The purpose of this study was to investigate the anxiety-like behaviors, circadian rhythms and sleep, and to elucidate the possible underlying mechanisms of the abnormal sleep behavior in Shank3 gene knockout (Shank3-KO) mice. The anxiety-like behaviors were detected by elevated plus-maze (EPM) test, open field test (OFT) and tail suspension test (TST). The circadian rhythms were detected by running wheel test. The electroencephalogram (EEG)/electromyogram (EMG) recordings were performed synchronically by polysomnograph. The distribution of SHANK3 in anterior cingulate cortex (ACC), paraventricular thalamus (PVT), nucleus accumbens (NAc), basolateral amygdala (BLA) and hippocampal CA2 region in wild type (WT) mice was detected by immunofluorescence assay. The protein expression of c-Fos in PVT, ACC and NAc was also detected by immunofluorescence assay during light cycle. The colocalization of c-Fos and vesicular glutamate transporter 2 (Vglut2, a marker for glutamatergic neurons) in the PVT was detected by immunofluorescence double labeling experiment. The results of EPM test showed that, compared with the WT mice, the Shank3-KO mice showed less time in open arms and less number of open arm entries. The results of OFT showed that the Shank3-KO mice showed less time in central area and less number of central area entries. The immobility time of Shank3-KO mice was increased in the TST. The results of running wheel rhythm test showed that the phase shift time of Shank3-KO mice in the continuous dark period was increased. The results of EEG/EMG recording showed that, compared with the WT mice, the duration of wakefulness in Shank3-KO mice was increased and the duration of non-rapid eye movement (NREM) sleep was decreased during light phase; The bout number of wakefulness was increased, the bout number of NREM sleep was decreased, NREM-wake transitions were increased, and wake-NREM transitions were decreased during light phase. SHANK3 was expressed in ACC, PVT, NAc and BLA in the WT mice. The expression of c-Fos in the PVT of Shank3-KO mice was up-regulated 2 h after entering the light phase, and majority of c-Fos was co-localized with Vglut2. These results suggest that the anxiety level of Shank3-KO mice is increased, the regulation of the internal rhythms is decreased, and the bout number of wakefulness is increased during light phase. The glutamatergic neurons in PVT may be involved in the regulation of abnormal sleep behavior in Shank3-KO mice during the light phase.
Animals ; Mice, Knockout ; Mice ; Neurons/metabolism* ; Nerve Tissue Proteins/physiology* ; Male ; Midline Thalamic Nuclei/cytology* ; Circadian Rhythm/physiology* ; Sleep/physiology* ; Anxiety/physiopathology* ; Proto-Oncogene Proteins c-fos/metabolism* ; Vesicular Glutamate Transport Protein 2/metabolism* ; Mice, Inbred C57BL ; Microfilament Proteins

OBJECTIVE: To elucidate the modulation mechanism of Suanzaoren Decoction (SZRD) on basolateral amygdala (BLA) neuronal activity to alleviate chronic restraint stress (CRS)-related behavioral deficits. METHODS: The male C57BL/6J mice were assigned to 4 groups using the complete randomization method, including control (CON, n=19), CRS (n=19), SZRD (n=21), and fluoxetine (Flu, n=22) groups. Mice were restrained for 6 h per day, over a 21-d period to establish CRS models. The CON group remained in their cages without food or water during the 6-h matching period. SZRD and Flu groups received intragastric administration of SZRD (4.68 g/kg) and Flu (20 mg/kg) daily, respectively, 30 min before restraint for 21 consecutive days. The therapeutic effects of SZRD were evaluated using behavioral tests including the tail suspension test, elevated plus maze test, and forced swimming test. The cellular Fletcher B. Judson murine osteosarcoma proto-oncogene (c-Fos) expression in the BLA was measured using immunofluorescence, while action potential (AP) firing and synaptic transmission in BLA pyramidal neurons were evaluated using whole-cell patch-clamp recordings. RESULTS: SZRD administration significantly increased time spent in the open arms and open-arm entries while reducing immobility time (P<0.05 or P<0.01). It downregulated CRS-induced c-Fos expression and AP firing of pyramidal neurons in the BLA (P<0.01). Additionally, SZRD selectively attenuated excitatory (P<0.01), but not inhibitory, synaptic transmission onto BLA pyramidal neurons. CONCLUSION SZRD alleviated CRS-induced anxiety- and depression-like behaviors in mice by modulating the excitability and synaptic transmission of BLA pyramidal neurons.
Animals ; Drugs, Chinese Herbal/therapeutic use* ; Depression/complications* ; Pyramidal Cells/pathology* ; Male ; Mice, Inbred C57BL ; Basolateral Nuclear Complex/pathology* ; Restraint, Physical ; Anxiety/complications* ; Behavior, Animal/drug effects* ; Stress, Psychological/physiopathology* ; Mice ; Proto-Oncogene Proteins c-fos/metabolism* ; Action Potentials/drug effects* ; Synaptic Transmission/drug effects*

OBJECTIVES: To investigate the mechanism by which the pyramidal neurons of the anterior cingulate cortex (ACC) modulate the effects of enriched environment (EE) for relieving anxiety-like behaviors in mice. METHODS: C57BL/6J mice were randomly divided into control group, restraint stress (RS) group, and RS+EE group (n=8). The mice in the latter two groups were subjected to RS for 2 h daily for 3 days, and those in RS+EE group were housed in an EE during modeling. Anxiety-like behaviors of the mice were evaluated using the elevated plus-maze tests (EPM) and open field test (OFT). Changes in c-Fos expression in the ACC of the mice were detected with immunofluorescence assay, and pyramidal neuron excitability in the ACC (Pyn^ACC) was measured using patch-clamp technique. The miniature excitatory and inhibitory postsynaptic currents (mEPSC and mIPSC, respectively) were analyzed to assess synaptic transmission changes. RESULTS: Behavioral tests showed obvious anxiety-like behaviors in RS mice, and such behavioral changes were significantly improved in RS+EE mice. Immunofluorescence staining revealed significantly increased c-Fos expression in the ACC in RS mice but lowered c-Fos expression in RS+EE group. Compared with the control mice, the RS mice showed increased action potential firing rate of Pyn^ACC, which was significantly reduced in RS+EE group. Compared with the RS mice, the RS+EE mice showed also decreased frequency of mEPSCs of Pyn^ACC, but the amplitude exhibited no significant changes. No obvious changes in the frequency or amplitude of mIPSCs were observed in RS+EE mice. CONCLUSIONS EE reduces excitability of Pyn^ACC to alleviate anxiety-like behaviors induced by RS in mice.
Animals ; Anxiety/physiopathology* ; Gyrus Cinguli ; Mice, Inbred C57BL ; Mice ; Pyramidal Cells/physiology* ; Restraint, Physical ; Stress, Psychological ; Proto-Oncogene Proteins c-fos/metabolism* ; Male ; Behavior, Animal ; Environment ; Excitatory Postsynaptic Potentials

Anxiety disorder is a major symptom of autism spectrum disorder (ASD) with a comorbidity rate of ~40%. However, the neural mechanisms of the emergence of anxiety in ASD remain unclear. In our study, we found that hyperactivity of basolateral amygdala (BLA) pyramidal neurons (PNs) in Shank3 InsG3680 knock-in (InsG3680^+/+) mice is involved in the development of anxiety. Electrophysiological results also showed increased excitatory input and decreased inhibitory input in BLA PNs. Chemogenetic inhibition of the excitability of PNs in the BLA rescued the anxiety phenotype of InsG3680^+/+ mice. Further study found that the diminished control of the BLA by medial prefrontal cortex (mPFC) and optogenetic activation of the mPFC-BLA pathway also had a rescue effect, which increased the feedforward inhibition of the BLA. Taken together, our results suggest that hyperactivity of the BLA and alteration of the mPFC-BLA circuitry are involved in anxiety in InsG3680^+/+ mice.
Animals ; Prefrontal Cortex/metabolism* ; Basolateral Nuclear Complex/metabolism* ; Mice ; Anxiety/metabolism* ; Nerve Tissue Proteins/genetics* ; Male ; Gene Knock-In Techniques ; Pyramidal Cells/physiology* ; Mice, Transgenic ; Neural Pathways/physiopathology* ; Mice, Inbred C57BL ; Microfilament Proteins

OBJECTIVE: To elucidate the specific mechanisms by which electroacupuncture (EA) alleviates anxiety and fear behaviors associated with posttraumatic stress disorder (PTSD), focusing on the role of lipocalin-2 (Lcn2). METHODS: The PTSD mouse model was subjected to single prolonged stress and shock (SPS&S), and the animals received 15 min sessions of EA at Shenmen acupoint (HT7). Behavioral tests were used to investigate the effects of EA at HT7 on anxiety and fear. Western blotting and enzyme-linked immunosorbent assay were used to quantify Lcn2 and inflammatory cytokine levels in the prefrontal cortex (PFC). Additionally, the activity of PFC neurons was evaluated by immunofluorescence and in vivo electrophysiology. RESULTS: Mice subjected to SPS&S presented increased anxiety- and fear-like behaviors. Lcn2 expression in the PFC was significantly upregulated following SPS&S, leading to increased expression of the proinflammatory cytokines tumor necrosis factor-α and interleukin-6 and suppression of PFC neuronal activity. However, EA at HT7 inhibited Lcn2 release, reducing neuroinflammation and hypoexcitability in the PFC. Lcn2 overexpression mitigated the effects of EA at HT7, resulting in anxiety- and fear-like behaviors. CONCLUSION EA at HT7 can ameliorate PTSD-associated anxiety and fear, and its mechanism of action appears to involve the inhibition of Lcn2-mediated neural activity and inflammation in the PFC. Please cite this article as: Yang YD, Zhong W, Chen M, Tang QC, Li Y, Yao LL, et al. Electroacupuncture alleviates behaviors associated with posttraumatic stress disorder by modulating lipocalin-2-mediated neuroinflammation and neuronal activity in the prefrontal cortex. J Integr Med. 2025; 23(5):537-547.
Electroacupuncture ; Stress Disorders, Post-Traumatic/metabolism* ; Animals ; Lipocalin-2/metabolism* ; Prefrontal Cortex/physiopathology* ; Male ; Mice ; Neurons/physiology* ; Disease Models, Animal ; Fear ; Behavior, Animal ; Mice, Inbred C57BL ; Neuroinflammatory Diseases/metabolism* ; Anxiety/therapy* ; Acupuncture Points

The nucleus accumbens (NAc) plays an important role in various emotional and motivational behaviors that rely on heightened wakefulness. However, the neural mechanisms underlying the relationship between arousal and emotion regulation in NAc remain unclear. Here, we investigated the roles of a specific subset of inhibitory corticotropin-releasing hormone neurons in the NAc (NAc^CRH) in regulating arousal and emotional behaviors in mice. We found an increased activity of NAc^CRH neurons during wakefulness and rewarding stimulation. Activation of NAc^CRH neurons converts NREM or REM sleep to wakefulness, while inhibition of these neurons attenuates wakefulness. Remarkably, activation of NAc^CRH neurons induces a place preference response (PPR) and decreased basal anxiety level, whereas their inactivation induces a place aversion response and anxious state. NAc^CRH neurons are identified as the major NAc projection neurons to the bed nucleus of the stria terminalis (BNST). Furthermore, activation of the NAc^CRH-BNST pathway similarly induced wakefulness and positive emotional behaviors. Taken together, we identified a basal forebrain CRH pathway that promotes the arousal associated with positive affective states.
Animals ; Septal Nuclei/metabolism* ; Nucleus Accumbens/physiology* ; Corticotropin-Releasing Hormone/metabolism* ; Wakefulness/physiology* ; Neurons/metabolism* ; Male ; Mice ; Mice, Inbred C57BL ; Neural Pathways/physiology* ; Anxiety/physiopathology* ; Reward

BACKGROUND: Depression and anxiety have been correlated with elevated risks for quality-of-life (QOL), adverse outcomes, and medical expenditure in patients with acute coronary syndrome (ACS). However, the relevant data are lacking for Chinese ACS populations, especially regarding different effects of major depression, anxiety, and comorbidity. The objective of this study was to evaluate the dynamic changes of depression and/or anxiety over 12 months and examine the effects of depression, anxiety, and comorbidity on QOL, adverse outcomes, and medical expenditure in Chinese patients with ACS. METHODS: For this prospective longitudinal study, a total of 647 patients with ACS were recruited from North China between January 2013 and June 2015. Among them, 531 patients (82.1%) completed 12-month follow-ups. Logistic regression model was utilized for analyzing the association of baseline major depression, anxiety, and comorbidity with 12-month all-cause mortality, cardiovascular events, QOL, and health expenditure. RESULTS: During a follow-up period of 12 months, 7.3% experienced non-fatal myocardial infarction (MI) and 35.8% cardiac re-hospitalization. Baseline comorbidity, rather than major depression/anxiety, strongly predicted poor 12-month QOL as measured by short-form health survey-12 (odds ratio [OR]: 1.77, 95% confidence interval [CI]: 1.22-2.52, P = 0.003). Regarding 12-month non-fatal MI and cardiac re-hospitalization, baseline anxiety (OR: 2.83, 95% CI: 1.33-5.89, P < 0.01; OR: 4.47, 95% CI: 1.50-13.00, P < 0.01), major depression (OR: 2.58, 95% CI: 1.02-6.15, P < 0.05; OR: 5.22, 95% CI: 1.42-17.57, P < 0.03), and comorbidity (OR: 6.33, 95% CI: 2.96-13.79, P < 0.0001, OR: 14.08, 95% CI: 4.99-41.66, P < 0.0001) were all independent predictors, and comorbidity had the highest predictive value. Number of re-hospitalization stay, admission frequency within 12 months and medical expenditure within 2 months were the highest in patients with ACS with comorbidity. CONCLUSIONS Major depression and anxiety may predict 12-month non-fatal MI and cardiac re-hospitalization. However, comorbidity has the highest predictive value with greater medical expenditure and worse QOL in Chinese patients with ACS. And depression with comorbid anxiety may be a new target of mood status in patients with ACS.
Acute Coronary Syndrome ; economics ; physiopathology ; Aged ; Anxiety ; physiopathology ; Depression ; physiopathology ; Female ; Humans ; Logistic Models ; Longitudinal Studies ; Male ; Middle Aged ; Myocardial Infarction ; economics ; physiopathology ; Prospective Studies ; Quality of Life

Medial orbitofrontal cortex (mOFC) abnormalities have been observed in various anxiety disorders. However, the relationship between mOFC activity and anxiety among the healthy population has not been fully examined. Here, we conducted a resting state functional magnetic resonance imaging (R-fMRI) study with 56 healthy male adults from the Nathan Kline Institute/Rockland Sample (NKI-RS) to examine the relationship between the fractional amplitude of low-frequency fluctuation (fALFF) signals and trait anxiety across the whole brain. A Louvain method for module detection based on graph theory was further employed in the automated functional subdivision to explore subregional correlates of trait anxiety. The results showed that trait anxiety was related to fALFF in the mOFC. Additionally, the resting-state functional connectivity (RSFC) between the right subregions of the mOFC and the precuneus was correlated with trait anxiety. These findings provided evidence about the involvement of the mOFC in anxiety processing among the healthy population.
Adult ; Aged ; Aged, 80 and over ; Anxiety/physiopathology* ; Brain/physiopathology* ; Brain Mapping ; Healthy Volunteers ; Humans ; Image Processing, Computer-Assisted ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Phenotype ; Prefrontal Cortex/physiopathology* ; Surveys and Questionnaires ; Young Adult

BACKGROUNDPolycystic ovary syndrome (PCOS) and its characteristic symptoms have been associated with physical and psychological issues in women of reproductive age. The current study was conducted in response to the dearth of systematic research related to psychological functioning and quality of life in patients with PCOS in Southwest China, and to determine whether patients with PCOS exhibit poorer mental health (MH) compared to healthy women of the same age and living in the same region, without a PCOS diagnosis.

METHODSWe enrolled 120 outpatients with PCOS and 100 healthy controls in this study. Standardized questionnaires were administered to assess general MH conditions (General Health Questionnaire-12-item version), anxiety (State-Trait Anxiety Inventory), depression (Beck Depression Inventory), as well as health-related quality of life (HRQoL) measured using the 36-item short-form health survey. The independent samples t-test was conducted for continuous study variables. For categorical variables, the Pearson Chi-square test, Fisher's exact test, and logistic regression were performed.

RESULTSThe prevalence of anxiety (13.3% vs. 2.0%) and depression (27.5% vs. 3.0%) was higher in patients with PCOS compared to the controls (both P< 0.05). Patients with PCOS had decreased HRQoL. Patients with PCOS who had fertility requirements were more likely to be anxious and depressed than those without fertility requirements (anxiety: 22.6% [12/53] vs. 5.9% [4/67], χ2 = 7.117, P = 0.008; depression: 37.7% (20/53) vs.19.4% (13/67), χ2 = 4.988, P = 0.026).

CONCLUSIONSPCOS and related symptoms may be risk factors for depression and anxiety. Professionals should be concerned with the MH of women with PCOS, and psychological therapy should be considered.

Adult ; Anxiety ; diagnosis ; etiology ; China ; Depression ; diagnosis ; etiology ; Female ; Humans ; Mental Disorders ; diagnosis ; etiology ; Polycystic Ovary Syndrome ; complications ; physiopathology ; psychology ; Psychiatric Status Rating Scales ; Quality of Life ; Surveys and Questionnaires ; Young Adult

OBJECTIVETo compare the effects of electroacupuncture (EA) and moxibustion therapies on patients with diarrhea-predominant irritable bowel syndrome (D-IBS).

METHODSA total of 60 D-IBS patients were randomly allocated to the EA group (30 cases) and moxibustion group (30 cases). Before and after treatment, the gastrointestinal symptoms and psychological symptoms were scored by Visual Analogue Scale, Bristol Stool Form Scale, Hamilton Anxiety Rating Scale (HAMA), and Hamilton Depression Rating Scale (HAMD); the expressions of 5-hydroxytryptamine (5-HT), 5-HT3 receptor (5-HT3R), and 5-HT4 receptor (5-HT4R) in the sigmoid mucosal tissue were measured by immunohistochemical staining. Additionally, the effects on the functional brain areas of the anterior cingulate cortex (ACC), insular cortex (IC) and prefrontal cortex (PFC) were observed by functional magnetic resonance imaging.

RESULTSCompared with before treatment, both EA and moxibustion groups reported significant improvements in abdominal pain and abdominal bloating after treatment (P<0.01 or P<0.05). The moxibustion group reported greater improvements in defecation emergency, defecation frequency, and stool feature than the EA group (P<0.01). Both HAMA and HAMD scores were significantly decreased in the moxibustion group than in the EA group (P<0.01). Both groups demonstrated significantly reduced expressions of 5-HT, 5-HT3R and 5-HT4R in the colonic mucosa after treatment (P<0.01), with a greater reduction of 5-HT in the moxibustion group (P<0.05). Finally, decreased activated voxel values were observed in the left IC, right IC and PFC brain regions of patients in the moxibustion group under stimulation with 150 mL colorectal distension after treatment (P<0.05 or P<0.01), while in the EA group only PFC area demonstrated a reduction (P<0.05).

CONCLUSIONMoxibustion can significantly improve the symptoms of D-IBS, suggesting that moxibustion may be a more effective therapy than EA for D-IBS patients.

Adult ; Anxiety ; Brain ; physiology ; Cerebral Cortex ; physiopathology ; Colon, Sigmoid ; chemistry ; Depression ; Diarrhea ; physiopathology ; Electroacupuncture ; Gastrointestinal Tract ; physiology ; Gyrus Cinguli ; physiopathology ; Humans ; Immunohistochemistry ; Intestinal Mucosa ; chemistry ; Irritable Bowel Syndrome ; physiopathology ; psychology ; therapy ; Magnetic Resonance Imaging ; Moxibustion ; Pain Measurement ; Prefrontal Cortex ; physiopathology ; Receptors, Serotonin, 5-HT3 ; analysis ; Serotonin ; analysis