Local Reactive Oxygen Species Research Articles

A theory of mechanical stress-induced H2O2 signaling waveforms in Planta

Recent progress in nanotechnology-enabled sensors that can be placed inside of living plants has shown that it is possible to relay and record real-time chemical signaling stimulated by various abiotic and biotic stresses. The mathematical form of the resulting local reactive oxygen species (ROS) wave released upon mechanical perturbation of plant leaves appears to be conserved across a large number of species, and produces a distinct waveform from other stresses including light, heat and pathogen-associated molecular pattern (PAMP)-induced stresses. Herein, we develop a quantitative theory of the local ROS signaling waveform resulting from mechanical stress in planta. We show that nonlinear, autocatalytic production and Fickian diffusion of H2O2 followed by first order decay well describes the spatial and temporal properties of the waveform. The reaction-diffusion system is analyzed in terms of a new approximate solution that we introduce for such problems based on a single term logistic function ansatz. The theory is able to describe experimental ROS waveforms and degradation dynamics such that species-dependent dimensionless wave velocities are revealed, corresponding to subtle changes in higher moments of the waveform through an apparently conserved signaling mechanism overall. This theory has utility in potentially decoding other stress signaling waveforms for light, heat and PAMP-induced stresses that are similarly under investigation. The approximate solution may also find use in applied agricultural sensing, facilitating the connection between measured waveform and plant physiology.

Reversible Regulation of the Reactive Oxygen Species Level Using a Semiconductor Heterojunction.

Here, we proposed a novel solution for reversible regulation of the reactive oxygen species (ROS) level using a semiconductor heterojunction. Two metal-based ROS scavengers containing n-type CeO2 nanoparticles and n-type Cu-doped diatom biosilica (Cu-DBs) were integrated by a hydrothermal method to form a typical n-n semiconductor heterojunction (Ce/Cu-DBs). Unlike the control of the ROS level by a single ROS scavenger or ROS-generating agent, Ce/Cu-DBs could quickly eliminate ROS by cascade catalytic reaction, which readily switched to ROS generation through a near-infrared (NIR)-triggered photocatalytic effect. This NIR mediated ROS regulation system provided a noninvasive strategy for reversible control of the ROS level in vitro and in vivo. The Ce/Cu-DBs could relieve cellular oxidative stress by clearing local excessive ROS while inhibiting bacterial growth by increasing ROS levels under NIR radiation. Benefiting from the reversible regulatory effect of Ce/Cu-DBs, programmable healing of infected wounds was realized via on-demand anti-infection and inflammation reduction. This work provided a general method with highly spatiotemporal resolution to a remote and sustainable control ROS level, which had great potential for the biomedical field and regulation of chemical reactions.

Logic-Based Diagnostic and Therapeutic Nanoplatform with Infection and Inflammation Monitoring and Microenvironmental Regulation Accelerating Wound Repair

Infectious cutaneous wounds are a thorny clinical problem. The microenvironment of the infectious wound is complicated and changes at different healing stages. Traditional treatments either have a single effect such as anti-inflammation, antibacteria, or angiogenesis or a simple mixture of several functions. They fail to deal with the change of the physiological healing process, leading to unsatisfactory outcomes. Herein, we have designed a logic-based smart nanoplatform (named as ZEM), aiming to self-monitor the wound microenvironment and accordingly react to the changes of the healing process, fitting multiple needs of physiological repair at different stages. ZEM was synthesized using zeolitic imidazolate framework-8 (ZIF-8) coated with an epigallocatechin gallate (EGCG)/Mg2+ complex. We characterized ZEM in the aspects of morphology, physical and chemical properties, and ion release pattern. At the initial stage, ZEM sensed the weakly acidic environment and responsively released a large number of zinc ions to eliminate bacterial infection. Then came the second inflammation stage, where ZEM responded to the oxidative stress of the local wound area with EGCG absorbing excessive reactive oxygen species (ROS), contributing to the downregulation of intracellular ROS. Meanwhile, local inflammation was alleviated by reducing the expression of proinflammatory M1 phenotype factors (IL-6, TNF-α, and IL-1β). Since the balance of local ROS had been achieved, the resulting disintegration of the EGCG/Mg2+ complex gave rise to the sustainable release of Mg2+ at the proliferation stage, promoting vascularized healing. In vivo animal experiments further proved the diagnostic and therapeutic functions of ZEM. All these results demonstrated that ZEM was a promising treatment strategy in soft tissue engineering.

ROS responsive nanoparticles loaded with lipid-specific AIEgen for atherosclerosis-targeted diagnosis and bifunctional therapy

Atherosclerosis, which is triggered by endothelial damage, progressive local inflammation and excessive lipid accumulation, is one of the most common cardiovascular diseases in recent years. Drug delivery systems have shown great potential for the accurate diagnosis and effective treatment of early atherosclerosis, but are accompanied by disadvantages such as poor stability, lack of active targeting and non-specific recognition capabilities, which still need to be further developed. In our work, a multifunctional nanoparticle (LFP/PCDPD) with reactive oxygen species (ROS) responsive drug release, lipid removal, and lipid-specific AIE fluorescence imaging was constructed. Cyclodextrin structure with lipid removal function and PMEMA blocks with ROS-response-mediated hydrophobic to hydrophilic conversion were simultaneously introduced into the structure of LFP/PCDPD to load the anti-inflammatory drug prednisolone (Pred) and lipid-specific AIEgen (LFP). The active targeting function of LFP/PCDPD was conferred by the high affinity of dextran to the vascular adhesion molecule-1 (VCAM-1) and CD44 receptor on the surface of broken endothelial cells. After intravenous injection into ApoE−/− mice, LFP/PCDPD actively enriched in the microenvironment of local ROS overexpression and rich lipids in atherosclerosis. Pred and LFP were released while lipids were removed, thus enabling proactive targeting of atherosclerosis and efficient “two-pronged” treatment.

A reversible mitochondrial complex I thiol switch mediates hypoxic avoidance behavior in C. elegans

C. elegans react to metabolic distress caused by mismatches in oxygen and energy status via distinct behavioral responses. At the molecular level, these responses are coordinated by under-characterized, redox-sensitive processes, thought to initiate in mitochondria. Complex I of the electron transport chain is a major site of reactive oxygen species (ROS) production and is canonically associated with oxidative damage following hypoxic exposure. Here, we use a combination of optogenetics and CRISPR/Cas9-mediated genome editing to exert spatiotemporal control over ROS production. We demonstrate a photo-locomotory remodeling of avoidance behavior by local ROS production due to the reversible oxidation of a single thiol on the complex I subunit NDUF-2.1. Reversible thiol oxidation at this site is necessary and sufficient for the behavioral response to hypoxia, does not respond to ROS produced at more distal sites, and protects against lethal hypoxic exposure. Molecular modeling suggests that oxidation at this thiol residue alters the ability for NDUF-2.1 to coordinate electron transfer to coenzyme Q by destabilizing the Q-binding pocket, causing decreased complex I activity. Overall, site-specific ROS production regulates behavioral responses and these findings provide a mechanistic target to suppress the detrimental effects of hypoxia.

Sorafenib-loaded silica-containing redox nanoparticles for oral anti-liver fibrosis therapy.

Liver fibrosis is a chronic disease resulting from repetitive or prolonged liver injury with limited treatment options. Sorafenib has been reported to be a potential antifibrotic agent; however, its therapeutic effect is restricted because of its low bioavailability and severe adverse effects in the gastrointestinal (GI) tract. In this study, we developed sorafenib-loaded silica-containing redox nanoparticles (sora@siRNP) as an oral nanomedicine to treat liver fibrosis. The designed siRNP were prepared by self-assembly of amphiphilic block copolymers, which possess antioxidant nitroxide radicals as a side chain of the hydrophobic segment and porous silica particles in the nanoparticle core. The silica moieties in the core formed a crosslink between the self-assembling block copolymers to afford stable drug absorption, which could be useful in harsh GI conditions after oral drug administration. Based on in vitro evaluation, sora@siRNP exerted antiproliferative and antifibrotic effects against hepatic stellate cells (HSCs) and low toxicity against normal endothelial cells. A pharmacokinetic study showed that siRNP significantly improved the bioavailability and distribution of sorafenib in the liver. In an in vivo study using a mouse model of CCl4-induced liver fibrosis, oral administration of sora@siRNP significantly suppressed the fibrotic area in comparison to free sorafenib administration. In mice with CCl4-induced fibrosis, free sorafenib administration did not suppress the expression of α-smooth muscle actin; however, mice treated with sora@siRNP showed significantly suppressed expression of α-smooth muscle actin, indicating the inhibition of HSC activation, which was confirmed by in vitro experiments. Moreover, oral administration of free sorafenib induced severe intestinal damage and increased leakage into the gut, which can be attributed to the generation of reactive oxygen species (ROS). Our antioxidant nanocarriers, siRNP, reduced the adverse effects of local ROS scavenging in the GI tract. Our results suggest that sora@siRNP could serve as a promising oral nanomedicine for liver fibrosis.

Multifunctional SGQDs-CORM@HA nanosheets for bacterial eradication through cascade-activated “nanoknife” effect and photodynamic/CO gas therapy

Infection associated with multidrug-resistant (MDR) bacteria has become a serious threat to public health, and there is an urgent demand of developing new antibiotics that offer combinatorial therapy to effectively combat MDR. Herein, a multifunctional two-dimensional nanoantibiotic was facilely designed and established on the basis of the covalent conjugation of CO-releasing molecule (CORM-401) and electrostatic adsorption of hyaluronic acid (HA) onto single-layered graphene quantum dots (SGQDs) to assemble SGQDs-CORM@HA nanosheets, abbreviated as SCH. Upon the enrichment of as-prepared nanoantibiotics in the community of targeted microbe, bacterial-secreted hyaluronidase (HAase) would cleave HA on SCH, and the sharp edges as well as the reactive sites on SGQDs-CORM nanosheets were exposed for cascade activation of synergistic antibacterial effects. Specifically, ultrathin SGQDs-CORM nanosheets can penetrate into bacterial cells deemed as the unique “nanoknife” effect. Under white light irradiation, SGQDs-CORM nanosheets can act as a high-efficient photosensitizer to generate cytotoxic singlet oxygen (1O2), as a well-recognized reactive oxygen species (ROS), to produce high oxidative stress and damage bacteria. As a complementary to photodynamic therapy (PDT), the accumulation of local ROS further triggered the release of CO to hinder the bacterial growth via causing plasma membrane damage and inducing metabolic disorders. Such synergistic treatment regimen arising from cascade-activated “nanoknife” effect and photodynamic/CO gas therapy, was devoted to outstanding on-demand antibacterial performance both in vitro and in vivo. Fascinatingly, the nanoplatform showed good biocompatibility toward both normal somatic cells and non-targeted bacteria. The remarkable antibacterial capability and admirable biocompatibility endow SCH with great potential to fight against MDR pathogens for in-coming clinical translations.

Homology and Immune Checkpoint Dual-Targeted Sonocatalytic Nanoagents for Enhancing Sonodynamic Tumor Therapy.

Sonocatalytic nanoagents (SCNs), a kind of sonosensitizers, could catalyze oxygen to generate abundant reactive oxygen species (ROS) under stimulations of noninvasive and deep-penetrating ultrasound (US), which is commonly used for sonodynamic therapy (SDT) of tumors such as malignant melanoma. However, poor bioavailability of most SCNs and fast quenching of extracellular-generating ROS from SDT limit further applications of SCNs in the SDT of tumors. Herein, we synthesized a new kind of TiO2-based SCN functionalized with the malignant melanoma cell membrane (B16F10M) and programmed cell death-ligand 1 antibody (aPD-L1) for homology and immune checkpoint dual-targeted and enhanced sonodynamic tumor therapy. Under US irradiation, the synthesized SCN can catalytically generate a large amount of 1O2. In vitro experiments validate that functionalized SCNs exhibit precise targeting effects, high tumor cell uptake, and intracellular sonocatalytic killing of the B16F10 cells by a large amount of localized ROS. Utilizing the melanoma animal model, the functionalized SCN displays visible long-term retention in the tumor area, which assists the homology and immune checkpoint synergistically dual-targeted and enhanced in vivo SDT of the tumor. We suggest that this highly bioavailable and dual-functionalized SCN may provide a promising strategy and nanoplatform for enhancing sonodynamic tumor therapies.

Photosynthesis-independent production of reactive oxygen species in the rice bundle sheath during high light is mediated by NADPH oxidase

When exposed to high light, plants produce reactive oxygen species (ROS). In Arabidopsis thaliana, local stress such as excess heat or light initiates a systemic ROS wave in phloem and xylem cells dependent on NADPH oxidase/respiratory burst oxidase homolog (RBOH) proteins. In the case of excess light, although the initial local accumulation of ROS preferentially takes place in bundle-sheath strands, little is known about how this response takes place. Using rice and the ROS probes diaminobenzidine and 2',7'-dichlorodihydrofluorescein diacetate, we found that, after exposure to high light, ROS were produced more rapidly in bundle-sheath strands than mesophyll cells. This response was not affected either by CO2 supply or photorespiration. Consistent with these findings, deep sequencing of messenger RNA (mRNA) isolated from mesophyll or bundle-sheath strands indicated balanced accumulation of transcripts encoding all major components of the photosynthetic apparatus. However, transcripts encoding several isoforms of the superoxide/H2O2-producing enzyme NADPH oxidase were more abundant in bundle-sheath strands than mesophyll cells. ROS production in bundle-sheath strands was decreased in mutant alleles of the bundle-sheath strand preferential isoform of OsRBOHA and increased when it was overexpressed. Despite the plethora of pathways able to generate ROS in response to excess light, NADPH oxidase-mediated accumulation of ROS in the rice bundle-sheath strand was detected in etiolated leaves lacking chlorophyll. We conclude that photosynthesis is not necessary for the local ROS response to high light but is in part mediated by NADPH oxidase activity.

Red Phosphorus Decorated TiO2 Nanorod Mediated Photodynamic and Photothermal Therapy for Renal Cell Carcinoma.

Clear cell renal cell carcinoma (ccRCC) is a serious and tenacious disease. Photodynamic therapy (PDT) and photothermal therapy (PTT) are effective means of cancer treatment. However, PDT combined with PTT has been rarely reported in ccRCC treatment. In the present study, by developing the core-shell structured TiO2 @red phosphorus nanorods (TiO2 @RP NRs) as a photosensitizer, the feasibility and effectiveness of synchronous PDT and PTT treatments for ccRCC are demonstrated. The core-shell structured TiO2 @RP NRs are synthesized to drive the PDT and PTT for ccRCC, in which the RP shell is the sensitizer even in the near-infrared (NIR) region. The optimized TiO2 @RP NRs can respond to NIR and produce local heat under irradiation. The NRs are estimated in ccRCC treatments via cell counting kit-8 assay, propidium iodide staining, qRT-PCR, and reactive oxygen species (ROS) probes in vitro, while terminal deoxynucleotidyl transferase dUTP nick-end labeling is conducted in vivo. After NIR irradiation, TiO2 @RP NRs can efficiently kill ccRCC cells by producing local heat and ROS and cause low injury to normal kidney cells. Furthermore, treatment with TiO2 @RP NRs and NIR can kill significant numbers of deep-tissue ccRCC cells in vivo. This work highlights a promising photo-driven therapy for kidney cancer.

Constructing biocompatible MSN@Ce@PEG nanoplatform for enhancing regenerative capability of stem cell via ROS-scavenging in periodontitis

The excessive local reactive oxygen species (ROS) level of periodontitis results in the quantity and function decline of periodontal ligament stem cells (PDLSCs), which aggravates the destruction and impedes the restoration of periodontal tissue. Therefore, regulating and restoring local ROS levels may promote stem cell survival and function recovery, improving periodontal tissue regeneration. Thus, protecting PDLSCs from oxidative stress injury and promoting regenerative capacity are both important. In this study, we synthesized a simple multi-functional nanocomposite by loading ceria oxide (CeO2) onto the surface of mesoporous silica (MSN@Ce), and then modified them with PEG (MSN@Ce@PEG) for better dispersion and biocompatibility, achieving simultaneous intracellular ROS-regulation and osteogenic differentiation promotion for PDLSCs against oxidative stress in periodontitis. We investigated the in-vitro regulatory effects of MSN@Ce@PEG NPs in hPDLSCs with H2O2-induced oxidative stress injury and the in-vivo therapy effects in a rat model of oxidative stress-induced periodontal disease, as well as the potential biotoxicity. Results have demonstrated that MSN@Ce@PEG NPs have good biocompatibility, and also showed MSN@Ce@PEG NPs not only efficiently regulated intracellular ROS, which protected cells from aging and improved the differentiative capacity of hPDLSCs, but also prevented inflammatory destruction of periodontal tissue. MSN@Ce@PEG NPs have great potential in ROS-regulation for safe and efficient periodontal therapy and are expected to be applied in other biomedical fields associated with redox metabolism imbalance.

Mechanism Insight into Rapid Photodriven Sterilization Based on Silver Bismuth Sulfide Quantum Dots.

Microbial contamination and the prevalence of resistant bacteria is considered a worldwide public health problem. Therefore, recently, great efforts have been made to develop photoresponsive platforms for the simultaneous photodynamic antibacterial (PDA) and photothermal antibacterial (PTA) therapy processes as mediated by specific light. However, owing to the absorption mismatches of the photothermal agents and photodynamic photosensitizers, it has been discovered that many synergistic photoresponsive antibacterial platforms cannot be excited by a single-wavelength light. In this study, silver bismuth sulfide quantum dots (AgBiS2 QDs) identified from the literature as a near-infrared light (NIR) that triggers bifunctional materials with simultaneous photodynamic and photothermal effects for photoresponsive bacterial killing were used. Specifically, AgBiS2 QDs were successfully synthesized via a bottom-up approach, using polyethylenimine (PEI) as an assistant molecule. With PEI wrapping, the attachment between the negatively charged membrane surfaces of the bacterial cells and AgBiS2 QDs was enhanced via the electrostatic interactions. The photodriven antibacterial activity of AgBiS2 QDs was then investigated against both S. aureus and E. coli. The results revealed a significant reduction in bacterial survival. The killing effect was found to be independent of the AgBiS2 QDs, and redox potentials controlled the photogenerated electrons that thermodynamically favored the formation of multiple reactive oxygen species (ROS). A possible phototriggered antibacterial mechanism was then proposed in which the AgBiS2 QDs are anchored first to the bacterial surface and then induce breaking on its outer membrane by high local heat and ROS under single 808 nm NIR laser illumination to finally induce bacterial death.

Copper ferrite heterojunction coatings empower polyetheretherketone implant with multi-modal bactericidal functions and boosted osteogenicity through synergistic photo/Fenton-therapy

Drug-free antibacterial therapy, such as photodynamic therapy and chemodynamic therapy, has become a mainstream in the treatment of implant-associated infection (IAI) due to its considerable potential on effective eradication of pathogenic bacteria via yielding overwhelming reactive oxygen species (ROS). However, superfluous ROS after therapeutical process may elicit a detrimental effect on subsequent tissue regeneration. To address this tough issue, we innovatively devise a photo-activated copper ferrite (CuFe2O4) heterojunction coating on bioinert polyetheretherketone (PEEK) implant through hydrothermal treatment and π-π-π stacking. The heterojunction coatings not only generate localized hyperthermia and ROS upon 808 nm NIR illumination, but also trigger Fenton(-like) reaction and glutathione (GSH) depletion in bacterial infectious microenvironment (IME), all leading to robust and cyclic antibacterial property through synergistic photothermal/photodynamic/chemodynamic therapy. Besides, the decorated implants are demonstrated to show superior cytocompatibility and osteogenic-promoting capability. The mice subcutaneous implantation further confirms the excellent in vivo photo-disinfection. The advantage of the heterojunction coatings lies on its enhancement in tissue generation and osteogenesis after antibacterial dynamic therapy because the liberation of micronutrient element (Cu, Fe) is significantly augmented exposed to NIR light, showing photo-enhanced osteogenicity. As envisaged, this work provides a new strategy to empower PEEK implants with an integrative feature of bacteriostasis and osteogenicity to combat intractable IAI through synergistic photo/Fenton-therapy.

Chloroplast-derived photo-oxidative stress causes changes in H2O2 and EGSH in other subcellular compartments.

Metabolic fluctuations in chloroplasts and mitochondria can trigger retrograde signals to modify nuclear gene expression. Mobile signals likely to be involved are reactive oxygen species (ROS), which can operate protein redox switches by oxidation of specific cysteine residues. Redox buffers, such as the highly reduced glutathione pool, serve as reservoirs of reducing power for several ROS-scavenging and ROS-induced damage repair pathways. Formation of glutathione disulfide and a shift of the glutathione redox potential (EGSH) toward less negative values is considered as hallmark of several stress conditions. Here we used the herbicide methyl viologen (MV) to generate ROS locally in chloroplasts of intact Arabidopsis (Arabidopsis thaliana) seedlings and recorded dynamic changes in EGSH and H2O2 levels with the genetically encoded biosensors Grx1-roGFP2 (for EGSH) and roGFP2-Orp1 (for H2O2) targeted to chloroplasts, the cytosol, or mitochondria. Treatment of seedlings with MV caused rapid oxidation in chloroplasts and, subsequently, in the cytosol and mitochondria. MV-induced oxidation was significantly boosted by illumination with actinic light, and largely abolished by inhibitors of photosynthetic electron transport. MV also induced autonomous oxidation in the mitochondrial matrix in an electron transport chain activity-dependent manner that was milder than the oxidation triggered in chloroplasts by the combination of MV and light. In vivo redox biosensing resolves the spatiotemporal dynamics of compartmental responses to local ROS generation and provides a basis for understanding how compartment-specific redox dynamics might operate in retrograde signaling and stress acclimation in plants.

A versatile nanocomposite based on nanoceria for antibacterial enhancement and protection from aPDT-aggravated inflammation via modulation of macrophage polarization

Antibacterial photodynamic therapy (aPDT) is of vital importance for the treatment of periodontal diseases due to its great potential on effective elimination of pathogenic bacteria via overwhelming reactive oxygen species (ROS) generation. However, the excessive ROS after the therapeutic process may impose an oxidative stress within periodontal pockets, consequently leading to an irreversible destroy in surrounding tissue and severely limit its biomedical applications. In this study, considering the contradiction between ROS in bacteriostasis and inflammation, the role of ROS in different temporal and spatial states has been fully studied. Accordingly, we have designed composite nanomaterials that can play ROS based aPDT and anti-inflammatory effect by eliminating ROS, taking account of different ratio of photosensitizer/ROS scavenger to realize a time-sequential manner. Herein, a simple multifunctional nanocomposite was fabricated by coating red light-excited photosensitizer chlorin e6 (Ce6) onto nanoceria, achieving simultaneous sterilization and inflammation elimination via a dual directional regulation effect. This nano-based platform could utilize the aPDT for antibacterial purpose in the first stage with red-light irradiation, and subsequently scavenge the residual ROS via nanoceria to modulate host immunity by down-regulating the M1 polarization (pro-inflammatory) of macrophages and up-regulating the M2 polarization (anti-inflammatory and regenerative) of macrophages. Moreover, the local ROS level induced by activated inflammation pathway can be adjusted in a very long time because of the charge conversion effect of CeO2. The regenerative potential of inflammatory surrounding tissues was improved in the animal model. Our strategy will open a new inspiration to fight against the defects of aPDT in the treatment of periodontal disease, even in the anti-infection therapy for the future clinical application.

TrpML-mediated astrocyte microdomain Ca2+ transients regulate astrocyte-tracheal interactions.

Astrocytes exhibit spatially-restricted near-membrane microdomain Ca2+transients in their fine processes. How these transients are generated and regulate brain function in vivo remains unclear. Here we show that Drosophila astrocytes exhibit spontaneous, activity-independent microdomain Ca2+ transients in their fine processes. Astrocyte microdomain Ca2+ transients are mediated by the TRP channel TrpML, stimulated by reactive oxygen species (ROS), and can be enhanced in frequency by the neurotransmitter tyramine via the TyrRII receptor. Interestingly, many astrocyte microdomain Ca2+ transients are closely associated with tracheal elements, which dynamically extend filopodia throughout the central nervous system (CNS) to deliver O2 and regulate gas exchange. Many astrocyte microdomain Ca2+ transients are spatio-temporally correlated with the initiation of tracheal filopodial retraction. Loss of TrpML leads to increased tracheal filopodial numbers, growth, and increased CNS ROS. We propose that local ROS production can activate astrocyte microdomain Ca2+ transients through TrpML, and that a subset of these microdomain transients promotes tracheal filopodial retraction and in turn modulate CNS gas exchange.

Food-borne Lactiplantibacillus plantarum protect normal intestinal cells against inflammation by modulating reactive oxygen species and IL-23/IL-17 axis

Food-associated Lactiplantibacillus plantarum (Lpb. plantarum) strains, previously classified as Lactobacillus plantarum, are a promising strategy to face intestinal inflammatory diseases. Our study was aimed at clarifying the protective role of food-borne Lpb. plantarum against inflammatory damage by testing the scavenging microbial ability both in selected strains and in co-incubation with normal mucosa intestinal cells (NCM460). Here, we show that Lpb. plantarum endure high levels of induced oxidative stress through partially neutralizing reactive oxygen species (ROS), whereas they elicit their production when co-cultured with NCM460. Moreover, pre-treatment with food-borne Lpb. plantarum significantly reduce pro-inflammatory cytokines IL-17F and IL-23 levels in inflamed NCM460 cells. Our results suggest that food-vehicled Lpb. plantarum strains might reduce inflammatory response in intestinal cells by directly modulating local ROS production and by triggering the IL-23/IL-17 axis with future perspectives on health benefits in the gut derived by the consumption of functional foods enriched with selected strains.

Vascular Bundles Mediate Systemic Reactive Oxygen Signaling during Light Stress.

Systemic signaling and systemic acquired acclimation (SAA) are essential for plant survival during episodes of environmental stress. Recent studies highlighted a key role for reactive oxygen species (ROS) signaling in mediating systemic responses and SAA during light stress in Arabidopsis (Arabidopsis thaliana). These studies further identified the RESPIRATORY BURST OXIDASE HOMOLOG D (RBOHD) protein as a key player in mediating rapid systemic ROS responses. Here, we report that tissue-specific expression of RBOHD in phloem or xylem parenchyma cells of the rbohD mutant restores systemic ROS signaling, systemic stress-response transcript expression, and SAA to a local treatment of light stress. We further demonstrate that RBOHD and RBOHF are both required for local and systemic ROS signaling at the vascular bundles of Arabidopsis. Taken together, our findings highlight a key role for RBOHD-driven ROS production at the vascular bundles of Arabidopsis in mediating light stress-induced systemic signaling and SAA. In addition, they suggest that the integration of ROS, calcium, electric, and hydraulic signals, during systemic signaling, occurs at the vascular bundles.

Membranous NOX5-derived ROS oxidizes and activates local Src to promote malignancy of tumor cells

Reactive oxygen species (ROS) localized at the precise subcellular compartments are essential for regulating the activity of signaling proteins. Furthermore, ROS are master regulators of tumor malignant progression that respond to a diverse set of environmental stress, especially hypoxia. NADPH oxidases (NOXs) appear to be activated within discrete subcellular compartments to facilitate local ROS production. However, the subcellular function of NOXs in hypoxic tumor is still unclear. In this study, we demonstrated that NOX5 was greatly upregulated in clinical esophageal squamous cell carcinoma (ESCC) tumors, ESCC cell lines or primary ESCC cells, and elevated NOX5 was correlated to malignancy of ESCC tumors and poor prognosis. NOX5 induced the malignant progression of ESCC by activating Src, especially under hypoxic condition. Mechanistically, we showed that hypoxia promoted the interaction between NOX5 and Pyk2 on cell membrane via facilitating Ca2+-mediated Pyk2 Tyr402 site phosphorylation. Subsequently, Pyk2 acted as a scaffold for c-Abl phosphorylating the catalytic domain of NOX5 Tyr476/478 sites, which in turn upregulated hydrogen peroxide (H2O2) inside the Pyk2/NOX5 complex to oxidize and activate local Src. These findings provide insights into the biological significance of NOX5 in the development of ESCC.

Cross-Talk between NADPH Oxidase and Mitochondria: Role in ROS Signaling and Angiogenesis.

Angiogenesis, a new vessel formation from the pre-existing ones, is essential for embryonic development, wound repair and treatment of ischemic heart and limb diseases. However, dysregulated angiogenesis contributes to various pathologies such as diabetic retinopathy, atherosclerosis and cancer. Reactive oxygen species (ROS) derived from NADPH oxidase (NOX) as well as mitochondria play an important role in promoting the angiogenic switch from quiescent endothelial cells (ECs). However, how highly diffusible ROS produced from different sources and location can communicate with each other to regulate angiogenesis remains unclear. To detect a localized ROS signal in distinct subcellular compartments in real time in situ, compartment-specific genetically encoded redox-sensitive fluorescence biosensors have been developed. Recently, the intercellular communication, “cross-talk”, between ROS derived from NOX and mitochondria, termed “ROS-induced ROS release”, has been proposed as a mechanism for ROS amplification at distinct subcellular compartments, which are essential for activation of redox signaling. This “ROS-induced ROS release” may represent a feed-forward mechanism of localized ROS production to maintain sustained signaling, which can be targeted under pathological conditions with oxidative stress or enhanced to promote therapeutic angiogenesis. In this review, we summarize the recent knowledge regarding the role of the cross-talk between NOX and mitochondria organizing the sustained ROS signaling involved in VEGF signaling, neovascularization and tissue repair.