Cyclooxygenase Pathway Research Articles

Aspirin modulates production of pro-inflammatory and pro-resolving mediators in endothelial cells.

Endothelial cells synthesize biochemical signals to coordinate a response to insults, resolve inflammation and restore barrier integrity. Vascular cells release a variety of vasoactive bioactive lipid metabolites during the inflammatory response and produce pro-resolving mediators (e.g., Lipoxin A4, LXA4) in cooperation with leukocytes and platelets to bring a halt to inflammation. Aspirin, used in a variety of cardiovascular and pro-thrombotic disorders (e.g., atherosclerosis, angina, preeclampsia), potently inhibits proinflammatory eicosanoid formation. Moreover, aspirin stimulates the synthesis of pro-resolving lipid mediators (SPM), so-called Aspirin-Triggered Lipoxins (ATL). We demonstrate that cytokines stimulated a time- and dose-dependent increase in PGI2 (6-ketoPGF1α) and PGE2 formation that is blocked by aspirin. Eicosanoid production was caused by cytokine-induced expression of cyclooxygenase-2 (COX-2). We also detected increased production of pro-resolving LXA4 in cytokine-stimulated endothelial cells. The R-enantiomer of LXA4, 15-epi-LXA4, was enhanced by aspirin, but only in the presence of cytokine challenge, indicating dependence on COX-2 expression. In contrast to previous reports, we detected arachidonate 5-lipoxygenase (ALOX5) mRNA expression and its cognate protein (5-lipoxygenase, 5-LOX), suggesting that endothelial cells possess the enzymatic machinery necessary to synthesize both pro-inflammatory and pro-resolving lipid mediators independent of added leukocytes or platelets. Finally, we observed that, endothelial cells produced LTB4 in the absence of leukocytes. These results indicate that endothelial cells produce both pro-inflammatory and pro-resolving lipid mediators in the absence of other cell types and aspirin exerts pleiotropic actions influencing both COX and LOX pathways.

Abstract 5175: DX-002, a dual EP2 and EP4 antagonist, alters the tumor microenvironment to enhance tumor immunogenicity and responses to immune checkpoint inhibitors

Abstract Prostaglandin E2 (PGE2) generated by the cyclooxygenase (COX) pathways regulates both inflammation and tumor microenvironment (TME). Among the four E-type prostanoid receptors EP1-4, it has been demonstrated that PGE2 exhibits its immune suppression and tumorigenic function through engagement with EP2 and EP4. Mechanistically, tumor-derived PGE2 acts on EP2 and EP4 in immune cells (e.g., NK cells and dendritic cells) and elicits immunosuppression in TME1,2. Importantly, the deletion of both EP2 and EP4, but not deletion of either receptor, is required to reverse PGE2-mediated immune evasion. Furthermore, high levels of PGE2-EP2/EP4 associated inflammatory signature have been shown in multiple human cancer types to correlate with poor prognosis and worse response to immune checkpoint blockade in the clinic1. Therefore, PGE2-EP2/EP4 signaling likely acts as a key regulatory node linking inflammation and immunosuppression in TME, and its inhibition may lead to a less immunosuppressive TME and enhance the response to immune checkpoint inhibitors. COX inhibitors have been used to block PGE2 synthesis but cause cardiovascular and gastrointestinal risks due to the inhibition of all prostanoids synthesis3. Thus, therapeutically targeting both EP2 and EP4 is an effective and more selective approach to limit the immunosuppressive functions of PGE2 while avoiding potential side effects associated with COX inhibition. Here we describe the discovery of a potent and selective EP2/EP4 dual antagonist DX-002. At 10 nM concentration, it displayed >90% inhibition of PGE2 binding to EP2 or EP4, whereas no significant inhibition was seen towards PGE2 binding to EP1 or EP3. In human NK cells DX-002 reversed PGE2-mediated inhibition of IFN-γ production more potently than selective EP2 or EP4 antagonist. DX-002 has excellent pharmacokinetics profiles across all animal species. In the mouse tumor model, treatment of DX-002 produced superior tumor regression as a monotherapy over selective EP2 or EP4 antagonists. In addition, DX-002 also provided significant synergistical effect for tumor regression in combination with anti-PD1 in both immune checkpoint inhibitor insensitive and sensitive models, in association with increased tumor infiltration of CD8+ T cells. Based on its in vitro and in vivo profiles along with other properties, DX-002 is being profiled as a development candidate as a potential cancer immunotherapeutic agent. Citation Format: Lintong Li, Guorui Yao, Wei Zheng, Yongshuai Chai, Zhongying Huang, Guojian Ma, Linlin Nie, Dengfeng Xu, Jiuquan Yang, Nanxin Li. DX-002, a dual EP2 and EP4 antagonist, alters the tumor microenvironment to enhance tumor immunogenicity and responses to immune checkpoint inhibitors. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5175.

Abstract 4159: The COX-2/PGE2 pathway as a mediator of resistance to anti-PD-1 therapy

Abstract Background: We previously found upregulation of the cyclooxygenase-2/prostaglandin E2 (COX-2/PGE2) pathway in the tumor microenvironment (TME) of cancer types that respond poorly to anti-PD-1 therapy, and showed that TME-resident cytokines such as IL-1B induce COX-2 expression in human tumor and myeloid cells. To investigate a potential role for COX-2/PGE2 in anti-PD-1 resistance, we studied PGE2-mediated effects on human myeloid and T cells and their dependence on prostaglandin (EP) receptor signaling. Methods: Monocytes (Monos) and T cells were enriched from normal donor peripheral blood by negative selection. PGE2 effects were tested on Monos activated with IFN-g (PD-L1 expression; FACS) or LPS (cytokine secretion; ELISA). PGE2 effects on monocytic dendritic cells (DCs) generated with GM-CSF+IL-4 and matured with CD40L were assessed by phenotyping for costimulatory molecules (CD80, CD86) and subpopulation markers (CD14, CD16, CD83). T cells activated with anti-CD3/CD28 +/- PGE2 for 2-6 days were assessed for expression of CD25, CD69, CD107a, CD137, OX40, GITR, LAG-3, PD-1, PD-1, PD-L1 and TIM3 (FACS); proliferation markers (Ki67, CFSE; FACS); cytokine secretion (IFN-g, IL-2, IL-10, TGF-b; ELISA); and metabolic function (Seahorse assay). In some experiments, cells were pre-incubated with inhibitors of EP2 (PF-04418948), EP4 (ONO-AE3-208), or the dual inhibitor TPST-1495 (Tempest Therapeutics). Results: Myeloid cell functions were suppressed by PGE2, which reduced IFNg-induced PD-L1 expression and modulated LPS-induced cytokine secretion. Furthermore, PGE2 inhibited DC maturation, evidenced by persistent CD14 and CD16 expression and a decreased CD83+ subset. In CD4 and CD8 T cells activated with anti-CD3/CD28, PGE2 reduced expression of CD25 and CD69 (early activation markers), CD107a (cytolytic potential), Ki67 (cell proliferation), co-stimulatory molecules (CD137, OX40, GITR) and checkpoint molecules (LAG-3, PD-1, PD-L1, TIM3) in a dose-dependent manner. PGE2 also reduced the CFSE-low (proliferating) population in activated CD4, CD8 and Treg cells. Secretion of proinflammatory cytokines IFN-g and IL-2, and inhibitory cytokines IL-10 and TGF-b from activated T cells was also suppressed by PGE2. These effects were specifically reversed by chemical inhibitors of EP2 and/or EP4. PGE2 exposure at the time of T cell activation was associated with depressed mitochondrial oxidation and cellular glycolysis, suggesting global effects on T cell metabolism. Conclusion: Understanding and preventing anti-PD-1 resistance is a critical goal in oncology. Our results suggest that the COX-2/PGE2 pathway exerts immunosuppressive effects on myeloid and T cells via the EP2 and EP4 receptors. COX-2 and PD-L1 are distinct immune checkpoints, providing a rationale to test anti-PD-1 in conjunction with COX-2 pathway inhibitors such as IL-1B antagonists and highly specific EP2 and EP4 inhibitors in patients with cancer. Citation Format: Shuming Chen, Seoho Lee, Tracee L. McMiller, Isaac Morales, Preethi Sankaran, Brian Francica, Robert D. Leone, Tom W. Dubensky, Suzanne L. Topalian. The COX-2/PGE2 pathway as a mediator of resistance to anti-PD-1 therapy. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4159.

Ge-Gen Decoction Exerts an Anti-Primary Dysmenorrhea Effect in Rats by Inactivating the HSP90/NLRP3/NF-κB/COX-2 Pathway.

Although Ge-Gen decoction (GGD) has beneficial effects on primary dysmenorrhea (PD), the underlying mechanisms remain poorly understood. Our previous proteomic data revealed decreased level of heat shockprotein 90 (HSP90) in uterine tissues of rats with PD after GGD treatment. However, the potential role of HSP90 in the anti-PD effect of GGD and the underlying mechanisms remain unexplored. This study investigated the potential role and mechanism of HSP90 in the anti-PD effect of GGD using a PD rat model. Wistar female rats were used to investigate the potential role of HSP90 in the anti-PD effect of GGD. The rat PD model was established by injecting estradiol benzoate and oxytocin. GGD, Terazosin (an agonist of HSP90) or GGD combined with Terazosin were orally administered to the PD rats. The expression levels of protein and cytokines, including HSP90, nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), nuclear factor kappa B (NF-κB) and cyclooxygenase-2 (COX-2) in the uterine tissue of rats in each group were detected by immunohistochemical assay or Western blot. GGD ameliorated the writhing response, suppressed the protein levels of HSP90 and inflammation-associated proteins, including NLRP3, NF-κB, and COX-2 in uterine tissues of rats with PD. Terazosin attenuated the anti-PD effect of GGD and reversed the effects of GGD on the protein levels of NLRP3, NF-κB and COX-2 in uterine tissues. GGD exerts an anti-PD effect and suppresses levels of HSP90 and some inflammation associated proteins in uterine tissues of rats.

Probiotic treatment with Kefir reduces vascular oxidative stress while suppressing COX2 mediated relaxation in intestinal arteries of an animal model of menopause

Functional foods such as probiotics are known to have benefits in various diseases including metabolic disorders and cardiovascular disease (CVD). In women, CVD has been shown to be linked with their gut microbiota and hormones. Here, we have evaluated the effects of chronic Kefir, a fermented milk beverage in Russia, Central Asia, Middle East and Eastern Europe, on mesenteric artery, using an animal model of menopause focusing on the superoxide anion and COX2 pathways. Two-month-old female Wistar rats were ovariectomized and treated by gavage with Kefir (5 % w/v, 3 mL/kg/day) or milk (Control) during two months. After this period, third-order mesenteric artery segments were isolated and mounted in a myograph system for evaluation of concentration-response curves to acetylcholine. We performed western blot analyses and measured oxidative stress through dihydroethidium (DHE) staining. Kefir reduced vascular oxidative stress, despite not changing SOD2 levels. COX2 levels were not changed by kefir, despite an apparent tendency towards reduction. However, in the functional experiments, under incubation with a COX2 inhibitor, a suppression of this pathway was observed in the kefir group, which suggests an interaction between inflammatory pathways and oxidative stress in this model. The effect of acute incubation with a superoxide anion scavenger on vascular responsiveness was equal in both groups. Kefir reduces vascular oxidative stress levels while suppressing COX2-mediated relaxation in mesenteric vessels in an animal model of menopause; which appears to involve an interplay between these two factors.

Linking dysbiosis to precancerous stomach through inflammation: Deeper than and beyond imaging.

Upper gastrointestinal endoscopy is considered the gold standard for gastric lesions detection and surveillance, but it is still associated with a non-negligible rate of missing conditions. In the Era of Personalized Medicine, biomarkers could be the key to overcome missed lesions or to better predict recurrence, pushing the frontier of endoscopy to functional endoscopy. In the last decade, microbiota in gastric cancer has been extensively explored, with gastric carcinogenesis being associated with progressive dysbiosis. Helicobacter pylori infection has been considered the main causative agent of gastritis due to its interference in disrupting the acidic environment of the stomach through inflammatory mediators. Thus, does inflammation bridge the gap between gastric dysbiosis and the gastric carcinogenesis cascade and could the microbiota-inflammation axis-derived biomarkers be the answer to the unmet challenge of functional upper endoscopy? To address this question, in this review, the available evidence on the role of gastric dysbiosis and chronic inflammation in precancerous conditions of the stomach is summarized, particularly targeting the nuclear factor-κB (NF-κB), toll-like receptors (TLRs) and cyclooxygenase-2 (COX-2) pathways. Additionally, the potential of liquid biopsies as a non-invasive source and the clinical utility of studied biomarkers is also explored. Overall, and although most studies offer a mechanistic perspective linking a strong proinflammatory Th1 cell response associated with, but not limited to, chronic infection with Helicobacter pylori, promising data recently published highlights not only the diagnostic value of microbial biomarkers but also the potential of gastric juice as a liquid biopsy pushing forward the concept of functional endoscopy and personalized care in gastric cancer early diagnosis and surveillance.

Do NSAIDs used prior to standard inferior alveolar nerve blocks improve the analgesia of mandibular molars with irreversible pulpitis? An umbrella review.

The aim of the present umbrella review was to answer the following question: "Does the use of NSAIDs as premedication increase the efficacy of the standard inferior alveolar nerve block on teeth with symptomatic irreversible pulpitis?" Systematic reviews with and without meta-analyses that evaluated the influence of premedication on anesthetic efficacy of the inferior alveolar nerve in symptomatic irreversible pulpitis of mandibular molars were searched in six electronic databases (MEDLINE/PubMed, Scopus, Web of Science, Cochrane Library, EMBASE, and Grey Literature Reports), without the restriction of language or year of publication. A Measurement Tool to Assess systematic Reviews (AMSTAR 2) was used to evaluate the quality of the included studies. Twelve systematic reviews were included. Only one did not perform a meta-analysis. The AMSTAR 2 overall confidence ranged from very low to high. In general, the main findings of the systematic reviews were that non-steroidal anti-inflammatory drugs (e.g., ibuprofen, oxicam, diclofenac, association of ibuprofen with acetaminophen, and ketorolac) increased the success rate of the inferior alveolar nerve block. From the "very low" to "high"-quality evidence available, this umbrella review concluded that NSAIDs as premedication acts through cyclooxygenase pathways and block the synthesis of specific prostaglandins that complicate the mechanism of action of the anesthesia, improving its success rate. Non-steroidal anti-inflammatory drugs can increase the success rate of the anesthetic technique of inferior alveolar nerve block efficacy in situations of mandibular molars with symptomatic irreversible pulpitis.

An integrated transcriptomic and metabolomic approach to investigate the heterogeneous Candida albicans biofilm phenotype

Candida albicans is the most prevalent and notorious of the Candida species involved in bloodstream infections, which is characterised by its capacity to form robust biofilms. Biofilm formation is an important clinical entity shown to be highly variable among clinical isolates. There are various environmental and physiological factors, including nutrient availability which influence the phenotype of Candida species. However, mechanisms underpinning adaptive biofilm heterogeneity have not yet been fully explored.Within this study we have profiled previously characterised and phenotypically distinct C. albicans bloodstream isolates. We assessed the dynamic susceptibility of these differing populations to antifungal treatments using population analysis profiling in addition to assessing biofilm formation and morphological changes. High throughput methodologies of RNA-Seq and LC-MS were employed to map and integrate the transcriptional and metabolic reprogramming undertaken by heterogenous C. albicans isolates in response to biofilm and hyphal inducing serum.We found a significant relationship between biofilm heterogeneity and azole resistance (P < 0.05). In addition, we observed that in response to serum our low biofilm forming (LBF) C. albicans exhibited a significant increase in biofilm formation and hyphal elongation. The transcriptional reprogramming of LBF strains compared to high biofilm forming (HBF) was distinct, indicating a high level of plasticity and variation in stress responses by heterogenous strains. The metabolic responses, although variable between LBF and HBF, shared many of the same responses to serum. Notably, a high upregulation of the arachidonic acid cascade, part of the COX pathway, was observed and this pathway was found to induce biofilm formation in LBF 3-fold.C. albicans is a highly heterogenous bloodstream pathogen with clinical isolates varying in antifungal tolerance and biofilm formation. In addition to this, C. albicans is capable of highly complex and variable regulation of transcription and metabolic pathways and heterogeneity across isolates further increases the complexity of these pathways. Here we have shown with a dual and integrated approach, the importance of studying a diverse panel of C. albicans isolates, which has the potential to reveal distinct pathways that can harnessed for drug discovery.

Brain metastasis in breast cancer: focus on genes and signaling pathways involved, blood-brain barrier and treatment strategies.

Breast cancer(BC) is one of the most prevalent types of cancer in women. Despite advancement in early detection and efficient treatment, recurrence and metastasis continue to pose a significant risk to the life of BC patients. Brain metastasis(BM) reported in 17-20 percent of BC patients is considered as a major cause of mortality and morbidity in these patients. BM includes various steps from primary breast tumor to secondary tumor formation. Various steps involved are primary tumor formation, angiogenesis, invasion, extravasation, and brain colonization. Genes involved in different pathways have been reported to be associated with BC cells metastasizing to the brain. ADAM8 gene, EN1 transcription factor, WNT, and VEGF signaling pathway have been associated with primary breast tumor; MMP1, COX2, XCR4, PI3k/Akt, ERK and MAPK pathways in angiogenesis; Noth, CD44, Zo-1, CEMIP, S0X2 and OLIG2 are involved in invasion, extravasation and colonization, respectively. In addition, the blood-brain barrier is also a key factor in BM. Dysregulation of cell junctions, tumor microenvironment and loss of function of microglia leads to BBB disruption ultimately resulting in BM. Various therapeutic strategies are currently used to control the BM in BC. Oncolytic virus therapy, immune checkpoint inhibitors, mTOR-PI3k inhibitors and immunotherapy have been developed to target various genes involved in BM in BC. In addition, RNA interference (RNAi) and CRISPR/Cas9 are novel interventions in the field of BCBM where research to validate these and clinical trials are being carried out. Gaining a better knowledge of metastasis biology is critical for establishing better treatment methods and attaining long-term therapeutic efficacies against BC. The current review has been compiled with an aim to evaluate the role of various genes and signaling pathways involved in multiple steps of BM in BC. The therapeutic strategies being used currently and the novel ones being explored to control BM in BC have also been discussed at length.

Alterations of pain pathways by experimental sleep disturbances in humans: central pain-inhibitory, cyclooxygenase, and endocannabinoid pathways.

There is strong evidence that sleep disturbances are an independent risk factor for the development of chronic pain conditions. The mechanisms underlying this association, however, are still not well understood. We examined the effect of experimental sleep disturbances (ESDs) on three pathways involved in pain initiation/resolution: (1) the central pain-inhibitory pathway, (2) the cyclooxygenase (COX) pathway, and (3) the endocannabinoid (eCB) pathway. Twenty-four healthy participants (50% females) underwent two 19-day long in-laboratory protocols in randomized order: (1) an ESD protocol consisting of repeated nights of short and disrupted sleep with intermittent recovery sleep; and (2) a sleep control protocol consisting of nights with an 8-hour sleep opportunity. Pain inhibition (conditioned pain modulation, habituation to repeated pain), COX-2 expression at monocyte level (lipopolysaccharide [LPS]-stimulated and spontaneous), and eCBs (arachidonoylethanolamine, 2-arachidonoylglycerol, docosahexaenoylethanolamide [DHEA], eicosapentaenoylethanolamide, docosatetraenoylethanolamide) were measured every other day throughout the protocol. The central pain-inhibitory pathway was compromised by sleep disturbances in females, but not in males (p < 0.05 condition × sex effect). The COX-2 pathway (LPS-stimulated) was activated by sleep disturbances (p < 0.05 condition effect), and this effect was exclusively driven by males (p < 0.05 condition × sex effect). With respect to the eCB pathway, DHEA was higher (p < 0.05 condition effect) in the sleep disturbance compared to the control condition, without sex-differential effects on any eCBs. These findings suggest that central pain-inhibitory and COX mechanisms through which sleep disturbances may contribute to chronic pain risk are sex specific, implicating the need for sex-differential therapeutic targets to effectively reduce chronic pain associated with sleep disturbances in both sexes. NCT02484742: Pain Sensitization and Habituation in a Model of Experimentally-induced Insomnia Symptoms. https://clinicaltrials.gov/ct2/show/NCT02484742.

Vaccination with celecoxib-treated dendritic cells improved cellular immune responses in an animal breast cancer model

PurposeProstaglandin E2 (PGE2), a product of cyclooxygenase (COX) pathway of arachidonic acid, exerts inhibitory impacts on dendritic cell (DC) activity to repress anti-tumor immune responses. Therefore, targeting COX during DC vaccine generation may enhance DC-mediated antitumor responses. We aimed to investigate the impacts of DC vaccine treated with celecoxib (CXB), a selective COX2 inhibitor, on some T cell-related parameters. Materials and methodsBreast cancer (BC) was induced in BALB/c mice, and then they received DC vaccine treated with lipopolysaccharide (LPS-mDCs), LPS with a 5 ​μM dose of CXB (LPS/CXB5-mDCs) and LPS with a 10 ​μM dose of CXB (LPS/CXB10-mDCs). The frequency of splenic Th1 and Treg cells and amounts of IFN-γ, IL-12 and TGF-β production by splenocytes, as well as, the expression of Granzyme-B, T-bet and FOXP3 in tumors were determined using flow cytometry, ELISA, and real-time PCR, respectively. ResultsCompared with untreated tumor group (T-control), treatment with LPS/CXB5-mDCs and LPS/CXB10-mDCs decreased tumor growth (P ​= ​0.009 and P ​< ​0.0001), escalated survival rate (P ​= ​0.002), increased the frequency of splenic Th1 cells (P ​= ​0.0872, and P ​= ​0.0155), increased the IFN-γ (P ​= ​0.0003 and P ​= ​0.0061) and IL-12 (P ​= ​0.001 and P ​= ​0.0009) production by splenocytes, upregulated T-bet (P ​= ​0.062 and P ​< ​0.0001) and Granzyme-B (P ​= ​0.0448 and P ​= ​0.4485), whereas decreased the number of Treg cells (P ​= ​0.0014, and P ​= ​0.0219), reduced the amounts of TGF-β production by splenocytes (P ​= ​0.0535 and P ​= ​0.0169), and reduced the expression of FOXP3 (P ​= ​0.0006 and P ​= ​0.0057) in comparison with T-control group. ConclusionsOur findings show that LPS/CXB-treated DC vaccine potently modulated antitumor immune responses in a mouse BC model.

The 5-lipoxygenase/cyclooxygenase-2 cross-over metabolite, hemiketal E2, enhances VEGFR2 activation and promotes angiogenesis

Consecutive oxygenation of arachidonic acid by 5-lipoxygenase and cyclooxygenase-2 yields the hemiketal eicosanoids, HKE2 and HKD2. Hemiketals stimulate angiogenesis by inducing endothelial cell tubulogenesis in culture; however, how this process is regulated has not been determined. Here, we identify vascular endothelial growth factor receptor 2 (VEGFR2) as a mediator of HKE2-induced angiogenesis invitro and invivo. We found that HKE2 treatment of human umbilical vein endothelial cells dose-dependently increased the phosphorylation of VEGFR2 and the downstream kinases ERK and Akt that mediated endothelial cell tubulogenesis. Invivo, HKE2 induced the growth of blood vessels into polyacetal sponges implanted in mice. HKE2-mediated effects invitro and invivo were blocked by the VEGFR2 inhibitor vatalanib, indicating that the pro-angiogenic effect of HKE2 was mediated by VEGFR2. HKE2 covalently bound and inhibited PTP1B, a protein tyrosine phosphatase that dephosphorylates VEGFR2, thereby providing a possible molecular mechanism for how HKE2 induced pro-angiogenic signaling. In summary, our studies indicate that biosynthetic cross-over of the 5-lipoxygenase and cyclooxygenase-2 pathways gives rise to a potent lipid autacoid that regulates endothelial cell function invitro and invivo. These findings suggest that common drugs targeting the arachidonic acid pathway could prove useful in antiangiogenic therapy.

GLUT10 is a novel immune regulator involved in lung cancer immune cell infiltration and predicts worse survival when transcriptionally downregulated

BackgroundGlucose transporter 10 (GLUT10) is encoded by the SLC2A10 gene. Our recent investigations have shown that GLUT10 is not only involved in glucose metabolism but also involved in the body’s immune response to cancer cells. However, the role of GLUT10 in tumor prognosis and in tumor immunity has not been reported. MethodsWe knocked down SLC2A10 and performed transcriptome sequencing to analyse the biological function of GLUT10 and found that GLUT10 may be involved in immune signaling. Then, we studied the expression level of SLC2A10 in cancers by the Oncomine database and Tumor Immune Estimation Resource (TIMER) site. We also evaluated the prognostic potential of SLC2A10 in different cancers using the Kaplan‒Meier plotter database and PrognoScan online software. The correlations between SLC2A10 expression and immune infiltrates were analysed by TIMER. In addition, correlations between SLC2A10 expression and gene marker sets of immune infiltrates were analysed by TIMER and Gene Expression Profiling Interactive Analysis (GEPIA). Immunofluorescence staining of cyclooxygenase-2 (COX-2) and GLUT10 in lung cancer tissue and adjacent tissue was performed to confirm our findings from the database research. ResultsKnocking down SLC2A10 widely activated immune and inflammatory signaling. SLC2A10 was abnormally expressed in several tumors. The expression level of SLC2A10 was closely correlated with cancer prognosis. Low SLC2A10 expression was related to poorer prognosis and increased malignancy of lung cancer. Lung cancer patients with low expression of SLC2A10 have a much shorter median survival time than patients with high expression of SLC2A10. SLC2A10 expression is closely related to the infiltration of different types of immune cells, particularly macrophages. Both database research and lung cancer sample research revealed that GLUT10 might modulate immune cell infiltration via the COX-2 pathway. ConclusionsBy transcriptome experiments, database studies, and human sample studies, we found that GLUT10 is a new immune signaling molecule involved in tumor immunity, especially in the immune cell infiltration of lung adenocarcinoma (LUAD). GLUT10 may modulate the immune cell infiltration of LUAD via the COX-2 pathway.

Synthesis and Biological Evaluation of Octahydroquinazolinones as Phospholipase A2, and Protease Inhibitors: Experimental and Theoretical Exploration

Phospholipase A2 (PLA2) promotes inflammation via lipid mediators and releases arachidonic acid (AA), and these enzymes have been found to be elevated in a variety of diseases, including rheumatoid arthritis, sepsis, and atherosclerosis. The mobilization of AA by PLA2 and subsequent synthesis of prostaglandins are regarded as critical events in inflammation. Inflammatory processes may be treated with drugs that inhibit PLA2, thereby blocking the COX and LOX pathways in the AA cascade. To address this issue, we report herein an efficient method for the synthesis of a series of octahydroquinazolinone compounds (4a-h) in the presence of the catalyst Pd-HPW/SiO2 and their phospholipase A2, as well as protease inhibitory activities. Among eight compounds, two of them exhibited overwhelming results against PLA2 and protease. By using FT-IR, Raman, NMR, and mass spectroscopy, two novel compounds were thoroughly studied. After carefully examining the SAR of the investigated compounds against these enzymes, it was found that compounds (4a, 4b) containing both electron-donating and electron-withdrawing groups on the phenyl ring exhibited higher activity than compounds with only one of these groups. DFT studies were employed to study the electronic nature and reactivity properties of the molecules by optimizing at the BLYP/cc-pVDZ. Natural bond orbitals helped to study the various electron delocalizations in the molecules, and the frontier molecular orbitals helped with the reactivity and stability parameters. The nature and extent of the expressed biological activity of the molecule were studied using molecular docking with human non-pancreatic secretory phospholipase A2 (hnps-PLA2) (PDB ID: 1DB4) and protease K (PDB ID: 2PWB). The drug-ability of the molecule has been tested using ADMET, and pharmacodynamics data have been extracted. Both the compounds qualify for ADME properties and follow Lipinski's rule of five.

Molecular Targets and Mechanisms of 6,7-Dihydroxy-2,4-dimethoxyphenanthrene from Chinese Yam Modulating NF-κB/COX-2 Signaling Pathway: The Application of Molecular Docking and Gene Silencing

Chinese yam (Dioscorea opposita) tuber has a significant effect of invigorating the intestine and improving the symptoms of long-term diarrhea according to the records of the Chinese Pharmacopoeia. Phenanthrene polyphenols from Chinese yam, with higher inhibition of cyclooxygenase-2 (COX-2) than anti-inflammatory drugs, are an important material basis in alleviating ulcerative colitis via nuclear factor kappa-B (NF-κB)/COX-2 pathway, based on our previous research. The present study further explored the target and molecular mechanisms of phenanthrenes' modulation of the NF-κB/COX-2 signaling pathway by means of molecular docking and gene silencing. Firstly, interleukin-8 (IL-8) and tumor necrosis factor-α (TNF-α) expression of 6-hydroxy-2,4,7-trimethoxyphenanthrene (PC2)/6,7-dihydroxy-2,4-dimethoxyphe-nanthrene (PC4) were compared on TNF-α induced human colon adenocarcinoma (Caco-2) cells. Secondly, molecular docking and dynamics simulation were implemented for PC2/PC4 and COX-2. Finally, COX-2 silencing was performed on TNF-α induced Caco-2 cells to confirm the target of PC4 on NF-κB/COX-2 pathway. Lower expression of IL-8 and TNF-α in PC4 treated Caco-2 cells indicated that PC4 had stronger anti-inflammatory activity than PC2. The binding of PC4 and COX-2 was stronger due to the hydrogen bond between hydroxyl group and Tyr385. No significant differences were found in phosphorylation nuclear factor kappa-B inhibitor alpha (pIkBα), phosphorylation NF-κB (pNF-κB) and phosphorylation extracellular signal-regulated kinase 1/2 (pERK1/2) expression between control and PC4 group after silencing, while these protein expressions significantly decreased in PC4 group without silencing, which confirmed that COX-2 was the important target for PC4 in alleviating ulcerative colitis. These findings indicate that PC4 was supposed to have inhibited NF-κB pathway mediated inflammation via suppression of positive feedback targeting COX-2.

COX-2/PGE2 upregulation contributes to the chromosome 17p-deleted lymphoma

Deletions of chromosome 17p, where TP53 gene locates, are the most frequent chromosome alterations in human cancers and associated with poor outcomes in patients. Our previous work suggested that there were p53–independent mechanisms involved in chromosome 17p deletions-driven cancers. Here, we report that altered arachidonate metabolism, due to the deficiency of mouse Alox8 on chromosome 11B3 (homologous to human ALOX15B on chromosome 17p), contributes to the B cell malignancy. While the metabolites produced from lipoxygenase pathway reduced, chromosome 11B3 deletions or Alox8 loss, lead to upregulating its paralleling cyclooxygenase pathway, indicated by the increased levels of oncometabolite prostaglandin E2. Ectopic PGE2 prevented the apoptosis and differentiation of pre-B cells. Further studies revealed that Alox8 deficiency dramatically and specifically induced Cox-2(Ptgs2) gene expression. Repressing Cox-2 by its shRNAs impaired the tumorigenesis driven by Alox8 loss. And, in turn, tumor cells with Alox8 or 11B3 loss were sensitive to the COX-2 inhibitor celecoxib. This correlation between COX-2 upregulation and chromosome 17p deletions was consistent in human B-cell lymphomas. Hence, our studies reveal that the arachidonate metabolism abnormality with unbalanced ALOX and COX pathways underlies human cancers with 17p deletions and suggest new susceptibility for this disease.

Circ_DLEU2 knockdown represses cell proliferation, migration and invasion, and induces cell apoptosis through the miR-582-5p/COX2 pathway in acute myeloid leukemia.

Acute myeloid leukemia (AML) is a malignant hematological neoplasm in adults. Researche indicates that circular RNAs (circRNAs) play paramount roles in the pathological process of AML. In this study, the role of circ_DLEU2 (circ_0000488) in AML is revealed. The expression of circ_DLEU2, microRNA-582-5p (miR-582-5p) and cyclooxygenase 2 (COX2) was determined by quantitative real-time PCR. Protein expression was detected by western blot. Cell proliferation was investigated by cell cycle, 5-Ethynyl-29-deoxyuridine and 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide (MTT) assays. Cell apoptosis was elucidated by apoptosis analysis assay. The targeting relationship between miR-582-5p and circ_DLEU2 or COX2 was predicted by the starbase online database, and identified by a dual-luciferase reporter assay. Circ_DLEU2 and COX2 expression were substantially up-regulated, while miR-582-5p was down-regulated in AML marrow samples and cells compared with control groups. Circ_DLEU2 knockdown suppressed cell proliferation, whereas it induced cell arrest at G0/G1 phase and cell apoptosis in AML; however, these effects were attenuated by miR-582-5p inhibitor. Additionally, circ_DLEU2 was associated with miR-582-5p, and miR-582-5p bound to COX2 in AML cells. Also, we found that circ_DLEU2 regulated COX2 expression by interacting with miR-582-5p. Circ_DLEU2 silencing hindered AML malignant progression via downregulating COX2 through sponging miR-582-5p. Our finding provides a theoretical basis for studying circRNA-directed therapy of AML.

Controlling Inflammation Improves Aging Skeletal Muscle Health.

Chronic inflammation is associated with a decline in aging skeletal muscle health. Inflammation also seems to interfere with the beneficial skeletal muscle adaptations conferred by exercise training in older individuals. We hypothesize that the cyclooxygenase pathway is partially responsible for this negative inflammatory influence on aging skeletal muscle health and plasticity.

Herbal Medicines - a Fruitful Approach to Periodic Illness Dysmenorrhoea: Evidence-based Review.

Dysmenorrhea (menstrual or periodic pain) is a discomfort that occurs during painful periods. It is the first and most prominent reason for female lower abdominal pain. Most adolescent girls consider it a curse due to the periodic occurrence of painful cramps and bleeding. The pathogenesis of painful periods is most likely because of increased prostanoids, notably prostaglandins, produced by the cyclooxygenase pathway (PGs). Misuse of synthetic medications leads to the development of medication resistance and deposits toxic residues in the body; thus, there is a critical need for safe and effective alternatives. In recent decades, herbal treatment approaches have found extensive applications in the treatment of various ailments. Herbal therapies are an alternate source, which include several bioactive chemicals, and recent improvements in our understanding of the value of herbal therapy methods have caused a sharp rise in their production. The main focus of this review was to study herbal treatment options; the recent studies conducted on herbal therapies and various experimental investigations on dysmenorrhea and herbal therapy methods have been studied, and randomized controlled trials and animal models have been discussed describing the anti-inflammatory properties of some potential herbal medicines that can be used as treatment options for dysmenorrhoea. This review aimed to present herbal treatments that can be used as alternative traditional synthetic medications and oral hormonal contraceptives in the treatment of painful menstruation.

Atorvastatin attenuates allergic inflammation by blocking prostaglandin biosynthesis in rats with allergic rhinitis.

Prostaglandins (PGs) are bioactive lipid mediators derived from the nuclear and plasma membranes via the cyclooxygenase (COX) pathway of arachidonic acid (AA) metabolism. PGs bridge the interactions between various immunomodulatory cells in allergic rhinitis (AR) and are considered key players in regulating pro-inflammatory and anti-inflammatory responses. AA conversion to PGs involves rate-limiting enzymes that may be blocked by statins. The mechanisms by which statins regulate these enzymes in AR remain unclear. We investigated the effects of oral atorvastatin on PGs production in AR. An ovalbumin-induced AR rat model was constructed and the changes in nasal symptom score and nasal mucosa histopathological characteristics of AR rats under different atorvastatin doses were assessed. qRT-PCR, western blotting, and immunofluorescence were used to detect the mRNA and protein expression levels of rate-limiting enzymes and downstream molecules of AA metabolism in the nasal mucosa and liver. Oral atorvastatin significantly alleviated symptoms and eosinophil infiltration in the nasal mucosa, inhibited goblet cell hyperplasia and mast cell recruitment, and decreased mucus secretion in AR rats. Increasing atorvastatin dose increased the anti-inflammatory effects. High-dose atorvastatin inhibited upregulation of the inflammatory mediator PGD2 in the nasal mucosa of AR rats. Compared to the control group, the mRNA and protein expression of the rate-limiting enzymes COX-2, PGDS, and PGES in AA metabolism in the AR group were upregulated but downregulated after the oral administration of high-dose atorvastatin. Atorvastatin also showed dose-dependent inhibition of ERK1/2 and downstream NF-κB phosphorylation in the nasal mucosa and liver of AR rats. Atorvastatin inhibited allergic inflammation and attenuated AR nasal symptoms by downregulating PGD2 and rate-limiting enzyme expression in PGD2 biosynthesis, possibly by blocking the RAS/ERK/NF-κB signaling pathway.