A1 Receptor Research Articles

Role of the Dorsal Cortex of the Inferior Colliculus in the Precedence Effect.

BACKGROUND The precedence effect (PE) is a physiological phenomenon for accurate sound localization in a reverberant environment. Physiological studies of PE have mostly focused on the central nucleus of the inferior colliculus (CNIC), which receives ascending and descending projections, as well as projections from the shell of the inferior colliculus (IC) and contralateral IC. However, the role of the dorsal cortex of the IC (DCIC), which receives ascending and descending projections to ensure sound information processing and conduction on PE formation, remains unclear. Therefore, this study aimed to understand the role, if any, of the DCIC on PE formation in male Sprague Dawley rats. MATERIAL AND METHODS In vivo, 16-channel electrophysiological recordings were performed in anesthetized rats to investigate neuronal responses in the CNIC, after inducing electrolytic lesions in the DCIC. In vitro, the expression of inhibitory gamma-aminobutyric acid (GABA)ergic receptors in the CNIC was analyzed by Western blot. RESULTS After inducing electrolytic lesions in the DCIC, normalized neural responses of the CNIC to lagging stimuli were significantly increased (P<0.05), half-maximal inter-stimuli delays were shortened (P<0.05), and the expression of GABA A receptor a1 and GABA B receptor 2 decreased (P<0.05). Furthermore, neurons in the CNIC showed a contralateral preference when paired sounds in the free field were presented. CONCLUSIONS Our study suggests that the DCIC could modulate PE formation in the CNIC, potentially involving inhibitory GABAergic mechanisms. This study showed the role of the DCIC on PE formation and proposed a potential structure for identifying likely mechanisms of the PE in the IC.

Screening potential diagnostic biomarkers for PLA2R‑associated idiopathic membranous nephropathy by WGCNA analysis and LASSO algorithm

Adult nephrotic syndrome is primarily caused by membranous nephropathy (MN), with idiopathic membranous nephropathy (IMN) being a prominent subtype. The onset of phospholipase A2 receptor (PLA2R1)-associated IMN is critically linked to M-type PLA2R1 exposure, yet the mechanism underlying glomerular injury remains unclear. In this study, membranous nephropathy datasets (GSE115857, GSE200828) were retrieved from GEO. Differential gene expression was analyzed using the ‘limma’ R package. WGCNA filtered PLA2R-related modules and intersected genes. LASSO regression, evaluated by ROC analysis, identified characteristic genes. Binomial logistic regression assessed their association with IMN. Validation was performed in the GSE133288 dataset. IHC and qRT-PCR detected characteristic gene expression in PLA2R-positive patients. This study identified elevated PLA2R expression in IMN patients among 117 DEGs. PPI analysis suggested enrichment in Golgi membranes, co-regulation, and glucocorticoid responsiveness, implicating the PPAR pathway by KEGG. WGCNA revealed a 440-gene brown module associated with IMN-PLA2R, with ECM1, SLC19A2, RASD1, FOSB, KDELR3, ZFP36, and ELF4 highlighted as diagnostic markers by ROC analysis. Clinical validation confirmed ECM1 upregulation increased IMN risk, while upregulation of SLC19A2, ZFP36, RASD1, and FOSB decreased it. ECM1 positively correlated with PLA2R, whereas SLC19A2, ZFP36, and FOSB negatively correlated. IHC analysis demonstrated consistent gene expression patterns in IMN tissues, with podocyte exposure to PLA2R-positive serum reducing viability and increasing apoptosis. Functional studies, prompted by RASD1 downregulation, revealed enhanced cell activity and reduced apoptosis upon RASD1 overexpression compared to the Serum + Ov-NC control. Collectively, this study identified diagnostic markers for PLA2R-related IMN, offering novel therapeutic targets for the treatment of IMN.

Detection and prognostic relevance of PLA2R epitopes in idiopathic membranous nephropathy: a simultaneous quantitative multiplex suspension array detection method

Abstract Objective This study aimed to develop a multiplex suspension assay for the simultaneous quantitative detection of anti-cysteine-rich domain (anti-CysR)/C-type lectin domain 1 (CTLD1)/C-type lectin domain 6–8 (CTLD678)-IgG4 antibodies of M-type phospholipase A2 receptor (PLA2R) in the serum samples of patients to evaluate the clinical application value of PLA2R epitope spreading in disease prognosis. Methods CysR, CTLD1, and CTLD678 antigen domains were coupled to three types of microspheres. The optimal dilution ratio of biotinylated anti-human IgG4 was identified, and a multiplex suspension assay was evaluated in terms of linearity, sensitivity, precision, specificity, and recovery rate. Lastly, the relationship between epitope spreading and the severity of idiopathic membranous nephropathy (IMN) was evaluated. Results The content of all three epitopes could be detected simultaneously within two hours. The intra-assay precision ranged from 4.74% to 9.48% , and the inter-assay precision was between 5.13% and 13.92% . Specific experiments showed that the human IgA antibody did not cause a cross-reaction. The recovery rates ranged between 90% and 100%. The cut-off values of epitopes CysR, CTLD1 and CTLD678 between healthy individuals and patients were 6.30, 12.38, and 10.06 Ru/mL, respectively. Among them, the positive rates of epitopes CysR, CTLD1 and CTLD678 were 100%, 34%, and 75%, respectively. In addition, group 3 (CTLD1 response) accounted for 73% of the twelve patients who were not in remission. Meanwhile, When the concentration of CTLD1 exceeds 42.76 Ru/mL, the patient's prognosis may be poorer. Conclusion A multiplex suspension assay was developed for the simultaneous quantitative detection of anti-CysR/CTLD1/CTLD678-IgG4 antibodies. The epitope migration sequence of PLA2R molecules during disease progression may not follow a simple linear rule. Among them, the epitope CTLD1 is likely to exert the most significant influence on patient remission.

Bioinformatic Analysis for Exploring Target Genes and Molecular Mechanisms of Cadmium-Induced Nonalcoholic Fatty Liver Disease and Targeted Drug Prediction.

Cadmium (Cd) is a widely available metal that has been found to have a role in causing nonalcoholic fatty liver disease (NAFLD). However, the detailed toxicological targets and mechanisms by which Cd causes NAFLD are unknown. Therefore, the present work aims to reveal the main targets of action, cellular processes, and molecular pathways by which cadmium causes NAFLD. As shown in the bioinformatics analysis, there were 74 main targets of action for cadmium-induced NAFLD, hemopoietic cell kinase (HCK), EPH receptor A2 (EPHA2), MYC proto-oncogene (MYC), lysyl oxidase (LOX), dipeptidyl peptidase 7 (DPP7), nuclear factor erythroid 2-related factor 2 (NFE2L2), dual specificity phosphatase 6 (DUSP6), CD2 cytoplasmic tail binding protein 2 (CD2BP2), notch receptor 3 (NOTCH3), and phospholipase A2 group IVA (PLA2G4A) were screened as core genes. Testing these core genes in other databases, three differentially expressed genes, HCK, MYC, and DUSP6 were verified and used as targets for drug prediction in DsigDB; decitabine and retinoic acid were screened as potential therapeutic drugs for NAFLD based on the p-value and the combined score. The results of molecular docking showed that the predicted drugs can bind well to the core targets. In conclusion, cadmium is associated with NAFLD; the identified cadmium-toxicity targets, HCK, MYC, and DUSP6, may serve as biomarkers for the diagnosis of NAFLD and predicted drugs, decitabine and retinoic acid may have a potential role in the treatment of NAFLD.

The C3/C3aR pathway exacerbates acetaminophen-induced mouse liver injury via upregulating podoplanin on the macrophage.

Acute liver failure (ALF) is a life-threatening condition that occurs when the liver sustains severe damage and rapidly loses its function. The primary cause of ALF is the overdose of acetaminophen (APAP), and its treatment is relatively limited. The involvement of the complement system in the development of ALF has been implicated. However, the related mechanisms remain poorly understood. Complement 3 (C3) knockout mice, complement 3a receptor (C3aR) knockout mice, platelet C-type lectin-like receptor 2 (Clec-2)-deficient mice, and myeloid cell podoplanin (Pdpn)-deficient mice were generated. Liver tissues were collected for histological analysis, RNA sequencing, confocal immunofluorescence, and immunoblot analyses. Our data demonstrated that APAP activated the C3/C3aR pathway, leading to intrahepatic hemorrhage, ultimately resulting in hepatocyte necrosis. Deletion of C3 or C3aR mitigated APAP-induced liver injury (AILI). C3/C3aR signaling upregulated the expression and phosphorylation of transcription factors STAT3 and c-Fos in hepatic Kupffer cells, which in turn increased PDPN expression, promoting platelet recruitment to the Kupffer cells via the interaction of PDPN and the CLEC-2 on platelets. Since the activation of platelets mediated by C3/C3aR occurs irrespective of the major hemostatic pathways, blocking the C3/C3aR pathway in ALF could be a coagulopathy-sparing and novel therapeutic approach. In summary, this study unveiled the critical roles of the C3/C3aR pathway in developing AILI, providing evidence that the C3/C3aR pathway could be an effective therapeutic target for AILI.

GabaAergic sedative prospection of sclareol-linalool co-treatment: An antagonistic intervention through in vivo and in silico studies

Sleep disturbance causes many health problems in humans worldwide. This study evaluated the effects and possible mechanisms of sclareol (SCL) and/or linalool (LIN) through in vivo and in silico studies. For this, young chicks SCL (5, 10, and 20 mg/kg) and/or LIN (50 mg/kg) were orally administered thirty minutes before to the thiopental sodium (TS)-induced chicks with or without the standard drug diazepam (DZP: 3 mg/kg). Incidence, onset, and duration of sleep were then noted. The results suggest that SCL dose-dependently increased the onset and decreased the duration of sleep in animals. In contrast, LIN50 significantly (p < 0.05) decreased onset and increased sleep duration. SCL20 combined with LIN50 and/or DZP3 modulated the sleep parameters in animals. In combination, LIN50 showed better effects with DZP3, where the percentage decrease in latency and increase in sleep duration were 54.20 and 168.65 %, respectively. SCL20 when combined with LIN50 + DZP3 also modulated the onset and duration of sleep in animals. Further, in silico studies suggest that SCL and LIN have binding affinities with the 6X3X protein of the GABAA receptor (α1 and β2 subunits) of −6.9 and −6.8 kcal/mol, respectively. The standard drug DZP showed a binding affinity of −5.0 kcal/mol. Taken together, SCL may exert an angiogenic-like effect and antagonize LIN and/or DZP-mediated sedative effects in TS-induced chicks, possibly through the GABAA receptor α1 and β2 subunits interaction pathway.

Updated diagnostic and therapeutic management for membranous nephropathy.

Pioneering contributions in membranous nephropathy over the last decade have greatly enhanced our comprehension of its pathogenesis, diagnosis, and treatments, igniting renewed interest in this entity. This review provides an updated perspective on the diagnosis and therapeutic management of membranous nephropathy. The identification of antiphospholipase A2 receptor (PLA2R) antibodies in 50-80% of membranous nephropathy patients was a key breakthrough. High or increasing PLA2R antibody levels are linked to persistent nephrotic syndrome and the need for targeted treatment. Given the high specificity of PLA2R antibodies, a kidney biopsy may not be required for pure nephrotic syndrome cases with no comorbidities. Over the years, various target antigens and associated conditions have been identified in membranous nephropathy patients, leading to a reclassification of membranous nephropathy. Treatment approaches vary based on baseline characteristics and changes in proteinuria and PLA2R titers. Rituximab has emerged as the first-line therapy for most patients without severe risk factors, with other emerging therapies under development. Advances in the diagnosis and treatment of membranous nephropathy have moved the management towards a more precision-based approach, though further studies and new therapies are needed for a comprehensive management strategy.

The receptor binding mechanism of mouse sPLA2 group IIE

Secreted phospholipase A2s (sPLA2s) participate in physiological function by their enzyme and receptor binding activity. Muscle-type phospholipase A2 receptor (M-type PLA2R) is the sPLA2 binding protein with the highest affinity so far, and also inhibits the enzyme activity of sPLA2. There is species specificity and pH dependence for the binding of M-type PLA2R to sPLA2. Mouse sPLA2 Group IIE (mGIIE) has been verified to have a high affinity for mouse M-type PLA2R (M-type mPLA2R) at the nanomolar scale. For further exploration of the receptor binding mechanism of GIIE, in this study, we use Alphafold Multimer to generate complex models of mGIIE with the M-type mPLA2R ectodomain, wild-type CTLD5 domain of mPLA2R, and three CTLD5 mutants, respectively. mPLA2R-mGIIE models exhibit heterogeneous extended mPLA2R conformations with uncovered sPLA2-binding surface of CTLD5 domain. Complexed models of mGIIE with wild-type and mutated mCTLD5 further confirm that helix α1 of mCTLD5, especially essential residues F838 and W842, interact with the substrate pocket of mGIIE and thus inhibit its enzyme activity. Peptides from helix α1 of mCTLD5 are verified to inhibit the enzymatic activity of mGIIE. This AI-guided research would substantially accelerate our understanding of the functional study of GIIE, and provide the lead-peptide for the further inhibitor design of sPLA2.

Effectiveness of a Novel PLA2R1 Knock-In Rat Model in Repairing Renal Function Damage.

Phospholipase A2 receptor 1 (PLA2R1) exists in many animals and plays an important role in membranous nephropathy. In this study, we aimed to evaluate a PLA2R1 knock-in rat model with repaired kidney function to study the molecular mechanisms of membranous nephropathy. We constructed the PLA2R1 knockout [PLA2R1(-)] model and PLA2R1 knock in [PLA2R1(+)] model in rats. Consistent complement C3 and IgA expression was confirmed through colocalization studies. Urinary biochemical indicators were performed using Automatic Biochemistry Analyzer. The complement C3, IgG, and Nephrin were detected by immunofluorescence assay. The expression levels of complement C3, IgA, and PLA2R1 were detected by western blot. The differential expression proteins (DEPs) between control and PLA2R1(+) models were detected by liquid chromatography with tandem mass spectrometry. The PLA2R1(-) model showed proteinuria, complement C3 aggregation, and IgA and IgG deposition in the glomerulus. Comparing with the PLA2R1(-) model, the PLA2R1(+) model, the deposition of complement C3 and IgA in the glomerulus did not completely disappear, and IgG expression weakened. Moreover, the absolute value of urinary protein was much lower in the PLA2R1(+) model than in the PLA2R1(-) model, and some of the humanized PLA2R1 gene fragments repaired some of the kidney functions. Humanized PLA2R1-insertion in rats can repair part of the renal function and reduce proteinuria, which will help in studying the molecular mechanisms of membranous nephropathy, as well as the entire membranous nephropathy-related system and complement activation signaling pathway.

New antiplatelet approach: inhibiting Pim kinase to reduce constitutive surface expression of thromboxane A2 receptor.

New antiplatelet approach: inhibiting Pim kinase to reduce constitutive surface expression of thromboxane A2 receptor.

Constitutive surface expression of the thromboxane A2 receptor is Pim kinase-dependent.

The thromboxane A2 receptor (TPαR) plays an important role in the amplification of platelet responses during thrombosis. Receptor activity is regulated by internalization and receptor desensitization. The mechanism by which constitutive surface expression of the TPαR is regulated is unknown. Recently, it has been demonstrated that proviral insertion in murine lymphoma (Pim) kinase inhibitors reduce platelet functional responses in a TPαR-dependent manner. To investigate whether Pim kinases regulate constitutive TPαR surface expression. TPαR surface expression was measured in platelets, and human embryonic kidney 293T (HEK293T) cells transfected with tagged TPαRs in the presence and absence of Pim kinase inhibitors using flow cytometry and confocal microscopy. TPαR-dependent calcium flux was assessed using Fluo-4 AM. Site prediction modeling and site-directed mutagenesis were used to identify the TPαR PIM kinase phosphorylation site. Surface expression of TPαR and calcium responses to U46619 were reduced in platelets and HEK293T cells following Pim kinase inhibition. Overexpression of kinase-dead Pim-1 also reduced TPαR surface expression on HEK293T cells. Reduced surface expression of the TPαR was found to be mediated by increased receptor internalization in a dynamin and β-arrestin-dependent manner. Four putative Pim kinase phosphorylation sites in the TPαR were mutated, and serine 57 in the first intracellular loop of TPαR was identified to be a novel regulatory site important for maintaining TPαR surface expression and thromboxane A2-dependent functional responses. Pim kinase inhibition may offer a novel therapeutic approach to limit cellular responses to thromboxane A2, independent of cyclooxygenase inhibition and direct antagonism of the receptor.

Clinical and preclinical evidence of psilocybin as antidepressant. A narrative review.

In the rapidly growing field of psychedelic research, psilocybin (and active metabolite psilocin) has been proposed as a promising candidate in the search for novel treatments for neuropsychiatric disorders. Clinical trials have revealed that psilocybin has a large, rapid, and persistent effect in the improvement of symptoms of depression and anxiety. The safety profile is considered favourable, with low toxicity and good tolerance. Several preclinical studies have also been carried out to determine the long-term mechanism of action of this drug. In this sense, preclinical studies in naïve animals as well as in animal models of disease have shown somewhat discrepant results in conventional tests for assessment of depression- and anxiety-like phenotype in response to psilocybin, but overall suggest positive outcomes. Additionally, several valuable assays in rodent models have been developed over the years to elucidate the neurochemical correlates of serotonin 2A receptor (5HT2AR) activation in the brain, primary molecular target of psilocin. This review aims to provide a general overview of the current and most recent literature in the therapeutic potential of psilocybin through a description of clinical trials of psilocybin-assisted psychotherapy, and to showcase the scene in the up-to-date preclinical research. A detailed description of preclinical rodent models and experimental approaches that have been used to study the neurobiological and behavioural actions of psilocybin is provided, and potential therapeutic mechanisms of action are discussed.

Serum BAFF levels are associated with the prognosis of idiopathic membranous nephropathy

Objective High serum levels of B-cell activation factor (BAFF) and a proliferation-inducing ligand (APRIL) have been observed in patients with idiopathic membranous nephropathy (iMN); however, their relationships with disease severity and progression remain unclear. Methods Patients with iMN diagnosed via renal biopsy were enrolled in this study. The concentrations of BAFF and APRIL were determined using ELISA kits. Proteinuria remission, including complete remission (CR) and partial remission (PR), and renal function deterioration were defined as clinical events. The Cox proportional hazards method was used to analyze the relationship between cytokine levels and disease progression. Results Seventy iMN patients were enrolled in this study, with a median follow-up time of 24 months (range 6–72 months). The serum levels of BAFF and APRIL were higher in iMN patients than in healthy controls but lower than those in minimal change disease (MCD) patients. The serum BAFF level was positively correlated with the serum APRIL level, serum anti-phospholipase A2 receptor (anti-PLA2R) antibody level, and 24-h proteinuria and negatively correlated with the serum albumin (ALB) level. However, no significant correlation was observed between the serum APRIL level and clinical parameters. According to the multivariate Cox proportional hazards regression model adjusted for sex, age, systolic blood pressure (SBP), estimated glomerular filtration rate (eGFR), immunosuppressive agent use, 24-h proteinuria, APRIL level, and anti-PLA2R antibody, only the serum BAFF level was identified as an independent predictor of PR (HR, 0.613; 95% CI, 0.405–0.927; p = 0.021) and CR of proteinuria (HR, 0.362; 95% CI, 0.202–0.648; p < 0.001). Conclusions A high serum BAFF level is associated with severe clinical manifestations and poor disease progression in patients with iMN.

Isorauhimbine and Vinburnine as Novel 5-HT2A Receptor Antagonists From Rauwolfia serpentina for the Treatment of Insomnia: An In Silico Investigation

Objective: This study aimed to identify and delineate the antagonistic effects of phytochemicals present in Rauwolfia serpentina (Indian snakehead) on the serotonin 2A receptor (5HT2AR) for insomnia treatment. Methods: Molecular docking and molecular dynamics simulations were performed to analyze the binding affinity and stability of protein–ligand complexes. The dynamic behavior of the complexes was studied via normal mode analysis performed via the iMODS server. The blood–brain barrier permeability of the phytochemicals was hence analyzed via the Brain or IntestinaL EstimateD permeation method (BOILED-Egg). The bioactivity of phytochemicals as “g-protein coupled receptor (GPCR) ligands” was analyzed via the MolInspiration server. Lipinski’s rule of five was used as a benchmark to assess the drug likeness of the phytochemicals. The SWISS ADME server was used to analyze the physiochemical properties of the selected phytocompounds. Furthermore, the preclinical efficacy and safety of the compounds were assessed via absorption, distribution, metabolism, excretion, and toxicity (ADME/T) analysis performed via the Deep-PK and ProTox 3.0 servers. Results: Cumulative results from various computational parameters yielded isorauhimbine and vinburnine as hit compounds because they are 5HT2AR antagonists. These phytochemicals exhibited promising docking scores, and their root mean square deviation, radius of gyration, and solvent accessible surface area confirmed their stability with the target protein. The ADME/T profile and drug-likeness values of these two compounds validated their safety, efficacy, and potential as drug candidates. Conclusion: We conclude that isorauhimbine and vinburnine stand out as potential phytochemicals from R. serpentina that could potentially serve as potential antagonistic drug candidates against 5HT2AR for insomnia treatment. However, these findings need to be validated through wet lab experiments to fully elucidate their therapeutic potential against insomnia.

Extracellular AMP Inhibits Pollen Tube Growth in Picea meyeri via Disrupted Calcium Gradient and Disorganized Microfilaments.

Adenosine monophosphate (AMP) is a hydrolysis product of adenosine triphosphate (ATP) and adenosine diphosphate (ADP). In mammalian cells, extracellular AMP functions as a signaling molecule by binding to adenosine A1 receptors, thereby activating various intracellular signaling pathways. However, the role of extracellular AMP in plant cells remains largely unclear, and homologs of A1 receptors have not been identified. Our previous studies have demonstrated that extracellular ATP (eATP) is crucial for the normal germination and growth of Picea meyeri pollen tubes. In the present study, we observed that the exogenous addition of ATP to a pollen culture medium could be degraded into AMP and adenosine. Furthermore, the addition of AMP and adenosine to the culture medium was found to inhibit pollen germination and tube elongation. Notably, the addition of an AMP receptor inhibitor into the culture medium mitigated the inhibitory effects of AMP on pollen tube growth. Through intracellular staining for Ca2+ and microfilaments, we discovered that high concentrations of AMP disrupt the Ca2+ concentration gradient and impair microfilament organization, ultimately resulting in inhibited pollen tube elongation. In conclusion, we propose that extracellular AMP, as a hydrolysis product of eATP, also plays a significant role in regulating P. meyeri pollen germination and tube growth in vitro.

The serotonergic psychedelic DOI impairs deviance detection in the auditory cortex.

Psychedelics are known to induce profound perceptual distortions, yet the neural mechanisms underlying these effects, particularly within the auditory system, remain poorly understood. In this study, we investigated the effects of the psychedelic compound 2,5-Dimethoxy-4-iodoamphetamine (DOI), a serotonin 2A receptor agonist, on the activity of neurons in the auditory cortex of awake mice. We examined whether DOI administration alters sound-frequency tuning, variability in neural responses, and deviance detection (a neural process reflecting the balance between top-down and bottom-up processing). Our results show that while DOI does not alter the frequency selectivity of auditory cortical neurons in a consistent manner, it increases trial-by-trial variability in responses and consistently diminishes the neural distinction between expected (standard) and unexpected (oddball) stimuli. This reduction in deviance detection was primarily driven by a decrease in the response to oddball sounds, suggesting that DOI dampens the auditory cortex's sensitivity to unexpected events. These findings provide insights into how psychedelics disrupt sensory processing and shed light on the neural mechanisms underlying the altered perception of auditory stimuli observed in the psychedelic state.

Synergistic Anxiolytic Effects of Linalool and Sesamol Co-Treatment on Swiss Albino Mice: A Potential GABAergic Intervention.

Sesamol (SES) and linalool (LIN) are aromatic compounds that have neuroprotective effects. The main purpose of this study is to evaluate the anxiolytic activity of LIN and SES co-treatment on Swiss albino mice and analyze its possible mechanism through in silico study. In this sense, the mice were given the gamma-aminobutyric acid type A receptors (GABAA) agonist diazepam (DZP; 3mg/kg, p.o.) as a positive control. A vehicle (10mL/kg) was served as control. The tested chemicals, single-dose LIN (50mg/kg) and SES (50mg/kg), as well as a combination (LIN+SES) and (DZP+LIN+SES), were administered orally in order to conduct several behavioral tests, including open-field, swings box, hole-crossing, and dark-resident time tests. Further, molecular docking studies of LIN, SES, and DZP were carried out through different software. The results showed that LIN and SES individually have significant anxiolytic-like activity in mice. Further, when LIN was combined with SES and with (SES+DZP), it exhibited a relatively lower locomotor activity in mice compared to individual treatment groups, indicating a synergistic action. In addition, the molecular docking analysis revealed that LIN and SES have a moderate binding affinity (-5.0 and -5.1kcal/mol) toward the GABAA receptor α3 subunit. In conclusion, our findings suggest that LIN and SES exerted synergistic anxiolytic activity on Swiss albino mice, possibly through the GABAergic interaction pathways.

CLEC7A Knockdown Alleviates Ischemic Stroke by Inhibiting Pyroptosis and Microglia Activation.

Ischemic stroke (IS) is the leading cause of mortality worldwide. Herein, we aimed to identify novel biomarkers and explore the role of C-type lectin domain family 7 member A (CLEC7A) in IS. Differentially expressed genes (DEGs) were screened using the GSE106680, GSE97537, and GSE61616 datasets, and hub genes were identified through construction of protein-protein interaction networks. An IS model was established by middle cerebral artery occlusion and reperfusion (MCAO/R). Neural function was assessed using triphenyl tetrazolium chloride, hematoxylin-eosin, and terminal deoxynucleotidyl transferase-mediated nick-end labeling. A cell counting kit was used to detect cell viability following oxygen-glucose deprivation/reperfusion (OGD/R). Inflammatory factors were detected using enzyme-linked immunosorbent assay. The mRNA and protein expression levels were detected using reverse transcription-quantitative polymerase chain reaction and western blotting, respectively. Fc fragment of Immunoglobulin G (IgG) receptor IIIa (FCGR3A), Fc fragment of Immunoglobulin E (IgE) receptor Ig (FCER1G), Complement component 5a receptor 1 (C5AR1), CLEC7A, Plasminogen activator, urokinase (PLAU), and C-C motif chemokine ligand 6 (CCL6) were identified as important hub genes, from which CLEC7A was selected as the primary subject of this study. The activation of microglia and pyroptosis were observed in MCAO/R model with increased levels of interleukin (IL)-1β, IL-18, tumor necrosis factor-α, and lactate dehydrogenase. CLEC7A knockdown was found to promote cell viability in BV2 cells and inhibiting pyroptosis in HT22 cells. CLEC7A knockdown in microglia also decreased infarct volume and neurological deficit scores, and alleviated injury and neuronal apoptosis in IS rats. CLEC7A knockdown inhibited pyroptosis and microglial activation in the MCAO/R model. A pyroptosis activator reversed the effect of CLEC7A knockdown on the viability of OGD/R-treated HT22 cells. CLEC7A is a promising biomarker of IS. CLEC7A knockdown alleviates IS by inhibiting pyroptosis and microglial activation.

Mechanistic Insights into the Adenosine A1 Receptor’s Positive Allosteric Modulation for Non-Opioid Analgesics

The adenosine A1 receptor (A1R) is a promising target for pain treatment. However, the development of therapeutic agonists is hampered by adverse effects, mainly including sedation, bradycardia, hypotension, or respiratory depression. Recently discovered molecules able to overcome this impediment are the positive allosteric modulator MIPS521 and the A1R-selective agonist BnOCPA, which are both potent and powerful analgesics with fewer side effects. While BnOCPA directly activates the A1R from the canonical orthosteric site, MIPS521 binds to an allosteric site, acting in concert with orthosteric adenosine and tuning its pharmacology. Given their overlapping profile in pain models but distinct mechanisms of action, we combined pharmacology and microsecond molecular dynamics simulations to address MIPS521 and BnOCPA activity and their reciprocal influence when bound to the A1R. We show that MIPS521 changes adenosine and BnOCPA G protein selectivity in opposite ways and propose a structural model where TM7 dynamics are differently affected and involved in the G protein preferences of adenosine and BnOCPA.

Cyperus rotundus Extract and Its Active Metabolite α-Cyperone Alleviates Paclitaxel-Induced Neuropathic Pain via the Modulation of the Norepinephrine Pathway.

Paclitaxel is a widely used anticancer drug for ovarian, lung, breast, and stomach cancers; however, its clinical use is often limited by the side effects of peripheral neuropathy. This study evaluated the effects of Cyperus rotundus (C. rotundus) extract and its active metabolite, α-cyperone, on paclitaxel-induced neuropathic pain. The oral administration of C. rotundus extract at doses of 500 mg/kg and intraperitoneal administration of α-cyperone at doses of 480 and 800 μg/kg prevented both the development of cold and mechanical pain. The gene and protein expressions of tyrosine hydroxylase and noradrenergic receptors (α1- and α2-adrenergic), which were upregulated by paclitaxel, were significantly downregulated in the C. rotundus extract-treated group. In the locus coeruleus region of the mouse brain, C. rotundus extract administration also reduced the elevated expression of tyrosine hydroxylase induced by paclitaxel. The concentration of α-cyperone in C. rotundus extract was quantified using high-performance liquid chromatography (HPLC). In the group treated with α-cyperone, at levels corresponding to its content in C. rotundus, both cold and mechanical allodynia were effectively prevented. This study suggests that α-cyperone shows potential as a preventive agent for paclitaxel-induced neuropathic pain.