress depression model and mouse tail suspension models.[Methods]Rat chronic stress model:except for the normal group,the rats in other groups were given corresponding chronic stress,and administered by gavage 1 h before modeling,for a total of 21 d.The changes of each indicator before and after the experiment were observed through the body weight change,the sugar water test,and open field test.The relevant hormone levels were detected by radioimmunoassay.Mouse tail suspension depression model:after continuous administration for 7 d,the activity times was recorded with the mouse automatic recorder,and the mouse immobility time was recorded after tail suspension,to explore the effects of each administration group on the tail suspension immobility time of mice.[Results]Chronic stress depression model:21 d after modeling,compared with the normal group,rats in the model group exhibited significantly reduced body weight,sucrose preference index,and horizontal and vertical movement scores(P<0.05).Compared with the model group,the low-dose Shenwei Ningyu Tablets group had significant differences in the sugar water test,horizontal and vertical movement scores(P<0.05).In addition,all three dose groups of Shenwei Ningyu Tablets could effectively reduce the content of CRF in chronic stress model rats,and the low dose group could significantly reduce the ACTH level in model rats(P<0.05).Mouse tail suspension depression model:the immobility time after tail suspension in each administration group was significantly different from that in the model group(P<0.05).[Conclusions]Shenwei Ningyu Tablets has a certain anti-depression effect on both the rat chronic stress depression model and the mouse tail suspension depression model.

Shuganjieyu capsule increases neurotrophic factor expression in a rat model of depression

Shuganjieyu capsule has been approved for clinical treatment by the State Food and Drug Ad-ministration of China since 2008. In the clinic, Shuganjieyu capsule is often used to treat mild to moderate depression. In the rat model of depression established in this study, Shuganjieyu capsule was administered intragastrically daily before stress. Behavioral results conifrmed that depressive symptoms lessened after treatment with high-dose （150 mg/kg） Shuganjieyu capsule. Immunohistochemistry results showed that high-dose Shuganjieyu capsule signiifcantly increased phosphorylation levels of phosphorylation cyclic adenosine monophosphate response element binding protein and brain-derived neurotrophic factor expression in the medial prefrontal cortex and hippocampal CA3 area. Overall, our results suggest that in rats, Shuganjieyu capsule effec-tively reverses depressive-like behaviors by increasing expression levels of neurotrophic factors in the brain.

Exploration of the mechanism by which icariin modulates hippocampal neurogenesis in a rat model of depression

Icariin(ICA) has a significant capacity to protect against depression and hippocampal injury,but it cannot effectively cross the bloodbrain barrier and accumulate in the brain.Therefore,the mechanism by which ICA protects against hippocampal injury in depression remains unclear.In this study,we performed proteomics analysis of cerebrospinal fluid to investigate the mechanism by which ICA prevents dysfunctional hippocampal neurogenesis in depression.A rat model of depression was established through exposure to chronic unpredictable mild stress for 6 weeks,after which 120 mg/kg ICA was administered subcutaneously every day.The results showed that ICA alleviated depressive symptoms,learning and memory dysfunction,dysfunctional neurogenesis,and neuronal loss in the dentate gyrus of rats with depression.Neural stem cells from rat embryonic hippocampi were cultured in media containing 20% cerebrospinal fluid from each group of rats and then treated with 100 μM corticosterone.The addition of cerebrospinal fluid from rats treated with ICA largely prevented the corticosterone-mediated inhibition of neuronal proliferation and differentiation.Fifty-two differentially expressed proteins regulated by chronic unpredictable mild stress and ICA were identified through proteomics analysis of cerebrospinal fluid.These proteins were mainly involved in the ribosome,PI3 K-Akt signaling,and interleukin-17 signaling pathways.Parallel reaction monitoring mass spectrometry showed that Rps4 x,Rps12,Rps14,Rps19,Hsp90 b1,and Hsp90 aa1 were up-regulated by chronic unpredictable mild stress and down-regulated by ICA.In contrast,Htr A1 was down-regulated by chronic unpredictable mild stress and up-regulated by ICA.These findings suggest that ICA can prevent depression and dysfunctional hippocampal neurogenesis through regulating the expression of certain proteins found in the cerebrospinal fluid.The study was approved by the Experimental Animal Ethics Committee of Guangzhou University of Chinese Medicine of China in March 2017.

Citrus aurantium L. extract alleviate depression by inhibiting gut microbiota-mediated inflammation in mice

Gut microbiota plays a crucial role in the pathophysiology of depression.This study aimed to explore the antidepressant effect of mature whole Citrus aurantium fruit extract(FEMC)in the chronic unpredictable mild stress(CUMS)model.The behavioral tests were applied to assess antidepressant effect and 16S rRNA sequencing was used to analyze the changes of gut microbiota.The results showed that the major components of FEMC were naringin and neohesperidin and significantly increased the sucrose preference index of the mice.FEMC also could reduce the feeding latency in an open field test and the rest time in a novelty suppressed feeding test.In addition,FEMC could increase CUMS-induced reduction in the levels of BDNF,PSD95,and SYN in the hippocampus.Moreover,FEMC intervention slightly decreased the ratio of Firmicutes to Bacteroidota.Meanwhile,FEMC reduced the abundance of the Prevotellaceae_Ga6A1_group,[Ruminococcus]_torques_group,which have been reported to be closely related to inflammation.Bioinformatics analysis revealed that mitogen-activated protein kinase(MAPK)signaling pathway and lipopolysaccharide biosynthesis were involved in the anti-inflammatory effect of FEMC in the CUMS animal model.Finally,the ELISA results showed that FEMC could significantly reduce the expression of pro-inflammatory cytokines IL-6 and TNF-αin the serum of depressive mice.Our results suggest FEMC can am eliorate depressive behavior by i nhibiting gut microbiota-mediated inflammation in mice.

Overexpression of brain-derived neurotrophic factor in the hippocampus protects against post-stroke depression

Post-stroke depression is associated with reduced expression of brain-derived neurotrophic factor （BDNF）. In this study, we evaluated whether BDNF overexpression affects depression-like behavior in a rat model of post-stroke depression. The middle cerebral artery was occluded to produce a model of focal cerebral ischemia. These rats were then subjected to isolation-housing combined with chronic unpredictable mild stress to generate a model of post-stroke depression. A BDNF gene lentiviral vector was injected into the hippocampus. At 7 days after injection, western blot assay and real-time quantitative PCR revealed that BDNF expression in the hippo- campus was increased in depressive rats injected with BDNF lentivirus compared with depressive rats injected with control vector. Furthermore, sucrose solution consumption was higher, and horizontal and vertical movement scores were increased in the open field test in these rats as well. These findings suggest that BDNF overexpression in the hippocampus of post-stroke depressive rats alleviates depression-like behaviors.

Establishment and evaluation of a mouse model of affective disorder combined with atherosclerosis

Objective:To establish a mouse model of affective disorder complicated with atherosclerosis(AS)by high fat feeding and chronic mild unpredictable stimulation(CUMS),and to provide an animal model for the later study of the prevention and treatment of affective disorder complicated with atherosclerosis by traditional Chinese medicine.Methods:10 C57BL/6J mice were used as blank group,and 20 ApoE-/-mice were randomly divided into AS group and AS+CUMS group.After one week of adaptive feeding,except for the blank group,the other two groups were fed with high fat diet.Meanwhile,the AS+CUMS group was given chronic unpredictable mild stress.The model was evaluated after 16 weeks of modeling.During the experiment,the body weight,food intake,excitability,hair color and other general morphology of mice in each group were observed and recorded.Behavioral indexes(Sucrose preference tests and Open Field test)were detected in each group.The levels of 5-hydroxytryptamine(5-HT)and hypothalamic-pituitary-adrenal(HPA)axis hormones,including adrenocorticotropin(ACTH)and corticosterone(CORT),were detected by ELISA.Serum lipid levels,including total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDL-C)and high density lipoprotein cholesterol(HDL-C),were detected by ELISA.HE staining was used to observe the pathological condition of aorta.Results:Compared with blank group,the excitability and food intake of AS+CUMS group were significantly decreased.There were no significant differences in sugar water consumption and activity capacity of mice in AS group,while sugar water consumption and activity capacity of mice in AS+CUMS group were significantly decreased(P<0.01).The serum 5-HT levels in AS group and AS+CUMS group were decreased,and the levels of ACTH and CORT in AS+CUMS group were significantly decreased(P<0.01),and the levels of ACTH and CORT in AS+CUMS group were significantly increased(P<0.01).The serum TC,TG and LDL-C levels of mice in AS group and AS+CUMS group were significantly increased(P<0.01),while the HDL-C level of mice in both groups was significantly decreased(P<0.01).HE staining results showed that the area and degree of plaques in the active vascular lumen of AS group and AS+CUMS group were larger and heavier.Conclusion:High fat feeding combined with CUMS was successful in establishing a mouse model of emotional disorder combined with atherosclerosis.

Effects of CUMS combined with CRS on hippocampal glial cells and synaptic plasticity in depressed mice

Objective:To explore the effects of CUMS combined with CRS on mouse hippocampal glial cells and synaptic plasticity-related proteins. Methods: Forty mice were randomly divided into normal group (n=20) and model group (n=20). The model group used CUMS combined with CRS to prepare a mouse model of depression for 7 weeks. The behavioral evaluation of the mice at 3 weeks and 7 weeks after modeling was performed by sugar water preference test, open field test and tail suspension test. After the experiment, the samples were collected, and the content of TNF-a in the hippocampus of mice was detected by enzyme-linked immunosorbent assay. Immunohistochemical method was used to detect the Iba-1 and GFAP MOD values of mouse hippocampal CA1 area, CA3 area and DG area. Western blot was used to detect the protein expression of Iba-1, GFAP, SYN1 and PSD-95 in the hippocampus. fluorescence quantitative PCR method was used to detect the expression of SYN1, PSD-95 mRNA in hippocampus. Results: At the 3rd week after modeling, the body weight, sugar water preference rate, total distance moved, number of standing uprights, and stay time in the central area of the mice in the model group were all lower than those in the normal group (P<0.05), and the tail suspension immobility time was longer than that in the normal group (P<0.01). After 7 weeks of modeling, the body weight, sugar water preference rate, total distance moved, number of erection times, central area residence time, and average movement speed of the mice in the model group were lower than those in the normal group (P< 0.05), the tail suspension immobility time was longer than that in the normal group (P<0.01). The contents of TNF-a in the hippocampus were higher than those in the normal group (P<0.05). The GFAP MOD value and the relative expression of GFAP protein in hippocampal CA1, CA3 and DG regions were significantly lower than those in the normal group (P<0.05). The Iba-1 MOD value and the relative expression of Iba-1 protein in hippocampal CA1, CA3 and DG regions were significantly higher than those in the normal group (P<0.05). The relative expression of SYN1 and PSD-95 protein and the relative expression of SYN1 and PSD-95 mRNA in the hippocampus were significantly lower than those in the normal group (P<0.05). Conclusion: After 3 weeks of CUMS and CRS modeling, the depression-like behavior of mice appeared, and the depression of mice was more obvious after 7 weeks of modeling. The depression mouse model made by CUMS combined with CRS method may be related to increased hippocampal inflammation, excessive activation of microglia, decreased number of astrocytes and decreased synaptic plasticity.

基于GABA能神经系统探究淫羊藿次苷Ⅱ的抗抑郁作用及机制

目的基于γ氨基丁酸(GABA)能神经系统研究淫羊藿次苷Ⅱ(ICSⅡ)的抗抑郁作用及潜在机制。方法将雄性昆明小鼠分为对照组(C组,10只)和造模组(50只),造模组小鼠以慢性不可预知温和应激法复制抑郁模型。刺激21 d后,将造模组小鼠随机分为抑郁模型组(NS组)、阳性对照药组[ECS组,草酸艾司西酞普兰15 mg/(kg·d)]和ICSⅡ低、中、高剂量组[ICSⅡ-L、ICSⅡ-M、ICSⅡ-H组,ICSⅡ10、20、30 mg/(kg·d)],每组10只。各药物组小鼠灌胃相应药物,每天1次,连续14 d。检测各组小鼠的糖水偏爱率、运动总距离、悬尾实验和强迫游泳实验的静止不动时间,观察其海马CA3区神经元及尼氏体形态,检测海马组织中GABA、谷氨酸(Glu)含量、GABA/Glu比值及GABA能神经系统相关蛋白(GABA A受体α1、GABA B受体1、GABA囊泡转运体、谷氨酸脱羧酶67、GABA膜转运体3)的表达情况。结果与C组比较,NS组小鼠糖水偏爱率显著降低,运动总距离显著缩短,悬尾实验和强迫游泳实验的静止不动时间均显著延长(P<0.05);海马CA3区的神经元形态不规则且尼氏体减少,结构完整的神经元数量显著减少(P<0.05);海马组织中Glu含量显著升高,GABA含量、GABA/Glu比值和GABA能神经系统相关蛋白表达水平均显著降低(P<0.05)。与NS组比较,各药物组大鼠的抑郁样行为均有所改善,上述指标普遍逆转(P<0.05)。结论ICSⅡ能够改善抑郁模型小鼠的抑郁样行为,其机制可能与调节GABA、Glu平衡,增加GABA的合成、转运、释放以及调节相关受体的表达进而改善GABA能神经系统功能有关。

开心散联合氟西汀对慢性压力应激抑郁小鼠肠道菌群与机体致炎物质表达的影响

目的评价开心散联合氟西汀对慢性压力应激(CUMS)抑郁小鼠肠道菌群与机体致炎物质表达的影响,探索其调控肠-脑轴抗抑郁作用机制。方法构建小鼠CUMS抑郁模型,通过检测糖水偏嗜率、悬尾不动时间与强迫游泳不动时间评价开心散联合氟西汀改善小鼠抑郁行为的效果。采用ELISA法测定受试动物大脑皮层、血清和小肠组织中致炎物质白介素6(IL-6)、白介素1β(IL-1β)、肿瘤坏死因子-α(TNF-α)与脂多糖(LPS)的表达。采用16S rRNA基因测序法对小鼠粪便中肠道菌群组成进行分析。采用Western blot法考察小鼠小肠组织中ZO-1、Occludin和Claudin 5等肠道屏障蛋白的表达情况。结果开心散联合氟西汀显著上调模型小鼠糖水偏嗜率(P<0.01),缩短悬尾不动时间与强迫游泳不动时间(P<0.05,P<0.01)。开心散与氟西汀中剂量联用的抗抑郁效用与单用氟西汀高剂量的趋势相似。联合用药还可显著抑制CUMS模型动物大脑皮层、血清和小肠致炎物质的上调(P<0.05,P<0.01)。肠道菌群分析显示,联合用药可改善模型动物肠道革兰氏阳性菌与阴性菌的比例,其对毛螺菌、双歧杆菌、瘤胃球菌、布劳特氏菌、优杆菌、丁酸肠杆菌、丹毒丝菌、另枝菌、脱硫弧菌、红椿菌-UCG-002等肠道菌相对丰度的改善与其缓解抑郁样行为与皮层致炎物质表达下调显著相关(P<0.05,P<0.01)。另外,联合用药还可显著上调肠道屏障蛋白表达(P<0.05,P<0.01)。结论开心散联合氟西汀可缓解CUMS模型动物的抑郁样行为并减少氟西汀单用的用量。两者连用调控肠道菌群组成,抑制肠道致炎物质表达,上调肠道屏障蛋白表达,从而减少血清与中枢的致炎物质的表达,可能是其调控肠-脑轴抗抑郁的作用环节。

基于CUMS模型探究抑郁症对细菌感染的影响及作用

【目的】抑郁症是一种常见的精神疾病,对患者的身体健康有着深远的影响。抑郁症与较高的细菌感染风险相关;然而,这是否是一种因果关系,以及抑郁症如何影响感染仍不清楚。因此,本研究旨在通过构建慢性不可预测轻度应激(CUMS)模型,探究抑郁表型在小鼠细菌感染中的作用。【方法】小鼠经CUMS诱导4周,通过行为学测试评估抑郁表型。随后,小鼠腹腔注射肺炎克雷伯菌模拟细菌感染,感染后48 h收集血清和腹腔组织。苏木精-伊红染色(HE)观察组织病理学改变,酶联免疫吸附试验(ELISA)检测炎性因子水平。此外,对感染前收集的小鼠粪便样本进行肠道菌群16S rDNA基因测序分析,并检测未感染小鼠结肠组织中NF-κB/NLRP3信号通路的表达水平。【结果】行为学测试结果显示,与对照组相比,CUMS小鼠体质量显著降低(P<0.0001,t=5.426),蔗糖偏好率显著降低(P<0.001,t=4.937),游泳静止时间显著增加(P<0.001,t=16.37),旷场中央区域停留时间显著减少(P<0.01,t=3.575)。生存分析显示,与对照组小鼠相比,感染后CUMS小鼠的生存率显著降低(P<0.05)。HE染色结果显示,CUMS小鼠肝脏(P<0.05,t=4.025)、肾脏(P<0.05,t=2.828)、肠系膜(P<0.01,t=5.367)组织损伤程度明显加重。ELISA结果显示,炎症因子IL-6(P<0.01,t=3.365)、IL-1β(P<0.01,t=4.061)、TNF-α(P<0.01,t=4.460)和LPS(P<0.0001,t=27.24)水平升高。16S rDNA测序结果显示,CUMS小鼠肠道菌群结构发生改变,与对照组小鼠明显不同,表现出菌群失调。与对照组相比,CUMS小鼠结肠组织中NF-κB(P<0.01,t=6.825)和NLRP3(P<0.001,t=9.561)的表达水平升高。【结论】CUMS小鼠发生更严重的细菌感染。CUMS诱导的抑郁表型可能因为破坏肠道菌群组成和激活NF-κB/NLRP3信号通路,增加了小鼠对细菌感染的易感性。

南方红豆杉果乙酸乙酯部位对抑郁症模型大鼠表型和炎症介质的影响

目的:探讨南方红豆杉果乙酸乙酯部位对抑郁症模型大鼠的影响及可能作用机制。方法:采用随机数字法将20只雌性斯泼累格·多雷(SD)大鼠随机分为对照组、模型组、红豆杉果组和氟西汀组,每组5只。除对照组外,其余各组采用慢性不可预知温和应激(CUMS)法4周制备抑郁症模型。造模后红豆杉果组给予南方红豆杉果乙酸乙酯提取物200 mg/(kg·d)灌胃,对照组和模型组给予等体积的生理盐水灌胃,共给药4周。在造模结束后对对照组和模型组进行糖水实验和旷场实验,给药结束后对各组大鼠进行糖水实验和旷场实验,检测各组大鼠血清中白细胞介素-1β(IL-1β)、IL-6、IL-18的含量。结果:在造模结束后与对照组比较,模型组大鼠的糖水消耗率明显减少(P<0.01),活动距离明显降低(P<0.01)。实验结束后,与模型组比较,红豆杉果组和氟西汀组大鼠的糖水消耗率明显增加(P<0.01),活动距离明显增加(P<0.01);与对照组比较,模型组血清中IL-1β、IL-6、IL-18的含量均明显升高(P<0.05,P<0.01);与模型组比较,红豆杉果组和氟西汀组血清中IL-1β、IL-6、IL-18的含量均明显下降(P<0.05,P<0.01)。结论:南方红豆杉果乙酸乙酯部位有抗抑郁的作用,同时有抗炎的作用,可能通过降低抑郁症大鼠炎症反应起到缓解大鼠抑郁样行为。

百合地黄汤对抑郁症大鼠肠道菌群的影响

目的:探究百合地黄汤对抑郁症大鼠肠道菌群的影响。方法:采用孤养和慢性不可预知性温和应激刺激制备大鼠抑郁症模型,经百合地黄汤给药后,采用糖水偏好试验和旷场试验进行药效学评价,并对大鼠的结肠进行病理学检测;收集大鼠粪便,应用16S rRNA高通量测序技术对大鼠肠道菌群多样性进行分析,并进行功能预测。结果:与正常组比较,模型组大鼠体质量明显降低;给药后,各剂量组大鼠体质量均有增加的趋势。糖水偏好实验结果显示,与正常组比较,模型组大鼠糖水偏爱度明显减弱(P<0.01);给药后,各剂量组糖水偏爱度均有增强的趋势,且中剂量组和低剂量组差异有统计学意义(P<0.01)。在旷场实验中,与正常组比较,模型组大鼠自主活动显著减弱(P<0.01);给药后,各剂量组大鼠自主活动均有增强的趋势,且低剂量组与模型组比较差异有统计学意义(P<0.05)。病理结果显示,模型组大鼠结肠黏膜遭到破坏,经百合地黄汤给药后,各给药组结肠黏膜损伤均有不同程度的恢复。在菌群多样性方面,经百合地黄汤治疗后,在门水平上,厚壁菌门的丰度明显增加,拟杆菌门、疣微菌门、变形菌门及蓝藻门的丰度明显降低,其中变形菌门和蓝藻门改变具有统计学意义(P<0.05)。在属水平上,降低了norank_f__Muribaculaceae、瘤胃球菌属及粪球菌属等菌属的丰度,升高了乳酸菌属、罗姆布茨菌属、Turicibacter及芽孢杆菌属等菌属的丰度,其中norank_f__Muribaculaceae、罗姆布茨菌属、Turicibacter、norank_f__Eubacterium_coprostanoligenes_group、Prevotellaceae_UCG-001及Prevotellaceae_UCG-001的改变有统计学意义(P<0.05)。京都基因和基因组百科全书(KEGG)功能预测显示,差异菌群的功能与氨基酸代谢、能量代谢、神经系统以及神经退行性疾病等相关。结论:百合地黄汤能够调节抑郁症大鼠的肠道菌群进而改善抑郁症大鼠的抑郁样行为。

人参皂苷Rg1和Rb1改善慢性不可预测应激致大鼠抑郁、焦虑样行为的作用比较

目的研究人参皂苷Rg1和Rb1改善慢性不可预测应激诱导大鼠的抑郁样和焦虑样行为的作用及比较。方法SPF级SD雄性大鼠70只,适应5 d后进行糖水偏爱实验检测,根据糖水偏爱指数将动物分为7组,即对照组、模型组、氟西汀组、人参皂苷Rg124 mg/kg剂量组、人参皂苷Rg148 mg/kg剂量组、人参皂苷Rb133 mg/kg剂量组、人参皂苷Rb167 mg/kg剂量组。除对照组外,其余大鼠每天随机接受1~2种不同的刺激,造模时间共35 d。于第36天进行糖水偏爱、旷场实验、新奇环境摄食抑制实验、大鼠高架十字迷宫实验、强迫游泳等行为学实验,考察其抗抑郁、抗焦虑作用。采用ELISA法测定大鼠血清和海马IL⁃1β、IL⁃6、TNF⁃α炎症因子水平,血清皮质酮(CORT)水平。结果与模型组相比,人参皂苷Rg1、Rb1组大鼠糖水偏爱指数提升,强迫游泳不动时间显著减少,人参皂苷Rg148 mg/kg剂量组新奇抑制摄食潜伏期显著减少,人参皂苷Rg1(24和48 mg/kg)剂量组开臂时间的比例显著上升,人参皂苷Rg1、Rb1两个剂量组血清中皮质酮的含量显著减少,人参皂苷Rg124 mg/kg剂量组血清中IL⁃1β和IL⁃6的水平显著降低,人参皂苷Rb133 mg/kg剂量组血清中TNF⁃α和IL⁃6的水平显著降低,人参皂苷Rg1(48 mg/kg)、Rb1(67 mg/kg)剂量组海马中IL⁃1β、IL⁃6和TNF⁃α的含量显著降低。结论两种人参皂苷均可能通过调节HPA轴、抑制神经炎症,从而改善慢性不可预测应激致大鼠抑郁、焦虑样行为,此外人参皂苷Rg1的抗焦虑作用优于Rb1。