Animal models and experimental medicine最新文献

Background: This study investigated the impacts and mechanisms of yunweiling in the management of Functional Constipation (FC) using network pharmacology and experimental research.

Methods: Using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), Genecard, and Online Mendelian Inheritance in Man (OMIM) databases, a potential gene target for yunweiling in treating FC was found. A pharmacological network was built and viewed in Cytoscape. A protein interaction map was created with STRING and Cytoscape. 'clusterProfiler' helped uncover its mechanism. Molecular docking was done with AutoDock Vina. In a constipation mouse model, Western blot was used to assess yunweiling's effectiveness.

Results: To investigate yunweiling's therapeutic effects on FC, we employed a loperamide-induced constipation model. Successful model establishment was confirmed by first black stool time, reduced stool output, and impaired gastrointestinal motility. Yunweiling treatment, especially at high and medium doses, significantly alleviated constipation symptoms by reducing first black stool time, increasing stool output, and enhancing gastrointestinal motility. HE staining revealed yunweiling's ability to restore colon tissue structure. Yunweiling modulated the expression of key proteins TP53, P-AKT, P-PI3K, RET, and Rai, implicating its involvement in the PI3K-Akt signaling pathway. Comparative analysis showed yunweiling to be more effective than its individual components (shionone, β-sitosterol, and daucosterol) in improving constipation. The combination of yunweiling with TP53 and PI3K-Akt inhibitors further enhanced its therapeutic effects, suggesting a synergistic mechanism.

Conclusions: The integration of network pharmacology and experimental investigations indicated the effectiveness of yunweiling in managing FC, offering essential theoretical support for clinical application.

Background: Alzheimer's disease (AD) and lung cancer are leading causes of mortality among the older population. Epidemiological evidence suggests an antagonistic relationship between them, whereby patients with AD exhibit a reduced risk of developing cancer and vice versa. However, the precise mechanism by which AD antagonizes lung cancer progression warrants further elucidation.

Methods: To this end, we established a co-morbidity model using 5xFAD transgenic mice induced with the carcinogen urethane. We visualized and quantified surface lung tumor colonies, assessed pathological parameters associated with lung cancer and AD using histopathological analysis, and employed single-cell sequencing and molecular pathological analyses to explore the mechanisms by which AD confers resistance to lung cancer.

Results: Our findings revealed a significant reduction in lung tumor incidence in the AD group compared with that in the wild-type (WT) group. The results indicated a close association between AD-induced inhibition of lung tumor progression and iron homeostasis imbalance and increased oxidative stress. Moreover, greater CD8⁺ T cytotoxic lymphocyte and effector natural killer cell infiltration in the lung tumor tissues of AD mice and enhanced CD8⁺ T cytotoxic lymphocyte-mediated killing of target cells may be the primary factors contributing to the inhibition of lung tumor growth in the presence of AD.

Conclusion: This study identified essential mechanisms through which AD suppresses lung tumorigenesis, thereby providing targets for potential therapeutic interventions in these diseases.

Background: Pregnancy affects learning and memory in women. Thus, to investigate the effects of pregnancy, the authors examined the brain electrophysiology of pregnant mice.

Methods: Using the whole-cell patch-clamp technique on isolated brain slices, we detected and compared the electrophysiological changes in the hippocampal CA1 (HIP CA1) region, medial prefrontal cortex (mPFC), and basolateral amygdala (BLA) among 15 pregnant and 15 nonpregnant mice.

Results: In pregnant mice, there was a trend toward an increase in the frequency of miniature excitatory postsynaptic currents (mEPSCs) (p = 0.092) and a trend toward a decrease in the amplitude of miniature inhibitory postsynaptic currents (mIPSC) (p = 0.071) in the HIP CA1. In the BLA, both the amplitudes of mEPSCs and mIPSCs were significantly reduced (p = 0.004 and 0.042, respectively). In the mPFC, the amplitudes of mEPSCs and hyperpolarization-activated currents (Ih), as well as the frequencies of mIPSCs, were higher compared to nonpregnant mice (p = 0.035, 0.009, and 0.038, respectively).

Conclusions: In pregnant mice, the electrophysiological change in neurons in the mPFC and BLA might contribute to the cognitive and emotional changes during pregnancy. A trend toward electrophysiological change in the HIP CA1 revealed that the mechanism of cognitive change during pregnancy might differ from that of other conditions.

Background: The composition of the intestinal flora and the resulting metabolites affect patients' sleep after surgery.

Methods: We intended to elucidate the mechanisms by which disordered intestinal flora modulate the pathophysiology of postoperative sleep disturbances in hosts. In this study, we explored the impacts of anesthesia, surgery, and postoperative sleep duration on the fecal microbiota and metabolites of individuals classified postprocedurally as poor sleepers (PS) and good sleepers (GS), as diagnosed by the bispectral index. We also performed fecal microbiota transplantation in pseudo-germ-free (PGF) rats and applied Western blotting, immunohistochemistry, and gut permeability analyses to identify the potential mechanism of its effect.

Results: Research finding shows the PS group had significantly higher postoperative stool levels of the metabolites tryptophan and kynurenine than the GS group. PGF rats that received gut microbiota from PSs exhibited less rapid eye movement (REM) sleep than those that received GS microbiota (GS-PGF: 11.4% ± 1.6%, PS-PGF: 4.8% ± 2.0%, p < 0.001). Measurement of 5-hydroxytryptophan (5-HTP) levels in the stool, serum, and prefrontal cortex (PFC) indicated that altered 5-HTP levels, including reduced levels in the PFC, caused sleep loss in PGF rats transplanted with PS gut flora. Through the brain-gut axis, the inactivity of tryptophan hydroxylase 1 (TPH1) and TPH2 in the colon and PFC, respectively, caused a loss of REM sleep in PGF rats and decreased the 5-HTP level in the PFC.

Conclusions: These findings indicate that postoperative gut dysbiosis and defective 5-HTP metabolism may cause postoperative sleep disturbances. Clinicians and sleep researchers may gain new insights from this study.

It is increasingly recognized that young, chow-fed inbred mice poorly model the complexity of human carcinogenesis. In humans, age and adiposity are major risk factors for malignancies, but most genetically engineered mouse models (GEMM) induce carcinogenesis too rapidly to study these influences. Standard strains, such as C57BL/6, commonly used in GEMMs, further limit the exploration of aging and metabolic health effects. A similar challenge arises in modeling periodontitis, a disease influenced by aging, diabesity, and genetic architecture. We propose using diverse mouse populations with hybrid vigor, such as the Collaborative Cross (CC) × Apc^Min hybrid, to slow disease progression and better model human colorectal cancer (CRC) and comorbidities. This perspective highlights the advantages of this model, where delayed carcinogenesis reveals interactions with aging and adiposity. Unlike Apc^Min mice, which develop cancer rapidly, CC × Apc^Min hybrids recapitulate human-like progression. This facilitates the identification of modifier loci affecting inflammation, diet susceptibility, organ size, and polyposis distribution. The CC × Apc^Min model offers a transformative platform for studying CRC as a disease of adulthood, reflecting its complex interplay with aging and comorbidities. The insights gained from this approach will enhance early detection, management, and treatment strategies for CRC and related conditions.

Background: The mechanisms underlying cardiac remodeling in aortic valvular (AoV) disease remain poorly understood, partially due to the insufficiency of appropriate preclinical animal models. Here, we present a novel murine model of aortic regurgitation (AR) generated by trans-apical wire destruction of the AoV.

Methods: Directed by echocardiography, apical puncture of the left ventricle (LV) was performed in adult male C57BL/6 mice, and a metal guidewire was used to induce AoV destruction. Echocardiography, invasive LV hemodynamic and histological examination were conducted to assess the degree of AR, LV function and remodeling.

Results: AR mice exhibited rapid aortic regurgitation velocity (424 ± 15.22 mm/s) immediately following successful surgery. Four weeks post-surgery, echocardiography revealed a 54.6% increase in LV diastolic diameter and a 55.1% decrease in LV ejection fraction in AR mice compared to sham mice. Pressure-volume catheterization indicated that AR mice had significantly larger LV end-diastolic volumes (66.2 ± 1.5 μL vs. 41.8 ± 3.4 μL), reduced LV contractility (lower dP/dt_max and Ees), and diminished LV compliance (smaller dP/dt_min and longer Tau) compared to sham mice. Histological examination demonstrated that AR mice had significantly larger cardiomyocyte area and more myocardial fibrosis in LV tissue, as well as a 107% and a 122% increase of heart weight/tibial length and lung weight/tibial length, respectively, relative to sham mice.

Conclusions: The trans-apex wire-induced destruction of the AoV establishes a novel and efficient murine model to develop AR, characterized by significant eccentric LV hypertrophy, heart failure, and pulmonary congestion.

Background: Qingyangshen (Cynanchum otophyllum C.K. Schneid) is a folk drug for treating depression and other mental disorders induced by social defeat stress. Neuroplasticity in the hippocampus is essential for the modulation of cognition and emotion, and its impairment may contribute to the development and progression of depression. Our previous studies have found that Qingyangshen glycosides (QYS) can improve depression-like behavior in social failure mouse models, mainly through PGC-1α/FNDC5/BDNF signaling pathways activation, but its effects and mechanisms on hippocampal neuroplasticity remain unknown.

Methods: Chronic social defeat stress (CSDS) was used to induce social defeat in mice. Morphological changes in the hippocampus were observed by H&E staining and Golgi staining. Immunofluorescence double staining was used to detect the expression of synaptophysin (SYN) and postsynaptic density protein-95 (PSD-95), while western blot was employed to evaluate PSD-95, SYN, and doublecortin (DCX) proteins. The pathological processing of social defeat and the therapeutic effects of QYS on it was confirmed through behavioral assessment associated with morphologic observation.

Results: During the whole study, the sucrose preference indices and OFT activity time of CSDS mice were significantly decreased (p ≤ 0.05), and the tail suspension immobility time was significantly increased (p ≤ 0.05), suggesting that the mice had significant depressive symptoms. Treatment with QYS (25, 50, and 100 mg/kg) significantly alleviated depressive symptoms in CSDS mice, which was demonstrated by significantly (p ≤ 0.05 or p ≤ 0.01) reducing the duration of tail-hanging immobility and increasing the tendency of sucrose preference indices and OFT activity time. QYS treatment also significantly increased the expression of DCX, PSD-95, and SYN proteins, which play a crucial role in depression.

Conclusions: QYS alleviated these symptoms by enhancing hippocampal neuroplasticity through upregulating the expression of synapse-associated proteins (SAPs). The therapeutic mechanism of QYS may involve modulating the neuroplasticity of hippocampus neurons by altering the expression of SAPs.

Acquired immune deficiency syndrome (AIDS) is the name used to describe several potentially life-threatening infections and disorders that happen when HIV has severely compromised the immune system. The primary effect of HIV is to decrease host immunity, exposing the host to external pathogens. The development of pharmaceutical drugs that directly cure the infection is crucial because of the current wide-ranging epidemic of HIV. Most therapeutic anti-HIV drugs are nucleosides. However, their high toxicity and potential for drug resistance restrict their use. Many of the most effective clinical drugs used to inhibit HIV, the activation of latent HIV, and AIDS have been obtained from natural sources. This review focuses on potential natural medicinal products for treating and managing HIV and AIDS. Notwithstanding, further clinical research studies are needed to understand the subject and its dynamics.

The laryngeal muscle evoked potential (LMEP) is a neurophysiological outcome parameter that guarantees integrity of the nerve-electrode interface during experiments with vagus nerve stimulation (VNS). This paper discusses a large series of minimally invasive LMEP recordings in 46 female Lewis rats, implanted with a custom-made VNS electrode around the left cervical vagus nerve. After a 3-week recovery, LMEPs were recorded twice in each animal, with swapping the anode and cathode positions of the VNS electrode (polarity inversion). A VNS-induced LMEP was identified as the initial negative peak wave post-stimulation artifact, consistently recorded in all sweeps at a given stimulation output current. Latency was defined as the time from stimulation onset to this negative peak, and stimulation threshold as the lowest current showing a clear and reproducible LMEP. An LMEP response was shown by 37/46 animals (80.4%), with stimulation intensity threshold of 0.37 ± 0.27 mA and latency of 2.39 ± 0.45 ms. Administering the cathodic pulse phase first at the caudal electrode contact resulted in the shortest LMEP latencies (MWU: p = 0.049. 2.36 ± 0.43 ms vs. 2.41 ± 0.47 ms). Minimally invasive LMEP recording provides a feasible and reliable means for checking electrode functioning and correct implantation.

Background: Reliable animal models are crucial to drug development for focal segmental glomerulosclerosis (FSGS), a rare kidney disease. Variability in success rates in literature and significant ethical concerns with animal welfare necessitate further optimization of adriamycin (ADR)-induced FSGS model developed on BALB/c mice.

Methods: High-performance liquid chromatography (HPLC) was used to assess ADR stability in water and upon light exposure. To identify the optimal ADR level, single intravenous ADR injections with dosing levels from 10 to 17 mg/kg body weight were administered to BALB/c mice to induce FSGS-like pathology. Body weight and proteinuria of FSGS mice were monitored and analyzed for FSGS model-associated morbidity. Animals were euthanized for hematological and kidney histological assessments 8 weeks post induction. To identify the suitable experiment time frame of the ADR-induced FSGS mouse model, a longitudinal study was performed, with an 11-week continuous monitoring of the symptoms.

Results: ADR was found to be unstable in aqueous media and light sensitive. A dosing level of 10.5 mg/kg of ADR was optimal for consistent FSGS mouse model induction on BALB/c strain, characterized by minimal mortality and sustained FSGS-like symptoms. Findings from the longitudinal study suggest that 6 weeks post ADR induction may represent the peak of FSGS pathology severity in this mouse model. This time frame may be used for FSGS drug development projects.

Conclusion: Based on the outcome from this study, we identified the optimal ADR dosing level and model testing duration. A standard operating procedure (SOP) for the ADR-induced FSGS mouse model was established to facilitate FSGS basic research and drug development.