Animal models and experimental medicine最新文献

Cynomolgus macaques, a species of Old World primate native to southeastern and eastern Asia and the island of Mauritius, are one of the most important nonhuman primate models for infectious disease. Although the closely related rhesus macaque is classified into subspecies based on geographic origin, no such subdivision exists for cynomolgus macaques, and they continue to be used interchangeably in infectious disease research, reducing the comparability of data produced from these studies. Research into the population genetics of cynomolgus macaques has found significant differences between macaques native to different areas, including their genetic diversity, with macaques from insular populations such as Mauritius and the Philippines exhibiting highly restricted heterozygosity compared to mainland populations native to Indonesia or Cambodia. In the context of infectious disease studies, research into pathogens, including Ebola virus, Crimean-Congo hemorrhagic fever virus, and Mycobacterium tuberculosis have found differences in study outcomes, survival times, and immune cell responses between different populations of macaques. This review provides an overview of the differences between cynomolgus macaque populations in the context of genetic diversity, and in response to infection, and highlights the need for clear reporting of geographic origin of primates used in research. This will improve data comparison between studies and help to further refine this important animal model.

Background: Atherosclerosis (AS), the leading cause of cardiovascular disease, involves complex molecular mechanisms that remain incompletely understood, particularly in the context of diet-induced vascular lesions.

Methods: We established an AS model in Bama miniature pigs using a high-cholesterol, high-fat diet (HCFD) and performed quantitative proteomic analysis on coronary artery tissues. Key proteins were identified using protein-protein interaction (PPI) network analysis and subsequently validated by histopathological evaluation in porcine and murine coronary arteries. The underlying molecular mechanisms were elucidated using Western blot analysis.

Results: The HCFD successfully induced an atherosclerotic phenotype characterized by significantly elevated serum lipid levels. Proteomic analysis identified 108 differentially expressed proteins (DEPs) between the AS and control groups. From four identified hub proteins, we focused on clusterin (CLU), which was markedly upregulated in atherosclerotic coronary tissues, particularly within endothelial cells (ECs) and smooth muscle cells (SMCs). Mechanistically, CLU upregulation activated the LRP1/AKT signaling pathway, thereby promoting atherogenesis.

Conclusion: Our study reveals that elevated CLU expression accelerates the process of AS by activating the LRP1/AKT pathway. These data elucidate a novel pro-atherogenic role for CLU and establish the CLU/LRP1/AKT axis as a promising therapeutic target for managing AS, particularly in pathologies driven by high-fat diets.

A lupus-like condition induced by intraperitoneal administration of pristane (2,6,10,14-tetramethylpentadecane) in mice is widely used as a model of systemic lupus erythematosus (SLE). Due to their phylogenetic distance from humans, murine models are not always suitable tool for studying the specific activity of therapeutic agents and the pathogenesis of SLE. In order to overcome species-specific limitations of murine models, this approach was tested in non-human primates-cynomolgus monkeys (Macaca fascicularis). Two intraperitoneal injections at a dose of 3.5 mL/kg, administered at weeks 1 and 23, recapitulated SLE features, including: production of antinuclear autoantibodies (ANA), membranoproliferative glomerulonephritis with immune complex (IC) deposition in the glomeruli. However, from week 27 five of eight pristane-treated monkeys developed progressive respiratory failure. Two of these died at week 28 and the remaining were euthanized at week 32. The histology of the monkey lungs suggested exogenous lipoid pneumonia. Thus, while pristane induced serological autoimmunity and characteristic renal manifestations in Macaca fascicularis, the consequent lipoid pneumonia limited the observation period and prevented comprehensive evaluation of SLE manifestations beyond 32 weeks.

The incidence of benign airway stenosis (BAS) is on the rise, and current treatment options are associated with a significant risk of restenosis. Therefore, there is an urgent need to explore new and effective prevention and treatment methods. Animal models serve as essential tools for investigating disease mechanisms and assessing novel therapeutic strategies, and the scientific rigor of their construction and validation significantly impacts the reliability of research findings. This paper systematically reviews the research progress and evaluation systems of BAS animal models over the past decade, aiming to provide a robust foundation for the optimized construction of BAS models, intervention studies, and clinical translation. This effort is intended to facilitate the innovation and advancement in BAS prevention and treatment strategies.

Background: Targeted delivery of biological macromolecules to the small intestine remains challenging due to their susceptibility to degradation in the hostile gastric environment.

Methods: This study introduces a minimally invasive, in situ injection technique for the murine small intestine that facilitates localized luminal delivery while circumventing gastric barriers. The procedure involves a small abdominal incision for direct injection into the duodenum near the pylorus. Postsurgical monitoring of physiological parameters, systemic inflammatory markers, liver function, and intestinal integrity was conducted over 72 h. Histopathological analysis was performed. The delivery of the functional protein TAT-EGFP (Tat protein fused to enhanced green fluorescent protein) to intestinal epithelial cells was evaluated and compared with oral gavage. As a proof of concept, single-cell RNA sequencing of the intestinal epithelium was performed after high-mobility group box 1 administration.

Results: Postsurgical monitoring indicated only transient, anesthesia-related hypothermia and minor behavioral alterations. No significant changes were observed over 72 h in body weight, core temperature, clinical severity scores, systemic inflammatory markers (C-reactive protein and leukocytes), liver function (alanine aminotransferase), or intestinal integrity. Histopathological analysis confirmed preserved tissue architecture and normal digestive, absorptive, and barrier functions. The model successfully delivered TAT-EGFP to intestinal epithelial cells, an outcome not achievable via oral gavage due to gastric degradation. Single-cell RNA sequencing of the intestinal epithelium after high-mobility group box 1 administration revealed inflammatory gene expression patterns in specific epithelial subpopulations.

Conclusions: Compared to traditional methods such as oral gavage or organoid culture, this technique offers precise, degradation-resistant delivery of macromolecules in a physiological context. The model's versatility makes it a powerful platform for intestinal research, with applications in drug delivery assessment, gene therapy evaluation, and host-microbiota interaction studies.

Background: The analgesic effects of multiple electroacupuncture (EA) sessions and single EA sessions differ significantly in pain management. Area 24b (A24b) of the anterior cingulate cortex (ACC) is crucial in pain processing. EA relieves pain by targeting and modulating the neuronal activity within this subregion. However, whether the cumulative effect of EA antinociception is connected to A24b mechanisms has remained unclear.

Methods: In our study, we used the Complete Freund's Adjuvant (CFA) model to induce inflammatory pain and the Spared Nerve Injury (SNI) model to induce neuropathic pain, and adult male C57BL/6, FosTRAP, and FosTRAP:Ai9 mice were used as experimental subjects to investigate the cumulative effect of EA antinociception and whether multiple EA sessions and a single EA session regulate different neuronal populations in the A24b.

Results: We observed that EA effectively alleviated pain in mice, with three EA sessions yielding superior analgesic effects compared to a single session. Using chemical genetics combined with FosCreER technology to activate EA-TRAPed cells in the A24b, we found that pain relief was more pronounced with three EA sessions. Moreover, chemogenetic inhibition of EA-TRAPed cells in the A24b reversed the analgesic effects of a single EA session but not those of three EA sessions. Fluorescent in situ hybridization results indicated that three EA sessions significantly increased the number of GABAergic neurons in the A24b compared with a single session. Additionally, retrograde tracing revealed that the A24b circuit that monosynaptically innervates EA-TRAPed cells included projections from the central lateral nucleus (CL), lateral mediodorsal thalamic nucleus (MDL), lateral habenula (LHb), dorsal raphe nucleus (DR), caudal linear nucleus of the raphe (CLi), dorsal tuberomamillary nucleus (DTM), periventricular hypothalamic nucleus (Pe) and hippocampal fields CA1, CA2, and CA3. These findings suggest that multiple EA sessions and single EA sessions activated different neuronal populations in the A24b. The enhanced analgesic effect of multiple EA sessions may be attributed to an increase in the proportion of GABAergic neurons within the A24b.

Conclusions: Multiple and single EA sessions recruit distinct neuronal populations in A24b, with the stronger analgesic effect of repeated EA linked to a higher proportion of GABAergic neurons in this region.

Background: This study aims to explore the establishment of an animal model of cardiac injury induced by trimethylamine-N-oxide (TMAO), a metabolite secreted by gut microorganisms, and to investigate its application in moderate-intensity continuous training (MICT) intervention.

Methods: C57BL6/J mice were randomly divided into four groups: normal mice (Nor, n = 15); mice administered TMAO (TMAO, n = 15); mice undergoing (Nor+MICT, n = 15); mice undergoing (MICT) and administered TMAO (TMAO+MICT, n = 15). Mice in the TMAO and TMAO+MICT groups received daily gavage of high-dose TMAO for 8 weeks, whereas those in the Nor+MICT and TMAO+MICT groups underwent MICT for 8 weeks (60 min per session, 5 days per week, at 50% maximal running capacity). Cardiac function was evaluated using ultrasound, myocardial histology was examined using hematoxylin and eosin (HE) staining, and nuclear magnetic resonance (NMR)-based metabolomics was employed for multivariate statistical and metabolic pathway analyses.

Results: Relative to the Nor group, TMAO-treated mice exhibited significant weight loss, elevated heart rate, and reduced ejection fraction and left ventricular fractional shortening, indicating cardiac impairment. Importantly, the TMAO+MICT group demonstrated significant improvements in these parameters compared to the TMAO group, alongside distinct alterations in myocardial metabolic profiles. TMAO altered five metabolic pathways relative to controls, whereas MICT induced significant changes in three pathways in TMAO-treated mice.

Conclusion: Eight weeks of high-dose TMAO administration induced significant cardiac dysfunction in mice, which was effectively mitigated by MICT intervention. Consequently, this animal model serves as a valuable tool for investigating the mechanisms underlying the impact of MICT on cardiovascular diseases.

Background: Chronic hyperuricemia is associated with complications such as gout and uric acid nephropathy, but uric acid also exhibits biological activities (e.g., antioxidant effects, potential neuroprotective properties against neurodegenerative diseases). Nonhuman primates are ideal models for studying neurodegenerative diseases; however, existing nonhuman primate hyperuricemia models cannot sustain long-term elevated serum uric acid levels, nor recapitulate the impaired uric acid excretion observed in clinical hyperuricemic patients.

Methods: First, we detected uricase expression in cynomolgus monkeys and compared it with that in mice. Then, we established a cynomolgus monkey hyperuricemia model by administering a mixture of potassium oxonate, hydrochlorothiazide, and adenine via fruits and vegetables. We further analyzed the regulatory effects of this model on uric acid metabolism (synthesis, degradation, and excretion) and the expression of uric acid transporter genes in the intestine and kidney.

Results: Cynomolgus monkeys express functional uricase, but at a lower level than mice. The established model maintained stable, long-term hyperuricemia by three mechanisms: increasing intestinal and renal uric acid excretion load, inhibiting hepatic uric acid degradation, and promoting uric acid synthesis. Additionally, the model downregulated the expression of intestinal/renal uric acid-secreting transporter genes, while upregulating uric acid-reabsorbing transporter genes.

Conclusions: This novel cynomolgus monkey hyperuricemia model provides a new tool for investigating the association between hyperuricemia and neurodegenerative diseases, and will help clarify the mechanism by which serum uric acid influences cognitive function.

Background: Ex vivo lung perfusion (EVLP) has emerged as a critical technique for lung preservation and evaluation prior to transplantation. While conventional rat EVLP systems utilize closed-loop dual cannulation of pulmonary artery (PA) and vein, the effect of the simplified model using single PA cannulation with passive venous drainage is unknown.

Methods: We developed two EVLP models in rats: a semi-closed circuit with PA-only cannulation and left atrial incision for passive venous drainage (SC-EVLP), and a closed circuit employing both arterial and venous cannulation (C-EVLP). Donor lungs were perfused for a defined duration and subsequently orthotopically transplanted. We evaluated pulmonary function parameters, histopathological injury scores, inflammatory cytokine levels, and apoptotic marker expression at the end of perfusion and posttransplantation.

Results: Compared to the conventional EVLP, the SC-EVLP group exhibited significantly lower PA pressure and improved dynamic lung compliance throughout perfusion. Although the levels of tumor necrosis factor-α in the perfusate were higher in the SC-EVLP group, other cytokine levels in the perfusate and bronchoalveolar lavage fluid exhibited no significant differences. Pulmonary edema was reduced in the SC-EVLP group, as indicated by a lower lung wet-to-dry ratio. After transplantation, lungs from the SC-EVLP group exhibited lower histological injury scores, reduced apoptosis, and decreased serum cytokine levels, suggesting attenuated inflammation and tissue damage.

Conclusions: In a rat model, single PA cannulation with passive venous drainage reduced pulmonary edema during EVLP and reduced lung injury and systemic inflammation after transplantation.

Background: Triptolide (TP) exhibits various pharmacological activities. Our previous studies have confirmed the efficacy of TP against lung adenocarcinoma (LUAD). However, the potent pharmacological activity of TP is underpinned by its complex mechanisms. Exploring its potential mechanisms is of great value for promoting the clinical application of TP and extending its clinical use.

Methods: Differentially expressed genes (DEGs) associated with LUAD were analyzed and acquired from the TCGA database, while DEGs related to TP were obtained through RNA sequencing. Hub genes were identified through LASSO and random forest models. The efficacy of TP against LUAD was validated using tumor-bearing mouse models and A549 cells. The validation of hub genes was conducted using RT-qPCR. The regulatory effect of hub genes on TP efficacy was validated through overexpression cell models. Furthermore, the potential mechanisms by which TP improves gemcitabine (GEM) resistance were explored using a GEM-resistant cell line in combination with the overexpression model.

Results: This study validated the therapeutic effect of TP against LUAD in vivo and in vitro. Bioinformatics revealed that the mechanism of TP's effect against LUAD might be associated with amino acid-related biological processes. Five hub genes were screened and identified by combining bioinformatics methods and experiments. The overexpression model validated that PSAT1 plays an effective role in the efficacy of TP and in alleviating GEM resistance.

Conclusion: This study preliminarily demonstrated that the anti-LUAD effect of TP was associated with the PSAT1-regulated serine biosynthesis pathway, and that TP effectively improves GEM resistance by inhibiting PSAT1 expression.