Animal models and experimental medicine最新文献

Background: The traditional method of heterotopic abdominal heart transplantation (HTx) involves cross-clamping the inferior vena cava, which inevitably leads to bilateral lower limb ischemia (LI). This study first aimed to investigate the impact of LI on renal function in rats subjected to unilateral nephrectomy (UNx). Second, a modified method utilizing renal vessel-assisted anastomosis in rats with left UNx was compared with the traditional method for abdominal HTx.

Methods: Male Sprague-Dawley rats were utilized as subjects for both experimental phases. In experiment 1, the animals were divided into four groups: sham operation group; LI group-rats undergoing occlusion of the abdominal aorta and vena cava below the renal vessels; UNx group-rats with left UNx; and LI + UNx group. All operated animals were monitored for up to 7 days for biochemical markers, renal histopathology, and survival rates. In experiment 2, we introduced the renal vessel-assisted method as the experimental group and compared it against the traditional method as the control within rat heterotopic HTx models. We assessed operative characteristics, echocardiography results, histological findings, and graft survival.

Results: First, LI resulted in acute kidney dysfunction characterized by a decrease in 7-day survival rates and creatinine clearance rates in both the LI and LI + UNx groups compared to the sham operation and UNx groups. Particularly, histopathological damage in the kidney and liver did not exhibit significant effects during this period. Second, the implementation of the renal vessel-assisted method significantly reduced bleeding volume at suture sites and enhanced the 7-day survival rate compared to the traditional method.

Conclusion: Acute kidney injury was induced by LI postoperation in treated rats. The renal vessel-assisted method demonstrated its effectiveness as a superior alternative that mitigates complications associated with the traditional method.

Background: The aim of the study was to develop a non-human primate model of metabolic dysfunction in Macaca fascicularis using chronic high-fat diet (HFD) to mimic clinical disease progression.

Methods: Thirty-five male macaques aged 10-15 years underwent an 18-month HFD intervention. Physiological parameters (BMI, BP, hematology), liver fat fraction (evaluated by ultrasound/MRI), cardiac function (assessed by echocardiography), and histopathology (using liver biopsy) were measured before and after the intervention. Serum proteomics with KEGG/STRING analyses identified molecular mechanisms.

Results: Within 6 months, HFD induced dyslipidemia (elevated TG, TCHO, HDL-C, LDL-C). After 18 months, metabolic dysfunction-associated steatohepatitis (MASH) was confirmed by histopathology in 57.14% (16/28) of macaques, diabetes (elevated FPG/HbA1c) in 17.86% (5/28), and myocardial hypertrophy (elevated LVMass/LAD) in 46.43% (13/28). Proteomics identified Bile acid-CoA: amino acid N-acyltransferase (BAAT) as a MASH hallmark protein, the level of which was inversely correlated with the degree of fibrosis. For diabetes, citrate synthase (CS) and malate dehydrogenase 1 (MDH1) impaired glucose oxidation via the TCA cycle, while hexose-6-phosphate dehydrogenase (H6PD) disrupted gluconeogenesis. Myocardial hypertrophy was associated with the downregulation of SRC proto-oncogene, non-receptor tyrosine kinase (SRC), mitogen-activated protein kinase 14 (MAPK14), emerin (EMD), and integrin subunit beta 1 (ITGB1).

Conclusions: An 18-month HFD successfully established a translational M. fascicularis model replicating key metabolic disorders (MASH, diabetes, cardiac hypertrophy). BAAT, CS/MDH1/H6PD, and SRC/MAPK14/EMD/ITGB1 were identified as mechanistic biomarkers for these conditions.

The rising incidence of dementia and associated neurodegenerative disorders poses a growing public health challenge. These conditions have traditionally been studied as isolated central nervous system disorders; however, emerging evidence suggests that broader systemic factors, including chronic inflammation, immune dysregulation, metabolic dysfunction, and genetic susceptibility, may also play a role. This review examines the interconnection between autoimmune diseases and metabolic syndromes in the pathogenesis and exacerbation of neurodegeneration. Conditions such as rheumatoid arthritis, systemic lupus erythematosus, and type 1 diabetes mellitus have been associated with a heightened risk of developing dementia through chronic immune activation, blood-brain barrier disruption, and neuroinflammatory signaling. Similarly, metabolic disorders such as diabesity promote insulin resistance and oxidative stress, accelerating cognitive decline. The review also discusses glaucoma as a neurodegenerative condition with autoimmune features, underscoring the need for expanded classification and treatment strategies. A key focus is the utilization of the Collaborative Cross (CC) mouse model, which enables the study of gene-environment interactions across genetically diverse backgrounds. Findings from CC mice reveal strain-dependent susceptibility to inflammation, cognitive impairment, and gut-brain axis dysfunction, providing a translational bridge to human variability. This review highlights the importance of integrating precision-based approaches to dementia research that consider systemic influences. Advancing our understanding of these multiorgan interactions holds potential for designing precision-based therapeutic approaches to postpone the onset or reduce the incidence of neurodegenerative conditions.

Background: The development of relevant and robust large animal models of hepatocellular carcinoma is needed to test new therapeutic strategies for this disease. Transgenic approaches hold promise in addressing this complex problem. One such model, the Oncopig, has been reported to develop tumors of up to 4 cm in diameter within 7-14 days at sites of in situ vector inoculation. However, the resulting lesions reportedly contained an extensive inflammatory component that has not been evaluated in detail.

Methods: Herein, we describe our results from multiparametric characterization of the lesions generated using liver biopsy cores incubated in vector solution and replaced in the tissue. The study consisted of 3 animals in 3 cohorts (total of 9 animals) that were evaluated at 14, 21, and 28 days. CT imaging, immunohistochemistry, multiplex immunofluorescence, and comprehensive blood analyses were used to quantify composition of the hepatic masses that developed following AdCre inoculation.

Results: The tumors were hypovascular on CT and predominantly composed of CD45+ cells with a strong lymphohistiocytic component, with no carcinomas identified. Ki-67 staining showed proliferation of CD45+ immune cells but no neoplastic component. To provide further insight, the results are evaluated in the context of tumor growth kinetics.

Conclusion: While progress has been made in generating targetable lesions, achieving a robust large animal model of liver cancer that faithfully recapitulates the human disease remains a challenging goal.

Background: Skeletal tuberculosis (TB) remains a persistent clinical and research challenge due to its chronic course, osteolytic destruction, and the limitations of existing animal models, which often require high-level biosafety containment or fail to replicate human skeletal pathology.

Methods: This study developed a biosafe, accessible, and versatile murine model of skeletal TB using Mycobacterium smegmatis, a fast-growing, nonpathogenic mycobacterial species with high genomic homology to Mycobacterium tuberculosis. Three infection routes-subperiosteal calvarial injection, intratibial injection, and intracardiac inoculation-were systematically evaluated for their ability to induce localized versus disseminated bone infection under standard biosafety level (BSL)-1 conditions.

Results: Subperiosteal calvarial and intratibial injection of M. smegmatis induced localized bone lesions characterized by osteolysis, sequestrum formation, granulomatous inflammation, and increased osteoclast activity. Intratibial infection additionally triggered compartment-specific immune responses, including neutrophil and macrophage expansion, transient B-cell depletion, and activation of interferon-γ⁺ (IFN-γ⁺) T cells, reflecting active immune remodeling at the infection site. Systemic dissemination via intracardiac injection reproducibly generated progressive vertebral and tibial bone destruction with organized granuloma formation and immune cell infiltration but without prominent sequestrum formation. Compared to intratibial infection, intracardiac delivery exhibited lower intragroup variability and more closely recapitulated the diffuse progression of extrapulmonary skeletal tuberculosis.

Conclusions: This M. smegmatis-based murine model provides a straightforward, reliable, and immunopathologically relevant platform for exploring host-pathogen dynamics, immune-driven bone destruction, and early-stage therapeutic testing in skeletal TB, all within standard BSL-1 laboratories. This model fills a critical gap by enabling BSL-1 research into skeletal TB mechanisms and drug development.

Background: In preclinical research, tumor growth inhibition in subcutaneous models is frequently employed to evaluate therapeutic efficacy; however, such models often lack clinical translatability.

Methods: To better approximate clinical reality, taking the case of doxorubicin treatment, we utilized an orthotopic transplant and resection (OtR) strategy to systematically assess the effects of neoadjuvant chemotherapy, adjuvant chemotherapy, and their combination on tumor growth, recurrence, and malignant progression.

Results: Surprisingly, none of the treatments improved mouse survival, with adjuvant therapy even shortening it. Although neoadjuvant chemotherapy delayed preoperative tumor growth, and all regimens reduced recurrence rates, none effectively prevented metastasis. Furthermore, all treatment groups exhibited weight loss, indicative of chemotherapy-induced cachexia.

Conclusions: Collectively, these findings demonstrate that reduced tumor growth in preclinical mouse models does not necessarily translate into overall survival benefit. Our results emphasize the critical importance of prioritizing metastasis prevention over tumor growth inhibition as a key efficacy endpoint in antitumor drug evaluation.

Pathological scarring, manifested in the form of hypertrophic scars (HTS) and keloid scars (KS), represents a major clinical challenge due to its aesthetic and functional implications for patients. Understanding the molecular mechanisms involved in these types of scars and developing effective treatments requires the use of controlled experimental models, especially animals, to overcome the limitations of clinical studies. The aim of this sistematic review is to critically analyze the animal models used in the last five years (2020-2025) for the study of pathological scars, highlighting their advantages, limitations and applicability in the development of new therapeutic strategies. Murine, rabbit and porcine models, as well as alternative models, offer varied perspectives on the formation and treatment of HTS and KS, with an emphasis on histological and molecular correlations with human pathology. By synthesizing recent data, the paper highlights the essential role of preclinical research in optimizing antifibrotic treatments and in advancing the translation of data into the clinical sphere. Overall, animal models remain essential for bridging mechanistic insights with clinical translation, supporting the development of more effective and personalized anti-scar therapies.

Background: Healthy non-pharmacological lifestyle factors, such as regular physical exercise and dietary supplementation, have been shown to significantly improve cognitive outcomes over time compared to a more sedentary lifestyle and poor diet. Furthermore, exercise may serve as a potential protective factor in attenuating the effects associated with cognitive decline that are characteristic of neurodegenerative disorders, such as Alzheimer's disease (AD). Evidence indicates that certain dietary interventions can also attenuate the effects of neurodegeneration and positively impact longevity. Supplementation with polyphenols such as ellagic acid (EA), which is abundant in pomegranate juice, may help provide neuroprotective and longevity benefits.

Methods: This study examined the potential protective potential of EA and exercise and provides insight into the combined use of a polyphenol-rich diet and exercise to enhance behavioral outcomes and lifespan in a transgenic Drosophila melanogaster (fruit fly) model of AD with the Aβ₄₂ gene.

Results: Fruit flies subjected to a 120-minute exercise regimen performed better on a climbing assay than flies that did not exercise. Conversely, flies that exercised for 30 min passed marginally more trials on a learning and memory assay using an aversive stimulus than flies that did not exercise, whereas both groups performed better than flies subjected to the more intense exercise condition.

Conclusion: These results suggest a hormetic effect of exercise regarding memory performance. Finally, flies fed a low dose of dietary EA (0.24 mg/mL) lived significantly longer than flies fed the control diet or higher concentrations of EA, again suggesting a hormetic effect of EA on longevity.

Background: Mastitis seriously affects the mammary health of humans and animals. Studies have found that inflammation and oxidative stress play key roles in the occurrence and development of mastitis. Therefore, in-depth research on related molecular mechanisms is of great significance.

Methods: Postpartum mice were anesthetized with pentobarbital and administered lipopolysaccharide to develop the mouse mastitis model. Proteomic analysis was performed to compare protein expression in mitochondria-associated endoplasmic reticulum membranes (MAM) from two mouse mammary gland groups. Western blot was used to detect the expression of MAM-related proteins in mitochondria. AlphaFold3 was used to predict the molecular structures of phosphofurin acidic cluster sorting protein 2 (PACS2) and mitofusin 2 (MFN2) and their interaction levels. The MFN2-PACS2 interaction was investigated using co-immunoprecipitation and small interfering RNA.

Results: The results showed that the inflammation level in the mammary gland tissue of mice with mastitis significantly increased, the total antioxidant capacity decreased, and the expression of MAM-related proteins MFN2 and PACS2 was significantly downregulated. In cell experiments, overexpression of MFN2 can inhibit inflammation and oxidative stress responses, and promote the interaction between MFN2 and PACS2 to affect the formation of MAMs.

Conclusion: In summary, this study suggests that mastitis can alter the expression of MAM-related proteins in mouse breast tissue. The interaction between MFN2 and PACS2 regulates the formation of MAMs. Overexpression of MFN2 can promote the formation of MAMs and inhibit inflammation and oxidative stress response in mammary epithelial cells. Our results provided a new theoretical basis and potential therapeutic targets for the prevention and treatment of mastitis.

Background: This study investigated the role of polydatin in regulating macrophage-epithelial cell (EC) interactions during asthma. An asthma model was induced in BALB/c mice using ovalbumin (20 μg).

Methods: The therapeutic effects of polydatin (20 and 40 mg/kg) were evaluated in this asthmatic mouse model. To assess the underlying mechanisms, Bronchial Epithelium Adenovirus 12-SV40 2B (BEAS-2B) cells were cocultured with Tohoku Hospital for Pediatrics-1 (THP-1) macrophages, in which toll-like receptor 4 (TLR4) was either overexpressed or knocked down, and subsequently stimulated with lipopolysaccharide (LPS) and ATP. THP-1 cells underwent a 1-h pretreatment with polydatin (50 and 100 μmol/L), Class Lipid Inhibitor-095 (CLI-095, TLR4 inhibitor, 1 μg/mL), or A438079 (P2X7R antagonist, 10 μmol/L) prior to LPS/ATP challenge.

Results: Findings from Western blotting, enzyme-linked immunosorbent assay, flow cytometry, real-time polymerase chain reaction, and immunofluorescence assays demonstrated that modulating TLR4 expression significantly altered interleukin-1β (IL-1β) secretion from THP-1 macrophages and mitochondrial reactive oxygen species (mtROS) production in BEAS-2B ECs. In the mouse asthma model, polydatin significantly alleviated airway inflammation, oxidative stress, and apoptosis, likely by interfering with TLR4/P2X7R-mediated signaling and suppressing the activation of the NOD-like receptor protein inflammasome. Additionally, polydatin significantly reduced IL-1β and IL-18 levels and inhibited the infiltration of macrophages and eosinophils. Correspondingly, polydatin significantly attenuated TLR4/P2X7R signaling in THP-1 cells stimulated with ATP and LPS, thereby reducing IL-1β and IL-18 secretion, calcium influx, mtROS production, and apoptosis in BEAS-2B ECs.

Conclusions: Polydatin is a promising therapeutic candidate for asthma, possibly by targeting macrophage-epithelium cross-talk via the TLR4/P2X7R axis. Future formulations as capsules or sprays may effectively alleviate airway inflammation and remodeling.