Oxidative Stress Modulation by Carnosine in Scaffold Free Human Dermis Spheroids Model: A Proteomic Study

Abstract

Carnosine is an endogenous β-alanyl-L-histidine dipeptide endowed with antioxidant and carbonyl scavenger properties, which is able to significantly prevent the visible signs of aging and photoaging. To investigate the mechanism of action of carnosine on human skin proteome, a 3D scaffold-free spheroid model of primary dermal fibroblasts from a 50-year-old donor was adopted in combination with quantitative proteomics for the first time. The label free proteomics approach based on high-resolution mass spectrometry, integrated with network analyses, provided a highly sensitive and selective method to describe the human dermis spheroid model during long-term culture and upon carnosine treatment. Overall, 2171 quantified proteins allowed the in-depth characterization of the 3D dermis phenotype during growth and differentiation, at 14 versus 7 days of culture. A total of 485 proteins were differentially regulated by carnosine at 7 days, an intermediate time of culture. Of the several modulated pathways, most are involved in mitochondrial functionality, such as oxidative phosphorylation, TCA cycle, extracellular matrix reorganization and apoptosis. In long-term culture, functional modules related to oxidative stress were upregulated, inducing the aging process of dermis spheroids, while carnosine treatment prevented this by the downregulation of the same functional modules. The application of quantitative proteomics, coupled to advanced and relevant in vitro scaffold free spheroids, represents a new concrete application for personalized therapies and a novel care approach.

L-carnosine Attenuates Bleomycin-Induced Oxidative Stress via NFκB Pathway in the Pathogenesis of Pulmonary Fibrosis

“…In Figure 5 A,B, we demonstrated the anti-fibrotic effect of Lcar in fibroblast. Except for this, L-carnosine can ameliorate fibroblast senescence and modulate various pathways such as TCA cycle, oxidative phosphorylation, apoptosis, and ECM reorganization [ 53 ]. Furthermore, L-carnosine treatment led to p53 protein reduction in mice ( Figure 5 B).…”

Polyphenols from Thinned Young Apples: HPLC-HRMS Profile and Evaluation of Their Anti-Oxidant and Anti-Inflammatory Activities by Proteomic Studies

“…Tryptic peptides were analyzed using a Dionex Ultimate 3000 nano-LC system (Sunnyvale, CA, USA) connected to an Orbitrap Fusion Tribrid Mass Spectrometer (Thermo Scientific, Bremen, Germany) equipped with a nano-electrospray ion source according to the procedure previously described [ 50 ].…”

“…The direct radical scavenging activity was evaluated by the DPPH assay and the results are reported in Table 1 and expressed as IC 50 . The lower the value obtained, the higher the direct radical scavenging activity.…”

The Effect of Carnosine on UVA-Induced Changes in Intracellular Signaling of Human Skin Fibroblast Spheroids

“…Following 72 h from transfer, the treated spheroid series were pre-incubated with 150 µM L-carnosine, previously observed as an efficacy dose to protect the dermis induced by aging [7], and added to the culture medium for 4 h. After pre-treatment, UVA series were prepared for UV exposure: the culture media with/or without treatment were discarded and DPBS1X was added to all tissues in order to avoid interference with UV exposure. ORA ® dermis spheroids were irradiated with UVA at a working distance of 12 cm, at 30 J/cm 2 (Xenon Lamp Oriel Solar Simulator, Filter WG 335 3 mm) aiming to induce a perturbation of protein assessment in scaffold-free dermis spheroids; the dose was defined considering the complex physiology of the model and according to internal validated data [10].…”

“…To date, several natural compounds endowed with antioxidants and carbonyl scavenger properties have been used to protect the skin against UV radiation [5,6]. Among them, carnosine, an endogenous β-alanyl-L-histidine dipeptide, has shown promising results in the prevention of Oxidative Stress modulation on the scaffold-free human dermis spheroids [7] and on UVA-irradiated nude mice skin [8]. Carnosine pretreatment in 3D-aged fibroblast resulted in modulation of important pathways such as free radical scavenging, apoptosis, mitochondrial function and ECM reorganization.…”