45 research outputs found
Ligand Bias and Its Association With Pro-resolving Actions of Melanocortin Drugs
Resolution Pharmacology identifies drugs developed on the biology of the resolution phase of inflammation, the complex molecular and cellular network of events that ensure the tight temporal and spatial control on the inflammatory response. As such, new anti-inflammatory and pro-resolving drugs could derive from pro-resolving mediators and receptors. To implement faithful screening programs, however, it is important to rely on predictive signaling pathway relevant for the ultimate bio-action of interest. Herein we performed an analysis with four prototypical melanocortin receptor (MC1,3,4,5) agonists. The choice fell on the natural agonist αMSH, the small molecule BMS-470539, and the synthetic derivatives [D-Trp8]-ÎłMSH and [Nle4,D-Phe7]-αMSH. We used human macrophages and quantified the effect of the four agonists on inhibition of cytokine release and promotion of efferocytosis. All agonists (1â10 ÎŒM) significantly inhibited cytokine release by LPS-stimulated cells whereas [D-Trp8]-ÎłMSH was the most effective in inducing efferocytosis (âŒ60% increase). To study the signaling profile, we monitored cAMP accumulation and ERK1/2 phosphorylation, and constructed biased plots that revealed a marked biased profile of [D-Trp8]-ÎłMSH toward phospho-ERK1/2. Correlation matrix analysis of all data pointed at phospho-ERK1/2 at any receptor as the most prominent pathway to attain pro-phagocytic actions, and MC1 receptor as the most relevant to drive anti-cytokine effects. In conclusion, the present study highlights the need to associate single-target signaling data with relevant functional outcomes. In this manner, we would increase our chances to optimize drug discovery programs during the early target validation and hit-to-lead phases
Mechanisms of endothelial cell dysfunction in cystic fibrosis
Although cystic fibrosis (CF) patients exhibit signs of endothelial perturbation, the functions of the cystic fibrosis
conductance regulator (CFTR) in vascular endothelial cells (EC) are poorly defined. We sought to uncover
biological activities of endothelial CFTR, relevant for vascular homeostasis and inflammation. We examined cells
from human umbilical cords (HUVEC) and pulmonary artery isolated from non-cystic fibrosis (PAEC) and CF
human lungs (CF-PAEC), under static conditions or physiological shear. CFTR activity, clearly detected in
HUVEC and PAEC, was markedly reduced in CF-PAEC. CFTR blockade increased endothelial permeability to
macromolecules and reduced transâendothelial electrical resistance (TEER). Consistent with this, CF-PAEC displayed
lower TEER compared to PAEC. Under shear, CFTR blockade reduced VE-cadherin and p120 catenin
membrane expression and triggered the formation of paxillin- and vinculin-enriched membrane blebs that
evolved in shrinking of the cell body and disruption of cell-cell contacts. These changes were accompanied by
enhanced release of microvesicles, which displayed reduced capability to stimulate proliferation in recipient EC.
CFTR blockade also suppressed insulin-induced NO generation by EC, likely by inhibiting eNOS and AKT
phosphorylation, whereas it enhanced IL-8 release. Remarkably, phosphodiesterase inhibitors in combination
with a ÎČ2 adrenergic receptor agonist corrected functional and morphological changes triggered by CFTR dysfunction
in EC. Our results uncover regulatory functions of CFTR in EC, suggesting a physiological role of CFTR
in the maintenance EC homeostasis and its involvement in pathogenetic aspects of CF. Moreover, our findings
open avenues for novel pharmacology to control endothelial dysfunction and its consequences in CF
Biological Effect of Licochalcone C on the Regulation of PI3K/Akt/eNOS and NF-ÎșB/iNOS/NO Signaling Pathways in H9c2 Cells in Response to LPS Stimulation
Polyphenols compounds are a group molecules present in many plants. They have antioxidant properties and can also be helpful in the management of sepsis. Licochalcone C (LicoC), a constituent of Glycyrrhiza glabra, has various biological and pharmacological properties. In saying this, the effect of LicoC on the inflammatory response that characterizes septic myocardial dysfunction is poorly understood. The aim of this study was to determine whether LicoC exhibits anti-inflammatory properties on H9c2 cells that are stimulated with lipopolysaccharide. Our results have shown that LicoC treatment represses nuclear factor-ÎșB (NF-ÎșB) translocation and several downstream molecules, such as inducible nitric oxide synthase (iNOS), intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). Moreover, LicoC has upregulated the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/endothelial nitric oxide synthase (eNOS) signaling pathway. Finally, 2-(4-Morpholinyl)-8-phenyl-1(4H)-benzopyran-4-one hydrochloride (LY294002), a specific PI3K inhibitor, blocked the protective effects of LicoC. These findings indicate that LicoC plays a pivotal role in cardiac dysfunction in sepsis-induced inflammation.The Italian Ministry for University and Research is acknowledged for financial support
Astaxanthin Treatment Reduced Oxidative Induced Pro-Inflammatory Cytokines Secretion in U937: SHP-1 as a Novel Biological Target
It has been suggested that oxidative stress activates various intracellular signaling pathways leading to secretion of a variety of pro-inflammatory cytokines and chemokines. SHP-1 is a protein tyrosine phosphatase (PTP) which acts as a negative regulator of immune cytokine signaling. However, intracellular hydrogen peroxide (H2O2), generated endogenously upon stimulation and exogenously from environmental oxidants, has been known to be involved in the process of intracellular signaling through inhibiting various PTPs, including SHP-1. In this study, we investigated the potential role of astaxanthin, an antioxidant marine carotenoid, in re-establishing SHP-1 negative regulation on pro-inflammatory cytokines secretion in U-937 cell line stimulated with oxidative stimulus. ELISA measurement suggested that ASTA treatment (10 ”M) reduced pro-inflammatory cytokines secretion (IL-1ÎČ, IL-6 and TNF-α) induced through H2O2, (100 ”M). Furthermore, this property is elicited by restoration of basal SHP-1 protein expression level and reduced NF-ÎșB (p65) nuclear expression, as showed by western blotting experiments
Headâneck melanoma: Clinical, histopathological and prognostic features of an Italian multicentric study
factor. As regards the scalp, some studies indicate a particularly aggressive biological behaviour for this anatomical localisation. Objectives: In this multicentric study, data regarding headâneck melanoma (HNM) have been revised. Methods: The design of the study included two main phases. In this retrospective study, data regarding HNM have been collected and analysed. Results: In summary, our data suggest that the posterior neck is the area most affected by thicker melanomas. Cheeks and neck melanoma are associated with reduced diseaseâfree years of life and overall survival compared with all other sites of HNM. Conclusions: This study provides useful information in defining the clinical features of HNM, thus improving diagnosis and treatment strategies
Proresolving Lipid Mediators and Receptors in Stem Cell Biology: Concise Review
Summary Accumulating evidence indicates that stem cells (SCs) possess immunomodulatory, antiâinflammatory, and prohealing properties. The mechanisms underlying these functions are being investigated with the final goal to set a solid background for the clinical use of SCs and/or their derivatives. Specialized proresolving lipid mediators (SPMs) are small lipids formed by the enzymatic metabolism of polyunsaturated fatty acids. They represent a leading class of molecules that actively and timely regulate the resolution of inflammation and promote tissue/organ repair. SC formation of these mediators as well as expression of their receptors has been recently reported, suggesting that SPMs may be involved in the immunomodulatory, proresolving functions of SCs. In the present review, we summarize the current knowledge on SPMs in SCs, focusing on biosynthetic pathways, receptors, and bioactions, with the intent to provide an integrated view of SPM impact on SC biology. Stem Cells Translational Medicine 2019;8:992â99
Combined Rehabilitation Protocol in the Treatment of Osteoarthritis of the Knee: Comparative Study of Extremely Low-Frequency Magnetic Fields and Soft Elastic Knee Brace Effect
The investigation of this observational caseâcontrol study aimed at determining the effectiveness of a combined treatment of extremely low-frequency electromagnetic fields (ELF) with a soft elastic knee brace versus ELF alone in knee osteoarthritis (KOA) with respect to a reduction in pain and functional recovery. We hypothesized that the combined use of ELF and a soft elastic knee brace may provide better results. Thirty-five patients (N = 35, divided into Group 1 = ELF and Group 2 = ELF with the soft elastic knee brace) were analyzed. The rehabilitative protocol consisted of 10 sessions of antiphlogistic and antiedema programs (first cycle) for 2 weeks, followed by twelve sessions of bone repair and connective tissue repair programs (second cycle) in patients with knee osteoarthritis (KOA) for 4 weeks. Patient evaluations were conducted at baseline (T0) and after 2 (T1) and 4 (T2) weeks of treatment. A follow-up evaluation was conducted 6 weeks after treatment (T3). The LIMFA© Therapy System was used to create multifrequency magnetoelectric fields with an intensity of 100 ”T and a low-frequency field. The Incrediwear Cred 40 knee sleeve (Incred) was used for alleviating knee pain. The Visual Analogue Scale (VAS), the Knee Injury and Osteoarthritis Outcome Score (KOOS), and the Lysholm score (Ls) were used as outcome measures. The results showed that pain at rest (Vr), pain in motion (Vm), KOOS, and Ls were significantly affected by ELF over time. In conclusion, Group 2 had a better response in terms of pain resolution at rest (p p < 0.05)