Disorders Of Homeostasis Research Articles

Zishenhuoxue decoction-induced myocardial protection against ischemic injury through TMBIM6-VDAC1-mediated regulation of calcium homeostasis and mitochondrial quality surveillance

BackgroundZishenhuoxue decoction (ZSHX), a Chinese herbal medicine, exhibits myocardial and vascular endothelial protective properties. The intricate regulatory mechanisms underlying myocardial ischemic injury and its association with dysfunctional mitochondrial quality surveillance (MQS) remain elusive. Hypothesis/purposeTo study the protective effect of ZSHX on ischemic myocardial injury in mice using a TMBIM6 gene-modified animal model and mitochondrial quality control-related experiments. Study designUsing model animals and myocardial infarction surgery-induced ischemic myocardial injury TMBIM6 gene-modified mouse models, the pharmacological activity of ZSHX in inhibiting ischemic myocardial injury and mitochondrial homeostasis disorder in vivo was tested. MethodsOur focal point entailed scrutinizing the impact of ZSHX on ischemic myocardial impairment through the prism of TMBIM6. This endeavor was undertaken utilizing mice characterized by heart-specific TMBIM6 knockout (TMBIM6CKO) and their counterparts, the TMBIM6 transgenic (TMBIM6TG) and VDAC1 transgenic (VDAC1TG) mice. ResultsZSHX demonstrated dose-dependent effectiveness in mitigating ischemic myocardial injury and enhancing mitochondrial integrity. TMBIM6CKO hindered ZSHX's cardio-therapeutic and mitochondrial protective effects, while ZSHX's benefits persisted in TMBIM6TG mice. TMBIM6CKO also blocked ZSHX's regulation of mitochondrial function in HR-treated cardiomyocytes. Hypoxia disrupted the MQS in cardiomyocytes, including calcium overload, excessive fission, mitophagy issues, and disrupted biosynthesis. ZSHX counteracted these effects, thereby normalizing MQS and inhibiting calcium overload and cardiomyocyte necroptosis. Our results also showed that hypoxia-induced TMBIM6 blockade resulted in the over-activation of VDAC1, a major mitochondrial calcium uptake pathway, while ZSHX could increase the expression of TMBIM6 and inhibit VDAC1-mediated calcium overload and MQS abnormalities. ConclusionsOur findings suggest that ZSHX regulates mitochondrial calcium homeostasis and MQS abnormalities through a TMBIM6-VDAC1 interaction mechanism, which helps to treat ischemic myocardial injury and provides myocardial protection. This study also offers insights for the clinical translation and application of mitochondrial-targeted drugs in cardiomyocytess.

Paxillin family proteins Hic-5 and LPXN promote lipid storage by regulating the ubiquitination degradation of CIDEC

Many metabolic diseases are caused by disorders of lipid homeostasis. CIDEC, a lipid droplet (LD)-associated protein, plays a critical role in controlling LD fusion and lipid storage. However, regulators of CIDEC remain largely unknown. Here, we established a homogeneous time resolved fluorescence (HTRF)-based high-throughput screening method and identified LPXN as a positive regulatory candidate for CIDEC. LPXN and Hic-5, the members of Paxillin family, are focal adhesion adaptor proteins that contribute to the recruitment of specific kinases and phosphatases, cofactors, and structural proteins, participating in the transduction of extracellular signals into intracellular responses. Our data showed that Hic-5 and LPXN significantly increased the protein level of CIDEC and enhanced CIDEC stability not through TAG synthesis and FAK signaling pathways. Hic-5 and LPXN reduced the ubiquitination of CIDEC and inhibited its proteasome degradation pathway. Furthermore, Hic-5 and LPXN enlarged lipid droplets and promoted lipid storage in adipocytes. Therefore, we identified Hic-5 and LPXN as novel regulators of CIDEC. Our current findings also suggest intervention with Hic-5 and LPXN might ameliorate ectopic fat storage by enhancing the lipid storage capacity of white adipose tissues.

Nanobodies as negative allosteric modulators for human calcium sensing receptor

Human calcium sensing receptor (CaSR) senses calcium ion concentrations in vivo and is an important class of drug targets. Mutations in the receptor can lead to disorders of calcium homeostasis, including hypercalcemia and hypocalcemia. Here, 127 CaSR-targeted nanobodies were generated from camels, and four nanobodies with inhibitory function were further identified. Among these nanobodies, NB32 can effectively inhibit the mobilization of intracellular calcium ions (Ca2+i) and suppress the G12/13 and ERK1/2 signaling pathways downstream of CaSR. Moreover, it enhanced the inhibitory effect of the calcilytics as a negative allosteric modulator (NAM). We determined the structure of complex and found NB32 bound to LB2 (Ligand-binding 2) domain of CaSR to prevent the interaction of LB2 domains of two protomers to stabilize the inactive state of CaSR.

Exosomes as a potential therapeutic approach in osteoimmunology.

Exosomes are natural extracellular vesicles that play a key role in inter- and intracellular communication. Currently they are considered as a promising therapeutic strategy for the treatment of various diseases. In osteoimmunology, exosomes can serve as biomarkers of bone homeostasis disorders and, at the same time, promising therapeutic agents with high stability in the biological environment, low immunogenicity and good bioavailability. In this review, we attempted to examine exosomes as natural mediators of intercellular communication, playing an essential role in the interaction of the immune system and bone tissue, based on an analysis of the PubMed database up to October 2023.

Evaluation of the effectiveness of third-generation lipid emulsion therapy for early pregnancy loss

BACKGROUND: Immune mechanisms play an important role in the pathogenesis of early pregnancy loss. To regulate disorders of immune homeostasis, immunoefferent therapy is used, such as immunocytotherapy, plasmapheresis, and intravenous administration of immunoglobulins and lipid emulsions, which may help to overcome infertility and recurrent miscarriage. Third-generation lipid emulsion (TGLE) is the modern “gold standard” of parenteral lipid therapy. It contains high concentrations of omega-3-polyunsaturated fatty acids, which can directly or indirectly affect the activity of natural killer (NK) cells. The literature comprises contradictory data on the efficacy of TGLE for repeated implantation failure. This may be due to the determination of biochemical parameters for its use and the selection of criteria for assessing the effectiveness of this therapy.
 AIM: The aim of this study was to evaluate the effectiveness of TGLE therapy in patients with a history of early pregnancy loss.
 MATERIALS AND METHODS: This prospective randomized controlled study enrolled 140 patients with early pregnancy loss (recurrent miscarriage and repeated implantation failure) at the Research Institute of Obstetrics, Gynecology and Reproducto logy named after D.O. Ott, Saint Petersburg, Russia from 2021 to 2023. Group 1 consisted of 50 women with TGLE therapy, and group 2 included 90 women without therapy. Patients of group 1 received lipid therapy according to the following regimen: TGLE 20% IV 6 ml per kilogram of body weight once a month for two months on end. Inclusion criteria: women from 18 to 45 years of age with two cases of reproductive failure – pregnancy loss up to 12 weeks of gestation and/or unsuccessful in vitro fertilization attempts (embryo transfers without effect) with normal karyotypes of the spouses. Exclusion criteria: women with miscarriage with an abnormal karyotype of the fetus, with anatomical abnormalities in the development of the reproductive apparatus, with an abnormal karyotype of any of the spouses, with hypersensitivity to omega-3 polyunsaturated fatty acids. We assessed the omega-3 index, functional characteristics of NK cells in the peripheral blood, and the effectiveness of lipid therapy based on biochemical and immunological parameters, as well as the clinical onset of pregnancy.
 RESULTS: The difference in the omega-3 index before and after treatment with TGLE was significant (p = 0.0002), this parameter being increased by 1.23 times compared to the initial level. The functional activity of NK cells in the blood was significantly different after the administration of TGLE: NK (CD3–CD(16+56+)) (p = 0.0041), NKT (CD3+CD(16+56+)) (p = 0.0498), and spontaneously activated NK (CD107a) (p = 0.0498). These parameters decreased by 1.63, 1.29, and 2.07 times relative to their initial levels. In group 1 of patients receiving TGLE therapy, clinical pregnancy occurred twice as often as compared to those who did not receive this therapy (56.41%, n = 22 vs. 31.25%, n = 20; p = 0.01).
 CONCLUSIONS: TGLE has shown effectiveness in pregnancy rates by increasing the omega-3 index and reducing the functional activity of NK cells in women with early pregnancy loss.

The Role of the Microbiome in Inflammation and Carcinogenesis.

Inflammation has been confirmed to exist in the tumor microenvironment, while the risk of cancer occurrence increases in cases of chronic inflammation. It is estimated that approximately 10% to 20% of cancers are associated with chronic infections and attendant inflammation. Bacteria, both pathogenic and commensal, viruses, and fungi actively participate in the development and maintenance of inflammation and tumor growth in humans. The exposome, which is a sum of human environmental exposures, such as industrial diet, consumed drugs, and toxins, affects the composition and function of the human microbiome, which could lead to dysbiosis and disorders in tissue homeostasis through different mechanisms, including the intensification of the immune response, activation and abnormal proliferation, and disruption to epithelial barrier integrity. Presently, science remains at the stage of revealing the complexity associated with the mechanisms involved in building relationships that cover the microbiome-inflammation-tumor, yet it is already known how important it is to care for microbial homeostasis of the organism.

Glycine recalibrates iron homeostasis of lens epithelial cells by blocking lysosome-dependent ferritin degradation

One of the major pathological processes in cataracts has been identified as ferroptosis. However, studies on the iron metabolism mechanism in lens epithelial cells (LECs) and the methods of effectively alleviating ferroptosis in LECs are scarce. Along these lines, we found that in the ultraviolet radiation b (UVB) induced cataract model in vitro and in vivo, the ferritin of LECs is over-degraded by lysosomes, resulting in the occurrence of iron homeostasis disorder. Glycine can affect the ferritin degradation through the proton-coupled amino acid transporter (PAT1) on the lysosome membrane, further upregulating the content of nuclear factor erythrocyte 2 related factor 2 (Nrf2) to reduce the damage of LECs from two aspects of regulating iron homeostasis and alleviating oxidative stress. By co-staining, we further demonstrate that there is a more sensitive poly-(rC)-binding protein 2 (PCBP2) transportation of iron ions in LECs after UVB irradiation. Additionally, this study illustrated the increased expression of nuclear receptor coactivator 4 (NCOA4) in NRF2-KO mice, indicating that Nrf2 may affect ferritin degradation by decreasing the expression of NCOA4. Collectively, glycine can effectively regulate cellular iron homeostasis by synergistically affecting the lysosome-dependent ferritin degradation and PCBP2-mediated ferrous ion transportation, ultimately delaying the development of cataracts.

Proteomic profiling of blood plasma in chronic experimental exposure to aluminum oxide as a tool for predicting adverse effects from critical human organs and systems

Introduction. Highly informative research methods, which include proteomic profiling, make it possible to predict the development of negative effects on the part of critical human organs and systems at the earliest stages of their formation. In combination with toxicological studies, these methods allow changes in the protein profile of blood plasma, identified in the experiment on biological models, to be extrapolated to humans. Determination of the expression, functional characteristics and tissue identity of altered proteins and peptides provides clarification of the mechanisms of homeostasis disorders at the cellular and molecular level associated with exposure to chemicals.
 
 The purpose of the work is to identify and evaluate changes in the proteomic profile of blood plasma during experimental exposure to aluminum oxide in order to predict negative effects on the part of critical human organs and systems.
 
 Materials and methods. When modelling inhalation exposure to aluminum oxide at a dose of 0.0025 mg/(kg·day), equivalent to the dose in natural exposure conditions with an annual averaging period, and taking into account interspecies dose transfer, in an experiment on Wistar rats, the content of this metal in urine and a comparative analysis of the proteomic profile of the blood plasma of exposed and unexposed individuals was carried out. Using the methods of statistical and bioinformatic analysis, the data obtained in the experiment were extrapolated to humans in order to predict negative effects on the part of critical organs and systems during chronic aerogenic exposure to aluminum.
 
 Results. With chronic inhalation intake of aluminum oxide at a dose of 0.0025 mg/(kg·day) in animals of the experimental group, the content of aluminum in the urine was 3.5 times higher than that in the control group. As a result of densitometric measurement and comparative analysis of proteomic maps of blood plasma of exposed and unexposed animals, 13 protein spots were found, the intensity of which significantly differs between groups, which was proved to be related to the aluminum content in the urine. When comparing the mass spectra of these protein spots, 8 proteins were identified that coincided with the proteins of the library mass spectrum (Actin-binding Rho-activating protein; Aldehyde dehydrogenase, cytosol 1; Apolipoprotein A-I; Sec22b protein transporting vesicles; Elongation factor 1-γ; Neurosecretory protein Vgf; Pumilio homologue 3; Teneurin-2). As a result of bioinformatics analysis, there were determined genes encoding the identified proteins, their participation in biological processes (cellular and metabolic processes, biological regulation processes) and tissues of organs in which they have an increased expression (liver, brain, heart, muscles, small intestine, spleen, uterus, testicles, adrenal glands). In humans, orthologues of established genes have been identified, which makes it possible to use the proteins identified in the experiment as markers of negative responses and to suggest the development of diseases of the cardiovascular system, liver, and brain under conditions of long-term aerogenic exposure to aluminum oxide.
 
 Limitations. The conducted study on Wistar rats simulated only chronic inhalation exposure to aluminum oxide.
 
 Conclusion. The transformation of the proteomic profile of blood plasma was established in an experiment on a biological model with chronic inhalation exposure to aluminum oxide at a dose of 0.0025 mg/(kg·day). Extrapolation of the data obtained and the detection of orthologous genes in humans suggests the development of negative effects in the form of diseases of the cardiovascular system, liver and brain with chronic inhalation intake of aluminum oxide into the body. The data obtained expand the theoretical understanding of the mechanisms of the toxic action of chemicals, including aluminum oxide compounds, at the cellular and molecular level to predict the negative effects of critical human organs and systems.
 
 Compliance with ethical standards. The study was approved by the local ethics committee of the Federal Research Center for Medical and Preventive Technologies of Public Health Risk Management of Rospotrebnadzor (minutes of the meeting No. 2 dated 11.02.2021), conducted in accordance with the generally accepted scientific principles of the Helsinki Declaration of the World Medical Association (ed. 2013).

Role of surfactant in the pathogenesis of bronchopulmonary pathology

Pulmonary surfactant is an important structure of the lungs, providing basic vital functions: reducing alveolar surface tension and facilitating breathing, preventing collapse of the airways and ensuring their patency, protection against infections and pathological environmental factors. Although disturbances of surfactant homeostasis are usually considered in the context of respiratory distress syndrome in preterm infants, it has been established that many lung diseases in adults are accompanied by significant disturbances of surfactant homeostasis. However, preclinical and clinical studies of bronchopulmonary pathology too often overlook the potential role of changes in the qualitative or quantitative composition of pulmonary surfactant in the pathogenesis and development of disease symptoms. In many lung diseases, whether these disturbances in homeostasis are cause or effect remains contravertial field. This review will examine current data on the composition, main functions of pulmonary surfactant and the role of its dysfunction in the pathogenesis of the most frequently encountered bronchopulmonary diseases in the practice of pulmonologists, therapists and rehabilitation specialists, in particular, such as acute respiratory distress syndrome in adults, including associated with a new coronavirus infection, pneumonia of various etiologies and their consequences, bronchial asthma, chronic obstructive pulmonary disease. The review presents the current best practices of replacement therapy with exogenous surfactant preparations and the prospects for pharmacological modulation of surfactant homeostasis disorders in the above nosological forms.

Brain Iron Homeostasis and Mental Disorders.

Iron plays an essential role in various physiological processes. A disruption in iron homeostasis can lead to severe consequences, including impaired neurodevelopment, neurodegenerative disorders, stroke, and cancer. Interestingly, the link between mental health disorders and iron homeostasis has not received significant attention. Therefore, our understanding of iron metabolism in the context of psychological diseases is incomplete. In this review, we aim to discuss the pathologies and potential mechanisms that relate to iron homeostasis in associated mental disorders. We propose the hypothesis that maintaining brain iron homeostasis can support neuronal physiological functions by impacting key enzymatic activities during neurotransmission, redox balance, and myelination. In conclusion, our review highlights the importance of investigating the relationship between trace element nutrition and the pathological process of mental disorders, focusing on iron. This nutritional perspective can offer valuable insights for the clinical treatment of mental disorders.

Vitamin D level and indicators of bone tissue metabolism in kazakh infants

BACKGROUND: Insufficiency and deficiency of vitamin D are recorded in a significant part of the population. Vitamin D, along with the formation of bone tissue, is included in all types of metabolism, determining the growth and development of the body, immunity, and the normal development of the nervous system. The consequences of deficiency can be long-term and in some cases irreversible. The problem of deficiency in infants is not well understood.
 AIM: To determine the content of vitamin D and indicators of bone metabolism in children of the first year of life of the Kazakh population.
 MATERIALS AND METHODS: 250 infants of the Kazakh population were examined. The content of vitamin D in the blood was determined. The state of bone metabolism was assessed by the results of a blood test to determine the serum concentration of calcium, phosphorus, parathyroid hormone, osteocalcin, calcitonin, and in the urine deoxypyridinoline.
 RESULTS: There is a wide prevalence (77.8%) of vitamin D insufficiency/deficiency in children under one year old in the Kazakh population. The greatest decrease in vitamin D was found in newborns. Statistically significant differences were found in the levels of bone metabolism indicators, such as total calcium, phosphorus, calcitonin and osteocalcin in different age groups. Indicators of bone metabolism, such as total calcium, phosphorus, parathyroid hormone, osteocalcin, depend on the degree of decrease in vitamin D levels. The most sensitive indicator that responds to changes in vitamin D levels is osteocalcin. The level of serum vitamin D can be used as a marker for the preclinical diagnosis of metabolic disorders of bone homeostasis in children.
 CONCLUSIONS: The study confirms the effect of vitamin D on metabolic processes in the skeletal system. The most sensitive indicator that responds to changes in vitamin D levels is osteocalcin. Research in this direction may be the basis for discussing the feasibility of vitamin D substitution and clarifying prenatal care schemes.

Genome editing without nucleases confers proliferative advantage to edited hepatocytes and corrects Wilson disease.

Application of classic liver-directed gene replacement strategies is limited in genetic diseases characterized by liver injury due to hepatocyte proliferation resulting in decline of therapeutic transgene expression and potential genotoxic risk. Wilson disease (WD) is a life-threating autosomal disorder of copper homeostasis caused by pathogenic variants in copper transporter ATP7B and characterized by toxic copper accumulation, resulting in severe liver and brain diseases. Genome editing holds promise for the treatment of WD, nevertheless to rescue copper homeostasis ATP7B function must be restored in at least 25% of the hepatocytes, which surpasses by far genome editing correction rates. We applied a liver-directed nuclease-free genome editing approach, based on adeno-associated viral vector (AAV) -mediated targeted integration of a promoterless mini-ATP7B cDNA into the Alb locus. Administration of AAV-Alb-mini-ATP7B in two WD mouse models resulted in extensive liver repopulation by genome edited hepatocytes holding a proliferative advantage over non-edited ones, and ameliorated liver injury and copper metabolism. Furthermore, combination of genome editing with a copper chelator currently used for WD treatment, achieved greater disease improvement compared to chelation therapy alone. Nuclease-free genome editing provided therapeutic efficacy and may represent a safer and longer lasting alternative to classic gene replacement strategies for WD.

ВЛИЯНИЕ ЭКСТРАКТА ЛИСТЬЕВ ГИНКГО ДВУЛОПАСТНОГО (GINKGO BILOBA L.) НА ПСИХОЭМОЦИОНАЛЬНЫЙ СТАТУС КРЫС НА ФОНЕ ОЖОГОВОЙ ТРАВМЫ КОЖИ

Abstract. The aim studying the effect of liquid extract leaf Ginkgo biloba on the psychoemotional status of rats under conditions of thermal skin injury. Materials and methods. The subject of the study was sexually mature male nonlinear white rats. Burn wounds were modeled in the interscapular region of the back. To study psychoemotional status, rat behavior studies were conducted using standard psychophysiological tests: Open Field and Porsolt. Results of the study. In the conditions of burn damage to the skin, signs of an alarming-depressive state, in particular disorientation in space, suppression of the psychoemotional state, and a change in the nature of behavioral reactions were noted. The study also found that the external use of Ginkgo biloba leaf extract contributed to a decrease in situational anxiety and the appearance of depressive-like behavioral reactions that arose under conditions of thermal skin injury. Conclusion. Thus, the revealed changes in the psychoemotional status of rats in burn skin damage are the result of functional disorders of the brain, activated during protective and adaptation reactions in burn injury. Results of assessment of psychoemotional state of animals in case of skin burn injury confirm involvement of nervous system in response to stress, reflex activity of which under stress conditions is manifested in change of behavioral reactions. Taking into account the psycho-corrective effect of Ginkgo bilobate leaf extract against the background of thermal skin injury, it is legitimate to argue that it is advisable to correct systemic homeostasis disorders accompanying the burn process.

N-Acetylneuraminic Acid Induces Vascular Endothelial Dysfunction through Targeting Mcu/SQSTM1 Pathway and Attenuates Atherosclerosis

Atherosclerosis (AS) is a major cause of various fatal diseases, such as stroke, and ischemic heart failure worldwide. The risk factors of AS include hypertension, diabetes mellitus, dyslipidemia, smoking and obesity. Recently, several investigations have shown that metabolic disorder within plaque was characterized as one of the main causes for those risk factors. The increase of N-acetylneuraminic acid (Neu5Ac), a metabolite produced by hexosamine-sialic acid pathway branching from glucose metabolism, in blood was identified as a metabolic biomarker for AS progression. Our previous study revealed that Neu5Ac could promote endothelial inflammatory response and attenuates AS plaque progression in apoE -/- mice. Mitochondrial dysfunction was suggested to be involved for endothelial injury after Neu5Ac accumulation in circulation. However, the underlaying mechanisms have not been fully understood yet. Therefore, the aim of the study was to understand the mechanism of mitochondrial injury induced by Neu5Ac accumulation in vivo using apoE-/- mouse model and in vitro using the endothelial cell line, HUVECs. Investigations have shown that impaired Ca 2+ homeostasis plays an interactive role in the progression of mitochondrial dysfunction. In our present study, we found that Neu5Ac induced Ca 2+ overload, increased ROS levels and promoted the expression of mitochondrial injury makers such as Parkin, TOMM20 and TIMM23. While pretreatment of HUVECs with BAPTA-AM in vitro could inhibit the increase of the intracellular Ca 2+ concentration and reversed the mitochondrial fragment induced by Neu5Ac, suggesting that Neu5Ac in circulation promoted mitochondrial Ca 2+ uptake and induced the mitochondrial homeostasis disorder. Based on the reported results that mitochondrial Ca 2+ uniporter (MCU) was the key regulator of impaired Ca 2+ homeostasis, we then detected the expression of MCU both in vivo and in vitro. We found an increase of MCU both at protein and mRNA levels in HUVEC cell lines. Meanwhile, we also observed that MCU expression was upregulated in aortic arch of apoE -/- mice after Neu5Ac treatment. Silencing MCU in vitro, mitochondrial Ca 2+ concentrations were downregulated and the ability of endothelial adhesion to monocytes was reversed as well. These results suggested that Neu5Ac specifically targeted MCU and induced Ca 2+ uptake. Targeting MCU might be a potential strategy for preventing AS progression induced by Neu5Ac. Furthermore, mitochondrial Ca 2+ uptake was considered as an important determinant of cell fate and mitophagy/autophagy pathway. Our previous study has confirmed that Neu5Ac could activate SQSTM1/p62-mediated excessive autophagy and subsequently induced endothelial injury. To investigate whether MCU silencing could attenuate the autophagy mediated AS progression, we investigated the SQSTM1/p62 expression after silencing MCU in HUVECs. The present results confirmed that Neu5Ac increased mitochondrion-dependent autophagy, and the effects of which were largely relieved by MCU silencing. In summary, impaired Ca 2+ homeostasis was involved in the endothelial injury induced by accumulated Neu5Ac in blood. Furthermore, MCU upregulation is responsible for Neu5Ac-induced pathological AS disorder. Our findings reveal a novel molecular mechanism regulating mitochondrial dysfunction and may open a new window for therapeutic targeting in the treatment of AS.

Zinc oxide nanoparticles-induced testis damage at single-cell resolution: Depletion of spermatogonia reservoir and disorder of Sertoli cell homeostasis

The widespread application of zinc oxide nanoparticles (ZnO NPs) in our daily life has initiated an enhanced awareness of their biosafety concern. An incredible boom of evidence of organismal disorder has accumulated for ZnO NPs, yet there has been no relevant study at the single-cell level. Here, we profiled > 28,000 single-cell transcriptomes and assayed > 25,000 genes in testicular tissues from two healthy Sprague Dawley (SD) rats and two SD rats orally exposed to ZnO NPs. We identified 10 cell types in the rat testis. ZnO NPs had more deleterious effects on spermatogonia, Sertoli cells, and macrophages than on the other cell types. Cell–cell communication analysis indicated a sharp decrease of interaction intensity for all cell types except macrophages in the ZnO NPs group than in the control group. Interestingly, two distinct maturation states of spermatogonia were detected during pseudotime analysis, and ZnO NPs induced reservoir exhaustion of undifferentiated spermatogonia. Mechanically, ZnO NPs triggered fatty acid accumulation in GC-1 cells through protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling and peroxisome proliferator-activated receptor alpha (PPARα)/acyl‐CoA oxidase 1 (Acox1) axis, contributing to cell apoptosis. In terms of Sertoli cells, downregulated genes were highly enriched for tight junction. In vitro and in vivo experiments verified that ZnO NPs disrupted blood-testis barrier formation and growth factors synthesis, which subsequently inhibited the proliferation and induced the apoptosis of spermatogonia. As for the macrophages, ZnO NPs activated oxidative stress of Raw264.7 cells through nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) pathway and promoted cell apoptosis through extracellular signal-regulated kinase (ERK) 1/2 pathway. Collectively, our work reveals the cell type-specific and cellularly heterogenetic mechanism of ZnO NPs-induced testis damage and paves the path for identifying putative biomarkers and therapeutics against this disorder.

Association of urinary arsenic metabolism with type 2 diabetes and glucose homeostasis: Cross-sectional and longitudinal associations.

BackgroundPrevious researches have assessed the relationships of urinary arsenic metabolism with type 2 diabetes (T2D) and glucose-insulin homeostasis, but the results were controversial, and potential mechanisms remain largely unclear. ObjectivesThis study aimed to investigate the cross-sectional and longitudinal associations of urinary arsenic metabolism with T2D prevalence and glucose changes in relatively higher arsenic exposure, and further to evaluate the underlying roles of oxidative damage in these relationships. MethodsWe included 796 participants at baseline, among them 509 participants were followed up after 2 years. Logistic regression model and leave-one-out approach were applied to evaluate the associations of arsenic metabolism with T2D prevalence. Linear mixed model was conducted to estimate the relationship of arsenic metabolism with glycemic changes over two years. The associations between arsenic metabolism and indicators of oxidative stress were assessed with a linear regression model. We further performed mediation analysis to investigate the role of oxidative stress in the associations of arsenic metabolism with 2-year change of glucose levels. ResultsHigher urinary MMA% increased T2D prevalence and baseline glucose levels. MMA% was positively associated with 2-year change of glucose levels. Moreover, we observed significant dose-response relationship between MMA% and 8-hydroxy-2-deoxyguanosine (8-OHdG). However, the mediating role of 8-OHdG in the association of MMA% and 2-year change of glucose levels was not observed in this population. ConclusionsIn this population exposure to relatively higher arsenic levels, higher MMA% contributed to increased T2D prevalence and glucose homeostasis disorder. Arsenic metabolism also affected oxidative stress levels, especially 8-OHdG. Further studies are required to investigate the potential mechanisms.

Some reflections on immunosuppressants and their delivery to the lungs by aerosol in the treatment of COVID-19

Disorders of immune homeostasis represent the key pathogenetic link of COVID-19, which often manifests as a hyperimmune response to the pathogen, leading to severe uncontrolled inflammation in lungs, followed by complications and death. Accordingly, a certain therapeutic potential is provided by different pharmacological drugs with distinct mechanisms of action, This class of drugs should, however, act in common direction by suppressing the immune response, thus being often classified as immunosuppressants (IS). Of them, the most promising are immunobiological preparations, which include monoclonal antibodies, as well as purinergic regulatory agents. There are several attempts to use the “classical” IS by a certain way, e.g., cytostatics and calcineurin inhibitors which found clinical application in transplantology and oncology. However, their usage for treatment of uncontrolled inflammation of respiratory tract was abandoned by the end of XX century. Meanwhile, the aerosol route of drug administration optimizes treatment, both in terms of their effectiveness, and the reduction of side effects thus promoting usage of IS for treatment of uncontrolled airway inflammation. Previously, the analysis of therapeutic opportunities for some IS delivered as aerosols to the lungs in COVID-19 therapy was not carried out, thus bein the purpose of our work. Methodological analysis was carried out using various databases of biomedical scientific information, including Index Medicus, PubMed, Embase, Cohrane Clinical Trials gov registry and patent databases.The efficiency of the impact of various IS subgroups in COVID-19, including their administration by inhalations into the respiratory ways, was assessed. The role of regulatory T cells considered the central regulator of immune response, in pathogenesis of COVID-19 was considered, and their therapeutic potential was characterized, dependent on phase and severity of the disease as well as drug dose dependence. Methods and approaches to the use of IP, advantages and disadvantages are discussed. The expediency and future prospects of their application are considered.One may conclude that the effectiveness of cytostatics and calcineurin inhibitors in the treatment of airway inflammation in COVID-19 remains unconfirmed and seems to be unpromising. Meanwhile, biological preparations, including monoclonal antibodies and purinergic regulatory agents, offer great promise in this respect.

The effect of inflammation on the course of experimental aseptic necrosis of femoral head

Aseptic necrosis of the femoral head is a staged process in which osteodestruction is replaced by the bone repair. The outcome of this disease may be characterized by severe discongruence of the hip joint area, disability of the patient. Recently, the research interest is drawn to molecular and cellular mechanisms of bone homeostasis disorders and ways of its correction. A number of studies have demonstrated the role of nonspecific inflammation in pathogenesis of aseptic necrosis. However, a more detailed study of dynamic changes in the activity of osteogenesis signaling pathways is required. The aim of this study was to assess the role of molecular patterns of inflammation and osteogenesis during aseptic necrosis of femoral head in experimental model. Surgical induction of aseptic necrosis of the femoral head was performed in 16 rats, which were removed biweekly from experiment (by 4 animals), for 8 weeks. The expression of genes encoding proteins involved in osteogenesis regulation was studied by qPCR with reverse transcription. Concentration of VCAM1, MMP9 proteins was assessed by immunoblotting. The results of our study demonstrated heterogenous dynamics of changes in molecular and cellular disorders associated with bone homeostasis regulation in pathogenesis of aseptic necrosis. For the first two weeks after surgical procedure, the expression of HIF1α and TNFα genes, as well as the concentration of MMP9 and VCAM1 proteins, were determined as predictor factors. After 1 month, VCAM1 protein concentration and TNFα gene expression acted as protector factors, whereas IL6 gene and MMP9 protein were considered predictive factors. After 6 weeks, the development of aseptic necrosis was promoted by expression of the IL4 gene, and after 8 weeks, by IL6 gene. Thus, an important role in regulation of osteoresorption belongs to nonspecific inflammation, which can be triggered by acute tissue hypoxia. A significant effect of the inflammation process persists up to 8 weeks after induction of avascular necrosis of femoral head. Pathogenesis of bone destruction is associated not only with an increased activity of osteoclastogenesis, but also with a decreased intensity of osteoblastogenesis. In general, the molecular and cellular pattern of bone homeostasis disorders varies depending on the stage of aseptic necrosis.

Effect of Supplementation with Curcuma longa and Rosmarinus officinalis Extract Mixture on Acute Phase Protein, Cathelicidin, Defensin and Cytolytic Protein Gene Expression in the Livers of Young Castrated Polish White Improved Bucks.

Goats are an excellent animal model for research on some physiological and pathophysiological processes in humans. The search for supplements that prevent homeostasis disorders and strengthen the immune system is necessary to reduce the risk of many diseases in both humans and animals. The aim of the study was to analyze the effect of supplementation with a mixture of dried extracts of Curcuma longa and Rosmarinus officinalis on the expression of acute-phase protein (SAA, HP, CRP, LALBA, AGP, CP, FGA, FGB, and FGG), cathelicidin (BAC5, BAC7.5, BAC3.4, MAP28, MAP34, and HEPC), beta-defensin-1 (GBD1, DEFB1), and beta-defensin-2, and cytolytic protein (LIZ and LF) genes in the livers of young castrated bucks of the Polish White Improved breed. The higher expression of LF in the control group suggests that it is important for the first line of hepatic immune defense and its expression is downregulated by the mixture of turmeric and rosemary extracts; thus, the spice-herb mixture mutes its activity. The lower expression of FGB and the higher expression of BAC5 genes in the livers of healthy, young castrated bucks who were administered the supplement suggest the silencing effects of the mixture on the acute-phase response and the stimulating effect on the antimicrobial activity of the immune system.

Oxidative stress sensor Keap1 recognizes HBx protein to activate the Nrf2/ARE signaling pathway, thereby inhibiting hepatitis B virus replication.

The Kelch-like ECH-associated protein 1 (Keap1)/NF-E2-related factor 2 (Nrf2)/antioxidant response element (ARE) signaling pathway is one of the most important defense mechanisms against oxidative stress. We previously reported that a cellular hydrogen peroxide scavenger protein, peroxiredoxin 1, a target gene of transcription factor Nrf2, acts as a novel HBV X protein (HBx)-interacting protein and negatively regulates hepatitis B virus (HBV) propagation through degradation of HBV RNA. This study further demonstrates that the Nrf2/ARE signaling pathway is activated during HBV infection, eventually leading to the suppression of HBV replication. We provide evidence suggesting that Keap1 interacts with HBx, leading to Nrf2 activation and inhibition of HBV replication via suppression of HBV core promoter activity. This study raises the possibility that activation of the Nrf2/ARE signaling pathway is a potential therapeutic strategy against HBV. Our findings may contribute to an improved understanding of the negative regulation of HBV replication by the antioxidant response.