1.Animal models of hemorrhage, parameters, and development of hemostatic methods
Gholamhossien DARYA ; Hamid MOHAMMADI ; Zeinab DEHGHAN ; Alireza NAKHAEI ; Amin DERAKHSHANFAR
Laboratory Animal Research 2025;41(1):20-33
Hemorrhage is a prevalent side effect of various injuries and can be life-threatening in certain instances. It is categorized into compressible and non-compressible types, each necessitating distinct modeling, laboratory assessments, and hemostatic approaches. This study utilized the keywords Hemorrhage, Bleeding, Animal Modeling, and Hemostat in reputable databases. The findings indicate that femoral artery hemorrhage and hepatic parenchymal hemorrhage are the predominant modeling techniques for compressible and noncompressible bleeding, respectively. Furthermore, it is noted that animal models of compressible hemorrhages are primarily situated in superficial body areas to investigate dressing or additive hemostats, while non-compressible hemorrhage models, typically located in visceral organs, are employed to examine adhesive or surgical instrumentbased hemostats.
2.An animal model of severe acute respiratory distress syndrome for translational research
Kuo‑An CHU ; Chia‑Yu LAI ; Yu‑Hui CHEN ; Fu‑Hsien KUO ; I.‑Yuan CHEN ; You‑Cheng JIANG ; Ya‑Ling LIU ; Tsui‑Ling KO ; Yu‑Show FU
Laboratory Animal Research 2025;41(1):81-92
Background:
Despite the fact that an increasing number of studies have focused on developing therapies for acute lung injury, managing acute respiratory distress syndrome (ARDS) remains a challenge in intensive care medicine.Whether the pathology of animal models with acute lung injury in prior studies differed from clinical symptoms of ARDS, resulting in questionable management for human ARDS. To evaluate precisely the therapeutic effect of trans‑ planted stem cells or medications on acute lung injury, we developed an animal model of severe ARDS with lower lung function, capable of keeping the experimental animals survive with consistent reproducibility. Establishing this animal model could help develop the treatment of ARDS with higher efficiency.
Results:
In this approach, we intratracheally delivered bleomycin (BLM, 5 mg/rat) into rats’ left trachea via a needle connected with polyethylene tube, and simultaneously rotated the rats to the left side by 60 degrees. Within sevendays after the injury, we found that arterial blood oxygen saturation (SpO2 ) significantly decreased to 83.7%, partial pressure of arterial oxygen (PaO2 ) markedly reduced to 65.3 mmHg, partial pressure of arterial carbon dioxide (PaCO2 )amplified to 49.2 mmHg, and the respiratory rate increased over time. Morphologically, the surface of the left lung appeared uneven on Day 1, the alveoli of the left lung disappeared on Day 2, and the left lung shrank on Day 7. A his‑ tological examination revealed that considerable cell infiltration began on Day 1 and lasted until Day 7, with a larger area of cell infiltration. Serum levels of IL-5, IL-6, IFN-γ, MCP-1, MIP-2, G-CSF, and TNF-α substantially rose on Day 7.
Conclusions
This modified approach for BLM-induced lung injury provided a severe, stable, and one-sided (left-lobe) ARDS animal model with consistent reproducibility. The physiological symptoms observed in this severe ARDS animal model are entirely consistent with the characteristics of clinical ARDS. The establishment of this ARDS animal model could help develop treatment for ARDS.
3.Measuring energy expenditure in Göttingen Minipigs using indirect calorimetry: validation and methodological considerations
Simon K. BREDUM ; Julie JACOBSEN ; Susanna CIRERA ; Berit Ø. CHRISTOFFERSEN
Laboratory Animal Research 2025;41(1):104-118
Background:
Obesity affects nearly a billion people globally and is associated with various health consequences.Current anti-obesity medications primarily target appetite, but drug candidates that modulate energy expenditure (EE) and substrate utilization based on respiratory exchange ratio (RER) are also essential to continuously improve the treatment modalities for people living with obesity. Selecting appropriate animal models and methods is crucial to improving translational value in preclinical research. While pig obesity models provide a relevant alternative to rodent models due to their similarities to humans, little is known about the assessment and translatability of EE in pigs. The aim of this study was to evaluate the translatability of minipigs for assessing the effect of EE-modulating drugs using indirect calorimetry and three positive control compounds that have known effects on EE and/or RER in humans. The study consisted of five sub-studies: Sub-study 1 assessed EE and RER based on sex (male/female) and diet (chow/high-fat diet) with and without correction for body composition; Sub-studies 2–4 evaluated changes in EE and RER after treatment with three positive control compounds: 2,4-dinitrophenol, DNP; a glucagon receptor agonist, GCG-RA; and a melanocortin receptor 4 agonist, MC4-RA; and sub-study 5 established three predictive equations for resting metabolic rate.
Results:
Sub-study 1 resulted in detectable differences in EE and RER based on diet/body sizes (P-value < 0.0001), while EE adjusted for body composition resulted in differences based on sex (P-value < 0.0001). Sub-studies 2–4 revealed that the three pharmacological interventions known to affect EE in humans, DNP, GCG-RA, and MC4-RA, showed similar effects in the Göttingen Minipigs by significantly increasing EE by 26.1% (P-value: 0.0014), 21.3% (P-value: 0.0491), and 25.4% (P-value: 0.0013), respectively, emphasizing the translational value of the model. In substudy 5, three predictive equations were established for RMR based on body composition, demographic and anthropometric measurements, and the most accurate equation based on all variables. All three equations demonstrated acceptable accuracy (adjusted R2 : 0.73–0.85).
Conclusions
The present study qualifies the use of Göttingen Minipigs for investigating EE in preclinical research and provides a framework for conducting such research.
4.Drug screening for ischemic stroke using larvae and adult zebrafish model: a review
Ni Made Dwi Mara Widyani NAYAKA ; I Ketut ADNYANA ; Kusnandar ANGGADIREDJA ; Indra WIBOWO
Laboratory Animal Research 2025;41(1):1-19
Ischemic stroke (IS) is the most recorded case of stroke that is caused by decreased blood flow to the brain. Nowadays, therapeutical agents for IS are limited and they have not shown maximum clinical results. Therefore, the exploration of new candidates for IS treatment continues to be done. Zebrafish as one of the animal models has its advantages and currently is being developed to be incorporated into the drug discovery pipeline of IS. This review explores the latest applications of the zebrafish model in screening potential therapeutic agents for IS. Key factors related to the experimental design such as developmental stage and strain, routes of drug administration, induction methods, and experimental parameters are also elaborated. Finally, this review offers future recommendations for the use of zebrafish in the pre-clinical study of IS. This review is beneficial as a reference for establishing drug screening protocols using the zebrafish IS model.
5.Carvacrol attenuated haloperidol-induced Parkinson’s disease via TNF/NFκβ-NLRP3-mediated pyroptosis
Faisal ALBAQAMI ; Khawaja Waqas AHMAD ; Fawad Ali SHAH
Laboratory Animal Research 2025;41(1):93-103
Background:
Parkinson’s disease is a debilitating and the second most common neurodegenerative disorder with a high prevalence. Parkinson’s disease has a multifaceted etiology characterized by an altered redox state and an excessive inflammatory response. In this study, we investigated the potential neuroprotective properties of carvacrol in a haloperidol-induced Parkinson’s model. In female Sprague-Dawley rats, the animal Parkinson model was induced by intraperitoneally administering 1 mg / kg of haloperidol once daily for fifteen days. Carvacrol was administered at a dose of 25 and 50 mg / kg once daily for fifteen days before haloperidol administration. In order to further illustrate the vital role of the tumor necrosis factor (TNF-α) pathway, we administered 50 mg / kg of the TNF-α inhibitor thalidomide once daily for 15 days.
Results:
Our results showed that haloperidol-induced motor deficits, changed endogenous antioxidant enzymes, along with higher levels of inflammasome (NLRP3) and other inflammatory mediators. Moreover, increased levels of lipid peroxidase (LPO) indicated a significant rise in oxidative stress due to haloperidol. Moreover, carvacrol reduced these effects by preventing pyroptosis mediated by the inflammasome (NLRP3) and TNF-α. The administration of thalidomide mitigated oxidative stress and suppresses inflammatory pathways through the augmentation of the intrinsic antioxidant system. Further, co-treatment of carvacrol with thalidomide synergized the neuroprotective effect of carvacrol as demonstrated by various immunoassays and histology analyses.
Conclusions
Taken together, our findings suggest that carvacrol mitigated haloperidol-induced Parkinson-like symptoms, partially through the downregulation of TNF-α and NLRP3.
6.Zebrafish as a model for human epithelial pathology
Ahmed ABU-SINIYEH ; Moayad KHATAIBEH ; Walid AL-ZYOUD ; Majed Al HOLI
Laboratory Animal Research 2025;41(1):34-56
Zebrafish (Danio rerio) have emerged as an influential model for studying human epithelial pathology, particularly because of their genetic similarity to humans and their unique physiological traits. This review explores the structural and functional homology between zebrafish and human epithelial tissues in organs, such as the gastrointestinal system, liver, and kidneys. Zebrafish possess significant cellular and functional homology with mammals, which facilitates the investigation of various diseases, including inflammatory bowel disease, nonalcoholic fatty liver disease, and polycystic kidney disease. The advantages of using zebrafish as a model organism include rapid external development, ease of genetic manipulation, and advanced imaging capabilities, allowing for the real-time observation of disease processes. However, limitations exist, particularly concerning the lack of organs in zebrafish and the potential for incomplete phenocopy of human conditions. Despite these challenges, ongoing research in adult zebrafish promises to enhance our understanding of the disease mechanisms and regenerative processes. By revealing the similarities and differences in epithelial cell function and disease pathways, this review highlights the value of zebrafish as a translational model for advancing our knowledge of human health and developing targeted therapies.
8.Effects of Cirsium japonicum var. maackii on avelliation of metabolic disease by improving insulin resistance
Hye-Bin YOON ; Yuseong JANG ; Hyeon-Gi PAIK ; Hwal CHOI ; Jihye CHOI ; Jungkee KWON
Laboratory Animal Research 2025;41(1):71-80
Background:
Metabolic syndrome (MetS) refers to a group of risk factors that cause health problems, such as obesity, diabetes, dyslipidemia, and hyperglycemia. MetS is characterized by insulin resistance, which leads to abnormal insulin sensitivity. Cirsium japonicum var. maackii (CJ) is perennial herbaceous species found in Asia that exhibits antioxidant, antidiabetic, antitumor, antifungal, and anti-inflammatory activities. In this study, we aimed to measure the effects of CJ on MetS by improving insulin resistance in a db/db type 2 diabetes mouse model. After administrating CJ extract (CJE) for db/db mouse for 6 weeks, we measured with the evaluation of Insulin resistance, lipid profiles, histological analysis of liver, damage of liver and kideny.
Results:
The results showed that CJE was effective in reducing body weight and fat mas and showed a positive effect on lowering blood glucose and improving insulin sensitivity. CJE improved dyslipidemia by increasing serum-HDL levels and decreasing serum-LDL levels. In addition, CJE reduced liver and kidney damage in histological analysis.
Conclusions
These results demonstrate the anti-diabetic effects of CJE and suggest its potential for improving MetS.Therefore, CJE may have potential values as a functional food material for managing MetS.
9.Regulatory roles of eugenol in paraquataltered SNCA/LZTS3/MAPT in the cerebellum of Wistar rats
Obinna Onwe UCHEWA ; Augustine Oseloka IBEGBU ; Samuel Okoronkwo OKAFOR ; Joseph Alo NWAFOR ; Ogugua Augustine EGWU
Laboratory Animal Research 2025;41(1):57-70
Background:
The Microtubules-associated protein tau (MAPT), alpha-synuclein (SNCA), and leucine zipper tumor suppressor 3 (LZTS3) genes are implicated in neurodegeneration and tumor suppression, respectively. This study investigated the regulatory roles of eugenol on paraquat-altered genes.
Results:
Forty male Wistar rats divided into five groups of eight rats were used. The control group received normal saline; the Paraquat (PQ)-untreated group received only Paraquat. The low dose of eugenol was 200 mg/kg, the medium dose of eugenol was 400 mg/kg, and the high dose of eugenol was 600 mg/kg. All groups except the control group received 10 mg/kg of PQ orally for 14 days at one-day intervals, allowing PQ in the rats for 28 days.Eugenol treatment started on the 29th and lasted 14 days. Motor impairments were determined using wire string and beam-walk; biomarkers were estimated using cerebellar homogenates, while frozen cerebellum was used to study LZTS3, MAPT, and SNCA gene expression. LZTS3 was significantly suppressed in the PQ-untreated group and highly expressed in the eugenol-treated group. The MAPT and SNCA genes were overexpressed in the PQ-untreated group compared to the control group. Eugenol significantly decreased the expression of these genes compared to that in the PQ-untreated group. Antioxidants were reduced considerably, and oxidative stress markers were increased significantly, which could have caused increased protein fibrillation and reduced limb functionality. Histology revealed that eugenol mitigated the alterations caused by Paraquat.
Conclusions
PQ can enhance tumor expression in addition to causing neurotoxicity, which decreases limb functionality, while eugenol, an antioxidant, can mitigate the effects of PQ.
10.Animal models of hemorrhage, parameters, and development of hemostatic methods
Gholamhossien DARYA ; Hamid MOHAMMADI ; Zeinab DEHGHAN ; Alireza NAKHAEI ; Amin DERAKHSHANFAR
Laboratory Animal Research 2025;41(1):20-33
Hemorrhage is a prevalent side effect of various injuries and can be life-threatening in certain instances. It is categorized into compressible and non-compressible types, each necessitating distinct modeling, laboratory assessments, and hemostatic approaches. This study utilized the keywords Hemorrhage, Bleeding, Animal Modeling, and Hemostat in reputable databases. The findings indicate that femoral artery hemorrhage and hepatic parenchymal hemorrhage are the predominant modeling techniques for compressible and noncompressible bleeding, respectively. Furthermore, it is noted that animal models of compressible hemorrhages are primarily situated in superficial body areas to investigate dressing or additive hemostats, while non-compressible hemorrhage models, typically located in visceral organs, are employed to examine adhesive or surgical instrumentbased hemostats.

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