Antioxidant protection by PECAM-targeted delivery of a novel NADPH-oxidase inhibitor to the endothelium in vitro and in vivo

Abstract Background: Ovulation is triggered by the preovulatory surge of the pituitary luteinizing hormone (LH). LH/hCG induction of reactive oxygen species (ROS) is required for successful ovulation. H2O2, one of ROS species, was shown to fully mimic the effect of LH/hCG in mice ovulation. However, the molecular process that generates H2O2 in the ovary during ovulation remains largely unknown. DUOX2, a member of the NOX/DUOX family of NADPH oxidase, is capable of generating H2O2. Results: Using global transcriptome RNAseq, we identified that DUOX2 is one of the transcripts that was markedly upregulated in granulosa cells during ovulation. Treatment with human chorionic gonadotropin (hCG), an ovulatory trigger, significantly increases the expression of DUOX2 mRNA and protein in human GCs both in vivo and in vitro. hCG-induced up-regulation of DUOX2 is mediated by the cAMP-PKA and the PKC pathway. A functional test reveals that DUOX2 chemical inhibitor, Diphenyleneiodonium (DPI), an NADPH oxidase inhibitor, decreased H2O2 levels in MGCs (Mural Granulosa Cells) treated with hCG. The inhibition of H2O2 by DPI suggests that DUOX2 activity is required for hCG-induced elevation of extracellular H2O2 in MGCs. In vivo treatment of mice with DPI significantly decreases the number of ovulated oocytes and markedly attenuates the expression of key ovulatory genes. These results support the putative role of DUOX2 in ovulation. Conclusions: DUOX2 is a ROS generator during the ovulatory process and is involved in the LH/hCG-induced signaling cascades leading to ovulation. Treatment with DUOX2 inhibitors may affect late folliculogenesis and ovulation and thus may serve for fertility control.

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Dextromethorphan Reduces Oxidative Stress and Inhibits Uremic Artery Calcification

International Journal of Molecular Sciences ◽

10.3390/ijms222212277 ◽

2021 ◽

Vol 22 (22) ◽

pp. 12277

Author(s):

En-Shao Liu ◽

Nai-Ching Chen ◽

Tzu-Ming Jao ◽

Chien-Liang Chen

Keyword(s):

Oxidative Stress ◽

Nadph Oxidase ◽

Vascular Calcification ◽

Wistar Rats ◽

Nicotinamide Adenine Dinucleotide Phosphate ◽

Oxidase Inhibitor ◽

In Vivo Studies ◽

Ros Production

Medial vascular calcification has emerged as a key factor contributing to cardiovascular mortality in patients with chronic kidney disease (CKD). Vascular smooth muscle cells (VSMCs) with osteogenic transdifferentiation play a role in vascular calcification. Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitors reduce reactive oxygen species (ROS) production and calcified-medium–induced calcification of VSMCs. This study investigates the effects of dextromethorphan (DXM), an NADPH oxidase inhibitor, on vascular calcification. We used in vitro and in vivo studies to evaluate the effect of DXM on artery changes in the presence of hyperphosphatemia. The anti-vascular calcification effect of DXM was tested in adenine-fed Wistar rats. High-phosphate medium induced ROS production and calcification of VSMCs. DXM significantly attenuated the increase in ROS production, the decrease in ATP, and mitochondria membrane potential during the calcified-medium–induced VSMC calcification process (p < 0.05). The protective effect of DXM in calcified-medium–induced VSMC calcification was not further increased by NADPH oxidase inhibitors, indicating that NADPH oxidase mediates the effect of DXM. Furthermore, DXM decreased aortic calcification in Wistar rats with CKD. Our results suggest that treatment with DXM can attenuate vascular oxidative stress and ameliorate vascular calcification.

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Pharmacokinetic Study of NADPH Oxidase Inhibitor Ewha-18278, a Pyrazole Derivative

Pharmaceutics ◽

10.3390/pharmaceutics11090482 ◽

2019 ◽

Vol 11 (9) ◽

pp. 482

Author(s):

Seul Gee Lee ◽

Jaeok Lee ◽

Kyung Min Kim ◽

Kee-In Lee ◽

Yun Soo Bae ◽

...

Keyword(s):

Statistical Significance ◽

Oxidase Inhibitor ◽

High Concentration ◽

Time Curve ◽

Suitable Candidate ◽

Nadph Oxidase Inhibitor ◽

Concentration Time ◽

Zero Time

In a previous study, the specific NOX1/2/4 inhibitor Ewha-18278 was confirmed as a possible treatment for osteoporosis both in vitro and in vivo. Here, we investigated the pharmacokinetics (PK) of the compound by intravenous (IV) and oral administrations to rats. Dimethyl sulfoxide (DMSO)-based and diazepam injection-based formulations were used to dissolve the compound. In the latter formulation applicable to humans, the changes in PK parameters were monitored at two different concentrations (1 mg/mL and 2 mg/mL). The area under the plasma concentration-time curve from zero time to infinity (AUCinf) of Ewha-18278 was highest in the DMSO-based formulation (2 mg/mL). Also, the concentration was increased 1.6-fold at the low concentration of the diazepam injection-based formulation compared to the high concentration. There was no statistical significance in the AUCinf of the compound between DMSO-based formulation (2 mg/mL) and diazepam injection-based formulation (1 mg/mL). These results suggest that Ewha-18278 can be delivered to humans by both IV and oral routes. In addition, the diazepam injection-based formulation of Ewha-18278 appears to be a suitable candidate for dosage development for future toxicity test and clinical trial.

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Adventitial application of the NADPH oxidase inhibitor apocynin in vivo reduces neointima formation and endothelial dysfunction in rabbits

Cardiovascular Research ◽

10.1016/j.cardiores.2007.06.005 ◽

2007 ◽

Vol 75 (4) ◽

pp. 710-718 ◽

Cited By ~ 29

Author(s):

E CHAN ◽

S DATLA ◽

R DILLEY ◽

H HICKEY ◽

G DRUMMOND ◽

...

Keyword(s):

Endothelial Dysfunction ◽

Nadph Oxidase ◽

Oxidase Inhibitor ◽

Neointima Formation ◽

Nadph Oxidase Inhibitor

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Perivascular gene transfer of NADPH oxidase inhibitor suppresses angioplasty-induced neointimal proliferation of rat carotid artery

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.00413.2004 ◽

2005 ◽

Vol 288 (2) ◽

pp. H946-H953 ◽

Cited By ~ 60

Author(s):

Hector M. Dourron ◽

Gary M. Jacobson ◽

James L. Park ◽

Jianhua Liu ◽

Daniel J. Reddy ◽

...

Keyword(s):

Carotid Artery ◽

Nadph Oxidase ◽

Targeted Delivery ◽

Neointimal Hyperplasia ◽

Fold Increase ◽

Oxidase Inhibitor ◽

Neointima Formation ◽

Nadph Oxidase Inhibitor ◽

Neointimal Proliferation ◽

Localized Delivery

Vascular stretch induces NADPH oxidase-derived superoxide anion (O2−), which has been implicated in hypertrophy and cell proliferation. We hypothesized that targeted delivery of an NADPH oxidase inhibitor to the adventitia would reduce stretch-induced vascular O2− and attenuate neointima formation. We designed a novel replication-deficient adenovirus containing a fibroblast-active promoter driving expression of NADPH oxidase inhibitory sequence gp91ds (Ad-PDGFβR-gp91ds/eGFP). 1) We characterized the specificity of this promoter using pPDGFβR-luciferase by showing induction of luciferase in cultured rat aortic fibroblasts but not in vascular smooth muscle cells. 2) Using RT-PCR, we observed expression of gp91ds and the reporter gene in fibroblasts after infection with Ad-PDGFβR-gp91ds/eGFP. 3) Using Ad-CMV-eGFP as a control, we delivered Ad-PDGFβR-gp91ds/eGFP to the adventitia of the rat common carotid artery (CCA). Immunohistochemistry confirmed localized delivery of the inhibitor to the adventitia. After CCAs were injured with an embolectomy catheter, we observed a significant increase in neointima-to-media area ratio in control CCAs, which was significantly attenuated in CCAs treated with the gp91ds-expressing virus. In a second group of rats, we detected a 10-fold increase in distension-stimulated O2−, which was significantly reduced in CCAs infected with gp91ds-expressing virus. These data demonstrate that localized adventitial delivery of an NADPH oxidase inhibitor is effective in reducing overall vascular O2− and neointima formation, suggesting that adventitial NADPH oxidase plays a functional role in development of neointimal hyperplasia.

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Internal and External Triggering Mechanism of “Smart” Nanoparticle-Based DDSs in Targeted Tumor Therapy

Current Pharmaceutical Design ◽

10.2174/1381612824666180510094607 ◽

2018 ◽

Vol 24 (15) ◽

pp. 1639-1651 ◽

Cited By ~ 2

Author(s):

Xian-ling Qian ◽

Jun Li ◽

Ran Wei ◽

Hui Lin ◽

Li-xia Xiong

Keyword(s):

Targeted Delivery ◽

Tumor Therapy ◽

Burst Release ◽

Tumor Site ◽

Chemotherapeutic Drugs ◽

Triggering Mechanism ◽

Triggering Factors ◽

Or Genes

Background: Anticancer chemotherapeutics have a lot of problems via conventional Drug Delivery Systems (DDSs), including non-specificity, burst release, severe side-effects, and damage to normal cells. Owing to its potential to circumventing these problems, nanotechnology has gained increasing attention in targeted tumor therapy. Chemotherapeutic drugs or genes encapsulated in nanoparticles could be used to target therapies to the tumor site in three ways: “passive”, “active”, and “smart” targeting. Objective: To summarize the mechanisms of various internal and external “smart” stimulating factors on the basis of findings from in vivo and in vitro studies. Method: A thorough search of PubMed was conducted in order to identify the majority of trials, studies and novel articles related to the subject. Results: Activated by internal triggering factors (pH, redox, enzyme, hypoxia, etc.) or external triggering factors (temperature, light of different wavelengths, ultrasound, magnetic fields, etc.), “smart” DDSs exhibit targeted delivery to the tumor site, and controlled release of chemotherapeutic drugs or genes. Conclusion: In this review article, we summarize and classify the internal and external triggering mechanism of “smart” nanoparticle-based DDSs in targeted tumor therapy, and the most recent research advances are illustrated for better understanding.

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Preparation and Optimization of Moxifloxacin Microspheres for Colon Targeted Delivery Using Quality by Design Approach: In Vitro and In Vivo Study

Current Drug Delivery ◽

10.2174/1567201813666160512145625 ◽

2016 ◽

Vol 13 (7) ◽

pp. 1021-1033 ◽

Cited By ~ 4

Author(s):

Bhumika Prajapati ◽

Prasant Kumar Jena ◽

Tejal Mehta ◽

Sriram Seshadri

Keyword(s):

Targeted Delivery ◽

Quality By Design ◽

Design Approach ◽

In Vivo Study ◽

Quality By Design Approach

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Hypoxia promotes the metastasis of pancreatic cancer through regulating NOX4/KDM5A-mediated histone methylation modification changes in a HIF1A-independent manner

Clinical Epigenetics ◽

10.1186/s13148-021-01016-6 ◽

2021 ◽

Vol 13 (1) ◽

Author(s):

Hongzhen Li ◽

Chunyan Peng ◽

Chenhui Zhu ◽

Shuang Nie ◽

Xuetian Qian ◽

...

Keyword(s):

Pancreatic Cancer ◽

Nadph Oxidase ◽

Western Blot ◽

Cancer Progression ◽

Histone Methylation ◽

Invasion And Metastasis ◽

Independent Manner ◽

Nadph Oxidase 4

Abstract Background Hypoxia is a characteristic of the tumor microenvironments within pancreatic cancer (PC), which has been linked to its malignancy. Recently, hypoxia has been reported to regulate the activity of important carcinogenic pathways by changing the status of histone modification. NOX4, a member of NADPH oxidase (NOX), has been found to be activated by hypoxia and promote cancer progression in several cancers. But whether it is involved in the epigenetic changes of tumor cells induced by hypoxia is still unclear, and its biological roles in PC also need to be explored. Methods A hypoxic-related gene signature and its associated pathways in PC were identified by analyzing the pancreatic cancer gene expression data from GEO and TCGA database. Candidate downstream gene (NOX4), responding to hypoxia, was validated by RT-PCR and western blot. Then, we evaluated the relationship between NOX4 expression and clinicopathologic parameters in 56 PC patients from our center. In vitro and in vivo assays were preformed to explore the phenotype of NOX4 in PC. Immunofluorescence, western blot and chromatin immunoprecipitation assays were further applied to search for a detailed mechanism. Results We quantified hypoxia and developed a hypoxia signature, which was associated with worse prognosis and elevated malignant potential in PC. Furthermore, we found that NADPH oxidase 4 (NOX4), which was induced by hypoxia and upregulated in PC in a HIF1A-independent manner, caused inactivation of lysine demethylase 5A (KDM5A), increased the methylation modification of histone H3 and regulated the transcription of EMT-associated gene_ snail family transcriptional repressor 1 (SNAIL1). This served to promote the invasion and metastasis of PC. NOX4 deficiency repressed hypoxia-induced EMT, reduced expression of H3K4ME3 and impaired the invasion and metastasis of PC cells; however, knockdown of KDM5A reversed the poor expression of H3KEME3 induced by NOX4 deficiency, thereby promoting EMT. Conclusions This study highlights the prognostic role of hypoxia-related genes in PC and strong correlation with EMT pathway. Our results also creatively discovered that NOX4 was an essential mediator for hypoxia-induced histone methylation modification and EMT in PC cells.

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Mechanisms and Pharmaceutical Action of Lipid Nanoformulation of Natural Bioactive Compounds as Efficient Delivery Systems in the Therapy of Osteoarthritis

Pharmaceutics ◽

10.3390/pharmaceutics13081108 ◽

2021 ◽

Vol 13 (8) ◽

pp. 1108

Author(s):

Oana Craciunescu ◽

Madalina Icriverzi ◽

Paula Ecaterina Florian ◽

Anca Roseanu ◽

Mihaela Trif

Keyword(s):

Drug Delivery ◽

Bioactive Compounds ◽

Targeted Delivery ◽

Drug Delivery Systems ◽

Delivery Systems ◽

Joint Disease ◽

Duration Of Action ◽

Immune System Activation

Osteoarthritis (OA) is a degenerative joint disease. An objective of the nanomedicine and drug delivery systems field is to design suitable pharmaceutical nanocarriers with controllable properties for drug delivery and site-specific targeting, in order to achieve greater efficacy and minimal toxicity, compared to the conventional drugs. The aim of this review is to present recent data on natural bioactive compounds with anti-inflammatory properties and efficacy in the treatment of OA, their formulation in lipid nanostructured carriers, mainly liposomes, as controlled release systems and the possibility to be intra-articularly (IA) administered. The literature regarding glycosaminoglycans, proteins, polyphenols and their ability to modify the cell response and mechanisms of action in different models of inflammation are reviewed. The advantages and limits of using lipid nanoformulations as drug delivery systems in OA treatment and the suitable route of administration are also discussed. Liposomes containing glycosaminoglycans presented good biocompatibility, lack of immune system activation, targeted delivery of bioactive compounds to the site of action, protection and efficiency of the encapsulated material, and prolonged duration of action, being highly recommended as controlled delivery systems in OA therapy through IA administration. Lipid nanoformulations of polyphenols were tested both in vivo and in vitro models that mimic OA conditions after IA or other routes of administration, recommending their clinical application.

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