Enhanced Membrane Permeability Research Articles

Therapeutic Potential of Two Visible Light Responsive Luminescent photoCORMs: Enhanced Cellular Internalization Driven by Lipophilicity

Herein we report the synthesis, characterization, and cellular internalization properties of two visible-light active luminescent Mn-based photoCORMs. The enhanced membrane permeability of the photoactive Mn carbonyl complex (photoCORM) derived from a designed lipophilic ligand namely, [Mn(CO)3(Imdansyl)(L1)](CF3SO3) (1) (where L1 = a diazabutadiene-based ligand containing two highly lipophilic adamantyl motifs, Imdansyl = dansylimidazole) promoted rapid internalization within human colorectal adenocarcinoma (HT-29) cells compared to [Mn(CO)3(Imdansyl)(L2)](CF3SO3) (2) (where L2 = a diazabutadiene ligand bearing two hydrophilic 1,3,5-triazaadamantyl group). Colocalization experiments using membrane stain indicate different extents of localization of the two CO complexes within the cellular matrix. Visible-light triggered CO release from the lipophilic photoCORM induced caspase-3/7 activation on HT-29 cells, which was detected using confocal microscopy. The rapid accumulation of the lipophilic photoCORM 1 in the cellular membrane resulted in more efficient CO-induced cell death compared to the hydrophilic analogue 2.

Next-generation unnatural monosaccharides reveal that ESRRB O-GlcNAcylation regulates pluripotency of mouse embryonic stem cells

Unnatural monosaccharides such as azidosugars that can be metabolically incorporated into cellular glycans are currently used as a major tool for glycan imaging and glycoproteomic profiling. As a common practice to enhance membrane permeability and cellular uptake, the unnatural sugars are per-O-acetylated, which, however, can induce a long-overlooked side reaction, non-enzymatic S-glycosylation. Herein, we develop 1,3-di-esterified N-azidoacetylgalactosamine (GalNAz) as next-generation chemical reporters for metabolic glycan labeling. Both 1,3-di-O-acetylated GalNAz (1,3-Ac2GalNAz) and 1,3-di-O-propionylated GalNAz (1,3-Pr2GalNAz) exhibit high efficiency for labeling protein O-GlcNAcylation with no artificial S-glycosylation. Applying 1,3-Pr2GalNAz in mouse embryonic stem cells (mESCs), we identify ESRRB, a critical transcription factor for pluripotency, as an O-GlcNAcylated protein. We show that ESRRB O-GlcNAcylation is important for mESC self-renewal and pluripotency. Mechanistically, ESRRB is O-GlcNAcylated by O-GlcNAc transferase at serine 25, which stabilizes ESRRB, promotes its transcription activity and facilitates its interactions with two master pluripotency regulators, OCT4 and NANOG.

Modulation of membrane permeability by carbon dioxide.

Promoting drug delivery across the biological membrane is a common strategy to improve bioavailability. Inspired by the observation that carbonated alcoholic beverages can increase the absorption rate of ethanol, we speculate that carbon dioxide (CO2 ) molecules could also enhance membrane permeability to drugs. In the present work, we have investigated the effect of CO2 on the permeability of a model membrane formed by 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine lipids to three drug-like molecules, namely, ethanol, 2',3'-dideoxyadenosine, and trimethoprim. The free-energy and fractional-diffusivity profiles underlying membrane translocation were obtained from μs-timescale simulations and combined in the framework of the fractional solubility-diffusion model. We find that addition of CO2 in the lipid environment results in an increase of the membrane permeability to the three substrates. Further analysis of the permeation events reveals that CO2 expands and loosens the membrane, which, in turn, facilitates permeation of the drug-like molecules. © 2019 Wiley Periodicals, Inc.

The impact and mechanism of quaternary ammonium compounds on the transmission of antibiotic resistance genes.

The emergence of antibiotic resistance genes (ARGs) in microbes can be largely attributed to the abuse and misuse of antibiotics and biocides. Quaternary ammonium compounds (QACs) have been used worldwide as common disinfectants and detergents; however, their potential impact on the spread and diffusion of ARGs is still unknown. In this study, we detected the QAC resistance gene (qacEΔ1), the 1 integron gene (intI1), and 12 ARGs (sul1, sul2, cfr, cml, fexA, tetA, tetG, tetQ, tetX, ermB, blaTEM, and dfrA1) in 48 water samples from three watersheds by quantitative PCR (qPCR). We investigated the evolution of bacterial antibiotic resistance under QAC and antibiotic environmental pressures by long-term continuous culture. In addition, five QACs were selected to investigate the effect of QAC on the efficiency of conjugation transfer. The changes in bacterial cell membrane and production of reactive oxygen species (ROS) were detected by flow cytometry, revealing the mechanism by which QAC affects the spread of antibiotic resistance. Our results showed that the QAC resistance gene was ubiquitous in watersheds and it had significant correlation with intI1 and seven ARGs (r = 0.999, p < 0.01). QACs could increase the resistance of bacteria to multiple antibiotics. Furthermore, all five QACs promoted the conjugation transfer of the RP4 plasmid; the optimal concentration of QACs was about 10-1-10-2mg/L and their transfer efficiencies were between 1.33 × 10-6 and 8.87 × 10-5. QACs enhanced membrane permeability of bacterial cells and stimulated bacteria to produce ROS, which potentially promoted the transfer of plasmids between bacteria. In conclusion, this study demonstrated that QACs may facilitate the evolution and gene transfer of antibiotic resistance gene among microbiome.

Metabolomic analysis of quorum sensing inhibitor hordenine on Pseudomonas aeruginosa.

Proton magnetic resonance-based metabolomics analysis was performed to determine the global metabolite changes in pathogenic bacterium Pseudomonas aeruginosa PAO1 following exposure to quorum sensing (QS) inhibitor hordenine. Pyocyanin inhibition assay confirmed that hordenine exhibited potent QS inhibitory activity. A total of 40 metabolites were assigned by PMR spectra. Hordenine treatment resulted in the destruction of QS system in P. aeruginosa PAO1 by downregulating the expressions of genes involved in QS. The synthesis of antioxidant enzymes was repressed and the oxidative stress was enhanced due to the dysfunctional QS system of P. aeruginosa PAO1. The enhanced oxidative stress induced by the dysfunctional QS system of P. aeruginosa PAO1 altered the membrane components, enhanced membrane permeability, and disturbed energy metabolism, amino acid metabolism, and nucleotide metabolism, and would ultimately attenuate the pathogenicity of P. aeruginosa PAO1. Hordenine may have promising potential for controlling nosocomial pathogens.

An Atomistic Simulation Study on POC/PIM Mixed-Matrix Membranes for Gas Separation

Although high flux membranes are indispensable to membrane-based gas separation technologies, enhancing membrane permeability proves to be difficult due to the trade-off between permeability and ga...

Bacterial membrane destabilization with cationic particles of nano-silver to combat efflux-mediated antibiotic resistance in Gram-negative bacteria

AimsWith the purpose of exploring combinatorial options that could enhance the bactericide efficacy of linezolid against Gram-negative bacteria, we assessed the extent of combination of nano-silver and linezolid. Main methodsIn this study, we selected Escherichia coli MTCC 443 as a model to study the combinatorial effect of nano-silver and linezolid to combat efflux-mediated resistance in Gram-negative bacteria. The acting mechanism of nano-silver on E. coli MTCC 443 was investigated by evaluating interaction of nano-silver with bacterial membrane as well as bacterial surface charge, morphology, intracellular leakages and biological activities of membrane bound respiratory chain dehydrogenase and deoxyribonucleic acids (DNA) of the cells following treatment with nano-silver. Key findingsThe alternation of zeta potential due to the interaction of nano-silver towards bacterial membrane proteins was correlated with enhancement of membrane permeability, which allows the penetration of linezolid into the cells. In addition, the binding affinity of nano-silver towards bacterial membrane depressed biological activities of membrane bound respiratory chain dehydrogenases and DNA integrity. SignificanceOur findings suggested that nano-silver could not only obstruct the activities of efflux pumps, but also altered membrane integrity at the same time and thus increased the cytoplasmic concentration of the linezolid to the effective level.

Activity and characterization of a pH-sensitive antimicrobial peptide

Antimicrobial peptides (AMPs) have been an area of great interest, due to the high selectivity of these molecules toward bacterial targets over host cells and the limited development of bacterial resistance to these molecules throughout evolution. Previous work showed that when Histidine was incorporated into the peptide C18G it lost antimicrobial activity. The role of pH on activity and biophysical properties of the peptide was investigated to explain this phenomenon. Minimal inhibitory concentration (MIC) results demonstrated that decreased media pH increased antimicrobial activity. Trichloroethanol (TCE) quenching and red-edge excitation spectroscopy (REES) showed a clear pH dependence on peptide aggregation in solution. Trp fluorescence was used to monitor binding to lipid vesicles and demonstrated the peptide binds to anionic bilayers at all pH values tested, however, binding to zwitterionic bilayers was enhanced at pH 7 and 8 (above the His pKa). Dual Quencher Analysis (DQA) confirmed the peptide inserted more deeply in PC:PG and PE:PG membranes, but could insert into PC bilayers at pH conditions above the His pKa. Bacterial membrane permeabilization assays which showed enhanced membrane permeabilization at pH 5 and 6 but vesicle leakage assays indicate enhanced permeabilization of PC and PC:PG bilayers at neutral pH. The results indicate the ionization of the His side chain affects the aggregation state of the peptide in solution and the conformation the peptide adopts when bound to bilayers, but there are likely more subtle influences of lipid composition and properties that impact the ability of the peptide to form pores in membranes.

Hybrids made from antimicrobial peptides with different mechanisms of action show enhanced membrane permeabilization

Combining two known antimicrobial peptides (AMPs) into a hybrid peptide is one promising avenue in the design of agents with increased antibacterial activity. However, very few previous studies have considered the effect of creating a hybrid from one AMP that permeabilizes membranes and another AMP that acts intracellularly after translocating across the membrane. Moreover, very few studies have systematically evaluated the order of parent peptides or the presence of linkers in the design of hybrid AMPs. Here, we use a combination of antibacterial measurements, cellular assays and semi-quantitative confocal microscopy to characterize the activity and mechanism for a library of sixteen hybrid peptides. These hybrids consist of permutations of two primarily membrane translocating peptides, buforin II and DesHDAP1, and two primarily membrane permeabilizing peptides, magainin 2 and parasin. For all hybrids, the permeabilizing peptide appeared to dominate the mechanism, with hybrids primarily killing bacteria through membrane permeabilization. We also observed increased hybrid activity when the permeabilizing parent peptide was placed at the N-terminus. Activity data also highlighted the potential value of considering AMP cocktails in addition to hybrid peptides. Together, these observations will guide future design efforts aiming to design more active hybrid AMPs.

Mechanisms of Fluoroquinolone and Aminoglycoside Resistance in Keratitis-Associated Pseudomonas aeruginosa.

Global emergence of multidrug resistant (MDR) strains limits therapeutic efficacy in Pseudomonas aeruginosa corneal ulcers. Identifying the primary causal factors of resistance shall improve clinical management. In this study, we sought to identify the underlying mechanisms of fluoroquinolone and aminoglycoside resistance in MDR, non-MDR, and drug susceptible P. aeruginosa (n = 19) strains obtained from keratitis patients. Phenotypic assays were performed to study the bacterial growth kinetics, efflux, permeability, and biofilm formation. Mutational alteration of target genes (DNA sequencing), relative expression of efflux system genes (real time PCR), and detection of aminoglycoside modifying enzyme (AME) genes (PCR) were done by molecular methods. We repeatedly found the mutations in quinolone resistance determining region of fluoroquinolone target genes, gyrA and parC, and the presence of AME genes, aph(3″)-I and aph(6)-I, in all MDR isolates. Furthermore, the MDR isolates were largely characterized by slower growth, cytotoxic type III secretion system genotype, better biofilm-forming ability, and the presence of additional AME genes. The non-MDR isolates were resensitized upon inhibition of active efflux or enhancement of membrane permeability. Altogether this study highlights target gene alteration and enzymatic drug modification as the major mechanisms of quinolone and aminoglycoside resistance in P. aeruginosa keratitis isolates.

Cu-ATCUN Derivatives of Sub5 Exhibit Enhanced Antimicrobial Activity via Multiple Modes of Action.

Antimicrobial peptides (AMPs) are short, amphipathic peptides that are typically cationic in sequence and display broad-spectrum activity against bacteria, fungi, and protists. Herein, we report the effect of appending the amino terminal copper and nickel binding motif (ATCUN) to Sub5. The Cu-ATCUN derivatives show a two- to three-fold increase in antimicrobial activity for a variety of microbes, relative to Sub5, with MICs as low as 0.3 ± 0.1 μM toward Enterococcus faecium. Sub5 and the ATCUN derivatives bind both plasmid DNA and 16s A-site rRNA with low micromolar affinity. Native Sub5 and the metallopeptide derivatives were shown to promote damage against DNA to similar extents in cellular studies against both Escherichia coli and Staphylococcus epidermidis, with an almost threefold higher activity against the latter organism. Liposome experiments show that the metallopeptides have a greater affinity for model membranes of E. coli and S. aureus relative to Sub5, which correlates with their enhanced antimicrobial activity. Sub5 and the metalloderivatives also display no cytotoxicity toward adult human dermal fibroblasts. Addition of the ATCUN motif conferred the ability to promote lipid oxidation toward E. coli and S. epidermidis and enhanced membrane permeability, as evidenced by the extent of ATP leaked from cellular membranes relative to Sub5 alone. These data suggest that Cu-ATCUN derivatives inhibit microbes through multiple modes of action, resulting in an enhancement in their overall potency.

Solubility and Permeability Improvement of Quercetin by an Interaction Between α-Glucosyl Stevia Nanoaggregates and Hydrophilic Polymer

The effect of composite formation between α-glucosyl stevia (Stevia-G) and hydrophilic polymers on solubility and permeability enhancement of quercetin hydrate (QUE) was evaluated. Polyvinylpyrrolidone K-30 (PVP), hydroxypropyl methylcellulose 2910-E (HPMC), and hydroxypropyl cellulose SSL (HPC) were selected as candidate hydrophilic polymers. Fluorescence studies with pyrene and curcumin suggested composite formation occurs between Stevia-G aggregate and polymers. Furthermore, the strength of interaction between Stevia-G aggregate and polymers was as follows: PVP > HPMC > HPC. Evaporated particles (EVPs) of QUE with Stevia-G and polymers showed synergic QUE solubility enhancement. Solubility of QUE from the EVPs was enhanced in the following order: Stevia-G/PVP > Stevia-G/HPMC > Stevia-G/HPC, in accordance with the degree of interaction. Enhanced membrane permeability of QUE from the EVPs of Stevia-G/PVP was confirmed using Caco-2 cells. The amount of QUE that permeated Caco-2 cells from the EVPs of Stevia-G/PVP was 13.7-, 4.7-, and 2.1-fold higher than that of the untreated QUE powder, EVPs of Stevia-G, and EVPs of PVP, respectively. These results indicated that the composite formed by Stevia-G and PVP can dramatically enhance the solubility and membrane permeability of QUE.

Emerging Peptide Science in Canada

Emerging Peptide Science in Canada

An acoustic strategy for gold nanoparticle loading in platelets as biomimetic multifunctional carriers.

In recent years, a wide variety of bioinspired colloidal particles with novel cell mimetic functions have been the subject of extensive research in materials science, chemistry, biology, physics, and engineering. However, most of the approaches are derived from natural cell membrane coatings, which are still too primitive compared with living cells. In this study, we have chosen gold nanoparticles (GNPs) to explore the bioactivity response of living platelets and nanoparticle loading efficiency under different ultrasonic intensity and frequency treatment conditions. The results show that GNPs with no surface modification could be easily loaded into intra-platelets by both incubation (30 min) and ultrasonic exposure (1 min) methods. The amount of GNP loading was (4.4 ± 0.9) × 10-3 and (5.8 ± 2.4) × 10-3 pg per platelet upon incubation and acoustic triggering (1 MHz, 0.25 W cm-2), respectively. Although the other US treatment intensities (0.75, 1.50 and 2.25 W cm-2) also promoted higher amounts of GNPs in the platelets, the higher US intensity might bring about partial damage of the platelet membrane. Compared with 1 MHz ultrasonic exposure, the change of the GNP loading amount was not significantly higher upon ultrasonic frequency treatment of 45, 80 or 100 kHz. Therefore, it has been found that an US intensity of 0.25 W cm-2 could facilitate the intra-platelet delivery efficacy of the GNPs without damaging the biological activity. Furthermore, two possible pathways of GNPs entering into platelets upon US treatment are presented: one is the endocytosis/open canalicular system (OCS), and the other is cell membrane permeability enhancement, which is proved by the SEM and TEM results. Finally, the GNP-loaded platelets have been demonstrated as useful probes for photoacoustic imaging (PAI) and dark-field microscopy (DFM)-based imaging, which might allow a wide range of potential applications in diagnostics and therapy of platelet-related diseases.

Fermentative Production of N-Methylglutamate From Glycerol by Recombinant Pseudomonas putida.

N-methylated amino acids are present in diverse biological molecules in bacteria, archaea and eukaryotes. There is an increasing interest in this molecular class of alkylated amino acids by the pharmaceutical and chemical industries. N-alkylated amino acids have desired functions such as higher proteolytic stability, enhanced membrane permeability and longer peptide half-lives, which are important for the peptide-based drugs, the so-called peptidomimetics. Chemical synthesis of N-methylated amino acids often is limited by incomplete stereoselectivity, over-alkylation or the use of hazardous chemicals. Here, we describe metabolic engineering of Pseudomonas putida KT2440 for the fermentative production of N-methylglutamate from simple carbon sources and monomethylamine. P. putida KT2440, which is generally recognized as safe and grows with glucose and the alternative feedstock glycerol as sole carbon and energy source, was engineered for the production of N-methylglutamate using heterologous enzymes from Methylobacterium extorquens. About 3.9 g L−1 N-methylglutamate accumulated within 48 h in shake flask cultures with minimal medium containing monomethylamine and glycerol. A fed-batch cultivation process yielded a N-methylglutamate titer of 17.9 g L−1.

Utilization of DES-Lignin as a Bio-Based Hydrophilicity Promoter in the Fabrication of Antioxidant Polyethersulfone Membranes.

Enhancement of membrane permeability at no detriment of its other performances, e.g., selectivity, is a goal-directed objective in membrane fabrication. A novel antioxidant DES-lignin (lignin extracted from birch wood by using a deep eutectic solvent) polyethersulfone (PES) membrane, containing 0–1 wt % DES-lignin, was fabricated with the phase inversion technique. The performance and morphology of the fabricated membranes were characterized by a pure water flux, polyethylene glycol (PEG) retention, Fourier transform infrared spectroscopy, scanning electron microscopy, and contact angle measurements. Membranes with less negative charge and better hydrophilicity were obtained when the DES-lignin content in the polymer solution was increased. With the highest dosage, the incorporation of DES-lignin in the membrane matrix improved the membrane permeability by 29.4% compared to a pure PES membrane. Moreover, no leakage of DES-lignin from the membrane structure was observed, indicating good compatibility of DES-lignin with the PES structure. It was also found that the improvement of both rejection and pure water flux could be achieved by using a small dosage of DES-lignin (0.25 wt %) in membrane fabrication. The membranes incorporated with DES-lignin showed higher DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)) scavenging activity compared to the pure membrane, where 2.6 and 1.1 times higher DPPH and ABTS scavenging activity was observed with the highest DES-lignin content (1 wt %). Thus, the results of this study demonstrate well the feasibility of utilizing DES-lignin as an antioxidant bio-based hydrophilicity promoter in the fabrication of ultrafiltration membranes.

Enhanced performance of polyhedral oligomeric silsesquioxanes /polysulfone nanocomposite membrane with improved permeability and antifouling properties for water treatment

Modification of polymeric membranes for generating membranes with enhanced permeability, rejection, and self-cleaning capability in water treatment has been the major goal of researchers in academia and industry recently. We report here a new polysulfone (PSF)/ polyhedral oligomeric silsesquioxanes (POSS) nanocomposite ultrafiltration membrane with enhanced permeability, rejection, and antifouling capability for treatment of feed water containing humic acid (HA) as a model foulant. The effects of incorporation of POSS nano-fillers on intrinsic properties of PSF membranes such as membrane surface charge, surface roughness, pore morphology, tortuosity, porosity, and consequently on membrane performance were evaluated. In general, the main effects of POSS nanoparticles at concentrations of less than 2% (wt.%) can be summarized as (1) increased hydrophilicity and negative electrical surface charge of the membranes which resulted in the enhanced self-cleaning and rejection properties and (2) a change in the membranes pores morphology towards formation of more pores on the top surface and interconnected long finger like pores in the membrane matrix which resulted in higher flux. The addition of 2% POSS into PSF membranes resulted in significantly enhanced membrane permeability, however reduced the rejection ability of the membranes. The concentration of 0.5% POSS was found to be the optimal concentration that increased the permeability as well as the antifouling capability of the membranes while the rejection capability of nanocomposite membranes for HA increased slightly as compared to pure PSF membranes.

Thermodynamics and kinetics of joint action of antiviral agent tilorone and DMSO on model lipid membranes

Individual and joint action of two water-soluble drugs, DMSO and tilorone, on model l-α-dipalmitoylphosphatidylcholine (DPPC) membranes were studied in equilibrium and kinetic regimes by differential scanning calorimetry (DSC). For equilibrium experiments, the drugs were introduced during preparation of the model membrane. In kinetic studies, one of the drugs was added to the DPPC membrane already containing the other drug, and the effects of drug-membrane interactions were monitored in real-time regime. It was found that tilorone and DMSO had opposite effects on the membrane melting temperature, which were non-additive under joint introduction of these drugs. Analysis of kinetics of DSC profiles under drugs introduction allowed us to discriminate two processes in drug-membrane interactions with different characteristic times, i.e., drug sorption onto the membrane (minutes) and drug diffusion through stacks of lipid bilayers (hours). It was established that 0.1 mol% DMSO effectively enhanced membrane penetration for tilorone with the rate of tilorone diffusion being dependent upon the scheme of drugs administration. A model was proposed describing how sorption of a dopant onto lipid membrane could affect the membrane permeability for other dopants. Conditions were determined for enhancement of membrane permeability, as it was observed for DPPC/DMSO/tilorone system.

Praziquantel-lipid nanocapsules: an oral nanotherapeutic with potential Schistosoma mansoni tegumental targeting.

PurposeLipid nanocapsules (LNCs) have shown potential to increase the bioavailability and efficacy of orally administered drugs. However, their intestinal translocation to distal target sites and their implication in pharmacokinetic (PK)–pharmacodynamic (PD) relationships are yet to be elucidated. In this study, the effect of LNCs on the PD activity and pharmacokinetics of praziquantel (PZQ), the mainstay of schistosomiasis chemotherapy, was investigated.Materials and methodsThe composition of LNCs was modified to increase PZQ payload and to enhance membrane permeability. PZQ–LNCs were characterized in vitro for colloidal properties, entrapment efficiency (EE%), and drug release. PD activity of the test formulations was assessed in Schistosoma mansoni-infected mice 7 days post-oral administration of a single 250 mg/kg oral dose. Pharmacokinetics of the test formulations and their stability in simulated gastrointestinal (GI) fluids were investigated to substantiate in vivo data.ResultsPZQ–LNCs exhibited good pharmaceutical attributes in terms of size (46–62 nm), polydispersity index (0.01–0.08), EE% (>95%), and sustained release profiles. Results indicated significant efficacy enhancement by reduction in worm burden, amelioration of liver pathology, and extensive damage to the fluke suckers and tegument. This was partly explained by PK data determined in rats. In addition, oral targeting of the worms was supported by the stability of PZQ–LNCs in simulated GI fluids and scanning electron microscopy (SEM) visualization of nanostructures on the tegument of worms recovered from mesenteric/hepatic veins. Cytotoxicity data indicated tolerability of PZQ–LNCs.ConclusionData obtained provide evidence for the ability of oral LNCs to target distal post-absorption sites, leading to enhanced drug efficacy. From a practical standpoint, PZQ–LNCs could be suggested as a potential tolerable single lower dose oral nanomedicine for more effective PZQ mass chemotherapy.

Differences in rutin effect on membrane phospholipids in skin fibroblasts irradiated with UVA and UVB

Chronic exposure of the skin to solar UV radiation induces a number of biological alterations, including a redox imbalance; therefore, there is a constant need for protective compounds, particularly natural antioxidants. The aim of this study was to determine the effect of rutin on interactions between membrane phospholipid and antioxidant proteins expression in UVA or UVB irradiated fibroblasts. Following exposure to UVA and UVB irradiation cells were incubated with rutin 12h before and/or up to 24h after irradiation, and the structural and metabolic changes were examined at selected time intervals. Rutin penetration through the fibroblast phospholipid bilayer was aided by UVA-induced bilitranslocase activity, while lipidomic analysis revealed that following UVB-irradiation rutin transport into cell is enhanced by changes in membrane permeability resulting from UVB-induced lipid peroxidation. Moreover, rutin partially prevented UVA/B-induced increase in phosphatidylethanolamine and phosphatidylcholine and decrease in sphingomyelin, as well as their membrane localization. However, the inhibition of phospholipase A2 activity and increase in ROS level resulted in protection against the reduction of phospholipids/free arachidonic and linoleic acids and the increase in the level of the lipid peroxidation product 4-HNE, which demonstrates the antioxidant proteins regulatory properties. The antioxidant activity of rutin, effectively prevented the enhanced ROS generation as well as antioxidant system destruction. Proteome analysis show that rutin treatment more strongly protects against UVA-induced rather than UVB-induced increases in the total expression of proteins involved in antioxidant response (such as SOD, TrxR, and Prxs 1/2). However, in the case of UVB-irradiated cells, rutin additionally enhances the levels of disulfide-isomerase – an enzyme that is responsible for the formation and breakage of disulfide bonds, what in the case of Nrf2/ARE pathway has significant meaning in the Nrf2 transcriptional activity level and leads to the synthesis of cytoprotective proteins. In conclusion, UVA and UVB radiation in partially different ways affect rutin interactions with the fibroblast biomembrane lipids as well as antioxidant proteins. Rutin membrane penetration is promoted by UVA-induced bilitranslocase activity, while UVB irradiation enhanced membrane permeability to facilitate the interaction of rutin with phospholipids. Moreover, rutin particularly influences UVA-induced antioxidant proteins expression and UVB-induced transcription signaling.