Development Of Pathology Research Articles

Connectome-based predictive modeling of brain pathology and cognition in Autosomal Dominant Alzheimer's Disease.

Autosomal Dominant Alzheimer's Disease (ADAD) through genetic mutations can result in near complete expression of the disease. Tracking AD pathology development in an ADAD cohort of Presenilin-1 (PSEN1) E280A carriers' mutation has allowed us to observe incipient tau tangles accumulation as early as 6 years prior to symptom onset. Resting-state functional Magnetic Resonance Imaging (fMRI) and Positron-Emission Tomography (PET) scans were acquired in a group of PSEN1 carriers (n=32) and non-carrier family members (n=35). We applied Connectome-based Predictive Modeling (CPM) to examine the relationship between the participant's functional connectome and their respective tau/amyloid-β levels and cognitive scores (word list recall). CPM models strongly predicted tau concentrations and cognitive scores within the carrier group. The connectivity patterns between the temporal cortex, default mode network, and other memory networks were the most informative of tau burden. These results indicate that resting-state fMRI methods can complement PET methods in early detection and monitoring of disease progression in ADAD.

Effect of diet-induced obesity and non-drug options of its correction on the functional and morphological liver characteristics in female Wistar rats

A nonalcoholic fatty liver disease is one of the unfavorable consequences of obesity. However, experimental studies on the mechanisms of this pathology development are being carried out mainly on male rodents, and the data on the development of fatty hepatosis in females are insufficient.The aim of the study was to investigate the effect of diet-induced obesity and non-drug options for its correction on the morphological liver characteristics and liver metabolic parameters in female Wistar rats. Experiments were conducted on sexually mature female Wistar rats and included the study of morphological and biochemical parameters of the liver functional state in animals kept on standard diet (StD, 16 weeks), on a high-calorie diet (HCD, 16 weeks) when switching to a standard diet (HCD/StD, 8/8 weeks), when the physical activity in the form of treadmill running was included (StD + running, HCD + running and HCD/StD + running, 8/8 weeks).Long-term use of HCD in female rats caused visceral obesity, liver fatty dystrophy with disruption of organ histoarchitectonics, shifts in hepatic metabolism and increased lipid peroxidation. Correction of diet-induced obesity by switching to StD led to a complete or partial normalization of the studied indices. In the “HCD + running” group, metabolic disorders are often more pronounced than in the “HCD” group. The transition to HCD/StD + running promoted the most complete restoration of metabolism and histostructure of the liver with regeneration signs of the organ.Thus, an optimal correction of visceral obesity, fatty liver dystrophy and its metabolism is impossible without diet normalization in female Wistar rats. An additional regenerating effect with respect to the liver histostructure is achieved with a combined variant of correction ‒ transition to a balanced diet and moderate physical activity.

Deep-ultraviolet Fourier ptychography (DUV-FP) for label-free biochemical imaging via feature-domain optimization

We present deep-ultraviolet Fourier ptychography (DUV-FP) for high-resolution chemical imaging of biological specimens in their native state without exogenous stains. This approach uses a customized 265-nm DUV LED array for angle-varied illumination, leveraging the unique DUV absorption properties of biomolecules at this wavelength region. We implemented a robust feature-domain optimization framework to overcome common challenges in Fourier ptychographic reconstruction, including vignetting, pupil aberrations, stray light problems, intensity variations, and other systematic errors. By using a 0.12 numerical aperture low-resolution objective lens, our DUV-FP prototype can resolve the 345-nm linewidth on a resolution target, demonstrating at least a four-fold resolution gain compared to the captured raw images. Testing on various biospecimens demonstrates that DUV-FP significantly enhances absorption-based chemical contrast and reveals detailed structural and molecular information. To further address the limitations of conventional FP in quantitative phase imaging, we developed a spatially coded DUV-FP system. This platform enables true quantitative phase imaging of biospecimens with DUV light, overcoming the non-uniform phase response inherent in traditional microscopy techniques. The demonstrated advancements in high-resolution, label-free chemical imaging may accelerate developments in digital pathology, potentially enabling rapid, on-site analysis of biopsy samples in clinical settings.

Clinical and translational mode of single-cell measurements: An artificial intelligent single-cell.

With rapid development and mature of single-cell measurements, single-cell biology and pathology become an emerging discipline to understand the disease. However, it is important to address concerns raised by clinicians as to how to apply single-cell measurements for clinical practice, translate the signals of single-cell systems biology into determination of clinical phenotype, and predict patient response to therapies. The present Perspective proposes a new system coined as the clinical artificial intelligent single-cell (caiSC) with the dynamic generator of clinical single-cell informatics, artificial intelligent analyzers, molecular multimodal reference boxes, clinical inputs and outs, and AI-based computerization. This system provides reliable and rapid information for impacting clinical diagnoses, monitoring, and prediction of the disease at the single-cell level. The caiSC represents an important step and milestone to translate the single-cell measurement into clinical application, assist clinicians' decision-making, and improve the quality of medical services. There is increasing evidence to support the possibility of the caiSC proposal, since the corresponding biotechnologies associated with caiSCs are rapidly developed. Therefore, we call the special attention and efforts from various scientists and clinicians on the caiSCs and believe that the appearance of the caiSCs can shed light on the future of clinical molecular medicine.

Image stitching algorithm for super-resolution localization microscopy combined with fluorescence noise prior.

Super-resolution panoramic pathological imaging provides a powerful tool for biologists to observe the ultrastructure of samples. Localization data can maintain the essential ultrastructural information of biological samples with a small storage space, and also provides a new opportunity for stitching super-resolution images. However, the existing image stitching methods based on localization data cannot accurately calculate the registration offset of sample regions with no or few structural points and thus lead to registration errors. Here, we proposed a stitching framework called PNanoStitcher. The framework fully utilizes the distribution characteristics of the background fluorescence noise in the stitching region and solves the stitching failure in sample regions with no or few structural points. We verified our method using both simulated and experimental datasets, and compared it with existing stitching methods. PNanoStitcher achieved superior stitching results on biological samples with no structural and few structural regions. The study provides an important driving force for the development of super-resolution digital pathology.

CAUSES OF MORBIDITY AND MORTALITY IN STELLER SEA LIONS (EUMETOPIAS JUBATUS) UNDER PROFESSIONAL CARE IN NORTH AMERICAN AQUARIUMS FROM 1979 TO 2021.

To date, published comprehensive pathology investigations documented in Steller sea lions (SSL; Eumetopias jubatus) are from free-ranging populations, whereas health data from those under professional care in aquariums are currently lacking. A retrospective review of gross and histopathologic reports of SSL under human care in North American aquariums from 1979 to 2021 (n = 20) was performed. Associations between age, sex, or birth origin (born in aquariums versus the wild) with cause of death (COD) and comorbidities were explored. Age was significantly associated with development of endocrine organ pathology (P = 0.011). A relationship between age and both cardiovascular and ocular disease was suggested by the data, but did not reach significance (P = 0.058). Ocular disease was significantly associated with being born in aquariums (P = 0.022). The most common COD was neoplasia (n = 10), which was significantly associated with aged animals (P = 0.038). Less frequent COD included sepsis (confirmed, n = 2; suspected, n = 3), cardiomyopathy (n = 1), clostridial enteritis (n = 1), Sarcocystis spp. (n = 1), complication secondary to sedation (n = 1), and unknown (n = 1). This is the first report documenting the high prevalence of neoplasia in SSL, with tumors found incidentally in three individuals, frequent metastasis (10/13, 77%), and many cases of multiple primary malignancies (6/13, 46%). These data expand upon the current understanding of disease in SSL, highlight this species' predisposition to neoplasia with increasing longevity, and underscore the need for heightened screening in aged animals, which may ultimately serve to elevate the care of SSL under professional care in aquariums.

Association Between Intracranial Aneurysms and Genes

Ruptured intracranial aneurysm(IA) leads to aneurysmal subarachnoid hemorrhage(aSAH), which is the most severe type of stroke. Besides age, sex, and alcohol consumption, family history is considered an important risk factor. Approximately 5% of the Japanese population is affected by unruptured cerebral aneurysms. However, their developmental processes and pathologies remain unclear. Familial cerebral aneurysms account for 10% of all cerebral aneurysm cases, and somatic gene mutations are believed to be involved in their development. This review summarizes the recent findings from family-based, genome-wide association, sequencing, and somatic gene mutation studies on IA.

The Role of Mast Cell-Neuron Tandem in the Regulation of Cardiac Function in Cardiovascular Pathologies

This review is consisted of new data about participation of mast cells (MCs) in the regulation of cardiac function in the development of some cardiovascular pathologies. In norm the number of MCs in the heart is very small compared to their number in barrier tissues. They are responsible for homeostasis of cardiac tissue and ensuring the interaction of the organ with the nervous and endocrine systems throughout the life of the organism. Pathological processes in the heart are accompanied by the development of inflammation reaction. MCs fulfil a significant role in the all stages of inflammation. The number of MCs significantly increases against the background of myocardial infarction development, fibrotic heart disease and vascular atherosclerosis. Their contribution to the pathogenesis of the disease is ambiguous, because on the one hand it is aimed at tissue adaptation to damage, and on the other hand - avalanche-like synthesis and release of mediators from activated MCs aggravates the course of the process. As a result, MCs change their secretory profile, interfere in the realization of cardiac functions against the background of inflammation, but at the same time modulate afferent information flow from the heart and efferent influences of the nervous system. The coordinated work of the system loses its stability, which can lead to severe consequences for the vital activity of the whole organism. The analysis of the current state of the problem indicates that the heart activity depends on the functional state of MCs, their complex interaction with the nervous system both in the norm and, especially, against the background of pathological processes development.

Investigation of pulmonary artery and circulating endothelin-1 expression in dogs with pulmonary hypertension secondary to myxomatous mitral valve disease.

Pulmonary hypertension (PH) is a condition characterized by abnormally elevated pressure in the pulmonary vasculature. It is a common complication of myxomatous mitral valve disease (MMVD) in dogs. Several vasoactive substances, including endothelin-1 (ET-1), have been suggested to contribute to pathological changes in the pulmonary arteries of patients with PH. This study aimed to examine the local and systemic expression of ET-1 in dogs with PH secondary to MMVD. Lung tissues were collected from 20 client-owned dogs during the first stage of the study and divided into three groups: normal dogs (n = 5), MMVD dogs (n = 8), and MMVD+PH dogs (n = 7). The expression of ET-1 and endothelin A receptor (ETAR) in the pulmonary arteries was determined using immunohistochemistry. Blood samples were collected from 61 client-owned dogs for the second stage of the study and divided into three groups: normal (n = 22), MMVD (n = 20), and MMVD+PH (n = 19). Plasma ET-1 concentration was measured using a sandwich enzyme-linked immunosorbent assay. There was no difference in ET-1 and ETAR expression in the pulmonary arteries among the three groups. Similarly, there was no difference in the plasma ET-1 concentration between the groups. In addition, no correlation was found between the immunohistochemical expression of ET-1 and ETAR and the thickness of the pulmonary arteries or between the plasma ET-1 level and echocardiographic variables. The lack of difference in the expression of ET-1 and ETAR in the pulmonary arteries and in the circulating ET-1 concentration among the studied groups suggests that ET-1 may not be related to the pathological development of PH secondary to MMVD in dogs. Due to the small sample size in this study, further research is needed to confirm these findings.

Apolipoprotein E in Alzheimer's Disease: Focus on Synaptic Function and Therapeutic Strategy.

Synaptic dysfunction is a critical pathological feature in the early phase of Alzheimer's disease (AD) that precedes typical hallmarks of AD, including beta-amyloid (Aβ) plaques and neurofibrillary tangles. However, the underlying mechanism of synaptic dysfunction remains incompletely defined. Apolipoprotein E (APOE) has been shown to play a key role in the pathogenesis of AD, and the ε4 allele of APOE remains the strongest genetic risk factor for sporadic AD. Itiswidelyrecognizedthat APOE4 accelerates the development of Aβ andtau pathology in AD. Recent studies have indicated that APOE affects synaptic function through a variety of pathways. Here, we summarize the mechanism of modulating synapses by various APOE isoforms and demonstrate the therapeutic potential by targeting APOE4 for AD treatment.

PRIMARY PREVENTION OF CARDIOVASCULAR DISEASES AND WAYS OF IMPROVING RISK ASSESSMENT OF THEIR DEVELOPMENT IN CLINICAL PRACTICE

Introduction. Cardiovascular diseases are a global problem and one of the main causes of death worldwide. Aim. To highlight and analyze modern approaches to the use in routine practice of risk factors for the development of cardiovascular pathology, which is appropriate for further clinical decision-making based on a meta-analysis of literary sources, clinical guidelines and the results of multicenter studies. Materials and methods. The search and selection of publications, systematic reviews and current recommendations is carried out using PubMed and Google Scholar databases, their detailed analysis and description. Results and discussion. The article provides data on primary prevention, which includes assessment and implementation of measures to reduce the overall risk of developing cardiovascular diseases in patients. Methods for assessing individual total or relative risk (use of SCORE and SCORE 2 scales, various risk calculators, etc.) taking into account the level of cholesterol and its various fractions are given. The importance of taking into account additional independent risk factors for the development of cardiovascular diseases is also considered. The possibility of taking into account the level of lipoprotein (a), high-density lipoprotein, and high-sensitivity C-reactive protein to improve the accuracy of predicting cardiovascular risk is discussed. Attention is paid to the possibility of using such "non-metabolic" factors as environmental pollution, psychosocial stress and stressful conditions, sleep disorders and obstructive sleep apnea syndrome, family history and genetic factors to change the risk category. Conclusions. The results of the conducted analysis prove that the given methods and importance of detection of subclinical atherosclerosis especially in patients with low or medium (marginal) cardiovascular risk. The use of both primary and secondary risk factors may be appropriate to inform decisions about changes in the calculated risk of such patients, the use and effectiveness of specific cardiac treatments, and the implementation of preventive measures such as statin therapy.

Evaluation of the effectiveness of a hyperlipidic-hyperglycemic diet in inducing weight gain and dyslipidemia in wistar rats

Hyperlipidic and hyperglycemic diets, called cafeteria diets, are important factors that favor the development of pathologies and comorbidities such as obesity, type 2 diabetes mellitus, neoplasms, and cardiovascular diseases. Thus, their use in experimental studies that aim to understand the factors that encompass the pathophysiology and treatment of obesity and the comorbidities involved has increased in recent years. Taking that into consideration, the aim of this study was to analyze the effectiveness of a high-fat-hyperglycemic (HHD) diet in inducing weight gain and dyslipidemia in Wistar rats. In this study, 22 rats were divided into 2 groups, and exposed to standard and high-fat-hyperglycemic diets for 11 weeks. After that, euthanasia was carried out along with collection of blood samples for analysis of the following parameters of the lipid profile: triglycerides (TG), total cholesterol (TC), low-density lipoprotein (LDL-c), high-density lipoprotein (HDL-c), and TG/HDL-c ratio. A more expressive weight gain was found in the rats that received HHD (p=0.006) in relation to the group maintained on the standard diet. Furthermore, no statistically detected alterations were found in the levels of TG, TC, LDL-c, HDL-c, and TG-HDL-c ratio. Analysis of the results indicated that HHD was effective in inducing weight gain in rats. However, there was no significant influence on the lipid profile.

Steroid profiling in human primary teeth via liquid chromatography-tandem mass spectrometry for long-term retrospective steroid measurement.

Steroid hormones are important modulators of many physiological processes, and measurements of steroids in blood, saliva, and urine matrices are widely used to assess endocrine pathologies and stress. However, these matrices cannot be used to retrospectively assess early-life stress and developmental endocrine pathologies, because they do not integrate steroid levels over the long term. A novel biological matrix in which to measure steroids is primary teeth (or "baby teeth"). Primary teeth develop early in life and accumulate various endogenous molecules during their gradual formation. Here, we developed and validated the first assay to measure steroids in human primary teeth using liquid chromatography-tandem spectrometry (LC-MS/MS). Our assay is highly sensitive, specific, accurate, and precise. It allows for the simultaneous quantification of 17 steroids in primary teeth (16 of which have not been examined previously in primary teeth). Overall, steroid levels in primary teeth were relatively low, and 8 steroids were quantifiable. Levels of dehydroepiandrosterone, cortisol, and progesterone were the highest of the 17 steroids examined. Next, we used this assay to perform steroid profiling in primary teeth from males and females. The same 8 steroids were quantifiable, and no sex differences were found. Levels of androgens (androstenedione and testosterone) were positively correlated, and levels of glucocorticoids (cortisol, cortisone, corticosterone, 11-dehydrocorticosterone) were also positively correlated. These data demonstrate that multiple steroids can be quantified by LC-MS/MS in human primary teeth, and this method potentially provides a powerful new way to retrospectively assess early-life stress and developmental endocrine pathologies.

Persistent Post COVID-19 Endothelial Dysfunction and Oxidative Stress in Women.

The assessment of endothelial dysfunction and free radical homeostasis parameters were performed in 92 women, aged 45 to 69 years, divided into the following groups: women without COVID-19 (unvaccinated, no antibodies, control); women with acute phase of COVID-19 infection (main group, COVID-19+); 12 months post COVID-19+; women with anti-SARS-CoV-2 IgG with no symptoms of COVID-19 in the last 12 months (asymptomatic COVID-19). Compared to the control, patients of the main group had lower glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities, decreased advanced glycation end products (AGEs) level, higher glutathione reductase (GR) activity, and higher glutathione S transferases pi (GSTpi), thiobarbituric acid reactants (TBARs), endothelin (END)-1, and END-2 concentrations (all p ≤ 0.05). The group with asymptomatic COVID-19 had lower 8-OHdG and oxidized glutathione (GSSG) levels, decreased total antioxidant status (TAS), and higher reduced glutathione (GSH) and GSH/GSSG levels (all p ≤ 0.05). In the group COVID-19+, as compared to the group without clinical symptoms, we detected lower GPx and SOD activities, decreased AGEs concentration, a higher TAS, and greater GR activity and GSTpi and TBARs concentrations (all p ≤ 0.05). The high content of lipid peroxidation products 12 months post COVID-19+, despite decrease in ENDs, indicates long-term changes in free radical homeostasis. These data indicate increased levels of lipid peroxidation production contribute, in part, to the development of free radical related pathologies including long-term post COVID syndrome.

Heparan sulfate proteoglycans in pathological protein aggregation and brain functionality

Proteoglycans are highly specialized proteins with specific interactive modules in their core proteins, allowing them to engage with a diverse range of cellular and structural proteins, contributing to many varied roles in cell processes. This review examines the roles of heparan sulfate (HS) proteoglycans (HSPGs) in the assembly of pathological protein aggregates in brain tissues, which negatively impact cognitive brain function. While HSPGs play roles in normal brain development, they also contribute to the abnormal accumulation and persistence of protein aggregates in Alzheimer’s and Parkinson’s diseases (AD and PD), prolonging their neurotoxic effects. HSPG-mediated effects on the innate immune system may also contribute to the development of neuroinflammation and pathogenesis of neurodegenerative changes in the central nervous system. These pathological developments lead to impaired cognition, permanent memory loss in dementia and AD, and an inability of the brain to exert normal neuromuscular control, affecting motor functions and body movement in PD. A deeper understanding of the properties of HS in these degenerative processes is essential for the development of therapeutic measures to treat these conditions. Significant improvements in neuroimaging, next-generation glycosaminoglycan analytical techniques, and functional glycomics hold great promise for elucidating the complexities of HS structure and function. These advancements may significantly aid in the development of HS biotherapeutics to treat these debilitating conditions, which are increasingly impacting the aging global population. While a number of promising therapeutic candidates have already emerged, further research is required to optimize their biotherapeutic applications in this challenging area of pathobiology, bioregulation, and the recovery of neural function.

Exploring tau protein and amyloid-beta propagation: A sensitivity analysis of mathematical models based on biological data

Alzheimer’s disease is the most common dementia worldwide. Its pathological development is well known to be connected with the accumulation of two toxic proteins: tau protein and amyloid-β. Mathematical models and numerical simulations can predict the spreading patterns of misfolded proteins in this context. However, the calibration of the model parameters plays a crucial role in the final solution. In this work, we perform a sensitivity analysis of heterodimer and Fisher–Kolmogorov models to evaluate the impact of the equilibrium values of protein concentration on the solution patterns. We adopt advanced numerical methods such as the IMEX-DG method to accurately describe the propagating fronts in the propagation phenomena in a polygonal mesh of sagittal patient-specific brain geometry derived from magnetic resonance images. We calibrate the model parameters using biological measurements in the brain cortex for the tau protein and the amyloid-β in Alzheimer’s patients and controls. Finally, using the sensitivity analysis results, we discuss the applicability of both models in the correct simulation of the spreading of the two proteins.Statement of significance: Alzheimer’s disease is related to the accumulation of tau protein and amyloid-β. Mathematical models to predict the spreading patterns require accurate parameter calibration. In this work, we perform a sensitivity analysis of heterodimer and Fisher–Kolmogorov models to evaluate the impact of the equilibrium values of protein concentration on the solution patterns obtained with advanced numerical simulations on patient-specific brain geometry derived from magnetic resonance images. By using biological measurements in the brain cortex for the proteins in Alzheimer’s patients and controls, we use sensitivity analysis to discuss the applicability of models in simulating protein spreading.

Classic Dietiology or Holistic Nutritional Approach: What Type of Nutrition are We Working Towards?

In order to describe the negative aspects produced by the industrialized production of food that we present in this article, I have coined a definition that summarizes all the negative contents of the current diet: "PESTIFOOD", which is the portmanteau of the words PESTICIDE and FOOD. After a brief premise on the importance that healthy food has for the survival of living things, I critically describe the intensive production of food and the negative impact it has had on the emergence of numerous metabolic pathologies. So that due to the lack of nutritional value of the foods that industrial production provides us, we must record the growing development of new pathologies linked mainly to junk food and Pestifood. In the current state of degradation of food resources, not even the adoption of fortified foods or supplements can solve the root problem, represented by the use of chemical substances to improve production and extend the expiry date of foods. The only solution to the food problem for the future will be that of eco-sustainable choices, to recover the value of food as "our true medicine" - Hippocrates.

CLINICAL AND PARACLINICAL ASPECTS OF INTESTINAL FUNCTIONAL DISORDERS IN INFANTS

The aim of the work – to conduct an analysis of risk factors, clinical characteristics andresults of paraclinical examination of disorders of the functional state of the intestine ininfants who had a history of perinatal pathology.Materials and methods. A comprehensive clinical and paraclinical examination of 60infants during the period of catamnetic observation was carried out. The І group consistedof 36 children aged 6-7 months with disorders of the functional state of the gastrointestinaltract, the ІІ group – 25 healthy children of the same age category. The functional state ofthe intestines was assessed by determining the content of albumin, alpha-1-antitrypsin,elastase, PMN-elastase and fecal calprotectin levels in stools using the enzyme- linkedimmunosorbent assay (ELISA), reagents of the Immundiagnostic AG (Germany) companybased on German- Ukrainian laboratory «BUKINMED» (Chernivtsi, Ukraine).The results. Symptoms of disorders of the functional state of the intestines in infants are:restlessness and prolonged crying, poor sucking, loss of appetite, refusal to eat, vomiting,tendency to loose stools or constipation, flatulence and intestinal colic, body weightdeficiency. In the coprofiltrate of children of the I group, who had the signs of the functionalstate disorders of the intestine, compared with the control group, an increase in the contentof fecal calprotectin, alpha-1-antitrypsin, PMN-elastase and albumin was found, which areinterdependent criteria of the digestive system dysfunction. Laboratory indicators can actas markers of food intolerance and indicate the presence of local neutrophilic inflammation. Conclusions. Violations of the functional state of the intestines in infants havea multifactorial etiology caused by a combination of adverse effects of ante- and perinatalfactors on the fetal body, the development of perinatal pathology in newborns, as well asthe presence of diseases of the neonatal period.

The therapeutic potential of traditional Chinese medicine in depression: focused on the modulation of neuroplasticity.

Depression, a mood disorder characterized by a persistent low mood and lack of enjoyment, is considered the leading cause of non-fatal health losses worldwide. Neuroplasticity refers to the brain's ability to adapt to external or internal stimuli, resulting in functional and structural changes. This process plays a crucial role in the development of depression. Traditional Chinese Medicine (TCM) shows significant potential as a complementary and alternative therapy for neurological diseases, including depression. However, there has been no systematic summary of the role of neuroplasticity in the pathological development of depression and TCM Interventions currently. This review systematically summarized recent literature on changes in neuroplasticity in depression and analyzed the regulatory mechanisms of active metabolites in TCM and TCM formulas on neuroplasticity in antidepressant treatment. Additionally, this review discussed the limitations of current research and the application prospects of TCM in regulating neuroplasticity in antidepressant research.

Mailuoning oral liquid ameliorates vasculitis in thromboangiitis obliterans rats via inactivating cGAS-STING-IRF3 and TLR4-MAPKs/NF-κB signaling pathways

Ethnopharmacological relevanceMailuoning oral liquid (MLN O), one traditional Chinese patent medicine, has a good therapeutic effect on thromboangiitis obliterans (TAO) in clinical practice. However, the underlying mechanism remains unclear. Aim of the studyThis study aimed to explore the effects and potential mechanisms of MLN O against TAO based on network pharmacology and experimental verification. Materials and methodsNetwork pharmacology was used to identify the intersectional targets and signaling pathways of MLN O and TAO. In vivo, the TAO model was established by injecting sodium laurate and dihydrotestosterone (DHT) into the femoral arteries of Wistar rats. Rats were given the indicated drugs by intragastric administration (i.g.), intravenous injection (i.v.), or subcutaneous injection (s.c.) per day for 21 days since a week before surgery. In vitro, HUVECs, RAW264.7, and THP-1 cells were stimulated by LPS and DHT to simulate the pathological changes of TAO. The anti-inflammatory, anticoagulant, and immunomodulatory effects of MLN O were evaluated by histological observation, blood biochemical indexes detection, H&E staining, immunohistochemistry, enzyme-linked immunosorbent assay (ELISA), qRT-PCR, western blotting and immunofluorescence assays. Furthermore, the vascular ring test was applied to explore the vasodilatory activity of MLN O. ResultsMLN O significantly improved the pathological signs in TAO rats through its excellent anti-inflammatory, anticoagulant, immunomodulatory, and vasodilatory effects. Specifically, MLN O alleviated the gangrene and reduced the thrombosis in TAO rats, meanwhile, suppressed the expressions of inflammatory factors and clotting factors, which is related to the inactivations of cGAS-STING-IRF3 and TLR4-MAPKs/NF-κB signaling pathways. However, the superphysiological dose of DHT deteriorated the pathological development of TAO in vitro and in vivo. Moreover, the results of network pharmacology are consistent with the experimental verification. ConclusionCollectively, this study indicates for the first time that MLN O could alleviate TAO by inhibiting cGAS-STING-IRF3 and TLR4-MAPKs/NF-κB signaling pathways, which sheds light on a novel clinical therapeutic strategy for TAO.