The Anti-Inflammatory Activity of Ferulic Acid on NF-κBDepends on Keap1

The process of inflammation leads to the onset of a state of oxidative stress and a series of cascade reactions that are associated..

Toxic mineral elements in Mytilus galloprovincialis from Sicilian coasts (Southern Italy)

We assessed the relationship between V, Cr, Mn, Hg, As, Cd, Sn, Sb and Pb concentrations in Mytilus galloprovincialis samples from the coasts of Sicily and the expression of metallothioneins. Toxic mineral elements assessment was carried out by A.A. Spectrometry and ICP-MS. The metallothioneins expression was performed by q-PCR method. Low metals' levels were found in the mussel samples examined, in comparison with what was reported in literature. The highest mean values of toxic mineral elements were found in Gela (Cr 0.178 ± 0.03 mg/Kg, Mn 4.325 ± 0.012 mg/Kg, As 3.706 ± 0.009 mg/Kg, Sn 0.148 ± 0.014 mg/Kg, Sb 0.009 ± 0.004 mg/Kg e Pb 0.364 ± 0.01 mg/Kg). Significant levels of Hg were found in samples from Catania (0.014 ± 0.005 mg/Kg). Only vanadium and lead concentrations showed significant differences between sampling areas (p < 0.05). Molecular analysis verified a basal expression of Mt1 and the absence of over-expression of Mt2, confirming the low mineral's concentrations found in the samples examined

Mitochondrial physiology

As the knowledge base and importance of mitochondrial physiology to evolution, health and disease expands, the necessity for harmonizing the terminology concerning mitochondrial respiratory states and rates has become increasingly apparent. The chemiosmotic theory establishes the mechanism of energy transformation and coupling in oxidative phosphorylation. The unifying concept of the protonmotive force provides the framework for developing a consistent theoretical foundation of mitochondrial physiology and bioenergetics. We follow the latest SI guidelines and those of the International Union of Pure and Applied Chemistry (IUPAC) on terminology in physical chemistry, extended by considerations of open systems and thermodynamics of irreversible processes. The concept-driven constructive terminology incorporates the meaning of each quantity and aligns concepts and symbols with the nomenclature of classical bioenergetics. We endeavour to provide a balanced view of mitochondrial respiratory control and a critical discussion on reporting data of mitochondrial respiration in terms of metabolic flows and fluxes. Uniform standards for evaluation of respiratory states and rates will ultimately contribute to reproducibility between laboratories and thus support the development of data repositories of mitochondrial respiratory function in species, tissues, and cells. Clarity of concept and consistency of nomenclature facilitate effective transdisciplinary communication, education, and ultimately further discovery

Mitochondrial physiology

As the knowledge base and importance of mitochondrial physiology to evolution, health and disease expands, the necessity for harmonizing the terminology concerning mitochondrial respiratory states and rates has become increasingly apparent. The chemiosmotic theory establishes the mechanism of energy transformation and coupling in oxidative phosphorylation. The unifying concept of the protonmotive force provides the framework for developing a consistent theoretical foundation of mitochondrial physiology and bioenergetics. We follow the latest SI guidelines and those of the International Union of Pure and Applied Chemistry (IUPAC) on terminology in physical chemistry, extended by considerations of open systems and thermodynamics of irreversible processes. The concept-driven constructive terminology incorporates the meaning of each quantity and aligns concepts and symbols with the nomenclature of classical bioenergetics. We endeavour to provide a balanced view of mitochondrial respiratory control and a critical discussion on reporting data of mitochondrial respiration in terms of metabolic flows and fluxes. Uniform standards for evaluation of respiratory states and rates will ultimately contribute to reproducibility between laboratories and thus support the development of data repositories of mitochondrial respiratory function in species, tissues, and cells. Clarity of concept and consistency of nomenclature facilitate effective transdisciplinary communication, education, and ultimately further discovery

Substance P Induces HO-1 Expression in RAW 264.7 Cells Promoting Switch towards M2-Like Macrophages.

Substance P (SP) is a neuropeptide that mediates many physiological as well as inflammatory responses. Recently, SP has been implicated in the resolution of inflammation through induction of M2 macrophages phenotype. The shift between M1-like and M2-like, allowing the resolution of inflammatory processes, also takes place by means of hemeoxygenase-1 (HO-1). HO-1 is induced in response to oxidative stress and inflammatory stimuli and modulates the immune response through macrophages polarisation. SP induces HO-1 expression in human periodontal ligament (PDL), the latter potentially plays a role in cytoprotection. We demonstrated that SP promotes M2-like phenotype from resting as well as from M1 macrophages. Indeed, SP triggers the production of interleukine-10 (IL-10), interleukine-4 (IL-4) and arginase-1 (Arg1) without nitric oxide (NO) generation. In addition, SP increases HO-1 expression in a dose- and time-dependent manner. Here we report that SP, without affecting cell viability, significantly reduces the production of pro-inflammatory cytokines and enzymes, such as tumor necrosis factor-alpha (TNF-α), interleukine-6 (IL-6), inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), and ameliorates migration and phagocytic properties in LPS-stimulated RAW 264.7 cells. M2-like conversion required retention of NF-κB p65 into the cytoplasm and HO-1 induced expression. Silencing of the HO-1 mRNA expression reversed the induction of pro-inflammatory cytokines in RAW 264.7 stimulated by LPS and down-regulated anti-inflammatory hallmarks of M2 phenotype. In conclusion, our data show that SP treatment might be associated with anti-inflammatory effects in LPS-stimulated RAW 264.7 cells by suppressing NF-κB activation and inducing HO-1 expression

Effects of SP on LPS-induced p65 transcription factor nuclear translocation.

<p>(A) RAW 264.7 cells were treated with or without SP (10 μM) and/or with LPS (100 ng/ml) for 4 hours and cytoplasmic and nuclear fractions were separated and analysed by western blotting for NF-κB p65 expression. We used β-actin (47 kDa) and lamin B1 (67 kDa) immunolabelling as loading controls for cytoplasmic and nuclear fractions, respectively. The numbers represent the fold difference of NF-κB p65 with untreated control (Ctrl) arbitrarily set at 1.0. The data shown represent two independent experiments with comparable outcomes. (B) RT-qPCR analysis of p65 (Rel A) mRNA in cells treated or untreated with SP (10 μM) for 4 hours. The data are mean ±SD of two separate experiments, each of which was performed in triplicate. **p < 0.01 versus untreated. (C) Immunofluorescence analysis of RAW 264.7 cells with NF-κB p65 (red) in cells treated with or without SP (10 μM) and LPS (100 ng/ml) for 4 hours. The nuclei of the cells have been counterstained with DAPI (blue). Scale bar 20 μm. Results are shown from two independent experiments with comparable outcomes.</p

SP induces M2-like phenotype.

<p>(A) RAW 264.7 cells were treated with SP (10 μM) and Arg1, IL-10 and IL-4 mRNA expression was evaluated with RT-qPCR. Data are the means ± SD of two experiments (each of which was performed in triplicate). *p<0.05 and **p<0.01 <i>versus</i> control. (B) RAW 264.7 cells were treated with SP (10 μM) for 24 hours and Arg1 (35 kDa) protein levels were determined by western blotting. Representative immunoblots of results obtained from three independent experiments are shown. (C) RAW 264.7 cells were treated with different doses of SP or stimulated with LPS 100 ng/ml for 24 hours. NO in the culture medium was measured as described in the methodology section. Data are expressed as nitric oxide concentration (μM) and are the means ± SD of three separated experiments, each of which was performed in triplicate. *p<0.05 versus untreated.</p

SP induces HO-1 mRNA and protein expression.

<p>RAW 264.7 cells were treated with the indicated concentrations of SP for (A) 4, (B) 6 and (C) 24 hours and mRNA expression levels were assessed by RT-qPCR. (B). The results shown are the means ± SD of two experiments (each of which was performed in triplicate). *p<0.05 and **p<0.01 <i>versus</i> each agent alone. (D) Cells were treated with SP (μM) for 24 hours. The induction of HO-1 protein expression (32 kDa) was analysed by western blotting. The numbers represent the fold of HO-1 difference with untreated control samples (Ctrl) arbitrarily set at 1.0. The data represent two independent representative experiments.</p

HO-1 induction promotes M2 macrophage polarisation.

<p>RAW 264.7 cells were transfected with siRNA HO-1 and siRNA NC (non-correlated). (A) RT-qPCR analysis of HO-1 mRNA in cells transfected 48 hours after transfection. Cells were treated with SP (10 μM) and/or LPS (100 ng/ml) for 24 hours and mRNA expression of selected genes was evaluated by RT-qPCR (B) IL-6 mRNA expression (C) TNF-α mRNA expression (D) Arg1 mRNA expression (E) IL-10 mRNA expression and (F) Rel A mRNA expression. The data are mean ±SD of two separate experiments, each of which was performed in triplicate. **p < 0.01 versus siRNA NC.</p