Rational design of Near-Infrared fluorescent probe for monitoring HNO in plants.

Nitroxyl (HNO), a reactive nitrogen species (RNS), is essential for plant growth. However, the action of HNO in plants has been difficult to understand due to the lack of highly sensitive and real-time in-situ monitoring tools. Herein, we presented a near-infrared fluorescent probe, DCI-HNO, based on dicyanoisophorone fluorophore, for real-time mapping HNO in plants. The introduction of a phosphine moiety as a specific HNO recognition unit can inhibit the intramolecular charge transfer (ICT) of probe DCI-HNO. However, in the presence of HNO, the ICT process occurred, leading to the emission at 665 nm. Probe DCI-HNO exhibited high sensitivity (97 nM), rapid response time (8 min), large Stokes shift (135 nm) for detection of HNO in plants. The novel developed probe has successfully imaged endogenous HNO produced during NO/H2S cross-talk in plant tissues. Additionally, the up-regulated in HNO levels during tobacco aging and in response to stress has been confirmed. Therefore, probe DCI-HNO has provided a reliable method for monitoring the NO/H2S cross-talk and revealing the role of HNO in plants.

Deciphering behaviors of 6:2 chlorinated polyfluorinated ether sulfonate (alternative-PFOS) on anammox processes: Nitrogen removal efficiency and microbial adaptability.

This study investigates the behaviors and effects of F-53B, an alternative to perfluorooctane sulfonate on anaerobic ammonium oxidation (anammox) processes. Results showed that the nitrogen removal efficiency (NRE) reached 83.8 % at a F-53B concentration of 0.5 mg·L-1, while NRE decreased to 66.9 % with 5 mg·L-1 of F-53B. The defluorination rates of 17.8 % (0.5 mg·L-1) and 9.3 % (5 mg·L-1) were observed, respectively, suggesting the occurrence of F-53B degradation. The relative abundance of Ca. Kuenenia decreased from 26.1 % to 16.2 % with the F-53B concentration increasing from 0.5 mg·L-1 to 5 mg·L-1. Meanwhile, Denitratisoma was selectively enriched with a relative abundance of 40.7 % at an F-53B concentration of 0.5 mg·L-1. Ca. Kuenenia could reduce reactive oxygen species induced by F-53B to maintain the balance of oxidative stress. This study gains insight into the behaviors and metabolic mechanisms of F-53B in anammox consortia, suggesting the feasibility of anammox processes for industrial wastewater.

Moisturizing and aroma-enhancing effects of low molecular weight fenugreek polysaccharides in cigarettes.

Polysaccharides possess excellent moisturizing effects due to their abundance of hydrophilic groups and film-forming properties. Additionally, they can produce a refreshing aroma during the pyrolysis process. However, there is scarce research on their application in the tobacco field. Herein, we investigated the effects of low molecular weight fenugreek polysaccharide (FP) obtained through ethanol fractionation and DEAE-52 cellulose column chromatography on moisture retention and aroma enhancement in tobacco. The moisture retention test revealed that the addition of FP increased the moisture retention index (MRI) of tobacco by 11.72 %-16.69 %, indicating that the hydrophilic nature of polysaccharides facilitated the migration of free water in tobacco to bound water, resulting in reduced water activity. Moreover, the contact angle between polysaccharide and tobacco was <90°, enabling better infiltration into tobacco and slowing down tobacco shrinkage caused by water loss. Among all the components, EFP-20 and EFP-40 demonstrated superior performance. Furthermore, FP exhibited excellent thermal stability below 200 °C and can decomposed to produce aromatic substances at high temperatures. It also demonstrated the ability to adsorb ethyl heptanoate and thermally decompose to produce a substantial amount of heptanoic acid. Consequently, the incorporation of FP in tobacco demonstrated favorable effects on both moisturization and aroma enhancement.

Multi-omics analysis reveals the collaboration and metabolisms of the anammox consortia driven by soluble/non-soluble Fe(III) as the sole iron element.

Iron is recognized as a physiological requirement for anammox bacteria (AnAOB), with Fe(II) considered to be the most effective form. However, Fe(III), instead of Fe(II) is the common iron form in natural and artificial ecosystems. In this study, the nitrogen removal performance and metabolic mechanisms in anammox consortia with soluble and non-soluble Fe(III) as the sole iron element were investigated. After the 150-day operation, the soluble (FeCl3) and insoluble (Fe2O3) Fe(III)-fed anammox systems reached nitrogen removal rates of 71.84 ± 0.80% and 50.20 ± 0.98%, respectively. AnAOB could survive with soluble (FeCl3) or insoluble (Fe2O3) Fe(III) as the sole iron element, reaching relative abundances of 18.49% and 13.16%, respectively. The results show that the formation of anammox core consortia can enable AnAOB's survival to adverse external conditions of Fe(II) deficiency. Metagenomic and metatranscriptomic analysis reveal that Ca. Kuenenia can only uptake Fe(II) into the cell for metabolisms either independently through the extracellular electron transfer or with the cross-feeding of symbiotic microbes. This study provides insight into the utilization and metabolic mechanisms of Fe(III) in Ca. Kuenenia-dominated consortia, and deepens the understanding of anammox core consortia in the nitrogen, carbon, and iron cycling, further promoting the practical applications of anammox processes.

Waste iron scraps promote anammox bacteria to resist inorganic carbon limitation.

The anaerobic ammonium oxidation (anammox) process is adversely affected by the limitation of inorganic carbon (IC). In this research, a new technique was introduced to assist anammox biomass in counteracting the adverse effects of IC limitation by incorporating waste iron scraps (WIS), a cheap and easily accessible byproduct of lathe cutting. Results demonstrated that reducing the influent IC/TN ratio from 0.08-0.09 to 0.04 resulted in a 20 % decrease in the nitrogen removal rate (NRR) for the control reactor, with an average specific anammox activity (SAA) of 0.65 g N/g VSS/day. Nevertheless, the performance of the WIS-assisted anammox reactor remained robust despite the reduction in IC supply. In fact, the NRR and SAA of the WIS-assisted reactor exhibited substantial improvements, reaching approximately 1.86 kg/(m3·day) and 0.98 g N/g VSS/day, respectively. These values surpassed those achieved by the control reactor by approximately 39 % and 51 %, respectively. The microbial analysis confirmed that the WIS addition significantly stimulated the proliferation of anammox bacteria (dominated by Candidatus Kuenenia) under IC limitation. The anammox gene abundances in the WIS-assisted anammox reactor were 3-4 times higher than those in the control reactor. Functional genes prediction based on the KEGG database revealed that the addition of WIS significantly enhanced the relative abundances of genes associated with nitrogen metabolism, IC fixation, and central carbon metabolism. Together, the results suggested that WIS promoted carbon dioxide fixation of anammox species to resist IC limitation. This study provided a promising approach for effectively treating high ammonium-strength wastewater using anammox under IC limitation.

Hydrogel-potassium humate composite alleviates cadmium toxicity of tobacco by regulating Cd bioavailability.

Cadmium (Cd) removal from soil to reduce Cd accumulation in plants is essential for agroecology, food safety, and human health. Cd enters plants from soil and affects plant growth and development. Hydrogels can easily combine with Cd, thereby altering its bioavailability in soil. However, few studies have evaluated the effects of hydrogel on the complex phytotoxicity caused by Cd uptake in plants and the microbial community structure. Herein, a new poly (acrylic acid)-grafted starch and potassium humate composite (S/K/AA) hydrogel was added to soil to evaluate its impact on tobacco growth and the soil microenvironment. The results indicate that the addition of S/K/AA hydrogel can significantly improve the biomass, chlorophyll (Chl) content, and photosynthetic capacity of tobacco plants during Cd stress conditions, and decrease Cd concentration, probably by affecting Cd absorption through the expression of Cd absorption transporters (e.g., NRAMP5, NRAMP3, and IRT1). Moreover, the application of S/K/AA hydrogel not only reduced the accumulation of reactive oxygen species (ROS), but also reduced the antioxidant activities of peroxidase (POD), superoxide dismutase (SOD), and catalase (CAT), suggesting that S/K/AA hydrogel alleviates Cd toxicity via a non-antioxidant pathway. Notably, we further analyzed the effectiveness of the hydrogel on microbial communities in Cd-contaminated soil and found that it increased the Cd-tolerant microbial community (Arthrobacter, Massilia, Streptomyces), enhancing the remediation ability of Cd-contaminated soil and helping tobacco plants to alleviate Cd toxicity. Overall, our study provides primary insights into how S/K/AA hydrogel affects Cd bioavailability and alleviates Cd toxicity in plants.

Fluorescent probes for lighting up ferroptotic cell death: A review.

Ferroptosis is a newly discovered form of regulated cellular demise, characterized by the accumulation of intracellular oxidative stress that is dependent on iron. Ferroptosis plays a crucial role not only in the development and treatment of tumors but also in the pathogenesis of neurodegenerative diseases and illnesses related to ischemia-reperfusion injury. This mode of cell death possesses distinctive properties that differentiate it from other forms of cell death, including unique morphological changes at both the cellular and subcellular levels, as well as molecular features that can be detected using specific methods. The use of fluorescent probes has become an invaluable means of detecting ferroptosis, owing to their high sensitivity, real-time in situ monitoring capabilities, and minimal damage to biological samples. This review comprehensively elucidates the physiological mechanisms underlying ferroptosis, while also detailing the development of fluorescent probes capable of detecting ferroptosis-related active species across various cellular compartments, including organelles, the nucleus, and the cell membrane. Additionally, the review explores how the dynamic changes and location of active species from different cellular compartments can influence the ignition and execution of ferroptotic cell death. Finally, we discuss the future challenges and opportunities for imaging ferroptosis. We believe that this review will not only aid in the elucidation of ferroptosis's physiological mechanisms but also facilitate the identification of novel treatment targets and means of accurately diagnosing and treating ferroptosis-related diseases.

Moisture Property and Thermal Behavior of Two Novel Synthesized Polyol Pyrrole Esters in Tobacco.

To overcome the shortcomings of high relative humidity and harmful oxidation products from traditional humectants, excellent humectants and flavor precursors were reported herein. Glucosamine hydrochloride was used as the starting material for the cyclization, oxidation, and alkylation processes that produced pyrrole acid. Then, esterification occurred with polyol catalyzed by EDC and DMAP to give the target compounds 2-(2,3-dihydroxypropyl) 4-methyl 5-methyl-1-propyl-1H-pyrrole-2,4-dicarboxylate (Gpe) and (2-hydroxypropyl) 4-methyl 5-methyl-1-propyl-1H-pyrrole-2,4-dicarboxylate (Ppe). Nuclear magnetic resonance (1H NMR, 13C NMR), infrared spectroscopy (IR), and high-resolution mass recorded spectrometry (HRMS) were used to confirm the two novel polyol pyrrole ester compounds. When Gpe and Ppe were added to the tobacco shred, low-field nuclear magnetic resonance (LF-NMR) imaging was applied to assess the hygroscopicity and moisturizing capacity. Furthermore, thermogravimetry (TG) and pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS) techniques were applied to study their thermal behaviors. These results showed that the target compounds (Gpe and Ppe) are good humectants with thermal properties of high-temperature stability and flavor release.

A new concept of waste iron recycling for the enhancement of the anammox process.

As a by-product of industry, waste iron scraps (WIS) are low-cost and widely available, which was potential for the development of iron-assisted anammox. In this study, the feasibility of adding WIS to enhance the nitrogen removal of the anammox process (also called WIS-assisted anammox) was demonstrated. Results indicated that the WIS-assisted anammox reactors performed a 15-35% higher nitrogen removal efficiency than that of the control. Compared to the sludge from the control, the sludge from the WIS-assisted anammox reactors had a higher iron content (78-113 g kg-1 SS) and a better specific anammox activity (10.8-15.5 mg N g-1 VSS h-1). The enhanced growth of the anammox bacteria (related to Ca. Kuenenia stuttgartiensis with 99% similarity) in the WIS-assisted anammox reactors was also confirmed by high-throughput sequencing and qPCR. Furthermore, the functional genes predicted by PICRUSt2 revealed a higher level of hydroxylamine oxidoreductase (hao)-like proteins expression of the biomass from the WIS-assisted anammox reactors, implying that the hydroxylamine-related anammox pathway was promoted. Additionally, the observation of cytoplasmic nitrate reductase (narG), copper-containing nitrite reductase (nirK), and nitric oxide reductase (norB) suggested that the introduction of WIS might promote the denitrification ability. This was correlated to the lower ΔNO3-/ΔNH4+ ratio observed in these WIS-assisted anammox reactors. Overall, the WIS-assisted anammox offers a sustainable nitrogen removal process for wastewater treatment with waste iron recycling.

Influence of Genotypic and Environmental Factors on Tobacco Leaves Based on Metabolomics.

Environmental factors affect plant metabolites, different climates, cultivation conditions, and biotic stresses and genotypes strongly affect their chemical composition and contents. Our aim is to examine the environmental and genetic interaction effects on tobacco metabolite composition. UPLC-QTOF MS/MS coupled with multivariate data analyses were applied for the metabolomics analysis of three tobacco cultivars from different planting regions in China. Principal component analysis (PCA) revealed that environmental factors have a greater effect on tobacco metabolism compared to genotypes. Twelve biomarkers were screened by orthogonal partial least squares discrimination analysis (OPLS-DA). Univariate analysis indicated that Malate, conjugated chlorogenic acid, chlorogenic acid, quercetin 3-rutinoside-7-glucoside, and unknown compound 5 were only influenced by environmental factors (independent of genotype). Quinate, neochlorogenic acid, and ouabagenin, taxezopidine K1, taxezopidine K2, and taxezopidine K3 in tobacco were influenced by the interaction of environmental factors and the genotype. Our results suggest that metabolomics based on UPLC-QTOF MS/MS could be used to analyze the ecological functions of biomarker metabolites and understand the mechanisms of plant adaption to the environment.

Pentoxifylline Enhances Antioxidative Capability and Promotes Mitochondrial Biogenesis in D-Galactose-Induced Aging Mice by Increasing Nrf2 and PGC-1α through the cAMP-CREB Pathway.

Aging is a complex phenomenon associated with oxidative stress and mitochondrial dysfunction. The objective of this study was to investigate the potential ameliorative effects of the phosphodiesterase inhibitor pentoxifylline (PTX) on the aging process and its underlying mechanisms. We treated D-galactose- (D-gal-) induced aging mice with PTX and measured the changes in behavior, degree of oxidative damage, and mitochondrial ultrastructure and content as well as the expression of nuclear factor erythroid 2-related factor 2- (Nrf2-) mediated antioxidant genes and peroxisome proliferator-activated receptor-gamma coactivator 1-alpha- (PGC-1α-) dependent mitochondrial biogenesis genes. The results demonstrated that PTX improved cognitive deficits, reduced oxidative damage, ameliorated abnormal mitochondrial ultrastructure, increased mitochondrial content and Nrf2 activation, and upregulated antioxidant and mitochondrial biogenesis gene expression in the hippocampus of wild-type aging mice. However, the above antiaging effects of PTX were obviously decreased in the brains of Nrf2-deficient D-gal-induced aging mice. Moreover, in hydrogen peroxide-treated SH-SY5Y cells, we found that cyclic adenosine monophosphate (cAMP) response element-binding protein (CREB) and Nrf2/PGC-1α act in a linear way by CREB siRNA transfection. Thus, PTX administration improved the aging-related decline in brain function by enhancing antioxidative capability and promoting mitochondrial biogenesis, which might depend on increasing Nrf2 and PGC-1α by activating the cAMP-CREB pathway.

Biochar for cadmium pollution mitigation and stress resistance in tobacco growth.

Pot experiments were conducted to investigate the influence of biochar addition and the mechanisms that alleviate Cd stress in the growth of tobacco plant. Cadmium showed an inhibitory effect on tobacco growth at different post-transplantation times, and this increased with the increase in soil Cd concentration. The growth index decreased by more than 10%, and the photosynthetic pigment and photosynthetic characteristics of the tobacco leaf were significantly reduced, and the antioxidant enzyme activity was enhanced. Application of biochar effectively alleviated the inhibitory effect of Cd on tobacco growth, and the alleviation effect of treatments is more significant to the plants with a higher Cd concentration. The contents of chlorophyll a, chlorophyll b, and carotenoids in the leaves of tobacco plants treated with biochar increased by 9.99%, 12.58%, and 10.32%, respectively, after 60 days of transplantation. The photosynthetic characteristics index of the net photosynthetic rate increased by 11.48%, stomatal conductance increased by 11.44%, and intercellular carbon dioxide concentration decreased to 0.92. Based on the treatments, during the growth period, the antioxidant enzyme activities of tobacco leaves comprising catalase, peroxidase, superoxide dismutase, and malondialdehyde increased by 7.62%, 10.41%, 10.58%, and 12.57%, respectively, after the application of biochar. Our results show that biochar containing functional groups can effectively reduce the effect of Cd stress by intensifying the adsorption or passivation of Cd in the soil, thereby, significantly reducing the Cd content in plant leaves, and providing a theoretical basis and method to alleviate soil Cd pollution and effect soil remediation.

Pentoxifylline enhances antioxidative capability and promotes mitochondrial biogenesis for improving age-related behavioral deficits.

Pentoxifylline (PTX) is a non-specific phosphodiesterase inhibitor with pleiotropic effects that is routinely used to treat peripheral vascular disease. In this study, we tested whether PTX could also counteract the detrimental effects of aging in the brain. To accomplish that, we treated aged rats with PTX and measured resulting behavioral alterations as well as changes in dopaminergic neurochemical levels, oxidative balance markers, mitochondrial function, nuclear factor erythroid 2-related factor 2 (Nrf2), peroxisome proliferator activated receptor-gamma coactivator 1-alpha (PGC-1α) and downstream gene expression, and cyclic adenosine monophosphate (cAMP) content in the brain. The results demonstrated that PTX improved motor and cognitive deficits and restored levels of dopamine and its metabolites in the brains of aged rats. PTX also reduced malondialdehyde levels and increased the GSH/GSSG ratio, mitochondrial ATP, nuclear Nrf2, and cAMP levels, and upregulated PGC-1α, nuclear respiratory factor 1, and mitochondrial transcription factor A expression in the substantia nigra and hippocampus of aged rats. Thus, increased nuclear Nrf2 levels and upregulation of PGC-1α, which enhance antioxidative capability and promote mitochondrial biogenesis, may be responsible for PTX-induced amelioration of behavioral deficits in aged rats.

Study on the preparation of geranyl acetone and ß-cyclodextrin inclusion complex and its application in cigarette flavoring.

ß-Cyclodextrin (ß-CD) inclusion complex containing geranyl acetone as a guest was prepared by saturated water solution method. Furthermore, the structure and properties of the inclusion complex were studied. The formation of the inclusion complex was demonstrated by. Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (X-RD), thermogravimetric analysis (TG) and differential scanning calorimetry (DSC). The thermodynamic analysis of the inclusion complex showed that the inclusion reaction is an endothermic spontaneous reaction. The average of △H, △S and △G is 11.66 kJ mol-1, 0.082 kJ mol-1 and - 14.49 kJ mol-1, respectively. Moreover, the kinetic analysis of thermal decomposition of the inclusion compound showed that the thermal decomposition reaction is a first-order reaction (the inclusion ratio is 1:1), the average activation energy of the reaction is 180.90 kJ mol-1, and the binding force in the inclusion compound is mainly Van der Waals force. The flavor test of cigarettes showed that the inclusion compound improved the stability of geranyl acetone and the sensory quality of cigarettes. This study improves the solubility and thermal stability of geranyl acetone, and provides theoretical support and technical guidance for expanding the application of geranyl acetone.

Novel environmental factors affecting microbial responses and physicochemical properties by sequentially applied biochar in black soil.

Biochar was increasingly used in agriculture soil amendment and has received widespread attention due to its potential to improve soil micro-ecological environment and crop growth. The raw material of the biochar used in this study is peanut shell, which is mixed with other organics and minerals to become a mineral-enhanced biochar under heating conditions (220 °C). When the third season crop is finished, we evaluated black soil physicochemical properties, microbial communities, and crop growth in long-term agricultural trials. Four treatments were set up: no amendment (control CK), nitrogen fertilizer only (70 kg ha-1 N), enhanced biochar only (5 t ha-1 B), and nitrogen fertilizer (70 kg ha-1) + enhanced biochar (5 t ha-1) (NB). The enhanced biochar promotes crop growth and increased the richness of the bacterial community, while reducing the richness of the fungal community. Nitrogen fertilizer + enhanced biochar increased soil microbial biomass carbon, nitrate nitrogen, and ammonium nitrogen by 43.75, 7.25, and 19.28%. In addition, we found changes in bacterial community were closely related to soil organic carbon, while changes in fungal community structure were closely related to soil carbon to nitrogen ratio. And the soil organic carbon and soil carbon to nitrogen ratio of biochar treatment were increased by 5.64 and 6.25% compared with fertilizer treatment, respectively. We concluded that enhanced biochar improved the soil more effectively and made the soil more conducive to crop growth. Regulating the microbial community by improving the physicochemical properties of soil was an important way to improve the stability and condition of the soil system with biochar. An enhanced biochar was of great significance for circular development of agriculture and soil improvement in Northeast China.

Testosterone enhances mitochondrial complex V function in the substantia nigra of aged male rats.

Deficits in coordinated motor behavior and mitochondrial complex V activity have been observed in aged males. Testosterone supplementation can improve coordinated motor behavior in aged males. We investigated the effects of testosterone supplementation on mitochondrial complex V function in the substantia nigra (a brain region that regulates motor activity) in aged male rats. These rats exhibited diminished ATP levels, attenuated mitochondrial complex V activity, and reduced expression of 3 of the 17 mitochondrial complex V subunits (ATP6, ATP8 and ATP5C1) in the substantia nigra. Testosterone supplementation increased ATP levels, mitochondrial complex V activity, and ATP6, ATP8 and ATP5C1 expression in the substantia nigra of the rats. Conversely, orchiectomy reduced mitochondrial complex V activity, downregulated ATP6 and ATP8 expression, and upregulated ATP5C1, ATP5I and ATP5L expression in the substantia nigra. Testosterone replacement reversed those effects. Thus, testosterone enhanced mitochondrial complex V function in the substantia nigra of aged male rats by upregulating ATP6 and ATP8. As potential testosterone targets, these two subunits may to some degree maintain nigrostriatal dopaminergic function in aged males.

Metabolic alterations associated with polycystic ovary syndrome: A UPLC Q-Exactive based metabolomic study.

BACKGROUND: The polycystic ovary syndrome (PCOS) is the most common endocrine and metabolic disorder syndrome of women in reproductive age. Metabolomic studies of the follicular fluid can reveal the potential metabolic pathways related to PCOS. The objection of this study was to explore the changes of metabolites in the follicular fluid of PCOS. METHODS: We collected follicular fluid samples of 35 patients with PCOS and 33 controls without PCOS for metabolomic analysis with UPLC Q-Exactive. The identified metabolites were annotated with KEGG and HMDB to determine the disturbances of metabolic pathways in PCOS. Based on the regression model, we conducted the ROC analysis to find the biomarker of PCOS in the follicular fluid. RESULTS: Metabolomic analysis identified 21 differential metabolites in PCOS, which revealed that the Vitamin B6 metabolism, phenylalanine metabolism and carnitine synthesis were the key changed pathways. We found that 7ß-Hydroxycholesterol was potential biomarker of PCOS based on the ROC analysis. CONCLUSION: We identified metabolic alterations and biomarker in the follicular fluid of PCOS, providing novel ways for the diagnosis and treatment of PCOS.

M10 peptide attenuates silica-induced pulmonary fibrosis by inhibiting Smad2 phosphorylation.

Silica-induced pulmonary fibrosis is a kind of worldwide occupational disease, and there is no effective treatment at present. Peptide therapy has attracted significant attention due to its simple structure, high selectiveness, strong bioactivity, relative safety, and high patient tolerance. In this study, we first confirmed that M10, a 10 amino acid peptide, has anti-fibrotic effects during the early and late stages of silica-induced fibrosis in mouse models and then partly explored the underlying mechanisms in vitro. M10 was detected in both the cell cytoplasm and nuclei. M10 showed no cytotoxicity to pulmonary epithelial cells and fibroblasts at the given concentrations. Functionally, M10 can reverse the silica-induced EMT process in epithelial cells and decrease TGF-ß1-stimulated fibroblast activation. Further mechanism investigations supported that M10 can block TGF-ß1 signalling by inhibiting phosphorylation of Smad2 protein in vitro and in vivo. All of the results indicate that M10 peptide may be a new method for the treatment of silica-induced pulmonary fibrosis.

miR-542-5p Attenuates Fibroblast Activation by Targeting Integrin α6 in Silica-Induced Pulmonary Fibrosis.

Silicosis is a very serious occupational disease and it features pathological manifestations of inflammatory infiltration, excessive proliferation of fibroblasts and massive depositions of the extracellular matrix in the lungs. Recent studies described the roles of a variety of microRNAs (miRNAs) in fibrotic diseases. Here, we aimed to explore the potential mechanism of miR-542-5p in the activation of lung fibroblasts. To induce a pulmonary fibrosis mouse model, silica suspension and the miR-542-5p agomir were administered to mice by intratracheal instillation and tail vein injection. We found that miR-542-5p was significantly decreased in mouse fibrotic lung tissues and up-regulation of miR-542-5p visually attenuated a series of fibrotic lesions, including alveolar structural damage, alveolar interstitial thickening and silica-induced nodule formation. The down-regulation of miR-542-5p was also observed in mouse fibroblast (NIH-3T3) treated with transforming growth factor ß1 (TGF-ß1). The proliferation and migration ability of NIH-3T3 cells were also inhibited by the transfection of miR-542-5p mimic. Integrin α6 (Itga6), reported as a cell surface protein associated with fibroblast proliferation, was confirmed to be a direct target of miR-542-5p. The knockdown of Itga6 significantly inhibited the phosphorylation of FAK/PI3K/AKT. In conclusion, miR-542-5p has a potential function for reducing the proliferation of fibroblasts and inhibiting silica-induced pulmonary fibrosis, which might be partially realized by directly binding to Itga6. Our data suggested that miR-542-5p might be a new therapeutic target for silicosis or other pulmonary fibrosis.

The CDR1as/miR-7/TGFBR2 Axis Modulates EMT in Silica-Induced Pulmonary Fibrosis.

Silicosis is one of the typical forms of pneumoconiosis characterized by abnormal proliferation of fibroblasts and deposition of extracellular matrix. Recent findings have shown that microRNAs and circular RNAs (circRNAs) are implicated in many diseases. However, the function of noncoding RNAs in pulmonary fibrosis remain to be elucidated. Here, miR-7 was found significantly decreased in silica-treated pulmonary epithelial cells as well as in fibrotic lung tissues of mice. Elevated expression of miR-7 via agomir injection relieved lung fibrosis in vivo. Further molecular study showed that miR-7 played its role against pulmonary fibrosis by blocking epithelial-mesenchymal transition (EMT) progression of human bronchial epithelial cells and A549 cells. Notably, transforming growth factor beta receptor 2 (TGFBR2) was identified as a target gene of miR-7 with bioinformatics tools, which was verified by dual luciferase receptor gene assay in human bronchial epithelial cells and A549 cells. Silica induced elevation of TGFBR2 could be abolished by exogenous expression of miR-7. Furthermore, bioinformatics software indicated that circRNA CDR1as had several binding sites for miR-7. The inhibitory effects of miR-7 on EMT and its target TGFBR2 were suppressed by circRNA CDR1as, which contributed to pulmonary fibrosis. Our studies also revealed overexpressed miR-7 could repress fibrogenesis of lung fibroblasts induced by TGF-ß1. Collectively, circRNA CDR1as stimulated by silica could sponge miR-7 to release TGFBR2, plays an important role during pulmonary fibrosis by promoting EMT process. These results indicated that the interaction between miR-7 and circRNA CDR1as may exert important functions and provide potential therapeutic targets in lung fibrotic diseases.