Effect of hydroxysafflor yellow A on depressive-like behavior and expression of GABAAR protein in hippocampus of chronic restraint stress model mice / 中华行为医学与脑科学杂志

Objective:To investigate the effects of hydroxysafflor yellow A (HSYA) on depressive-like behavior and expression of type A γ-aminobutyric acid receptor(GABAAR)in hippocampus of chronic restraint stress model mice.Methods:The SPF grade male C57BL/6C mice were divided into Control group, HSYA group, Model group, Model + HSYA group and Model + fluoxetine group according to random number table method, with 12 mice in each group.Mice model of depression was established by chronic restraint stress.Mice in HSYA group and Model+ HSYA group were intraperitoneally injected with HSYA(20 mg/kg), mice in Model+ fluoxetine group were injected intraperitoneally with fluoxetine (10 mg/kg), and mice in Control group and Model group administered with 0.9% sodium chloride solution intraperitoneally once a day for 14 days.Then, the forced swimming test (FST) and tail suspension test (TST) were performed to evaluate the depressive-like behavior of mice, and the protein expression levels of different subtypes of GABAAR in the hippocampus of mice were determined by Western blot.SPSS 19.0 and GraphPad Prism 8.0 software were used for data statistical analysis and mapping.One-way ANOVA was used for comparison among groups, and Tukey-HSD test was used for further pairwise comparison.Results:(1) In the behavioral tests, there were significant differences in swimming immobility time of FST and tail suspension immobility time of TST among the five groups ( F=21.59, 20.81, both P<0.05). The swimming immobility time ((143.91±9.97) s) and tail suspension immobility time (( 107.00±6.54) s) in Model group were higher than those in Control group ((52.92±6.70) s, ( 43.50±5.96) s, both P<0.05). There were no significant difference in swimming immobility time and tail suspension immobility time between Model+ HSYA group ((26.17±7.69)s, ( 20.17±7.89)s) and Model+ fluoxetine group ((61.60±16.22)s, (34.14±10.74)s)(both P>0.05), but the swimming immobility time and tail suspension immobility time in these two groups were lower than those in Model group (both P<0.05). (2) The Western blot results showed that there were significant differences in the expression of GABAARβ1 and GABAARβ2 protein in hippocampus among the four groups ( F=12.21, 11.40, both P<0.05). The expression levels of GABAARβ1(45.60±10.76) and GABAARβ2 (46.27±4.82) protein in hippocampus of Model group were lower than those in Control group ((100.00±3.44), (100.00±3.26), both P<0.05). Compared to Model group, the expression of GABAARβ1 (79.91±5.00) and GABAARβ2 (79.08±5.53) protein in hippocampus of Model+ HSYA group were higher (both P<0.05). In addition, the expression of GABAARα1 and GABAARγ1 proteins in hippocampus were not significantly different among the four groups( F=0.23, 0.10, both P>0.05). Conclusion:HSYA can effectively alleviate depressive-like behavior in depression model mice, which may be related with the upregulation of GABAARβ1 and GABAARβ2 of hippocampus tissue.

Expression and role of STING-TBK1-IRF3 signaling pathway in the brain of mice with chronic stress / 西安交通大学学报(医学版)

【Objective】 To explore the expression and role of stimulator of interferon gene (STING)-TANK-binding kinase 1 (TBK1)-interferon regulatory factor 3 (IRF3) signaling pathway in the brain of chronic stress mice. 【Methods】 Mice were divided into control (CON) group and chronic restraint stress (RST) group. Mice in the RST group were given chronic restraint stress stimulation (6 hours per day, 14 days). After 14 days, the mRNA expressions of pro-inflammatory cytokines CCL2, CXCL10, IL-1β, IL-6, IL-10, and TNFα in the brain were detected and analyzed by qRT-PCR; protein expression of STING, TBK1, p-TBK1, IRF3, and p-IRF3 were detected and analyzed by immunofluorescence staining and Western blotting. 【Results】 Compared to the CON group, the mRNA expressions of pro-inflammatory cytokines in the RST group were significantly increased (P<0.05). STING and microglia marker Iba-1 were highly co-located and the expression of STING was decreased as detected by immunofluorescence staining. Moreover, the protein expressions of STING, p-TBK1, and p-IRF3 were significantly decreased (all P<0.01). 【Conclusion】 Chronic restraint stress triggers a neuroinflammatory response and the STING-TBK1-IRF3 pathway in the brain of the RST mice is significantly inhibited.

Effects of Chronic Restraint Stress on Apoptosis of Amygdala Cells in Rats / 法医学杂志

OBJECTIVES@#To explore the damage effects of chronic restraint stress (CRS) on amygdala cells through the rat CRS model.@*METHODS@#The rat CRS model was established, and the changes in body weight and adrenal mass in control group and CRS group were monitored at 1 d, 7 d, 14 d and 21 d. The behavior changes were evaluated by the percentage of retention time of open arms and open arm entries using the elevated plus maze (EPM). ELISA was used to detect the concentrations of rat's corticotropin releasing hormone (CRH), adrenocorticotropic hormone (ACTH) and cortisol. The changes of expression of glucocorticoid receptor (GR) and glial fibrillary acidic protein (GFAP) in amygdala were determined by immunohistochemistry and Western blotting. Ultrastructure changes of glial cell were observed by transmission electron microscopy. The apoptosis rate of amygdala was measured by flow cytometry.@*RESULTS@#Compared with the control group at the same time points, body weight of CRS 1 d, 7 d, 14 d and 21 d groups increased slowly, but adrenal mass increased significantly; the serum level of CRH, cortisol and ACTH increased significantly at 7 d, 14 d and 21 d respectively; the expression of GR in amygdala was increased while that of GFAP was decreased; EPM test suggested that the percentage of retention time of open arms and open arm entries decreased significantly after 14 d. The CRS group showed different degrees of glial cell damage in amygdala, and the apoptosis rate of glial cell was significantly increased in 21 d group.@*CONCLUSIONS@#This study successfully established a CRS model in rats, and anxiety-like behavioral changes in model rats may be caused by apoptosis of amygdala astrocytes.

Effect of Shenwei Ningyu Pills on Immunoinflammatory Response Mediated by TLR4/MyD88 Signaling Pathway in Hippocampus of Rats Exposed to Chronic Restraint Stress / 中国实验方剂学杂志

Objective:To explore the effects of Shenwei Ningyu pills （SNP）， a new Chinese medicine for depression， on the immunoinflammatory response mediated by Toll-like receptor 4 （TLR4）/myeloid differentiation factor 88 （MyD88） signaling pathway in the hippocampus of rats exposed to chronic restraint stress （CRS）. Method:Forty-four male Sprague Dawley rats were randomly enrolled into a normal group， a model group， an escitalopram group， and an SNP group. Except for the rats in the normal group， all rats were exposed to CRS and isolated rearing for 21 days continuously. Rats in the escitalopram group and the SNP group were administered with escitalopram （30 mg·kg^-1） and SNP （18 mg·kg^-1） one hour prior to CRS， respectively. The changes in body weight， sucrose preference index， horizontal movement scores， and vertical movement scores were observed by body weight assessment， sucrose preference test， and open field test. The expression of hippocampal TLR4 and MyD88 was detected by Western blot. The content of serum interleukin-1<italic>β</italic> （IL-1<italic>β</italic>）， IL-10， and tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>） was detected by enzyme-linked immunosorbent assay （ELISA）. Result:The results of the behavioral assessment showed that there was no significant difference in the changes of behavioral baselines among the groups before intervention. However， significant differences were found among the groups following different interventions. The body weight， sugar preference index， horizontal movement score， and vertical movement score of rats in the model group decreased after CRS for 21 days as compared with those in the normal group （<italic>P</italic><0.01）. The above indicators in the SNP<italic> </italic>group and the escitalopram group were higher than those in the model group （<italic>P</italic><0.01）， which indicated that SNP<italic> </italic>exerted an obvious antidepressant effect. The results of Western blot and ELISA showed that compared with the normal group， the model group showed elevated levels of hippocampal TLR4 and MyD88 and serum IL-1<italic>β</italic> and TNF-<italic>α </italic>（<italic>P</italic>˂0.01） and dwindled serum IL-10 （<italic>P</italic>˂0.01）， while SNP<italic> </italic>and escitalopram reversed the conditions in the model group （<italic>P</italic>˂0.01） except for TNF-<italic>α</italic>. Conclusion:The present study indicated that the antidepressant effect of SNP was presumedly achieved by inhibiting the immunoinflammatory response mediated by the TLR4/Myd88 signaling pathway in CRS rats.

Changes of Microglia in Hippocampus of Mice Induced by Maternal Separation with Restraint Stress and Regulatory Effect of Wenyang Jieyu Prescription / 中国实验方剂学杂志

Objective:To observe the activation of microglia in hippocampus of depressed and anxious mice induced by maternal separation with acute restraint stress and the expression of interleukin-1<italic>β</italic>（IL-1<italic>β</italic>），interleukin-6（IL-6），tumor necrosis factor-<italic>α</italic>（TNF-<italic>α</italic>）， investigating the mechanism of Wenyang Jieyu prescription in treating anxiety and depression. Method:Eighty four male C57BL offspring were randomly divided into control group， acute restraint stress group and model group on postnatal day 0（PD0）. Maternal separation combined with acute restraint stress was used to prepare anxious and depressed model mice， dividing the model mice into model group， Wenyang， Jieyu， Wenyang Jieyu and fluoxetine group according to random number table method. During the period of PD21-PD90， the control， acute restraint stress and model mice were fed with normal diet， with the other groups fed with corresponding medicine mixed diet. The Wenyang， Jieyu and Wenyang Jieyu groups were given 5.85， 12.03 and 16.71 g·kg^-1·d^-1 respectively. The fluoxetine group was given 2.60 mg·kg^-1·d^-1. Open field， zero maze test and social interaction tests were used to evaluate the anxiety and depression of model mice. The expression of Iba-1 in hippocampal microglia was detected by immunohistochemistry（IHC）. The mRNA expression of IL-1<italic>β</italic>， IL-6， TNF-<italic>α</italic>， Iba-1 and glucocorticoid receptor（GR）were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. Result:Compared with the control group， total movement distance and time spent in central zone in 5 min of the model mice significantly decreased（<italic>P</italic><0.01）， time spent in opened arm and total movement distance decreased significantly（<italic>P</italic><0.05，<italic>P</italic><0.01）， investigation time during testing and training increased significantly（<italic>P</italic><0.01）. The expression of Iba-1 protein and mRNA，IL-1<italic>β</italic>，IL-6，TNF-<italic>α</italic> mRNA significantly increased（<italic>P</italic><0.01）， the expression levels of GR mRNA significantly decreased（<italic>P</italic><0.01）. The result of IHC staining showed that microglia were over activated. Compared with the model group， total movement distance and time spent in central zone in 5 min of mice in the Wenyang Jieyu and fluoxetine group significantly increased（<italic>P</italic><0.05，<italic>P</italic><0.01）.Time spent in opened arm significantly increased（<italic>P</italic><0.01）. Investigation time during testing and training significantly decreased（<italic>P</italic><0.05，<italic>P</italic><0.01）. The expression of Iba-1 protein and mRNA，IL-1<italic>β</italic>，IL-6，TNF-<italic>α</italic> mRNA significantly decreased（<italic>P</italic><0.05，<italic>P</italic><0.01）. The expression of GR mRNA increased significantly（<italic>P</italic><0.05，<italic>P</italic><0.01）. IHC staining showed the microglia recovered. Time spent in opened arm of mice in the Wenyang group and Jieyu group significantly increased （<italic>P</italic><0.01）， time spent investigating during testing decreased significantly （<italic>P</italic><0.05）， the expression levels of Iba-1 protein and mRNA，IL-6 mRNA significantly decreased （<italic>P</italic><0.05，<italic>P</italic><0.01）. The expression of GR mRNA of mice in the Wenyang group significantly increased （<italic>P</italic><0.05）， the expression of TNF-<italic>α </italic>mRNA significantly decreased （<italic>P</italic><0.05）. Total movement distance of mice in the Jieyu group increased significantly （<italic>P</italic><0.01）， time spent investigating during training decreased significantly （<italic>P</italic><0.05），the expression level of IL-1<italic>β </italic>mRNA significantly decreased （<italic>P</italic><0.05）. IHC staining showed that microglia recovered partly in both groups. Conclusion:The comprehensive curative effect and pharmacological action of Wenyang Jieyu prescription were better than Wenyang prescription and Jieyu prescription. Wenyang Jieyu prescription can treat anxiety and depression in maternal separation and acute restraint stress mice， its possible mechanism may be related to the decreased activation of microglia， down-regulation of IL-1<italic>β</italic>，IL-6，TNF-<italic>α</italic> expression and up-regulation of GR expression.

The Effect of Restraint Stress on the Reactivation of Colitis in Acetic Acid-Induced Ulcerative Colitis Rat

Aim: To evaluate the effect of restraint stress on the reactivation of ulcerative colitis in rats that have been chemically-induced with ulcerative colitis after allowing progression of natural healing to take place. Study Design: 36 animals were used for this study, and were divided into three groups; control (group 1), colitis alone (group 2) and colitis + restraint stress (group 3). Methodology: 36 healthy female wistar rats of weight of 170±20 were used. Colitis was induced by intra-rectal administration of 1 mL/100 g body weight of 7% acetic acid. Fourteen days post colitis induction, colitis + restraint stress group animals were exposed to restraint stress for six consecutive days while animals in colitis alone group were not. Colon levels of Malondialdehyde (MDA), Glutathione (GSH) concentration, Superoxide dismutase (SOD), Catalase (CAT) and Myeloperoxidase (MPO) activities were measured spectrophometrically. Histological assessment was also done on the colon tissues. Place and Duration of Study: Department of Physiology, Ladoke Akintola University of Technology, Ogbomoso, Oyo State, Nigeria, between April 2019 and August 2019. Results: Colitis caused significant increase in the levels of GSH, SOD, CAT and MPO on the 3rd day. On the 14th day, group 2 showed lowered level of antioxidant activities (SOD and CAT) when compared with the 3rd day, while on the 20th day, group 3 animals exhibited increased levels of antioxidant activities (as well as increase in MDA and MPO) when compared to group 2 and group 1. Histological examination revealed cyst in the mucosal layer, infiltration of inflammatory cells anddilation of vessels in the colitis + restraint stress on the 20th day post colitis induction. Results are considered significant when p≤0.05. Conclusion: The results of this study showed that restraint stress is capable of reactivating and inhibiting colitis healing in acetic acid-induced ulcerative colitis rats

Antidepressant-like effects of albiflorin involved the NO signaling pathway in rats model of chronic restraint stress / 中国天然药物

The depressant-like effects of albiflorin (AF) were studied on stressed chronic restraint stress (CRS) rats. Experimental rats were subjected to immobilization stress for a daily 6 h-restraining in a plastic restrainer for continuous 21 d and were treated with 30 or 15 mg·kg

Resveratrol Reverses Brain Glutathione System Involved Neuronal Loss After Immobilization Stress

Background: This study evaluated the antioxidant effects of Resveratrol on stress-induced neuronal loss in ratbrain involving brain glutathione system.Materials and Methods: The control rats received vehicle while another set of rats received 21 days restraintstress. The third and fourth group received similar intensity of restraint stress as well as either 10 or 20mg/kgdose of resveratrol respectively. The cognitive test included passive avoidance test. This was followed by estimationof reduced glutathione and glutathione reductase enzyme levels in brain homogenate and histomorphologicalstudy of hippocampus and medial prefrontal cortex.Results: Restraint stress has resulted in poor retrieval of learning behaviour and resveratrol has enhancedretrieval of learning behaviour in stressed condition in passive avoidance task. Both reduced glutathione andglutathione reductase levels were reduced after restraint stress and resveratrol at both the doses has normalizedtheir levels. Restraint stress has affected CA3, CA2 and dentate regions of the hippocampus and also medialprefrontal cortex. In all these areas resveratrol has minimized neuronal loss which were due to chronic stress.Conclusion: From the results of the present study we conclude that stress induced oxidative damage involvesbrain glutathione system and which in turn could be one of the causes for neuronal loss and resveratrol suggeststo protect the brain against stress in rat model.

An experimental investigation of the effects of chronic stress on bone-to-implant contact

OBJECTIVES: This study aimed to investigate the effects of chronic restraint stress on the osseointegration of titanium implants. MATERIALS AND METHODS: Twenty adult male Wistar albino rats were used in the study. After surgical insertion of titanium implants into the metaphyseal part of the tibial bone, rats were randomly divided into two groups: a control group (CNT group) and an experimental restraint stress group (RS group). In the CNT group, titanium implants were inserted surgically, and rats received no further treatment during the 47-day experimental period. In the RS group, restraint stress was applied for 3 hours per day for 45 days, beginning 2 days after implant surgery. Weight of the rats was measured prior to surgery and at the end of the study to analyze the effects of stress. At the end of the experimental period, rats were euthanized, and implants and surrounding bone tissues were used for undecalcified histological analysis. Serum cortisol levels were assessed in cardiac blood samples from the rats following centrifugation. RESULTS: Average weight of rats in the RS group was lower than that of rats in the CNT group after the experimental protocol had been completed (P0.05). CONCLUSION: The data analyzed in this study suggest that chronic restraint stress did not adversely affect rats during a 45-day osseointegration period.

The role of Nox-4 in gastric mucosal inflammation induced by chronic restraint stress in mice / 中华行为医学与脑科学杂志

Objective To investigate the expression of NADPH oxidase Nox-4 induced by stress in gastric mucosa and its role in inflammation.Methods Twenty male SPF Kunming mice were randomly divided into chronic restraint stress group(stress group) and control group.Stress mice were restrained in selfmade restraint device for 2 hours each day.The rest of the time,the mice in the two groups had free access to food and water normally,experiment lasted 14 days.The histopathological changes of gastric mucosa were assessed by HE staining under light microscope.The expression of Nox-4 in gastric mucosa of mice was carried out by immunohistochemical method.The relative expression levels of Nox-4,antioxidant protein (Mn-SOD,GSH,Catalase) and inflammatory factors(IL-8,IL-1β,TNF-α) in gastric mucosa were detected by real-time quantitative RT-PCR and ELISA.Results Basal cell proliferation,neutrophil,eosinophil and plasma cell infiltration and inflammatory changes were observed in the lamina propria and glandular epithelium of stress mice,while no obvious abnormalities were found in control mice.The expression of Nox-4 in stress group was deeper and more abundant than that in control group,mainly expressed in lamina propria and glandular epithelium.The mRNA expression levels of Nox-4 in gastric mucosa of stress group was(2.42±0.51) times higher than that of control group,and blood concentration of stress group was(2.23±0.67) times higher than that of control group(t=-46.32,P＜0.001).The RT-PCR of antioxidant proteins in gastric mucosa showed that the transcription levels of Mn SOD,GSH and Catalase in stress group were significantly lower than that of control group (Mn-SOD:0.59± 0.10,GSH:0.58± 0.11,Catalase:0.57± 0.09),and there were significant differences between the two groups(t=13.57,11.67,15.01,P＜0.01).RT-PCR results showed that the transcription levels of IL-8,IL-1β,TNF-α in stress group were significantly higher than those in control group (IL-8:1.47±0.34,IL-1β:1.48 ± 0.42,TNF-α:1.51 ± 0.37),and there were significant differences in two groups(t=-18.45,-19.14,-20.85,P＜0.01).ELISA results showed that the serum levels of inflammatory factors in stress group were significantly higher than those in control group(2.25±0.37,3.59±0.45,3.41±0.34),and the differences were statistically significant(t=-47.11,-79.36,-96.32,P＜0.01).Pearson correlation analysis showed that there was a positive correlation between serum concentration of Nox-4 and inflammatory factors(IL-8,IL-1β,TNF-αt) in stress group(r=0.97,0.99,0.98,P＜0.01).Spearman rank correlation analysis showed that the grade of gastric mucosal inflammation was positively correted with serum levels of Nox-4 and inflammatory factors (IL-8,IL-1β,TNF-α) (r =0.96,0.92,0.91,0.94,all P＜ 0.01)Conclusion Stress may lead to gastric mucosal lesion by overexpression of proinflammatory factors through destroying the balance of oxidation/antioxidant system in gastric mucosa.

Establishment and comparison of anxiety models by uncertain empty water bottle stimulation and restraint stress in mice / 中国药理学与毒理学杂志

OBJECTIVE To screen a mouse anxiety model with good reproducibility, little stimulation and user-friendliness by comparing the uncertain empty bottle drinking water stimulation with restraint stress to induce anxiety-like emotion in mice in order to find a suitable pharmacodynamic evaluation model for anti-anxiety drugs which provides an experimental basis. METHODS A mouse anxiety model was established using the uncertain empty bottle drinking water stimulation method. The mice were divided into freely drinking water group, regular drinking water group, physiological stress group and empty bottle stimulation group, with eight mice in each group. The freely drinking water group was free to eat and drink all day, the regular drinking water group mice were given free water for 2 periods per day, while the regular drinking water group mice were given water for 10 min at 2 fixed time with a 12 h interval every day. At the same time, physiological stress group mice had access to drinking water for 10 min once a day, but at another time they were given neither water nor empty bottles. The empty bottle stimulation group mice were given empty bottle stimulation randomly once or twice every day so that they drank water 12 times and empty bottle stimulation 16 times. The experiment lasted 14 d. The mouse anxiety model was established with restraint stress method. The experimental mice were divided into normal control group and restraint stress group, with eight mice in each group. The normal control group was normally reared, while the restraint stress group mice were confined to the tube at 9:00 every morning, headed towards the bottom of the centrifuge tube and were restrained for 2 h on the first day, and then the daily binding strength was extended for 2 h until 8 h every day for 21 d. The changes of body mass in mice after 7 and 14 d of empty bottle stimulation and after 7, 14, and 21 d of restraint stress were observed. At the end of the modeling, the mice were tested for autonomous activities and exploration behaviors through the open field test. The elevated plus maze test was used to detect the number of times they entered the open arms and the time spent in the open arms. The plasma corticosterone (CORT) content and the content of serotonin (5-HT) in the hippocampus of mice were determined by ELISA after 21 d of restraint stress. RESULTS Uncertain empty bottle drinking water stimulation method: compared with the freely drinking water group, the regular drinking water group showed a significant decrease in the residence time in the open arm (P<0.05). Compared with the regular drinking water group, the body mass of the mice decreased significantly after 7 d of empty bottle stimulation (P<0.05). There was no significant change in the behavioral indicators. Restraint stress method: compared with the normal control group, the body mass of mice was significantly decreased after 14 d of restraint stress (P<0.05). There was no significant change in the autonomous activity or exploration behavior of mice. The number of times the mice entered the open arms and the residence time in the open arms were significantly decreased (P<0.05). After 21 d of restraint stress, the body mass of the mice decreased significantly (P<0.05), and plasma CORT content (P<0.01) and 5-HT content in hippocampus (PO.01, P<0.05) of mice were significantly reduced. CONCLUSION The mouse anxiety model established with the uncertain empty bottle drinking water stimulation method results in poor stability and sensitivity, with more uncontrollable factors. The mouse anxiety model was not successfully established until 14 d of stimulation. The restraint stress for 14 d could successfully a establish mouse anxiety model, and the mechanism may be related to HPA axis dysfunction and 5-HT metabolic abnormalities.

Persistent hyperglycemia induced by chronic restraint stress in rat is associated with nucleus tractus solitarius injury / 解剖学报

Objective: To investigate the role of anterior part of commissural subnucleus of nucleus tractus solitarius (acNTS) injury in insulin-resistant hyperglycemia during chronic restraint stress (CRS). Methods: We produced the CRS models (n = 20, a 7-day restraint followed by a 3-day free moving procedure for 40 days) in rats, and detected the parameters related to glucose metabolism. Results: The CRS induced a moderate (not higher than 11 mmol/L) and irreversible insulin-resistant hyperglycemia in about 1/3 (n = 7) of the individuals. CRS-hyperglycemic rats showed a condensed staining of acNTS neurons, and Caspase-3 immunostaining and TUNEL also showed positive, indicating apoptotic changes of acNTS neurons. After acNTS mechanical damage (n= 6), the blood glucose level rised gradually, which also led to insulin-resistant hyperglycemia. The characteristics of hyperinsulinemia, increased islet volume, and serum corticosterone levels in acNTS mice were consistent with those of CRS mice. Conclusion: The result indicates that during CRS, injury (apoptosis) of glucose-sensitive acNTS neurons causes dysregulation of blood glucose. Restraint stress model has value as a potential application in the study of stress-induced hyperglycemia.

Acute Restraint Stress Augments 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine Neurotoxicity via Increased Toxin Uptake into the Brain in C57BL/6 Mice / 神经科学通报·英文版

As an environmental risk factor, psychological stress may trigger the onset or accelerate the progression of Parkinson's disease (PD). Here, we evaluated the effects of acute restraint stress on striatal dopaminergic terminals and the brain metabolism of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), which has been widely used for creating a mouse model of PD. Exposure to 2 h of restraint stress immediately after injection of a low dose of MPTP caused a severe loss of striatal dopaminergic terminals as indicated by decreases in the dopamine transporter protein and dopamine levels compared with MPTP administration alone. Both striatal 1-methyl-4-phenylpyridinium ion (MPP) and MPTP concentrations were significantly increased by the application of restraint stress. Striatal monoamine oxidase-B, which catalyzes the oxidation of MPTP to MPP, was not changed by the restraint stress. Our results indicate that the enhanced striatal dopaminergic terminal loss in the stressed mice is associated with an increase in the transport of neurotoxin into the brain.

Effects of chronic restraint stress on abilities of spatial learning and memory and levels of excitatory amino acids in hippocampal dentate gyrus in old rats / 吉林大学学报(医学版)

Objective:To explore the effects of chronic restraint stress (CRS) on the abilities of spatial learning and memory and the levels of excitatory amino acids in the hippocampal dentate gyrus (DG) in the old rats,and to investigate the neurochemical mechanism of CRS in affecting the spatial learning and memory abilities.Methods:Sixteen male SD rats (18 months old) were randomly divided into control group (n =8) and CRS group (n=8),and the rats in CRS group received CRS 2 h every day for 30 d.And then the spatial learning and memory abilities of rats were measured by Morris water maze (MWM) test,and the extracellular levels of excitatory amino acids including asparate (Asp) and glutamate (Glu) in the DG were simultaneously determined by in vivo microdialysis and HPLC.The levels of corticosterone (CORT) and epinephrine (EPI) in serum of the rats wereexamined by ELISA assay.Results:In CRS group,the escape latencies on the 2nd-4th days were significantly increased and the percentage of time spent in target quadrant on the 5th day was markedly decreased in MWM test compared with control group (P＜0.05).Compared with before training,the extracelluar level of Asp in the DG in control group was significantly increased on the 2nd day in MWM test;compared with control group,the extracelluar level of Asp in the DG in CRS group was significantly decreased on the 3rd day in MWM test (P＜0.05).Compared with before training,the Glu levels in the DG in MWM test in both control and CRS groups were markedly increased (P＜0.05),but there was no significant difference between two groups (P＞0.05).Compared with control group,the levels of CORT and EPI in the serum of the rats in CRS group were significantly increased (P＜0.05).Conclusion:CRS impairs the spatial learning and memory abilities in the old rats,which may be related to the decrease of Asp level in the hippicampal DG of the rats.

Repeated restraint stress promotes hippocampal neuronal cell ciliogenesis and proliferation in mice / 한국실험동물학회지

Stress severely disturbs physiological and mental homeostasis which includes adult neurogenesis in hippocampus. Neurogenesis in hippocampus is a key feature to adapt to environmental changes and highly regulated by multiple cellular signaling pathways. The primary cilium is a cellular organelle, which acts as a signaling center during development and neurogenesis in adult mice. However, it is not clear how the primary cilia are involved in the process of restraint (RST) stress response. Using a mouse model, we examined the role of primary cilia in repeated and acute RST stress response. Interestingly, RST stress increased the number of ciliated cells in the adult hippocampal dentate gyrus (DG). In our RST model, cell proliferation in the DG also increased in a time-dependent manner. Moreover, the analysis of ciliated cells in the hippocampal DG with cell type markers indicated that cells that were ciliated in response to acute RST stress are neurons. Taken together, these findings suggest that RST stress response is closely associated with an increase in the number of ciliated neurons and leads to an increase in cell proliferation.

Acute toxicity of total polymethoxyflavones from Bauhinia championii in mice and its effects on gastric ulcer / 中草药

Objective To study the acute toxicity of total polymethoxyflavones from Bauhinia championii (PMFBC) in mice and its effect on experimental gastric ulcer. Methods The acute toxicity of PMFBC was investigated by Bliss method. Three kinds of gastric ulcer model including indomethacin, ethanol, and water immersion-restraint stress were used to observe the effect of PMFBC in mice. Results The LD50 value of ig administration PMFBC on mice was 1 252.319 mg/kg, which showed acute toxicity; PMFBC had protective effect on three gastric ulcer models, which could significantly reduce the ulcer area of the mice, and had a high inhibition rate. Conclusion PMFBC has obvious inhibition effect on the fomation of gastric ulcer in mice, which can be safty used in a certain dose range.

Effect of Acupuncture on Expression of Glial Fibrillary Acidic Protein in Hippocampus and Prefrontal Cortex and Serum Interleukin-10 in Chronic Restraint Stress Depression Rats / 针刺研究

OBJECTIVE: To observe the effect of acupuncture on the expression of glial fibrillary acidic protein (GFAP) in hippocampus and prefrontal cortex and the level of serum interleukin 10 (IL-10) in rats with depression, so as to explore its mechanism underlying improvement of depression. METHODS: Thirty-two male SD rats were randomly divided into four groups: control, model, acupuncture, and medication (Fluoxetine, Flu) (n=8 rats in each). The depression model was established by using chronic restraint stress (constraint, fasting, water deprivation, etc.) combined with solitary raising for 28 days. Acupuncture was applied to "Baihui" (GV 20) and "Yintang" (GV 29), and bilateral "Sanyinjiao" (SP 6) for 20 min, once daily for 28 days. Fluoxetine (1.8 mg/kg) was given to rats of the medication group by gavage once every day for 28 days. Sucrose consumption test and open field test (crossing and rearing locomotion) were carried out to evaluate the behavioral changes. Western blot was used to detect the expression of GFAP in the hippocampus and prefrontal cortex, and the content of serum IL-10 was detected by enzyme linked immunosorbent assay (ELISA). RESULTS: After modeling, the sucrose consumption, the crossing numbers and rearing times, hippocampal GFAP protein expression and serum IL-10 content were significantly decreased and prefrontal GFAP protein expression was up-regulated markedly in the model group relevant to the control group (P0.05) and in down-regulating prefrontal GFAP protein expression (P>0.05 ) except up-regulation of IL-10 level. CONCLUSION: Acupuncture intervention plays a positive role in anti-depression in rats, which may be related to its effects in regulating the expression of GFAP in the hippocampal and prefrontal astrocytes, and in increasing the content of serum anti-inflammatory cytokine IL-10.

Effects of chronic restraint stress on abilities of spatial learning and memory and levels of excitatory amino acids in hippocampal dentate gyrus in old rats / 吉林大学学报(医学版)

Objective: To explore the effects of chronic restraint stress (CRS) on the abilities of spatial learning and memory and the levels of excitatory amino acids in the hippocampal dentate gyrus (DG) in the old rats, and to investigate the neurochemical mechanism of CRS in affecting the spatial learning and memory abilities. Methods: Sixteen male SD rats (18 months old) were randomly divided into control group (n=8) and CRS group (n=8), and the rats in CRS group received CRS 2 h every day for 30 d. And then the spatial learning and memory abilities of rats were measured by Morris water maze (MWM) test, and the extracellular levels of excitatory amino acids including asparate (Asp) and glutamate (Glu) in the DG were simultaneously determined by in vivo microdialysis and HPLC. The levels of corticosterone (CORT) and epinephrine (EPI) in serum of the rats were examined by ELISA assay. Results: In CRS group, the escape latencies on the 2nd-4th days were significantly increased and the percentage of time spent in target quadrant on the 5th day was markedly decreased in MWM test compared with control group (P0. 05). Compared with control group, the levels of CORT and EPI in the serum of the rats in CRS group were significantly increased (P<0. 05). Conclusion: CRS impairs the spatial learning and memory abilities in the old rats, which may be related to the decrease of Asp level in the hippicampal DG of the rats.

Study on mechanism of nourishing blood and smoothing liver effects of Paeoniae Radix Alba based on metabolomics information / 中草药

Objective: To investigate the nourishing blood and smoothing liver effects of Paeoniae Radix Alba (PRA) based on metabolomics information. Methods: The stagnation of liver qi and blood deficiency syndrome rats model was established by chronic restraint stress combined with radiation. Using LC/MS method as the core technology, the principal component analysis (PCA) and partial least squares discriminate analysis (PLS-DA) as the main data analysis methods, endogenous metabolites were screened in the model rats to study the intervention mechanism of PRA. Results: Through the impact of phosphatidylcholine, lysophosphatidylcholine, ceramide, deoxycytidine, betaine, and other 21 kinds of small molecular metabolites, PRA had a certain callback effect on the disturbance of metabolic trajectory, which can improve the state of stagnation of liver qi and blood deficiency induced by external stimulating factors (such as irradiation, restraint, and loneliness). Conclusion: The nourishing blood and smoothing liver effects of PRA may be related to the sphingolipid metabolism, glycerophospholipid metabolism, linoleic acid metabolism, and alpha-linolenic acid metabolism.

Effects of caspase-1 inhibitor VX765 on cold-restraint stress-induced acute gastric ulcer in mice / 中华烧伤杂志

Objective@#To investigate the protective effects of caspase-1 inhibitor VX765 on gastric mucosa of mice with cold-restraint stress-induced acute gastric ulcer.@*Methods@#Twenty-four specific pathogen free male C57BL/6J mice were divided into normal control group (NC), cold restrain group (CR), VX765 pre-treatment+ cold restrain group (VCR), and rabeprazole pre-treatment+ cold restrain group (RCR) according to the random number table, with 6 mice in each group. Mice in group NC were injected intraperitoneally with solution of 10 mL/kg dimethylsulfoxide (DMSO) and phosphate buffer solution (PBS). Mice in group CR were inflicted with acute gastric ulcer induced by cold-restraint stress 30 minutes after intraperitoneal injection of solution of DMSO and PBS. Mice in groups VCR and RCR were inflicted with acute gastric ulcer as above 30 minutes after intraperitoneal injection of solution of DMSO and PBS with dose of 12.5 μmol/kg containing 10 mg VX765 and 40 mg/kg containing 20 mg rabeprazole, respectively. Four hour after cold-restraint stress, serum content of tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6)was determined by enzyme-linked immunosorbent assay. Gross condition of gastric tissue was observed. Ulcer index was evaluated. Pathological change of gastric tissue was observed with HE staining. The relative expression of IL-1β, IL-18, and cleaved-caspase-1 in gastric tissue were detected by Western blotting. Mice in group NC were detected as above at the same time point. Data were processed with one-way analysis of variance and Bonferroni test.@*Results@#The serum content of TNF-α and IL-6 and the relative expression of cleaved-caspase-1, IL-1β, and IL-18 in gastric tissue of mice in group NC were significantly lower than those in group CR (with P values below 0.01). The content of the above-mentioned inflammatory indexes in serum and gastric tissue of mice in group VCR was significantly lower than that in group CR (with P values below 0.01). There were no statistically significant differences in content of the above-mentioned inflammatory indexes in serum and gastric tissue of mice between groups RCR and CR (with P values above 0.05). The content of the above-mentioned inflammatory indexes in serum and gastric tissue of mice in group VCR was significantly lower than that in group RCR (with P values below 0.01). Surface of gastric mucosa was smooth and morphology of mucosal cells was normal with clear structure of mice in group NC. Multiple hemorrhage of gastric mucosa, disorderly arrangement of mucosal cells, and large number of inflammatory cell infiltration around necrotic tissue were observed in mice of group CR. Decreased number of gastric mucosa bleeding, intact mucosal structure, and small amount of inflammatory cell infiltration around necrotic tissue were observed in mice of groups VCR and RCR. The ulcer indexes of mice in groups NC, CR, VCR, and RCR were 0, 18.7±1.1, 6.3±1.5, and 8.2±1.3, respectively. The ulcer index of mice in group NC was significantly lower than that in the other 3 groups (with P values below 0.05). The ulcer indexes of mice in groups VCR and RCR were close (P>0.05), which were significantly lower than ulcer index of mice in group CR (with P values below 0.05).@*Conclusions@#VX765 can effectively inhibit the activation of caspase-1, reduce production of inflammatory factor, and alleviate inflammatory response, which have protective effects on gastric mucosa of mice with cold-restraint stress-induced acute gastric ulcer.