Triticum vulgare Extract Modulates Protein-Kinase B and Matrix Metalloproteinases 9 Protein Expression in BV-2 Cells: Bioactivity on Inflammatory Pathway Associated with Molecular Mechanism Wound Healing.

Matrix metalloproteinases (MMPs) are a large family of ubiquitously expressed zinc-dependent enzymes with proteolitic activities. They are expressed in physiological situations and pathological conditions involving inflammatory processes including epithelial to mesenchymal transition (EMT), neuronal injury, and cancer. There is also evidence that MMPs regulate inflammation in tumor microenvironment, which plays an important role in healing tissue processes. Looking at both inflammatory and neuronal damages, MMP9 is involved in both processes and their modulation seems to be regulated by two proteins: tumor necrosis factor-alpha (TNF-alpha) and interleukin 6 (IL-6). However other important genes are involved in molecular regulation of transcription factors, protein-kinase B (AKT), and p65. In addition, Triticum vulgare extract (TVE) modulated the biological markers associated with inflammatory processes, including p65 protein. While there are no evidence that TVE might be involved in the biological modulation of other inflammatory marker as AKT, we would like to assess whether TVE is able to (1) modulate phosphorylation of AKT (pAKT) as an early marker of inflammatory process in vitro and (2) affect MMP9 protein expression in an in vitro model. The BV-2 cells (microglial of mouse) have been used as an in vitro model to simulate both inflammatory and neuronal injury pathologies. Here, MMP9 seems to be involved in cellular migration through inflammatory marker activation. We simulate an inflammatory preclinical model treating BV-2 cells with lipopolysaccharide (LPS) to induce proinflammatory activation affecting pAKT and p65 proteins. TVE is revealed to restore the native expression of AKT and p65. Additionally, TVE extract modulates also the protein concentration of MMP9. Nevertheless, immunofluorescence confocal analyses revealed that both AKT and MMP9 are regulated together, synchronously. This work seems to demonstrate that two important genes can be used to monitor the beginning of an inflammatory process, AKT and MMP9, in which TVE seems able to modulate their expression of inflammation-associated molecules.

A Triticum vulgare Extract Exhibits Regenerating Activity During the Wound Healing Process.

INTRODUCTION: Chronic skin lesions represent a problem of increasing occurrence, mostly due to the global ageing of the world population. Research in skin care and dermatology is constantly looking for new non-invasive solutions, preferably those based on the use of natural certified products, able to accelerate the spontaneous skin repair mechanisms and without altering the skin normal appearance and functionality. The wound healing process in the skin is finely regulated by several factors and orchestrated mechanisms, which modulate the progression and the fitting of different consequent phases, including haemostasis, inflammation, cell proliferation and tissue remodelling. It was previously shown that a patented Triticum vulgare aqueous extract was able to trigger the skin repair process by stimulating new tissue growth and reducing the expression levels of inflammatory mediators, such as IL-6, TNFα, prostaglandin E2, and nitric oxide. METHODS: Scratch assay was performed in Human Dermal Fibroblasts (HDF). The production of fibronectin was measured by gene expression, protein quantification and localization using specific antibodies in HDF. The polymerization of actin was measured using rhodamin-phalloidin in HDF. The epidermal lipid content was estimated in HaCaT (human spontaneously immortalized keratinocytes) using Nile Red staining and the increasing GBA gene expression and activity was demonstrated by RT-PCR and enzymatic activity assay. RESULTS: In the present study, it was demonstrated that the T. vulgare extract enhanced cell migration inducing the synthesis of fibronectin, new actin polymerization and stimulating the expression of the Hyaluronan Synthase 2. Furthermore it improved the restoration of the epidermal barrier stimulating lipid synthesis. CONCLUSION: In conclusion, we demonstrated that the T. vulgare extract possessed promising potential to be developed as a wound healing promoting agent in skin care and dermatology.

Antioxidant Capacity of Rigenase®, a Specific Aqueous Extract of Triticum vulgare.

Reactive species of oxygen (ROS), responsible for oxidative stress, accumulate in various tissues damaged by burns, decubitus ulcers, and vascular lesions. Antioxidants play an important and well-documented role in healing of chronic and acute wounds. Rigenase®, a specific extract of Triticum vulgare manufactured by Farmaceutici Damor, is employed in products used for the regeneration of tissue injuries. In this work, we show that Rigenase® exhibits a scavenging effect toward free radicals, thus pointing to its relevant antioxidant activity.

Triticum vulgare extract exerts an anti-inflammatory action in two in vitro models of inflammation in microglial cells.

Triticum vulgare has been extensively used in traditional medicine thanks to its properties of accelerating tissue repair. The specific extract of Triticum vulgare manufactured by Farmaceutici Damor (TVE-DAMOR) is already present in some pharmaceutical formulations used in the treatment of decubitus ulcers, skin lesions and burns. It has been recently suggested that this Triticum vulgare extract may possess potential anti-inflammatory properties. In the light of these premises the aim of the present paper was to verify the anti-inflammatory role of TVE, using the LPS-stimulated microglia model of inflammation. In particular the effect of different concentrations of TVE on the release of several mediators of inflammation such as nitric oxide, IL-6, PGE2 and TNF alpha was evaluated. More important, the anti-inflammatory effect of TVE was confirmed also in primary rat microglia cultures. The results of the present study show that TVE exerts anti-inflammatory properties since it reduces the release of all the evaluated markers of inflammation, such as NO, IL6, TNF alpha and PGE2 in LPS-activated BV2 microglial cells. Intriguingly, TVE reduced microglia activation and NO release also in primary microglia. Indeed, to verify the pathway of modulation of the inflammatory markers reported above, we found that TVE restores the cytoplasmic expression of p65 protein, kwown as specific marker associated with activation of inflammatory response. The evidence for an inhibitory activity on inflammation of this specific extract of Triticum vulgare may open the way to the possibility of a therapeutical use of the Triticum vulgare extract as an anti-inflammatory compound in certain pathological states such as burns, decubitus ulcers, folliculitis and inflammation of peripheral nerve.