A Cross‐Linked anti‐TNF ‐α aptamer for neutralization of TNF ‐α‐induced cutaneous Shwartzman phenomenon: A simple and novel approach for improving aptamers' affinity and efficiency

Background: Endothelial dysfunction is an independent predictor of poor prognosis in patients with heart failure (HF). Using a novel approach of endovascular endothelial cell (EC) sampling, we previously showed that ECs collected from conduit veins using J-wires are activated in patients with HF as evidenced by increased expression of pro-inflammatory genes including inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). The underlying cause(s) of this endothelial activation and dysfunction is not known. In this study, we tested the hypothesis that the HF state itself and the vascular strain associated with congestion can both account for these findings. Methods: We studied 6 normal (NL) dogs (LV ejection fraction >50%, central venous pressure (CVP) ~ 8 mmHg) and 6 dogs with HF (ejection fraction ~ 30%, CVP ~9 mmHg) produced by intracoronary microembolizations. This model of HF manifests the disease syndrome in the absence of other co-morbid conditions often present in patients with CHF. NL dogs were studied at baseline and 1 hr after rapid fluid load (500 ml Dextran-40) resulting in CVP>20 mmHg. ECs were scraped from jugular veins. mRNA was analyzed by RT-PCR and quantified in densitometric units (du). Results: Data are shown in the table . Endothelial iNOS and COX-2 were increased in HF dogs compared to NL. Tissue necrosis factor (TNF)-α, early growth response gene (EGR-1), receptor for advanced glycation end products (RAGE) and glutathione peroxidase (GPx) were also up-regulated in HF. In NL dogs, fluid load increased iNOS, COX-2, TNF-α, Egr-1, RAGE and GPx to levels that approached those of HF. Conclusions: The HF state itself, in the absence of other co-morbid conditions, and vascular stretch are both sufficient to activate venous ECs in dogs in a manner consistent with that seen in patients with HF. As such, molecular measures of EC dysfunction may potentially serve as “biomarkers” of severity of HF and possibly of impending decompensation.

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Gut-derived norepinephrine plays a critical role in producing hepatocellular dysfunction during early sepsis

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.2000.279.6.g1274 ◽

2000 ◽

Vol 279 (6) ◽

pp. G1274-G1281 ◽

Cited By ~ 46

Author(s):

Shaolong Yang ◽

Douglas J. Koo ◽

Mian Zhou ◽

Irshad H. Chaudry ◽

Ping Wang

Keyword(s):

Critical Role ◽

Cecal Ligation ◽

Plasma Norepinephrine ◽

Novel Approach ◽

Tnf Α ◽

Liver Preparation ◽

Icg Clearance ◽

Early Sepsis ◽

20 Nm

Although plasma norepinephrine (NE) increases and hepatocellular function is depressed during early sepsis, it is unknown whether gut is a significant source of NE and, if so, whether gut-derived NE helps produce hepatocellular dysfunction. We subjected rats to sepsis by cecal ligation and puncture (CLP), and 2 h later (i.e., early sepsis) portal and systemic blood samples were collected and plasma levels of NE were assayed. Other rats were enterectomized before CLP. Hepatocellular function was assessed with an in vivo indocyanine green (ICG) clearance technique, systemic levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6 were determined, and the effect of NE on hepatic ICG clearance capacity was assessed in an isolated, perfused liver preparation. Portal levels of NE were significantly higher than systemic levels at 2 h after CLP. Prior enterectomy reduced NE levels in septic animals. Thus gut appears to be the major source of NE release during sepsis. Enterectomy before sepsis also attenuated hepatocellular dysfunction and downregulated TNF-α, IL-1β, and IL-6. Perfusion of the isolated livers with 20 nM NE (similar to that observed in sepsis) significantly reduced ICG clearance capacity. These results suggest that gut-derived NE plays a significant role in hepatocellular dysfunction and upregulating inflammatory cytokines. Modulation of NE release and/or hepatic responsiveness to NE should provide a novel approach for maintaining hepatocellular function in sepsis.

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Regulation of dendritic cell function improves survival in experimental sepsis through immune chaperone

Innate Immunity ◽

10.1177/1753425919840423 ◽

2019 ◽

Vol 25 (4) ◽

pp. 235-243 ◽

Cited By ~ 2

Author(s):

Pengfei Li ◽

Ran Zhao ◽

Kevin Fan ◽

Stephen Iwanowycz ◽

Hongkuan Fan ◽

...

Keyword(s):

Cell Function ◽

Critical Role ◽

Cecal Ligation ◽

Experimental Sepsis ◽

Adaptive Immune ◽

Sepsis Model ◽

Novel Approach ◽

Tnf Α ◽

Heat Shock Protein Gp96 ◽

Dc Maturation

Dendritic cells (DCs) are professional Ag-presenting cells that play a critical role in both innate and adaptive immune responses. DCs recognize and respond to bacteria through multiple PRRs, including TLRs. Heat shock protein gp96/grp94 is a master essential chaperone for TLRs in the endoplasmic reticulum. We generated DC-specific gp96-knockout (KO) mice and showed that gp96 KO DCs were unable to respond to multiple TLR ligands. TLR-mediated hyperinflammatory response can lead to sepsis. However, the roles of neither DCs nor the DC-intrinsic gp96 in the process are completely understood. In a LPS-induced sepsis model, we hereby found that deletion of gp96 in DCs significantly reduced serum TNF-α levels and improved survival. Furthermore, using the well-defined polymicrobial sepsis model of cecal ligation and puncture, we found that DC-specific ablation of gp96 improved survival with significantly attenuated liver and renal injuries, decreased circulating inflammatory cytokines, altered DC maturation and activation, and increased serum Ig. Collectively, we demonstrate that deletion of gp96 in DCs is beneficial in protecting mice against sepsis induced by both endotoxemia and polymicrobial infections. We conclude that targeting gp96 in DCs may provide a potential novel approach for reducing the morbidity and mortality of sepsis.

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Disruption of Nox2 and TNFRp55/p75 eliminates cardioprotection induced by anisomycin

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.00306.2012 ◽

2012 ◽

Vol 303 (10) ◽

pp. H1263-H1272 ◽

Cited By ~ 4

Author(s):

Ting C. Zhao ◽

Ling Zhang ◽

Jun T. Liu ◽

Tai L. Guo

Keyword(s):

Infarct Size ◽

Myocardial Injury ◽

Left Ventricular ◽

Kinase Assay ◽

Protective Effects ◽

Perfused Heart ◽

Transient Activation ◽

Novel Approach ◽

Tnf Α ◽

A Subunit

Transient activation of p38 through anisomycin is demonstrated to precondition the heart against myocardial injury. However, it remains unknown whether specific TNF-α receptor (TNFR) p55/p75 and Nox2, a subunit of NADPH oxidase, are involved in this event. We sought to investigate whether the genetic disruption of TNFRp55/p75 and Nox2 eliminated cardioprotection elicited by anisomycin and whether p38-dependent activation of Nox2 stimulated TNFR to ultimately achieve protective effects. Adult wild-type and TNFR p55/p75−/−and Nox2−/−mice received intraperitoneal injections of anisomycin (0.1 mg/kg), a potent activator of p38. The hearts were subjected to 30 min myocardial ischemia/30 min reperfusion in the Langendorff perfused heart after 24 h. Left ventricular function was measured, and infarct size was determined. Myocardial TNF-α protein, Nox2, and superoxides releases were detected. Gel kinase assay was employed to detect the effect of p38 on Nox2 phosphorylation. Activation of p38 through anisomycin produces marked improvements in left ventricular functional recovery, and the reduction of myocardial infarction, which were abrogated by disruption of Nox2 and TNFR p55/p75. Disruption of Nox2 and TNFR p55/p75 abolished the effect of anisomycin-induced reduction of infarct size. Anisomycin induced the production of TNF-α, which was abrogated in Nox2−/−mice and by treatment with SB203580, but not by disruption of p55/p75. Anisomycin treatment resulted in an increase in Nox2 protein and the phosphorylation of Nox2, which was blocked by inhibition of p38. Taken together, these results indicate that stimulation of the Nox2 and TNFR p55/p75 pathway is a novel approach to anisomycin-induced cardioprotection.

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CircRNA_103765 acts as a proinflammatory factor via sponging miR-30 family in Crohn’s disease

Scientific Reports ◽

10.1038/s41598-020-80663-w ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Yulan Ye ◽

Liping Zhang ◽

Tong Hu ◽

Juan Yin ◽

Lijuan Xu ◽

...

Keyword(s):

Crohn’S Disease ◽

Crohn's Disease ◽

Cell Apoptosis ◽

Mononuclear Cells ◽

Circular Rnas ◽

Infliximab Treatment ◽

Peripheral Blood Mononuclear ◽

Novel Approach ◽

Tnf Α

AbstractIncreasing evidence suggests that circular RNAs (circRNAs) play critical roles in various pathophysiological activities. However, the role of circRNAs in inflammatory bowel disease (IBD) remains unclear. Here we report the potential roles of hsa_circRNA_103765 in regulating cell apoptosis induced by TNF-α in Crohn’s disease (CD). We identify that CircRNA_103765 expression was significantly upregulated in peripheral blood mononuclear cells (PBMCs) of patients with active IBD. A positive correlation with TNF-α significantly enhanced circRNA_103765 expression in CD, which was significantly reversed by anti-TNF-α mAb (infliximab) treatment. In vitro experiments showed that TNF-α could induce the expression of circRNA_103765, which was cell apoptosis dependent, while silencing of circRNA_103765 could protect human intestinal epithelial cells (IECs) from TNF-α-induced apoptosis. In addition, circRNA_103765 acted as a molecular sponge to adsorb the miR-30 family and impair the negative regulation of Delta-like ligand 4 (DLL4). Collectively, CircRNA_103765 is a novel important regulator of the pathogenesis of IBD via sponging miR-30 family-mediated DLL4 expression changes. Blockade of circRNA_103765 could serve as a novel approach for the treatment of IBD patients.

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Targeting Ameloblatoma into Apoptosis

The Indonesian Biomedical Journal ◽

10.18585/inabj.v10i1.354 ◽

2018 ◽

Vol 10 (1) ◽

pp. 35

Author(s):

Ferry Sandra

Keyword(s):

Tumor Cells ◽

Fas Ligand ◽

Squamous Metaplasia ◽

Death Receptor ◽

Novel Approach ◽

Tnf Α ◽

Mitogen Activated Protein ◽

Cellular Aspect ◽

Peripheral Cells ◽

Slow Growing

BACKGROUND: Generally ameloblastoma is a locally aggressive, slow growing, non-metastatic epithelial odontogenic benign tumor. However, rarely some ameloblastoma can metastasize in spite of a benign histologic appearance. Targeting ameloblastoma by inducing it into apoptosis could be a beneficial strategy, since many ameloblastoma cases were reported recurrent after surgical therapy.CONTENT: To investigate ameloblastoma in cellular aspect,cytological pattern of ameloblastoma was divided intoouter layer/peripheral and inner layer/central cells. Tumor necrosis factor (TNF)-α, Fas ligand (FasL), TNF receptor (TNFR)1/death receptor (DR)1, TNFR2/DR2, DR4, DR5andFas were highly expressed in central than peripheral cells. Despite inducing apoptosis, TNF-α can induce PI3K leading to Akt and p44/42 mitogen-activated protein kinases (MAPK) activation in AM-1 cells, which later induce cell survival and proliferation. Therefore apoptotic induction in ameloblastoma should be suggested in higher TNF-α concentration. Expression of FasL and Fas are closely associated with squamous metaplasia and granular transformation of the tumor cells, suggesting that apoptosis induced by FasL may play a role in the terminally differentiated or degenerative ameloblastoma cells. TNF-related apoptosis-inducing ligand (TRAIL) has emerged as an apoptotic inducing anticancer agent in tumor cells specifically. TRAIL induced activation of caspases, lowering mitochondrial membrane potential, high number of apoptotic cells in ameloblastoma cells. Therefore, TRAIL could be a potential agent for targeting ameloblastoma, although further study should be explored.SUMMARY: Targeting ameloblastoma by inducing it into apoptosis could be achieved effectively, although some criteria should be considered. Therefore understanding the underlying apoptosis signaling pathways are necessary for inducing ameloblasotma into apoptosis. Investigations on other apoptosis-related molecules, potential apoptosis-inducing natural products, and novel approach in reprogramming, are important in the future for a better anagement of ameloblastoma.KEYWORDS: ameloblastoma, apoptosis, TNF, Fas, TRAIL, Akt, MAPK, caspase

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Abstract 146: Disruption of Nox2 and TNFRp55/p75 Eliminates Anisomycin-Induced Preconditioning Effect in the Heart

Circulation Research ◽

10.1161/res.111.suppl_1.a146 ◽

2012 ◽

Vol 111 (suppl_1) ◽

Author(s):

Ting C Zhao ◽

Ling Zhang ◽

Tai-Liang Guo

Keyword(s):

Infarct Size ◽

Diastolic Pressure ◽

Left Ventricular ◽

Kinase Assay ◽

Protective Effects ◽

Triphenyltetrazolium Chloride ◽

Transient Activation ◽

Novel Approach ◽

Tnf Α ◽

A Subunit

Background: Transient activation of p38 through anisomycin is demonstrated to precondition the heart against myocardial injury. However, it remains unknown whether specific TNF-α receptor (TNFR) p55/p75 and Nox2, a subunit of NADPH-oxidase are involved in this event. Objective: We sought to investigate whether the genetic disruption of TNFRp55/p75 and Nox2 eliminates cardioprotection elicited by anisomycin and whether p38-dependent activation of Nox2 stimulates TNFR to ultimately achieve protective effects. Methods: Adult wild type and p55/p75 -/- and Nox2 -/- mice received intraperitoneal injections of anisomycin (0.1mg/kg), a potent activator of p38. The hearts were subjected to 30 min myocardial ischemia /30 min reperfusion in the Langendorff perused heart after twenty four hours. Left ventricular function was measured and infarct size was determined by triphenyltetrazolium chloride. Myocardial TNF-α protein, Nox2 and superoxides releases were detected. Gel kinase assay was employed to detect the effect of p38 on Nox2 phosphorylation. Results: Activation of p38 through anisomycin produces marked improvements in the recovery of left ventricular end diastolic pressure, rate pressure products, and the reduction of myocardial infarction, which were completely abrogated by disruption of Nox2 and TNFR p55/p75. Genetic disruption of Nox2 and TNFR p55/p75 abolished the effect of anisomycin-induced reduction of infarct size. Ansiomycin induced the production of TNFα, which was abrogated in Nox2 -/- mice. Notably, activation of p38 resulted in the phosphorylation of Nox2. Conclusion: Taken together, these results indicate that stimulation of the Nox2 and TNFR p55/p75 pathway is a novel approach to anisomycin-induced cardioprotection.

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Mebendazole augments sensitivity to sorafenib by targeting MAPK and BCL-2 signalling in n-nitrosodiethylamine-induced murine hepatocellular carcinoma

Scientific Reports ◽

10.1038/s41598-019-55666-x ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 9

Author(s):

Nancy S. Younis ◽

Amal M. H. Ghanim ◽

Sameh Saber

Keyword(s):

Hepatocellular Carcinoma ◽

Drug Resistance ◽

Kinase Inhibitor ◽

Mapk Pathway ◽

Hepatic Inflammation ◽

Novel Approach ◽

Tnf Α ◽

Mapk Signalling ◽

Cancer Types ◽

Tgf Β1

AbstractSorafenib (SO) is a multi-kinase inhibitor that targets upstream signals in the MAPK pathway. Drug resistance and transient survival benefits are the main obstacles associated with SO treatment in Hepatocellular carcinoma (HCC) patients. Mebendazole (MBZ), an anthelmintic agent, has demonstrated activity against various cancer types. Therefore, we aimed to investigate the possible mechanisms of MBZ other than its anti-tubulin activity. MBZ (100 mg/kg/day, P.O.) was administered to N-nitrosodiethylamine-induced HCC mice as a monotherapeutic agent or in combination with SO. Our results revealed that MBZ decreased AFP levels, improved liver function and histology and increased survival in HCC mice, particularly when administered in combination with SO. MBZ also reduced hepatic inflammation and fibrogenesis as evidenced by reductions in TNF-α and TGF-β1 levels, respectively. Increased hepatic caspases-3 and -9 and decreased BCL-2 levels suggest induced-cell death. In addition, MBZ demonstrated anti-angiogenic, anti-metastatic, and anti-proliferative effects, as indicated by reduced VEGF levels, MMP-2:TIMP-1 ratios, and reduced cyclin D1 levels and Ki67 immunostaining, respectively. Our main finding was that MBZ targeted downstream signal of the MAPK pathway by inhibiting ERK1/2 phosphorylation. Targeting downstream MAPK signalling by MBZ and upstream signalling by SO is a novel approach to minimizing resistance and prolonging survival.

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AKT-AMPKα-mTOR-dependent HIF-1α Activation is a New Therapeutic Target for Cancer Treatment: A Novel Approach to Repositioning the Antidiabetic Drug Sitagliptin for the Management of Hepatocellular Carcinoma

Frontiers in Pharmacology ◽

10.3389/fphar.2021.720173 ◽

2022 ◽

Vol 12 ◽

Author(s):

Eslam E. Abd El-Fattah ◽

Sameh Saber ◽

Mahmoud E. Youssef ◽

Hanan Eissa ◽

Eman El-Ahwany ◽

...

Keyword(s):

Gene Expression ◽

Hepatocellular Carcinoma ◽

Ki 67 ◽

Tissue Invasion ◽

Novel Approach ◽

Promising Target ◽

Tnf Α ◽

Key Factor ◽

Potential Basis ◽

Α Level

HIF-1α is a key factor promoting the development of hepatocellular carcinoma (HCC). As well, AKT-AMPKα-mTOR signaling is a promising target for cancer therapy. Yet, the AKT-AMPKα-mTOR-dependent activation of HIF-1α has not been studied in livers with HCC. In addition, the mechanisms underlying the potential antineoplastic effects of sitagliptin (STGPT), an antidiabetic agent, have not yet been elucidated. For that purpose, the N-nitrosodiethylamine (NDEA)-induced HCC mouse model was used in the present study using a dose of 100 mg/kg/week, i.p., for 8 weeks. NDEA-induced HCC mice received STGPT 20, 40, or 80 mg/kg starting on day 61 up to day 120. The present study revealed that STGPT inhibited HIF-1α activation via the interference with the AKT-AMPKα-mTOR axis and the interruption of IKKβ, P38α, and ERK1/2 signals as well. Accordingly, STGPT prolonged the survival, restored the histological features and improved liver function. Additionally, STGPT inhibited angiogenesis, as revealed by a significant downregulation in the VEGF and mRNA expression of CD309 with concomitant inhibition of tissue invasion was evident by an increased ratio of TIMP-1/MMP-2. STGPT exhibited apoptotic stimulatory effect as indicated upon calculating the BCL-2/Bax ratio and by the gene expression of p53. The decrease in AFP and liver index calculation, gene expression of Ki-67 confirmed the antiproliferative activity of STGPT. The anti-inflammatory potential was revealed by the decreased TNF-α level and the downregulation of MCP-1 gene expression. Moreover, an antifibrotic potential was supported by lower levels of TGF-β. These effects appear to be GLP1R-independent. The present study provides a potential basis for repurposing STGPT for the inhibition of HCC progression. Since STGPT is unlikely to cause hypoglycemia, it may be promising as monotherapy or adjuvant therapy to treat diabetic or even normoglycemic patients with HCC.

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