Simultaneous inhibition of peripheral CB1R and iNOS mitigates obesity-related dyslipidemia through distinct mechanisms

Diabetic dyslipidemia (DD), characterized by increased plasma triglycerides (TGs) and decreased high-density lipoprotein cholesterol (HDL) levels, is a major factor contributing to non-alcoholic steatohepatitis (NASH) and cardiovascular risk in type-2 diabetes. Activation of both the cannabinoid-1 receptor (CB1R) and inducible nitric oxide synthase (iNOS) are associated with NASH progression. Here, we tested whether dual-targeting inhibition of hepatic CB1R and iNOS improves DD in diet-induced obese (DIO) mice. DIO mice were treated for 14 days with (S)-MRI-1867, a peripherally-restricted hybrid inhibitor of CB1R and iNOS. <a>(R)-MRI-1867, the CB1R-inactive stereoisomer which retains iNOS inhibitory activity and JD-5037, a peripherally-restricted CB1R antagonist</a> were used to assess the relative contribution of the two targets to the effects of (S)-MRI-1867. (S)-MRI-1867 reduced hepatic steatosis, the rate of hepatic VLDL secretion, upregulated hepatic LDLR expression and reduced the circulating levels of the proprotein convertase subtilisin/kexin type 9 (PCSK9). The decrease in VLDL secretion could be attributed to CB1R blockade while the reduction of PCSK9 levels and the related increase in LDLR resulted from iNOS inhibition via a mTORC1-dependent mechanism. In conclusion, this approach based on the concomitant inhibition of CB1R and iNOS represents a promising therapeutic strategy for the treatment of dyslipidemia.

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Search for Potential Inducible Nitric Oxide Synthase Inhibitors with Favorable ADMET Profiles for the Therapy of Helicobacter pylori Infections

Current Topics in Medicinal Chemistry ◽

10.2174/1568026619666191112105650 ◽

2020 ◽

Vol 19 (30) ◽

pp. 2795-2804 ◽

Cited By ~ 2

Author(s):

Ricardo Pereira Rodrigues ◽

Juliana Santa Ardisson ◽

Rita de Cássia Ribeiro Gonçalves ◽

Tiago Branquinho Oliveira ◽

Vinicius Barreto da Silva ◽

...

Keyword(s):

Nitric Oxide ◽

Helicobacter Pylori ◽

Nitric Oxide Synthase ◽

Inducible Nitric Oxide Synthase ◽

Crucial Role ◽

Chemical Space ◽

Target Selection ◽

Oxide Synthase ◽

Inos Inhibition ◽

Inducible Nitric Oxide

Background: Helicobacter pylori is a gram-negative bacterium related to chronic gastritis, peptic ulcer and gastric carcinoma. During its infection process, promotes excessive inflammatory response, increasing the release of reactive species and inducing the production of pro-inflammatory mediators. Inducible Nitric Oxide Synthase (iNOS) plays a crucial role in the gastric carcinogenesis process and a key mediator of inflammation and host defense systems, which is expressed in macrophages induced by inflammatory stimuli. In chronic diseases such as Helicobacter pylori infections, the overproduction of NO due to the prolonged induction of iNOS is of major concern. Objective: In this sense, the search for potential iNOS inhibitors is a valuable strategy in the overall process of Helicobacter pylori pathogeny. Method: In silico techniques were applied in the search of interesting compounds against Inducible Nitric Oxide Synthase enzyme in a chemical space of natural products and derivatives from the Analyticon Discovery databases. Results: The five compounds with the best iNOS inhibition profile were selected for activity and toxicity predictions. Compound 9 (CAS 88198-99-6) displayed significant potential for iNOS inhibition, forming hydrogen bonds with residues from the active site and an ionic interaction with heme. This compound also displayed good bioavailability and absence of toxicity/or from its probable metabolites. Conclusion: The top-ranked compounds from the virtual screening workflow show promising results regarding the iNOS inhibition profile. The results evidenced the importance of the ionic bonding during docking selection, playing a crucial role in binding and positioning during ligand-target selection for iNOS.

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PAR-4/Ca2+-calpain pathway activation stimulates platelet-derived microparticles in hyperglycemic type 2 diabetes

Cardiovascular Diabetology ◽

10.1186/s12933-021-01267-w ◽

2021 ◽

Vol 20 (1) ◽

Author(s):

Alessandra Giannella ◽

Giulio Ceolotto ◽

Claudia Maria Radu ◽

Arianna Cattelan ◽

Elisabetta Iori ◽

...

Keyword(s):

Type 2 Diabetes ◽

Normal Glucose Tolerance ◽

Calpain Inhibitor ◽

Pathway Activation ◽

Normal Glucose ◽

Circulating Levels ◽

Dependent Mechanism

Abstract Background Patients with type 2 diabetes (T2DM) have a prothrombotic state that needs to be fully clarified; microparticles (MPs) have emerged as mediators and markers of this condition. Thus, we investigate, in vivo, in T2DM either with good (HbA1c ≤ 7.0%; GGC) or poor (HbA1c > 7.0%; PGC) glycemic control, the circulating levels of MPs, and in vitro, the molecular pathways involved in the release of MPs from platelets (PMP) and tested their pro-inflammatory effects on THP-1 transformed macrophages. Methods In 59 T2DM, and 23 control subjects with normal glucose tolerance (NGT), circulating levels of CD62E+, CD62P+, CD142+, CD45+ MPs were determined by flow cytometry, while plasma levels of ICAM-1, VCAM-1, IL-6 by ELISA. In vitro, PMP release and activation of isolated platelets from GGC and PGC were investigated, along with their effect on IL-6 secretion in THP-1 transformed macrophages. Results We found that MPs CD62P+ (PMP) and CD142+ (tissue factor-bearing MP) were significantly higher in PGC T2DM than GGC T2DM and NGT. Among MPs, PMP were also correlated with HbA1c and IL-6. In vitro, we showed that acute thrombin exposure stimulated a significantly higher PMP release in PGC T2DM than GGC T2DM through a more robust activation of PAR-4 receptor than PAR-1 receptor. Treatment with PAR-4 agonist induced an increased release of PMP in PGC with a Ca2+-calpain dependent mechanism since this effect was blunted by calpain inhibitor. Finally, the uptake of PMP derived from PAR-4 treated PGC platelets into THP-1 transformed macrophages promoted a marked increase of IL-6 release compared to PMP derived from GGC through the activation of the NF-kB pathway. Conclusions These results identify PAR-4 as a mediator of platelet activation, microparticle release, and inflammation, in poorly controlled T2DM.

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Uncoupling of Vascular Endothelial Growth Factor (VEGF) and Inducible Nitric Oxide Synthase (iNOS) in Gingival Tissue of Type 2 Diabetic Patients

Inflammation ◽

10.1007/s10753-015-0288-9 ◽

2015 ◽

Vol 39 (2) ◽

pp. 632-642 ◽

Cited By ~ 6

Author(s):

Guendalina Lucarini ◽

Giacomo Tirabassi ◽

Antonio Zizzi ◽

Giancarlo Balercia ◽

Alexia Quaranta ◽

...

Keyword(s):

Nitric Oxide ◽

Gingival Tissue ◽

Diabetic Patients ◽

Vascular Endothelial ◽

Type 2 Diabetic Patients ◽

Oxide Synthase ◽

Endothelial Growth Factor ◽

Type 2 Diabetic ◽

Inducible Nitric Oxide

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Effects of chronic inhibition of inducible nitric oxide synthase in hyperthyroid rats

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.00279.2004 ◽

2005 ◽

Vol 288 (6) ◽

pp. E1252-E1257 ◽

Cited By ~ 16

Author(s):

Isabel Rodríguez-Gómez ◽

Rosemary Wangensteen ◽

Juan Manuel Moreno ◽

Virginia Chamorro ◽

Antonio Osuna ◽

...

Keyword(s):

Nitric Oxide ◽

Nitric Oxide Synthase ◽

Inducible Nitric Oxide Synthase ◽

Control Group ◽

Hemodynamic Variable ◽

Male Wistar Rats ◽

Oxide Synthase ◽

Inos Inhibition ◽

Inducible Nitric Oxide

We hypothesized that nitric oxide generated by inducible nitric oxide synthase (iNOS) may contribute to the homeostatic role of this agent in hyperthyroidism and may, therefore, participate in long-term control of blood pressure (BP). The effects of chronic iNOS inhibition by oral aminoguanidine (AG) administration on BP and morphological and renal variables in hyperthyroid rats were analyzed. The following four groups ( n = 8 each) of male Wistar rats were used: control group and groups treated with AG (50 mg·kg−1·day−1, via drinking water), thyroxine (T4, 50 μg·rat−1·day−1), or AG + T4. All treatments were maintained for 3 wk. Tail systolic BP and heart rate (HR) were recorded weekly. Finally, we measured BP (mmHg) and HR in conscious rats and morphological, plasma, and renal variables. T4 administration produced a small BP (125 ± 2, P < 0.05) increase vs. control (115 ± 2) rats. AG administration to normal rats did not modify BP (109 ± 3) or any other hemodynamic variable. However, coadministration of T4 and AG produced a marked increase in BP (140 ± 3, P < 0.01 vs. T4). Pulse pressure and HR were increased in both T4- and T4 + AG -treated groups without differences between them. Plasma NOx (μmol/l) were increased in the T4 group (10.02 ± 0.15, P < 0.05 vs. controls 6.1 ± 0.10), and AG reduced this variable in T4-treated rats (6.81 ± 0.14, P < 0.05 vs. T4) but not in normal rats (5.78 ± 0.20). Renal and ventricular hypertrophy and proteinuria of hyperthyroid rats were unaffected by AG treatment. In conclusion, the results of the present paper indicate that iNOS activity may counterbalance the prohypertensive effects of T4.

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Inhibitory effects of tilianin on the expression of inducible nitric oxide synthase in low density lipoprotein receptor deficiency mice

Experimental & Molecular Medicine ◽

10.1038/emm.2006.52 ◽

2006 ◽

Vol 38 (4) ◽

pp. 445-452 ◽

Cited By ~ 22

Author(s):

Ki-Hoan Nam ◽

Jae-Hoon Choi ◽

Yun-Jeong Seo ◽

Young-Mi Lee ◽

Yong-Sung Won ◽

...

Keyword(s):

Nitric Oxide ◽

Inducible Nitric Oxide Synthase ◽

Low Density Lipoprotein ◽

Density Lipoprotein ◽

Low Density ◽

Lipoprotein Receptor ◽

Inhibitory Effects ◽

Density Lipoprotein Receptor ◽

Oxide Synthase ◽

Inducible Nitric Oxide

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Diyabetik sıçanların karaciğer ve böbrekleri üzerinde Caralluma tuberculata’nın hipoglisemik ve hipolipidemik etkisi ve güvenliği / Liquidambar orientalis Mill. reçine ekstraktlarının antioksidan, sitotoksik ve iNOS aktiviteleri

Turkish Journal of Biochemistry ◽

10.1515/tjb-2016-0030 ◽

2016 ◽

Vol 41 (3) ◽

Author(s):

Ayşe Nalbantsoy ◽

Mert Karış ◽

Leyla Karakaya ◽

Yurdanur Akgül

Keyword(s):

Nitric Oxide ◽

Antioxidant Activity ◽

Nitric Oxide Synthase ◽

Inducible Nitric Oxide Synthase ◽

No Production ◽

Raw 264.7 ◽

Water Extracts ◽

Oxide Synthase ◽

Inos Inhibition ◽

Inducible Nitric Oxide

AbstractObjective: The aim of this study is to investigate the in vitro cytotoxicity and iNOS (inducible nitric oxide synthase) inhibitory, and antioxidant activity in order to assess the traditional usage of Liquidambar orientalis Mill resin extract.Methods: Different solvent extracts of Liquidambar orientalis Mill resin were prepared. The cytotoxicity of extracts was determined using MTT (3-(4,5-dimethyl-2-thiazolyl)- 2,5-diphenyl-2H-tetrazoliumbromide) assay. HeLa (Human cervix adenocarcinoma), A-549 (Human alveolar adenocarcinoma), MCF-7 (Human breast adenocarcinoma), CaCo-2 (Human colon colorectal adenocarcinoma), mPANC96 (Human pancreas adenocarcinoma), PC-3 (Human prostate adenocarcinoma), U87MG (Human glioblastoma- astrocytoma) and as a normal cell line HEK293 (Human embryonic kidney cells) and Vero (African green monkey kidney epithelial cells) were used for testing cytotoxicity. RAW 264.7 (murine macrophage cell lines) was used to determine the inhibition levels of inducible nitric oxide synthase (iNOS). HL-60 (human acute myeloid leukemia) was used to determine antioxidant activity as DCF production per cent.Results: Hexane, dichloromethane, methanol and water extracts were prepared, and their iNOS inhibitory, cytotoxic and antioxidant activity were investigated. The estimated IC50 values of extracts varied from 6.68 to 48.90 μg/ ml after treatment with different doses of extracts for 48 h. Inhibition of the hexane, dichloromethane, methanol, and water extracts on LPS-induced NO production in RAW 264.7 macrophage were showed that the all extracts inhibited NO production in activated RAW 264.7 cells, except methanol extract. Hexane, dichloromethane and methanol extracts inhibited NO production with ICConclusion: This study is the first report showing the potential of Liquidambar orientalis Mill resin extracts for cytotoxicity, iNOS inhibition and the antioxidant activity as an alternative therapeutic approach for traditional uses. The results demonstrated that Liquidambar orientalis dichloromethane resin extracts showed strongest cytotoxic effect while all extracts except methanolic extracts exhibited moderate iNOS inhibition. All extracts other than hexane have a potent antioxidant effect in the cellular-based assay. In conclusion, further studies should focus on the purification of the active components of this extracts and to investigate the possible mode of action to obtain a better understanding of their potential use as cytotoxic, anti-inflammatory and antioxidant agents.

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Differential Changes of Aorta and Carotid Vasodilation in Type 2 Diabetic GK and OLETF Rats: Paradoxical Roles of Hyperglycemia and Insulin

Experimental Diabetes Research ◽

10.1155/2012/429020 ◽

2012 ◽

Vol 2012 ◽

pp. 1-16 ◽

Cited By ~ 8

Author(s):

Mei-Fang Zhong ◽

Wei-Li Shen ◽

Masaki Tabuchi ◽

Kyoko Nakamura ◽

Yi-Chen Chen ◽

...

Keyword(s):

Insulin Level ◽

Insulin Injection ◽

Diabetic Rats ◽

Endothelium Dysfunction ◽

Mild Hyperglycemia ◽

Oxide Synthase ◽

Type 2 Diabetic ◽

Inducible Nitric Oxide

We investigated large vessel function in lean Goto-Kakizaki diabetic rats (GK) and Otsuka Long-Evans Tokushima Fatty diabetic rats (OLETF) with possible roles of hyperglycemia/hyperosmolarity and insulin. Both young and old GK showed marked hyperglycemia with normal insulin level and well-preserved endothelium-dependent and endothelium-independent vasodilation in aorta and carotid artery. There were significant elevations in endothelial/inducible nitric oxide synthase (eNOS/iNOS) and inducible/constitutive heme oxygenase (HO-1/HO-2) in GK. The endothelium-dependent vasodilation in GK was inhibited partly by NOS blockade and completely by simultaneous blocking of HO and NOS. In contrast, OLETF showed hyperinsulinemia and mild hyperglycemia but significant endothelium dysfunction beginning at early ages with concomitantly reduced eNOS. Insulin injection corrected hyperglycemia in GK but induced endothelium dysfunction and intima hyperplasia. Hyperglycemia/hyperosmolarityin vitroenhanced vessel eNOS/HO. We suggest that hyperinsulinemia plays a role in endothelium dysfunction in obese diabetic OLETF, while hyperglycemia/hyperosmolarity-induced eNOS/HO upregulation participates in the adaptation of endothelium function in lean diabetic GK.

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