Animal models and experimental medicine最新文献

Background: In a previous study, we found that Atractylodes macrocephala and Paeoniae radix (AM-PR) was useful for the alleviation of functional constipation (FC). However, the precise mechanism underlying the compatibility between AM and PR in the treatment of FC remains uncertain. This study aims to analyze the pharmacokinetic differences in the active ingredients in the blood of rat models with FC when applied individually and in combination with AM-PR. It also seeks to compare the changes in the content of the active ingredient when applied individually and in combination with in vitro AM-PR, further in-depth investigation into its material foundation in terms of pharmacokinetics, as well as the composition of the substance.

Methods: Blood microdialysis samples were collected using microdialysis technology. These samples from rats with FC were compared after administration of AM, PR, and AM-PR. The concentration of the main active ingredients was determined using the Ultra Performance Liquid Chromatography-Tunable Ultraviolet (UPLC-TUV) method. The concentration of the main active ingredients of the decoction compatibility before and after combining AM-PR was also determined using the UPLC-TUV method.

Results: Our findings reveal that upon combination, the time to maximum concentration (T_max) of isochlorogenic acid A (ICA-A) and ataridolide II (ATR-II) T_max was prolonged, terminal elimination half-life (T_1/2) was reduced, and maximum plasma concentrations (C_max) increased. The T_max of ataridolide III (ATR-III) remained consistent, whereas its T_1/2 and C_max were significantly reduced. Conversely, for peoniflorin (PAE), T_max occurred sooner, T_1/2 was shortened, and C_max increased. The T_max for albiflorin (ALB) remained consistent, whereas T_1/2 and C_max witnessed significant increases. The area under the moment curve (AUMC) (0-t) and AUMC (0-∞) of PAE, ALB, ICA-A, ATR-II experienced an increase after AM-PR administration in rats, attributable to the heightened C_max. In comparison to individual herb administration, the T_max of ALB was advanced in combination, the T_max of PAE remained unchanged, and the T_max of ICA-A and ART-II was delayed, with an increased area under the concentration-time curve (AUC) (0-t), indicating enhanced C_max and bioavailability. Furthermore, the dissolution rates of PAE, ICA-A, and ATR-II significantly improved after compatibility.

Conclusions: This study partially clarifies the rationale and compatibility of AM-PR in treating FC and offers a new perspective on the pharmacokinetic interactions of AM-PR in FC treatment.

Background: The aim of the study was to explore a feasible method for alleviating limb ischemia-reperfusion injury (LI/RI) through the use of a high-concentration citrate solution (HC-A solution) for limb perfusion (LP).

Methods: Eighteen pigs were divided into three groups: the Sham group, LI/RI group, and HCA group. The Sham group underwent exposure of the iliac artery and vein. The LI/RI group underwent tourniquet placement and clamping of the iliac artery and vein to simulate LI/RI. The HCA group received HC-A solution LP for 30 min through the left iliac artery below the level of blood flow occlusion based on the LI/RI group. Oxidative stress markers and inflammatory response markers were compared among the three groups.

Results: Compared to the LI/RI group, the HCA group showed significantly lower levels of serum creatine kinase (CK), lactate dehydrogenase (LDH), malondialdehyde (MDA), tumor necrosis factor-α (TNF-α), aspartate aminotransferase (AST), and alanine aminotransferase (ALT), and significantly greater activities of serum superoxide dismutase (SOD) (p < 0.05). There were no significant differences in serum interleukin-6 (IL-6) or in muscle MDA, SOD, TNF-α, and IL-6 between the HCA group and the LI/RI group (p > 0.05). Compared to the LI/RI group, MDA, TNF-α, and IL-6 levels in the liver were significantly lower in the HCA group (p < 0.05), while SOD activities were not significantly different (p > 0.05). Histopathological examination revealed reduced skeletal muscle and liver damage in the HCA group compared to the LI/RI group.

Conclusions: HC-A solution LP can alleviate liver damage caused by LI/RI in pigs.

Background: Qi pi pill (QPP), which contains Renshen, Baizhu, Fuling, Gancao, Chenpi, Shanyao, Lianzi, Shanzha, Liushenqu, Maiya, and Zexie, was recommended for preventing and treating COVID-19 in Shandong Province (China). However, the mechanism by which QPP treats infectious diseases remains unclear. This study aims to investigate the therapeutic effect of QPP in vitro and on acute influenza infection in mice, exploring its mechanism of action against influenza A virus (IAV).

Methods: The in vitro activity of QPP was assessed using dose-response curve analysis and titer reduction assay, and its antiviral mechanism was identified in vitro by real-time polymerase chain reaction (PCR), time-of-addition, and enzymatic assays. The antiviral efficacy of QPP was further evaluated in vivo using BALB/c mice infected with IAV. At the same time, each single Chinese herbal medicine in QPP was evaluated to preliminarily identify those with antiviral effects.

Results: In vitro results showed that QPP exhibited a higher potency antiviral effect against both influenza A and B viruses, inhibiting viral RNA replication and release by targeting RNA-dependent RNA polymerase and neuraminidase. Additionally, QPP significantly decreased the expression of inflammatory cytokines in A549 cells. In vivo study revealed that QPP significantly reduced the lung index and viral load in lung tissue of mice infected with IAV. Renshen, Gancao, Zexie, and Lianzi were the Chinese herbal medicines from QPP that showed anti-IAV activity.

Conclusion: The antiviral activity of QPP targets IAV replication and release, cytokine modulation in host cells, and provides protection in mice with acute influenza infection.

The high morbidity and mortality of colorectal cancer (CRC) is a major challenge in clinical practice. Although a series of alternative research models of CRC have been developed, appropriate orthotopic animal models that reproduce the specific clinical response as well as pathophysiological immune features of CRC are still lacking. In the current study, we constructed a CRC orthotopic xenograft model by implanting the tumor tubes at the colorectum of mice and monitored the model development using bioluminescence imaging. This model successfully recapitulates the clinical chemotherapy efficacy, including reduced total flux, tumor weight, and the expression of Ki67 after treatment of the first-line chemotherapy regime of CRC (FOLFOX: oxaliplatin and 5-fluorouracil/calcium folinate). The model also reproduced the immunosuppressive effect of FOLFOX, indicated by decreased infiltration of macrophages and increased Treg cells in tumor. Additionally, the orthotopic xenograft approach may be applied in immunodeficient NCG/NSG mice for constructing patient-derived xenografts, and being used in clinical precision medicine and drug evaluation. We believe the current model is a successful surgical orthotopic xenograft approach for cancer research and deserves to be popularized, which will provide a convenient and efficient platform for in-depth mechanism exploration of CRC and preclinical drug evaluation.

Background: To investigate the toxicity of N-n-butyl haloperidol iodide (F2), a quaternary ammonium salt derivative of haloperidol, in mice for potential therapeutic purposes.

Methods: The acute median lethal dose (LD₅₀) of F2 was determined using the Bliss method following intravenous administration in mice. Routine surface electrocardiograms (ECGs) and arterial blood pressures (aBPs) were recorded under general anesthesia in untreated and pharmacologically vagotomized mice injected with F2. Sublethal doses of F2 were tested for their effects on aBP, heart rate, and biochemical parameters such as lactate dehydrogenase (LDH), blood urea nitrogen (BUN), and serum lactate levels. Histopathological changes in the heart, lungs, liver, and kidneys were evaluated after F2 administration.

Results: The acute LD₅₀ of F2 was determined to be 5.11 mg/kg. A 10 mg/kg dose of F2 caused severe hypotension, second-degree atrioventricular block, progressive prolongation of Pmurr intervals, and death due to cardiac asystole. Similar ECG and aBP changes were observed in atropine-pretreated mice, indicating that cholinergic effects do not play a major role in F2-induced toxicity. Sublethal doses of F2 (1.2 and 2.4 mg/kg) caused dose-dependent decreases in aBP and increases in heart rate. F2 induced significant, dose-dependent increases in LDH, BUN, and serum lactate levels. Histopathological analysis revealed acute lung lesions at 10 mg/kg, with no significant changes observed in the heart, liver, or kidneys.

Conclusion: Acute intravenous injection of F2 exhibits dose-dependent cardiopulmonary toxicity, characterized by severe hypotension, arrhythmias, and biochemical changes. These findings highlight the potential risks of F2 and the need for further evaluation of its safety profile for therapeutic use.

Background: Makorin ring finger protein 3 gene (MKRN3) gene mutation is the most common genetic cause of central precocious puberty (CPP) in children. Due to the lack of ideal MKRN3-modified animal model (MKRN3-modified mice enter puberty only 4-5 days earlier than normal mice), the related research is limited.

Methods: Therefore, the MKRN3-modified rabbit was developed using CRISPR (clustered regularly interspaced short palindromic repeats) gene editing technology. The genotype identification and phenotype evaluation of MKRN3-modified rabbits were carried out.

Results: The first estrus of MKRN3-modified female rabbits was observed ~27 days earlier than that of wild-type female rabbits, with a typical CPP phenotype. This study found increased gonadotropin releasing hormone (GnRH) and decreased gonadotropin inhibiting hormone (GnIH) in the hypothalamus of the CPP rabbit model with MKRN3 gene mutation. Although this study failed to fully clarify the pathogenesis of CPP caused by MKRN3 mutation, it found some differentially expressed genes and potential pathways through transcriptome sequencing.

Conclusions: This study established a novel CPP model: paternal MKRN3 gene-modified rabbit. It is hoped that the establishment of this model will help researchers better understand, treat, and prevent CPP in the future.

Background: Quantifying the rich home-cage activities of tree shrews provides a reliable basis for understanding their daily routines and building disease models. However, due to the lack of effective behavioral methods, most efforts on tree shrew behavior are limited to simple measures, resulting in the loss of much behavioral information.

Methods: To address this issue, we present a deep learning (DL) approach to achieve markerless pose estimation and recognize multiple spontaneous behaviors of tree shrews, including drinking, eating, resting, and staying in the dark house, etc. RESULTS: This high-throughput approach can monitor the home-cage activities of 16 tree shrews simultaneously over an extended period. Additionally, we demonstrated an innovative system with reliable apparatus, paradigms, and analysis methods for investigating food grasping behavior. The median duration for each bout of grasping was 0.20 s.

Conclusion: This study provides an efficient tool for quantifying and understand tree shrews' natural behaviors.

Background: Scientific evidence to guide clinicians on the use of different antiseizure drugs in combination therapy is either very limited or lacking. In this study, the impact of lacosamide and perampanel alone and in combination was tested in corneal kindling model in mice, which is a cost-effective mechanism for screening of antiseizure drugs.

Methods: The impact of lacosamide (5 mg/kg) and perampanel (0.125 mg/kg) alone and their combination was tested in corneal kindling process (3-mA current for 3 s applied twice daily for consecutive 12 days) in male BALB/c mice. Post-kindling, mice were subjected to a battery of behavioral tests assessing anxiety, memory, and depression-like behaviors. Brain tissues were then harvested for analysis of oxidative stress biomarkers.

Results: Our results showed that the combination therapy of lacosamide and perampanel was more effective in reducing seizure progression than monotherapy of these drugs. Animals treated with combination therapy showed significant behavioral improvements, as reduced anxiety and depression were noticed, and their cognitive abilities were notably better compared to animals of all other groups. Moreover, biochemical assays of isolated brains from combination-treated group revealed lesser amount of oxidative stress. In addition, outcomes of dual regime were comparable to the phenytoin in seizure control but showed superior benefits in mitigation of kindling-prompted behavioral dysfunction and oxidative stress.

Conclusions: This study suggests that the lacosamide and perampanel combination therapy worked noticeably better in halting the corneal kindling process in mice and improved the epilepsy-associated psychiatric disorders that might be due to antioxidant effects of both drugs.

Background: This study evaluates the efficacy of gabexate mesylate thermosensitive in-situ gel (GMTI) in the treatment of beagle grade III pancreatic trauma (PT) with the assistance of contrast-enhanced ultrasound (CEUS) and investigates its mechanism of action.

Methods: A grade III PT model consisting of 15 beagle dogs with severed main pancreatic ducts was created and treated with cephalic vein injection of gabexate mesylate (GM) (1.54 mL/10 kg, TID) and peripancreatic injection of GMTI (4.63 mL/10 kg, QD) guided by CEUS within 24 h post-surgery. Ascites and serum levels of amylase (AMY), lipase (LPS), C-reactive protein (CRP), interleukin (IL)-6, tumor necrosis factor (TNF)-α, and urinary trypsinogen activating peptide (TAP) were detected by ELISA. Histopathological changes in the canine pancreas were observed by Hematoxylin and Eosin staining.

Results: CEUS accurately displayed pancreatic lesions and guided catheterisation. Compared to the control group, the ascites was significantly reduced after treatment (p < 0.01). AMY and LPS ascites significantly decreased on post-operative 1st and 2nd day (p < 0.01). The levels of AMY, LPS, CRP, IL-6, and TNF-α in serum were decreased (p < 0.05 or p < 0.01). Urinary TAP was decreased 1 and 2 days after treatment (p < 0.05 or p < 0.01, respectively). In the control group, pancreatic tissue necrosis was evident in the wound area. Normal glandular cell structures and fibrous tissue hyperplasia were observed in the wound area after GMTI treatment. The GMTI group performed better than the GM group in improving pancreatic histology and reducing AMY levels in the early post-operative period.

Conclusion: Guided by CEUS, daily peripancreatic injections of GMTI in Beagles effectively inhibit pancreatic enzyme activity and aid in the adjuvant treatment of pancreatic trauma.

Background: The inability of damaged neurons to regenerate and of axons to establish new functional connections leads to permanent functional deficits after spinal cord injury (SCI). Although astrocyte reprogramming holds promise for neurorepair in various disease models, it is not sufficient on its own to achieve significant functional recovery.

Methods: A rat SCI model was established using a spinal cord impactor. Seven days postsurgery, adeno-associated virus were injected to overexpress the transcription factors NeuroD1 and Neurogenin-2 (Ngn2) in the spinal cord. The rats were then trained to walk on a weight-supported treadmill for 4 weeks, starting 14 days after modeling. The effects of these interventions on motor and sensory functions, as well as spinal cord tissue repair, were subsequently evaluated.

Results: The combination of NeuroD1 and Ngn2 overexpression with weight-supported exercise training significantly improved gait compared to either intervention alone. The group receiving the combined intervention exhibited enhanced sensitivity in sensory assessments. Immunofluorescence analysis revealed increased colocalization of astrocytes and microtubule-associated protein 2-positive neurons in the injury area. These effects were more pronounced than those observed with spinal cord tissue repair alone. Additionally, the combined intervention significantly reduced glial scarring and the size of the injury area.

Conclusion: Exercise intervention enhances the reprogramming effects of astrocytes and restores motor function, yielding better results than either intervention alone.