This study aimed to investigate the effect of quercetin on atherosclerosis(AS) in apolipoprotein E knockout(ApoE~(-/-))mice and the mechanism underlying the regulation of cholesterol efflux mediated by adenosine triphosphate-binding cassette transporter A1/G1(ABCA1/G1) pathway and neutrophil extracellular traps(NETs) formation. An AS model was established by feeding Apo E~(-/-)mice with a high-fat diet. The mice were divided into a blank control group, a model group, an atorvastatin calcium group, and lowand high-dose quercetin groups. Serum levels of total cholesterol(TC), triglyceride(TG), low-density lipoprotein cholesterol(LDLC), high-density lipoprotein cholesterol(HDL-C), interleukin(IL)-6, tumor necrosis factor-α(TNF-α), and IL-1β were measured using biochemical methods. Biliary cholesterol excretion rate was determined by acute bile duct cannulation. Hepatic pathology and aortic plaque area were observed by hematoxylin-eosin(HE) and oil red O staining. Collagen and elastic fiber changes in plaques were evaluated by Masson and Verhoeff's Van Gieson(VVG) staining. CD3 and CD68 protein expressions in the aortic root and the colocalization of citrullinated histone H3(Cit-H3) and myeloperoxidase(MPO) were detected by immunohistochemistry and immunofluorescence double staining, respectively. ABCA1 and ABCG1 protein expressions in liver and aortic tissues were detected by Western blot. The results showed that in the model group, serum levels of TC, TG, LDL-C, IL-6, TNF-α, and IL-1β were significantly elevated, hepatic lipid deposition was pronounced, aortic plaque area was enlarged, collagen deposition and elastic fiber rupture were aggravated, inflammatory cell infiltration and NET formation in plaques were enhanced, whereas serum HDL-C level,biliary cholesterol excretion rate, and ABCA1 and ABCG1 protein expressions decreased. Following quercetin intervention, all these abnormal indicators were significantly improved, as evidenced by reduced blood lipid and inflammatory cytokine levels, alleviated hepatic lipid deposition, decreased aortic plaque area, stabilized collagen and elastic fiber architecuture in plaques, suppressed inflammatory cell infiltration and NET formation, accelerated biliary cholesterol excretion, and upregulated ABCA1 and ABCG1 protein expressions. The effects of quercetin were dose-dependent, and the high-dose group showed comparable efficacy to atorvastatin calcium. In conclusion, quercetin may effectively attenuate the progression of AS in Apo E~(-/-)mice by upregulating ABCA1/G1-mediated cholesterol efflux, inhibiting NET formation, and alleviating inflammatory responses.

This study aims to investigate the antidepressant effects of essential oil from Schizonepeta tenuifolia and its mechanism of action on intervening the 5-hydroxytryptamine 1A receptor(5-HT1AR)/cyclic adenosine monophosphate(cAMP)/protein kinase A(PKA) signaling pathway in the depression mouse models induced by chronic unpredictable mild stress( CUMS). The 5-HT1AR agonist 8-OH-DPAT(1. 5 mg·kg~(-1)) and 5-HT2A receptor agonist DOI(1 mg·kg~(-1)) were administered to induce 5-hydroxytryptamine(5-HT) syndrome-like behaviors in mice, and the effects of essential oil from S. tenuifolia on the 5-HT system were observed. CUMS depression mouse models were employed, and after six weeks of continuous modeling, mice were divided into the model group, the vortioxetine group(10 mg·kg~(-1)), and the low-dose and high-dose(50, 100 mg·kg~(-1)) essential oil from S. tenuifolia groups based on the sucrose preference test( SPT) results, with a blank group as control. Mice in each group received continuous intragastric administration for four weeks, with the modeling procedure continuing during the treatment period. After four weeks of administration, the SPT, tail suspension test( TST), forced swimming test( FST), and open field test( OFT) were conducted to evaluate antidepressant-like effects of the drugs. Nissl staining was employed to assess hippocampal neuronal damage in mice. Enzyme-linked immunosorbent assay(ELISA) was used to measure the levels of tryptophan(Trp) and 5-HT in the serum and hippocampal tissue. Immunohistochemistry(IHC) was used to detect 5-HT1AR expression levels in the hippocampus. The RNA-Seq technique was used to analyze differentially expressed genes in the mice's hippocampus, and the potential action targets were analyzed via GO and KEGG enrichment analyses. Subsequently, RT-qPCR and Western blot were employed to detect m RNA and protein expression levels of key target molecules in the 5-HT1AR/cAMP/PKA signaling pathway in the hippocampal tissue. RESULTS:: show that essential oil from S. tenuifolia significantly increases the times of mice's body squats, tremors, and hindlimb abduction induced by 8-OH-DPAT and reduces DOI-induced count of head twitching episodes of mice. The sucrose preference rate and central zone residence time in the OFT of CUMS model mice were significantly increased. In contrast, the immobility time in the FST and TST was markedly shortened. After using the essential oil from S. tenuifolia, the number of Nissl bodies in the hippocampal CA3 and DG regions of CUMS model mice, as well as the levels of Trp in the serum and hippocampus and 5-HT in the hippocampus, were increased. In contrast, 5-HT1AR expression level in hippocampal CA3 and DG regions was upregulated. RNA-Seq detection results suggest that the antidepressant effects of essential oil from S. tenuifolia may be highly associated with the cAMP/PKA signal transduction pathway. Validation experiments demonstrate that essential oil from S. tenuifolia significantly upregulates the m RNA and protein expression levels of tryptophan hydroxylase 2(TPH2), 5-HT1AR, PKA, cAMP response element-binding protein( CREB), brain-derived neurotrophic factor(BDNF), tropomyosin receptor kinase B( TrkB), synaptophysin( SYP), and postsynaptic density protein 95(PSD95) in the hippocampus, the cAMP protein level was also increased. In summary, essential oil from S. tenuifolia exerts antidepressant effects by activating the 5-HT1AR/cAMP/PKA signaling pathway in the hippocampus of CUMS-induced depressive-like model mice, thereby influencing 5-HT system function and elevating central 5-HT concentrations.

Excessive neuroinflammation caused by activated microglia contributes to the progression of neurodegenerative diseases such as Alzheimer's and Parkinson's diseases. Microglia, the immune cells of the central nervous system, play vital roles in brain development, immune defense, and maintaining the neural environment. Cordycepin from Cordyceps militaris exhibits diverse biological activities, including anti-inflammatory, antioxidant, and hypoglycemic effects. This study investigated the effects of ethanol extract of C. militaris fermented from Gryllus bimaculatus (EECM) and cordycepin on lipopolysaccharide (LPS)-induced neuroinflammation in BV2 microglial cells. EECM significantly reduced the levels of nitric oxide (NO), interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α) elevated by LPS. Similarly, cordycepin decreased the secretion of NO, TNF-α, and IL-6. Additionally, LPS-induced expression of toll-like receptor 4 (TLR4) and inflammatory proteins (iNOS, COX-2, TNF-α, and IL-6) was significantly suppressed by cordycepin. Molecular docking revealed that cordycepin binds to the TLR4/MD2 complex (-6.2 kcal/mol) at Lys341, Ser364, and Phe365, suggesting competitive inhibition at the LPS-binding site. These findings indicate that cordycepin attenuates LPS-induced neuroinflammation by modulating the TLR4 signaling pathway, highlighting its therapeutic potential for neuroinflammatory conditions.

To overcome poor adenylyl cyclase (AC) selectivity and weak cyclic adenosine monophosphate (cAMP) agonistic activity of isoforskolin (ISOF), eleven derivatives were synthesised using ISOF as the parent compound. cAMP agonistic activity suggested that compound 10 possessed cAMP activation potency and preferential activation towards AC5 (92.8%), similar to the 96.8% of the positive control forskolin (FSK) and higher than the 90.0% of ISOF. Dose-response curves further demonstrated that the EC50 value of compound 10 was 1.85 μM, slightly higher than the 1.05 μM of FSK. Preliminary structure-activity suggested that substitution at the C7 position contributed to AC5-preferential activation. Network pharmacology and molecular docking predicted that compound 10 was associated with cAMP signalling, calcium signalling, and adrenergic signalling in cardiomyocytes, and formed a putative complex with AC5 (-9.21 kJ/mol) via hydrogen bonds, π-alkyl stacking, and hydrophobic interactions. Overall, compound 10 preferentially activates AC5, representing a promising candidate for developing AC5-preferential agonists.

Polyamines are ubiquitous polycations involved in diverse cellular functions, including proliferation, differentiation, and survival. While endogenous biosynthesis can maintain the intracellular levels of polyamines, uptake becomes critical during high metabolic demand. Despite the evidence for ATP-dependent polyamine uptake systems in the plasma membrane, their molecular characteristics remain unclear. We found that two human P5B ATPases-ATP13A3 and ATP13A4-were localized to the plasma membrane and directly mediated the uptake of putrescine and spermidine in an ATP-dependent manner. Using ATP13A3 and ATP13A4 heterologously expressed in COS-7 cells, we demonstrated that they mediate the saturable, ATP-dependent uptake of polyamines with distinct kinetic profiles, consistent with the characteristics of known uptake systems in mammalian cells. Site-directed mutagenesis of the conserved ATPase motif abolished uptake, confirming the requirement of ATP hydrolysis. The immunofluorescence analysis in COS-7 cells transiently expressing FLAG-tagged ATP13A3 or ATP13A4 revealed that both transporters localize to the plasma membrane. RNA interference in HepG2 cells to suppress endogenous ATP13A3 and ATP13A4 reduced polyamine uptake, supporting the physiological relevance of these transporters. It was also found that ATP13A3 is ubiquitously expressed and has moderate affinity for polyamines, whereas ATP13A4 is more specific tissue-restricted and has higher affinity for polyamines, particularly spermidine. These findings establish ATP13A3 and ATP13A4 as the first P-type ATPases shown to transport small organic solutes for cellular uptake at the plasma membrane. This study elucidates the roles of these P-type ATPases in polyamine homeostasis. Furthermore, given the emerging associations of ATP13A3 and ATP13A4 with neurological and metabolic disorders, these transporters are promising therapeutic targets in polyamine-related diseases.

The mechanisms by which pancreatic β cells sense and communicate environmental cues to mitochondria to maintain insulin secretion remain poorly understood. We identified that β cells transduce metabolic stress, such as during obesity or glucolipotoxicity, through the innate immune signaling adaptor TRAF6 to regulate mitochondrial quality control. TRAF6 was dispensable for β-cell function at baseline yet was essential for glucose-stimulated insulin secretion in mouse or human β cells following nutrient excess. TRAF6 was required for the recruitment of components of both ubiquitin- and receptor-mediated mitophagy pathways to clear dysfunctional mitochondria following metabolic stress. Interestingly, concomitant loss of the ubiquitin-mediated mitophagy effector Parkin restored glucose homeostasis and mitophagy flux in TRAF6-deficient β cells by relieving impediments in receptor-mediated mitophagy. Together, we found that TRAF6 forms a molecular relay linking environmental signals to multiple mitophagy pathways to equip β cells with the means to adapt to metabolic demand and prevent diabetes.Abbreviations: ADP: adenosine diphosphate; ATP: adenosine triphosphate; BNIP3: BCL2 interacting protein 3; GSIS: glucose-stimulated insulin secretion; HFD: high-fat diet; MFN2: mitofusin 2; PRKN: parkin RBR E3 ubiquitin protein ligase; RNF41/NRDP1: ring finger protein 41; SQSTM1/p62: sequestosome 1; T2D: type 2 diabetes; TAX1BP1: Tax1 binding protein 1; TLR: Toll-like receptor; TRAF6: TNF receptor associated factor 6.

ZBP1 senses foreign nucleic acids in the unusual, left-handed Z-conformation via binding through its Zα1 and Zα2 domains and initiates cell death pathways that enable viral clearance. dsDNA and dsRNA can adopt Z-conformations; however, both typically require chemical modifications or protein binding to induce Z-form conversion and stabilization. ZBP1 has been previously shown to bind and convert B-DNA to the Z-conformation and was assumed to be able to convert A-RNA as well, despite the lack of experimental validation. Here, we use NMR and other biophysical and biochemical experiments to characterize the Z-DNA and Z-RNA binding properties of ZBP1's Zα1 and Zα2 domains. While ZBP1's Zα domains are able to convert and stabilize unmodified dsDNA in the Z-conformation, they are incapable of flipping unmodified (CpG)n repeat dsRNA. ZBP1's Zα domains require dsRNAs with Z-promoting chemical modifications to be able to bind and stabilize the Z-conformation. These results contrast with ADAR1's Zα domain, which can bind and flip both dsDNA and dsRNA into the Z-conformation, potentially indicating finely tuned competition between ADAR1 and ZBP1 for pro-survival and pro-death outcomes, respectively. This work highlights the functional variability of Zα domains and narrows down potential physiological substrates of ZBP1 in infection and disease.

Mitochondria are a semi-autonomous organelle due to their unique structural organization and independent genetic system, and they play diverse roles in maintaining cellular homeostasis. The liver is the central metabolic organ and performs myriad functions, including energy production, metabolism, and detoxification. Liver diseases account for approximately two million mortalities per year. Mitochondrial dysfunction and damage are implicated in promoting the pathogenesis of different liver diseases. Mitochondria and mitochondrial nucleic acids, namely, mitochondrial DNA (mtDNA) and mitochondrial RNA (mtRNA), have emerged as regulators of innate immunity. Here, we first review the structural organization of mitochondria and the molecular mechanisms governing mitochondrial biogenesis, replication, and transcription. Next, we describe the mechanistic details by which mitochondrial dysfunction causes the release of mtDNA and mtRNA into the cytosol and extracellular milieu. Some of the mechanisms are through the opening of mitochondrial permeability transition pore (mPTP), voltage-dependent anion channel (VDAC) oligomerization, prohibitin (PHB1/2), mitochondria-derived vesicles (MDV), dysfunction of the mtRNA degradosomes, and senescence-associated secretory phenotype (SASP). Cell death pathways such as necrosis, necroptosis, and pyroptosis, in addition to perpetuating inflammation, also release mtDNA and mtRNA into the extracellular milieu. The released mtDNA and mtRNA act as damage associated molecular patterns (DAMPs) that engage various cytosolic DNA and RNA sensors and pattern recognition receptors (PRRs) such as cyclic guanosine-monophosphate-adenosine-monophosphate synthase (cGAS), toll-like receptor 9 (TLR9), toll-like receptor 3 (TLR3), Z-DNA-binding protein 1 (ZBP1), NOD-like receptor protein 3 (NLRP3), absent in melanoma 2 (AIM2), retinoic acid-inducible gene I (RIG-I), melanoma differentiation-associated protein 5 (MDA5), protein kinase R (PKR) and activate different signaling cascades downstream. This review provides a contemporary, comprehensive analysis of the available literature on the roles of mtDNA and mtRNA in liver diseases, including metabolic dysfunction-associated steatotic/steatohepatitis (MASLD/MASH), alcohol-associated liver disease (ALD), drug-induced liver injury (DILI), viral hepatitis, and hepatocellular carcinoma (HCC). Thus, the review highlights the potential of both mtDNA and mtRNA to serve as biomarkers of the pathological conditions of the liver, and the way the mitigation of mitochondrial dysfunction and the release of mtDNA or mtRNA can serve as potential therapeutic targets.

Melatonin has demonstrated antitumor activity in various cancer models, but its effects may depend on the therapeutic dose and cellular environment. We found that melatonin at millimolar concentrations significantly downregulated transglutaminase 2 (TGM2), histone serotonylation (H3K4me3Q5ser), and YTHDF1/m[6]A (N6-methyladenosine) in a colorectal cancer model. Cross-interference between TGM2 and YTHDF1 supports a link between these regulatory processes, while molecular docking, pharmacological interventions, and co-immunoprecipitation experiments suggest the existence of a signaling pathway involving melatonin and the HTR2B receptor. A combined analysis of CHIP-seq and meRIP-seq identified salt-induced kinase 1 (SIK1) as a potential downstream target, and SIK1 interference affected the proliferation and migration of colorectal cancer. Collectively, these findings support a working model in which melatonin-induced tumor suppression is associated with coordinated changes in TGM2/H3K4me3Q5ser and YTHDF1-associated m[6]A regulation, with SIK1 emerging as a potential convergent downstream effector.

Hyperpigmentation is a dermatological condition characterized by excessive melanin production, resulting in skin darkening. Intrinsic and extrinsic mediators, including ultraviolet (UV) radiation, hormonal changes, inflammation, and genetic predisposition, contribute to hyperpigmentation pathogenesis. The current review thoroughly examines the biological processes involved in hyperpigmentation, with particular emphasis on its signaling pathways, particularly the cyclic adenosine monophosphate (cAMP), wingless-related integration site (Wnt)/β-catenin, phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt), and mitogen-activated protein kinase (MAPK), thereby providing mechanistic insights that may facilitate the development of effective treatments. The review also evaluates the safety, toxicity, and therapeutic efficacy of TYR inhibitors, arbutin, azelaic acid, ellagic acid, kojic acid, resveratrol, and the melanosome inhibitor niacinamide, for managing hyperpigmentation. Additionally, the review summarizes the in vitro and in vivo studies of these agents and other emerging plant extracts. Recent advances in the use of plant extracts for hyperpigmentation have shown promising efficacy; nevertheless, their translation into clinical and commercial applications remains limited. This limited translation might be attributed to the challenges related to standardization and the paucity of clinical evidence regarding long-term safety. Moreover, ongoing studies on natural anti-hyperpigmentation agents have highlighted the critical role of patent analyses in the advancement of biomedical research. This review provides a strategic direction for researchers to assess the safety, efficacy, and toxicity profiles of natural anti-hyperpigmentation agents.

The Coronary Vasomotor Disorders International Study group (COVADIS) developed the diagnostic criteria for vasospastic angina and microvascular angina more than 8 years ago. This article critically reappraises these criteria (Latin: quid fecisti - what has been done) and proposes the strategies for their advancement (Latin: Quo Vadis - where are you are going?). Although the criteria have some limitations, they remain relevant considering their multifaceted clinical perspective including the presence of symptoms, myocardial ischaemia and objective evidence of coronary vasomotor dysfunction. To improve the criteria, the following strategic directions (Quo Vadis) are proposed, (1) optimising invasive functional coronary angiography assessment - the benchmark diagnostic investigation for coronary vasomotor disorders, (2) diagnosing mixed macrovascular and microvascular coronary vasomotor disorders, (3) advancing non-invasive coronary vasomotor assessment methods, (4) linking coronary vasomotor disorder criteria to clinical outcomes, (5) evaluating coronary vasomotor disorders in the presence of obstructive coronary artery disease, and (6) global partnering of coronary vasomotor disorder initiatives in national networks and patient forums. These strategies will improve the diagnostic approaches to coronary vasomotor disorders thereby facilitating the evolution of the COVADIS Diagnostic Criteria and thus management of these elusive disorders.