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Chinese Journal of Biotechnology ; (12): 177-189, 2024.
Article in Chinese | WPRIM | ID: wpr-1008088


Iron is an essential element for living organisms that plays critical roles in the process of bacterial growth and metabolism. However, it remains to be elucidated whether piuB encoding iron-uptake factor is involved in iron uptake and pathogenicity of Xanthomonas axonopodis pv. glycines (Xag). To investigate the function of piuB, we firstly generated a piuB deletion mutant (ΔpiuB) by homologous recombination. Compared with the wild-type, the piuB mutant exhibited significantly reduced growth and virulence in host soybean. The mutant displayed markedly increased siderophore secretory volume, and its sensitivity to Fe3+, Cu2+, Zn2+ and Mn2+ was significantly enhanced. Additionally, the H2O2 resistance, exopolysaccharide yield, biofilm formation, and cell mobility of ΔpiuB were significantly diminished compared to that of the wild-type. The addition of exogenous Fe3+ cannot effectively restore the above characteristics of ΔpiuB. However, expressing piuB in trans rescued the properties lost by ΔpiuB to the levels in the wild-type. Taken together, our results demonstrated that PiuB is a potential factor for Xag to assimilate Fe3+, and is necessary for Xag to be pathogenic in host soybean.

Iron , Glycine max , Virulence , Xanthomonas axonopodis/genetics , Hydrogen Peroxide
Braz. j. biol ; 84: e252735, 2024. tab, graf
Article in English | LILACS, VETINDEX | ID: biblio-1355873


Abstract Growth of plants is severely reduced due to water stress by affecting photosynthesis including photosystem II (PSII) activity and electron transport. This study emphasised on comparative and priority targeted changes in PSII activity due to progressive drought in seven populations of Panicum antidotale (P. antidotale) collected from Cholistan Desert and non-Cholistan regions. Tillers of equal growth of seven populations of P. antidotale grown in plastic pots filled with soil were subjected progressive drought by withholding water irrigation for three weeks. Progressive drought reduced the soil moisture content, leaf relative water content, photosynthetic pigments and fresh and dry biomass of shoots in all seven populations. Populations from Dingarh Fort, Dingarh Grassland and Haiderwali had higher growth than those of other populations. Cholistani populations especially in Dingarh Grassland and Haiderwali had greater ability of osmotic adjustment as reflected by osmotic potential and greater accumulation of total soluble proteins. Maximum H2O2 under water stress was observed in populations from Muzaffargarh and Khanewal but these were intermediate in MDA content. Under water stress, populations from Muzaffargarh and Dingarh Fort had greater K+ accumulation in their leaves. During progressive drought, non-Cholistani populations showed complete leaf rolling after 23 days of drought, and these populations could not withstand with more water stress condition while Cholistani populations tolerated more water stress condition for 31 days. Moreover, progressive drought caused PSII damages after 19 days and it became severe after 23 days in non-Cholistani populations of P. antidotale than in Cholistani populations.

Resumo O crescimento das plantas é severamente reduzido devido ao estresse hídrico, afetando a fotossíntese, incluindo a atividade do fotossistema II (PSII) e o transporte de elétrons. Este estudo enfatizou as mudanças comparativas e prioritárias na atividade do PSII devido à seca progressiva em sete populações de Panicum antidotale (P. antidotale) coletadas no Deserto do Cholistão e regiões fora do Cholistão. Perfilhos de igual crescimento de sete populações de P. antidotale cultivadas em vasos de plástico cheios de solo foram submetidos à seca progressiva, retendo a irrigação com água por três semanas. A seca progressiva reduziu o teor de umidade do solo, teor de água relativo nas folhas, pigmentos fotossintéticos e biomassa fresca e seca dos brotos em todas as sete populações. Populações de Dingarh Fort, Dingarh Grassland e Haiderwali tiveram maior crescimento do que as de outras populações. As populações de Cholistani, especialmente em Dingarh Grassland e Haiderwali, apresentaram maior capacidade de ajuste osmótico, refletido pelo potencial osmótico e maior acúmulo de proteínas solúveis totais. H2O2 máximo sob estresse hídrico foi observado em populações de Muzaffargarh e Khanewal, mas estas foram intermediárias no conteúdo de MDA. Sob estresse hídrico, as populações de Muzaffargarh e Dingarh Fort tiveram maior acúmulo de K+ em suas folhas. Durante a seca progressiva, as populações não cholistanesas mostraram rolagem completa das folhas após 23 dias de seca, e essas populações não conseguiram suportar mais condições de estresse hídrico, enquanto as populações cholistani toleraram mais condições de estresse hídrico por 31 dias. Além disso, a seca progressiva causou danos ao PSII após 19 dias e tornou-se severa após 23 dias em populações não cholistanesas de P. antidotale do que em populações cholistanesas.

Panicum , Photosynthesis , Plant Leaves , Desiccation , Droughts , Hydrogen Peroxide
Braz. j. oral sci ; 23: e241938, 2024. ilus
Article in English | LILACS, BBO | ID: biblio-1532506


Aim: To evaluate changes in the surface roughness and morphology of a nanofilled composite following toothbrushing with a whitening (WT) or regular toothpaste (RT), alone or combined with 35% hydrogen peroxide bleaching (HP). Methods: Seventy disc-shaped nanofilled composite (Filtek Z350XT) specimens were randomly divided into groups (n=10): WT, RT, TB (without toothpaste ­ control) or the combinations WT/ HP, RT/HP, TB/HP and HP. All groups underwent toothbrushing simulation (60,000 cycles) and bleaching treatment (4 sessions). Mean surface roughness (Ra, µm) was measured before (T0) and after treatments (TB). Surface morphology was assessed by scanning electron microscopy (SEM) at TB. Mean Ra was analyzed using general mixed models and multiple comparisons by the Tukey-Kramer test (α=5%). Results: HP caused no surface roughness changes on the nanofilled composite after treatment (p>0.05). RT toothbrushing, combined or not with HP, increased the surface roughness (p<0.05). WT and WT/ HP protocols had no effect on the surface roughness of the composite (p>0.05). The nanofilled composite submitted to RT toothbrushing combined with HP (RT/HP) presented substantial surface alterations under SEM, showing deep depressions and round-shaped defects. Toothbrushing with RT combined with the bleaching agent increased exposure of the inorganic fillers. Conclusion: WT toothbrushing, regardless of HP combination, or the single HP protocol had no effect on the surface roughness of the nanofilled composite. However, RT combined with HP negatively affected surface roughness and presented the most noticeable surface changes among groups

Tooth Bleaching , Toothpastes , Microscopy, Electron, Scanning , Composite Resins , Hydrogen Peroxide
Pesqui. bras. odontopediatria clín. integr ; 24: e220171, 2024. tab, graf
Article in English | LILACS, BBO | ID: biblio-1529141


Abstract Objective: To compare the clinical effect of two desensitizing agents used before the application of a bleaching gel based on 35% hydrogen peroxide (HP). Material and Methods: 30 patients were selected, and two desensitizing agents with different mechanisms of action were applied: Fluorine Neutral 2% (FN), which acts by blocking dentinal canaliculi while Potassium Nitrate 5% with 2% Sodium Fluoride (PN/SF) that acts in nerve transmission and blockade. Desensitizers were used before the application of 35% HP. For whitening, three clinical sessions were performed, with an interval of seven days, with three applications of the bleaching gel for 15 minutes, totaling 45 minutes/session. Tooth sensitivity (TS) was assessed with the numerical analog scale, and a spectrophotometer was used to obtain the color variation (ΔE). ΔE were submitted to ANOVA and Tukey test (p<0.05), and TS data were submitted to a two-way ANOVA analysis. Results: For sensitivity experience, the Tukey test indicated differences between PN/SF and the placebo I, but there was no statistically significant difference between FN and the placebo II. The TS was lower when the desensitizing gel was used during the bleaching procedure compared to after treatment, regardless of the desensitizing agents. Conclusion: PN/SF before in-office tooth bleaching can reduce TS intensity, and the use of desensitizing gel before bleaching did not affect the bleaching efficacy.

Humans , Male , Female , Adult , Dentin Desensitizing Agents/adverse effects , Hydrogen Peroxide/adverse effects , Sodium Fluoride/adverse effects , Spectrophotometers , Analysis of Variance , Fluorine
Rev. Ciênc. Plur ; 9(3): 32620, 26 dez. 2023. tab, ilus, graf
Article in Portuguese | LILACS, BBO | ID: biblio-1524464


Introdução:Sabe-se que a busca pela estética é algo cada vez mais crescente. Dentre os procedimentos mais procurados na odontologia estética, destaca-se o clareamento dental de consultório. Diante disso, ainda são poucos os estudos que avaliam os efeitos dos agentes clareadores de diferentes pHs na efetividade clareadora e na sensibilidade dentária.Objetivo:Avaliar a sensibilidade dentária e a eficácia clareadora de géis clareadores à base de peróxido de hidrogênio a 35% com diferentes pHs.Metodologia:Trata-se de um relato de três casos, descritivo e observacional, do tipo boca dividida (split-mouth) para cada estratégia clareadora (gel clareador com pH básico e gel clareador com pH ácido). Foram avaliados três pacientes de25, 26e 27anos de idade.Realizou-se registro de cor por meio da escala VITAClassical e avaliação da sensibilidade dentária pela escala visual analógica. Resultados:Todos os pacientes avaliados apresentaram cor A3 no registro de cor inicial e, após o clareamento dental,atingiram a cor A1. Todos relataram uma maior sensibilidade no hemiarco direito (local onde foi aplicada o gel clareador Whiteness HP que possui pH ácido. Dois pacientes relataram sensibilidade dentária no hemiarco esquerdo em que foi aplicado o gel clareador de pH básico. Conclusões:Observa-se a eficácia clínica dos géis clareadores de consultório à base de peróxido de hidrogênio a 35% na estabilidade de cor após o tratamento clareador, independente do seu pH. Ademais, nota-se que o gel clareador de pH básico promoveu menor sensibilidade pós-operatória (AU).

Introduction:It is known that the search for aesthetics is something increasingly growing. Among the most sought-after procedures in cosmetic dentistry, in-office tooth bleaching stands out. Therefore, there are still few studies that evaluate the effects of bleaching agents ofdifferent pHs on bleaching effectiveness and tooth sensitivity.Objective:To evaluate tooth sensitivity and bleaching efficacy of 35% hydrogen peroxide-based tooth bleaching gels with different pHs.Methodology:This is a report of three cases, descriptive and observational, of the split-mouth type for each bleaching strategy (bleaching gel with basic pH and bleaching gel with acidic pH). Three patients aged 25, 26 and 27 years were evaluated. Color registration was performed using the VITA Classical scale and tooth sensitivity was evaluated using the visual analogue scale.Results:All evaluated patients presented color A3 in the initial color registration and, after tooth bleaching, reached color A1. All reported greater sensitivity in the right hemi-arch (place where the Whiteness HP bleaching gel with an acid pH was applied. Two patients reported tooth sensitivity in the left hemi-arch where the basic pH bleaching gel was applied.Conclusions:The clinical efficacy of in-office tooth bleaching gels based on 35% hydrogen peroxide in terms of color stability after bleaching treatment is observed, regardless of its pH. In addition, it is noted that the basic pH bleaching gel promoted less postoperative sensitivity (AU).

Introducción: Se sabe que la búsqueda de la estética es algo cada vez más creciente. Entre los procedimientos más populares en odontología estética, se destaca el blanqueamiento dental en consultorio. Ante esto, aún existen pocos estudios que evalúen los efectos de agentes blanqueadores de diferentes pHs sobre la efectividad del blanqueamiento y la sensibilidad dental.Objetivo: Evaluar la sensibilidad dental y la eficacia blanqueadora de geles blanqueadores a base de peróxido de hidrógeno al 35 % con diferentes pH. Metodología: Este es un reporte de tres casos, descriptivo y observacional, del tipo boca partida para cada estrategia de blanqueamiento (gel blanqueador con pH básico y gel blanqueador con pH ácido). Se evaluaron tres pacientes de 25, 26 y 27 años. El registro de color se realizó con la escala clásica VITA y la sensibilidad dental se evaluó con la escala analógica visual.Resultados: Todos los pacientes evaluados presentaron color A3 en el registro de color inicial y, después del blanqueamiento dental, alcanzaron el color A1. Todos refirieron mayor sensibilidad en la hemiarcada derecha (lugar donde se aplicó el gel blanqueador de pH ácido Whiteness HP). Dos pacientes refirieron sensibilidad dental en la hemiarcadaizquierda donde se aplicó el gel blanqueador de pH básico.Conclusiones: Se observala eficacia clínica de los geles blanqueadores de consultorio a base de peróxido de hidrógeno al 35% en cuanto a la estabilidad del color tras el tratamiento blanqueador, independientemente de su pH. Además, se observa que el gel blanqueador de pH básico promovió una menor sensibilidad postoperatoria (AU).

Humans , Adult , Color , Dentin Sensitivity/complications , Tooth Bleaching Agents/adverse effects , Hydrogen Peroxide , Treatment Outcome , Observational Study , Hydrogen-Ion Concentration
Rev. Asoc. Med. Bahía Blanca ; 33(1): 14-19, jun. 2023.
Article in Spanish | LILACS, UNISALUD, BINACIS | ID: biblio-1436046


La pandemia de COVID-19 originada por el virus SARS-CoV-2 ha impactado en la atención profesional de los pacientes en la práctica odontológica. Se generan bioaerosoles por el odontólogo o por el propio paciente, que aumentan la posibilidad de diseminación del virus. Ante la urgente necesidad de establecer protocolos estrictos y efectivos de control de infecciones, decidimos investigar la efectividad de dos enjuagues bucales en la saliva de pacientes con diagnóstico positivo de SARS-CoV-2.

The COVID-19 pandemic caused by the SARS-CoV-2 virus has impacted on the professional care of patients. In dentistry, the generation of bioaerosols generated by the dentist or by the patient himself increases the possibility of the spread of the virus. Given the urgent need to establish strict and effective infection control protocols, we decided to investigate the effectiveness of two mouthwashes in the saliva of positive patients.

SARS-CoV-2 , Chlorhexidine , Dentistry , Hydrogen Peroxide
Rev. Ciênc. Plur ; 9(1): 29634, 27 abr. 2023. ilus
Article in Portuguese | LILACS, BBO | ID: biblio-1428369


Introdução:O aprimoramento das resinas compostas nos últimosanos em associação com a difusão de informações nas redes sociais tornou as facetas diretas tratamentos populares na dentística restauradora. No entanto, são procedimentos que exigem ampla destreza manual e conhecimento técnico. O fluxo digital através doescaneamento, enceramento digital e prototipagem 3D para construção de guias tem se tornado uma excelente alternativa para aumentar a previsibilidade e aumentar a longevidade destes trabalhos. Objetivo:Descrever o protocolo de confecção de facetas diretas em resina composta, através de um relato de caso, utilizando como auxílio o planejamento digital para confecção de modelo 3D, guia de silicone e paredes palatinas. Descrição do Caso:Paciente do gênero masculino, 43 anos, queixava-se do formato dos seus dentes. Ao exame clínico percebeu-se desgaste dental nos incisivos centrais e linha do sorriso levemente invertida. Após duas sessões de clareamento de consultório com Peróxido de hidrogênio (35%) e mockup direto com resina composta, foi realizada a moldagem e escaneamento do modelo de gesso no laboratório. O enceramento digital foi aprovado, o modelo 3D foi impresso para confecção da guia de silicone. Com auxílio da guia foram executadas facetas diretas nos elementos 13, 12, 11, 21, 22 e 23. Conclusão:O fluxo digital pode ser uma alternativa viável para minimizar as falhas na confecção de facetas diretas em resina composta (AU).

Introduction:The improvement of composite resins in recent years, together with information disseminated on social media, has made direct veneers popular treatments in restorative dentistry. However, these procedures require significant manual dexterity and technical knowledge. Digital work flow using scanning, digital wax-up and 3D prototyping for the construction of guides has become an excellent alternative to increase predictability and the longevity of these procedures. Objective:Describe the manufacturing protocol for direct composite resin veneers, using a case report and digital to construct the 3D model, silicone guide and palatine walls. Case description:Male patient, 43 years old, complained of the shape of his teeth. Clinical examination revealed tooth wear on the central incisors and a slightly inverted smile line. After two whitening sessions with hydroigen peroxide (35%) and direct mockup with composite resin, the plaster model was molded and scanned in the laboratory. Digital wax-up was approved, and the 3D model was printed to manufacture the silicone guide. With the help of the guide, the direct veneers were applied to elements 13, 12, 11, 21, 22 and 23.Conclusions:Digital flow may be a feasible alternative to minimize manufacturing flaws in direct composite resin veneers (AU).

Introducción: La mejora de las resinas compuestas en los últimos años, y la difusión de información en las redes sociales, ha popularizado las facetas directas en los tratamientos en odontología restauradora. Sin embargo, son procedimientos que requieren demasiado destreza manual y conocimientos técnicos. El flujo digital usando escaneo, encerado digital y prototipado 3D para la construcción de guías se ha convertido en una excelente alternativa para aumentar la previsibilidad y la longevidad de estos procedimientos. Objetivo: Describir el protocolo para la realización de carillas directas en resina compuesta, a través de un reporte de caso, utilizando el planeo digital como ayuda para la realización de un modelo 3D, guía de silicona y paredes palatinas. Descripción del caso: Paciente masculino, 43 años, se quejó de la forma de sus dientes. El examen clínico reveló desgaste dental en los incisivos centrales y una línea de sonrisa levemente invertida. Después de dos sesiones de blanqueamiento en consultorio con peróxido de hidrógeno (35%) y maqueta directa con resina compuesta, el modelo de yeso fue moldeado y escaneado en el laboratorio. El encerado digital fue aprovado, el modelo 3D fue impreso para hacer la guía de silicona. Con la ayuda de la guía se realizaron carillas directas en los elementos 13, 12, 11, 21, 22 y 23. Conclusiones: El fluxo digital puede ser una alternativa viable para minimizar fallas en la fabricación de carillas directas en resina compuesta (AU).

Humans , Male , Adult , Computer-Aided Design/instrumentation , Composite Resins/chemistry , Dental Veneers , Esthetics, Dental , Photography, Dental/instrumentation , Hydrogen Peroxide/chemistry
Chinese Journal of Cellular and Molecular Immunology ; (12): 626-632, 2023.
Article in Chinese | WPRIM | ID: wpr-981909


Objective To investigate the effect of H2O2-induced oxidative stress on autophagy and apoptosis of human bone marrow mesenchymal stem cells (hBMSCs). Methods hBMSCs were isolated and cultured. The cells were divided into control group, 3-MA group, H2O2 group, H2O2 combined with 3-MA group. DCFH-DA staining was used to analyze the level of reactive oxygen species (ROS). hBMSCs were treated with 0, 50, 100, 200, 400 μmol/L H2O2, and then the cell viability was detected by CCK-8 assay. The level of autophagy was detected by monodansylcadaverine (MDC) staining and LysoTracker Red staining. The cell apoptosis was detected by flow cytometry. Western blotting was used to detect the expression of beclin 1, mTOR, phosphorylated mTOR (p-mTOR), cleaved caspase-3(c-caspase-3) and caspase-3 proteins. Results Compared with the control group and 3-MA group, ROS level and autophagosomes were increased and the proliferation and apoptosis were decreased in H2O2 group. The protein expression of beclin 1, mTOR, c-caspase-3 was up-regulated, while the p-mTOR was down-regulated. Compared with the 3-MA group, the H2O2 combined with 3-MA group also had an increased ROS level and autophagosomes, but not with significantly increased apoptosis rate; The protein expression of beclin 1, mTOR, c-caspase-3 was up-regulated, and the p-mTOR was down-regulated. Conclusion H2O2 can induce hMSCs to trigger oxidative stress response. It enhances the autophagy and inhibits the proliferation and apoptosis of hBMSCs.

Humans , Beclin-1/metabolism , Caspase 3/metabolism , Reactive Oxygen Species/metabolism , Hydrogen Peroxide/pharmacology , Apoptosis , TOR Serine-Threonine Kinases/metabolism , Oxidative Stress , Autophagy , Mesenchymal Stem Cells/metabolism , Cell Proliferation
Chinese Journal of Biotechnology ; (12): 2070-2080, 2023.
Article in Chinese | WPRIM | ID: wpr-981190


5-aminovalanoic acid (5AVA) can be used as the precursor of new plastics nylon 5 and nylon 56, and is a promising platform compound for the synthesis of polyimides. At present, the biosynthesis of 5-aminovalanoic acid generally is of low yield, complex synthesis process and high cost, which hampers large-scale industrial production. In order to achieve efficient biosynthesis of 5AVA, we developed a new pathway mediated by 2-keto-6-aminohexanoate. By combinatory expression of L-lysine α-oxidase from Scomber japonicus, α-ketoacid decarcarboxylase from Lactococcus lactis and aldehyde dehydrogenase from Escherichia coli, the synthesis of 5AVA from L-lysine in Escherichia coli was achieved. Under the initial conditions of glucose concentration of 55 g/L and lysine hydrochloride of 40 g/L, the final consumption of 158 g/L glucose and 144 g/L lysine hydrochloride, feeding batch fermentation to produce 57.52 g/L of 5AVA, and the molar yield is 0.62 mol/mol. The new 5AVA biosynthetic pathway does not require ethanol and H2O2, and achieved a higher production efficiency as compared to the previously reported Bio-Chem hybrid pathway mediated by 2-keto-6-aminohexanoate.

Nylons , Lysine/metabolism , Hydrogen Peroxide/metabolism , Metabolic Engineering , Plastics/metabolism , Fermentation , Escherichia coli/metabolism , Aminocaproates/metabolism
West China Journal of Stomatology ; (6): 190-196, 2023.
Article in English | WPRIM | ID: wpr-981111


OBJECTIVES@#This study aimed to observe the color rebound and rebound rates of non-pulp discolored teeth within 1 year after routine internal bleaching to guide clinical practice and prompt prognosis.@*METHODS@#In this work, the efficacy of bleaching was observed in 20 patients. The color of discolored teeth was measured by using a computerized colorimeter before bleaching; immediately after bleaching; and at the 1st, 3rd, 6th, 9th, and 12th months after bleaching. The L*, a*, and b* values of the color of cervical, mesial, and incisal parts of the teeth were obtained, and the color change amounts ΔE*, ΔL*, Δa*, and Δb* were calculated. The overall rebound rate (P*) and the color rebound velocity (V*) were also analyzed over time.@*RESULTS@#In 20 patients following treatment, the average ΔE* of tooth color change was 14.99. After bleaching, the neck and middle of the teeth ΔE* and ΔL* decreased in the 1st, 3rd, 6th, 9th, and 12th months, and the differences were statistically significant. Meanwhile, from the 9th month after bleaching, the rebound speed was lower than that in the 1st month, and the difference was statistically significant. The incisal end of the tooth ΔE* and ΔL* decreased in the 6th, 9th, and 12th months after bleaching, and the differences were statistically significant. No significant difference was found in the rebound speed between time points. However, this rate settled after the 9th month, with an average color rebound rate of 30.11% in 20 patients.@*CONCLUSIONS@#The results indicated that internal bleaching could cause a noticeable color change on pulpless teeth. The color rebound after bleaching was mainly caused by lightness (L*), which gradually decreased with time, and it was slightly related to a* and b*. The color of the teeth after internal bleaching rebounded to a certain extent with time, but the color rebound speed became stable from the 9th month. Clinically, secondary internal bleaching can be considered at this time according to whether the colors of the affected tooth and the adjacent tooth are coordinated and depending on the patient's needs.

Humans , Tooth Bleaching/methods , Tooth, Nonvital/drug therapy , Color , Tooth Discoloration/drug therapy , Tooth , Hydrogen Peroxide/therapeutic use , Tooth Bleaching Agents/therapeutic use
Journal of Southern Medical University ; (12): 552-559, 2023.
Article in Chinese | WPRIM | ID: wpr-986961


OBJECTIVE@#To evaluate the regulatory effect of berberine on autophagy and apoptosis balance of fibroblast-like synoviocytes (FLSs) from patients with in rheumatoid arthritis (RA) and explore the mechanism.@*METHODS@#The inhibitory effect of 10, 20, 30, 40, 50, 60, 70, and 80 μmol/L berberine on RA-FLS proliferation was assessed using CCK-8 method. Annexin V/PI and JC-1 immunofluorescence staining was used to analyze the effect of berberine (30 μmol/L) on apoptosis of 25 ng/mL TNF-α- induced RA-FLSs, and Western blotting was performed to detect the changes in the expression levels of autophagy- and apoptosis-related proteins. The cells were further treated with the autophagy inducer RAPA and the autophagy inhibitor chloroquine to observe the changes in autophagic flow by laser confocal detection of mCherry-EGFP-LC3B. RA-FLSs were treated with the reactive oxygen species (ROS) mimic H2O2 or the ROS inhibitor NAC, and the effects of berberine on ROS, mTOR and p-mTOR levels were observed.@*RESULTS@#The results of CCK-8 assay showed that berberine significantly inhibited the proliferation of RA-FLSs in a time- and concentration-dependent manner. Flow cytometry and JC-1 staining showed that berberine (30 μmol/L) significantly increased apoptosis rate (P < 0.01) and reduced the mitochondrial membrane potential of RA-FLSs (P < 0.05). Berberine treatment obviously decreased the ratios of Bcl-2/Bax (P < 0.05) and LC3B-II/I (P < 0.01) and increased the expression of p62 protein in the cells (P < 0.05). Detection of mCherry-EGFP-LC3B autophagy flow revealed obvious autophagy flow block in berberine-treated RA-FLSs. Berberine significantly reduced the level of ROS in TNF-α-induced RA-FLSs and upregulated the expression level of autophagy-related protein p-mTOR (P < 0.01); this effect was regulated by ROS level, and the combined use of RAPA significantly reduced the pro-apoptotic effect of berberine in RA-FLSs (P < 0.01).@*CONCLUSION@#Berberine can inhibit autophagy and promote apoptosis of RA-FLSs by regulating the ROS-mTOR pathway.

Humans , Synoviocytes , Berberine/metabolism , Reactive Oxygen Species/metabolism , Tumor Necrosis Factor-alpha/metabolism , Hydrogen Peroxide/metabolism , Sincalide/metabolism , Cell Proliferation , Arthritis, Rheumatoid/metabolism , Signal Transduction , TOR Serine-Threonine Kinases/metabolism , Apoptosis , Fibroblasts , Autophagy , Cells, Cultured
Journal of Peking University(Health Sciences) ; (6): 975-981, 2023.
Article in Chinese | WPRIM | ID: wpr-1010156


OBJECTIVE@#To investigate the regulatory effect of interferon-α (IFN-α) on the apoptosis and killing function of CD56dimCD57+ natural killer (NK) cells in systemic lupus erythematosus (SLE) patients, and to explore the specific mechanism.@*METHODS@#A total of sixty-four newly treated SLE patients and sixteen healthy controls (HC) enrolled in the Second Hospital of Dalian Medical University were selected as the research subjects. And the gene expression levels of molecules related to NK cell-killing function were detected by real-time quantitative polymerase chain reaction. CD56dimCD57+ NK cells were co-cultured with the K562 cells, and the apoptotic K562 cells were labeled with Annexin-Ⅴ and 7-amino-actinomycin D. Peripheral blood mononuclear cells were treated with 20, 40, and 80 μmol/L hydrogen peroxide (H2O2), and treated without H2O2 as control, the expression level of perforin (PRF) was detected by flow cytometry. The concentration of IFN-α in serum was determined by enzyme linked immunosorbent assay. The expression levels of IFN-α receptors (IFNAR) on the surface of CD56dimCD57+ NK cells were detected by flow cytometry, and were represented by mean fluorescence intensity (MFI). CD56dimCD57+ NK cells were treated with 1 000 U/mL IFN-α for 24, 48 and 72 h, and no IFN-α treatment was used as the control, the apoptosis and the expression levels of mitochondrial reactive oxygen species (mtROS) were measured by flow cytometry and represented by MFI.@*RESULTS@#Compared with HC(n=3), the expression levels of PRF1 gene in peripheral blood NK cells of the SLE patients (n=3) were decreased (1.24±0.41 vs. 0.57±0.12, P=0.05). Compared with HC(n=5), the ability of peripheral blood CD56dimCD57+ NK cells in the SLE patients (n=5) to kill K562 cells was significantly decreased (58.61%±10.60% vs. 36.74%±6.27%, P < 0.01). Compared with the control (n=5, 97.51%±1.67%), different concentrations of H2O2 treatment significantly down-regulated the PRF expression levels of CD56dimCD57+ NK cells in a dose-dependent manner, the 20 μmol/L H2O2 PRF was 83.23%±8.48% (n=5, P < 0.05), the 40 μmol/L H2O2 PRF was 79.53%±8.56% (n=5, P < 0.01), the 80 μmol/L H2O2 PRF was 76.67%±7.16% (n=5, P < 0.01). Compared to HC (n=16), the serum IFN-α levels were significantly increased in the SLE patients (n=45) with moderate to high systemic lupus erythematosus disease activity index (SLEDAI≥10) [(55.07±50.36) ng/L vs. (328.2±276.3) ng/L, P < 0.001]. Meanwhile, compared with HC (n=6), IFNAR1 expression in peripheral blood CD56dimCD57+ NK cells of the SLE patients (n=6) were increased (MFI: 292.7±91.9 vs. 483.2±160.3, P < 0.05), and compared with HC (n=6), IFNAR2 expression in peripheral blood CD56dimCD57+ NK cells of the SLE patients (n=7) were increased (MFI: 643.5±113.7 vs. 919.0±246.9, P < 0.05). Compared with control (n=6), the stimulation of IFN-α (n=6) significantly promoted the apoptosis of CD56dimCD57+ NK cells (20.48%±7.01% vs. 37.82%±5.84%, P < 0.05). In addition, compared with the control (n=4, MFI: 1 049±174.5), stimulation of CD56dimCD57+ NK cells with IFN-α at different times significantly promoted the production of mtROS in a time-dependent manner, 48 h MFI was 3 437±1 472 (n=4, P < 0.05), 72 h MFI was 6 495±1 089 (n=4, P < 0.000 1), but there was no significant difference at 24 h of stimulation.@*CONCLUSION@#High serum IFN-α level in SLE patients may induce apoptosis by promoting mtROS production and inhibit perforin expression, which can down-regulate CD56dimCD57+ NK killing function.

Humans , Interferon-alpha/metabolism , Perforin/metabolism , Leukocytes, Mononuclear/metabolism , Hydrogen Peroxide/metabolism , Interferon-gamma/metabolism , CD56 Antigen/metabolism , Killer Cells, Natural/metabolism , Lupus Erythematosus, Systemic
Journal of Zhejiang University. Medical sciences ; (6): 296-305, 2023.
Article in English | WPRIM | ID: wpr-982047


OBJECTIVES@#To explore the physicochemical characteristics and biocompatibility of calcium peroxide (CPO)-loaded polycaprolactone (PCL) microparticle.@*METHODS@#The CPO/PCL particles were prepared. The morphology and elemental distribution of CPO, PCL and CPO/PCL particles were observed with scanning electron microscopy and energy dispersive spectroscopy, respectively. Rat adipose mesenchymal stem cells were isolated and treated with different concentrations (0.10%, 0.25%, 0.50%, 1.00%) of CPO or CPO/PCL particles. The mesenchymal stem cells were cultured in normal media or osteogenic differentiation media under the hypoxia/normoxia conditions, and the amount of released O2 and H2O2 after CPO/PCL treatment were detected. The gene expressions of alkaline phosphatase (ALP), Runt-associated transcription factor 2 (RUNX2), osteopontin (OPN) and osteocalcin (OCN) were detected by realtime RT-PCR. SD rats were subcutaneously injected with 1.00% CPO/PCL particles and the pathological changes and infiltration of immune cells were observed with HE staining and immunohistochemistry at day 7 and day 14 after injection.@*RESULTS@#Scanning electron microscope showed that CPO particles had a polygonal structure, PCL particles were in a small spherical plastic particle state, and CPO/PCL particles had a block-like crystal structure. Energy dispersive spectroscopy revealed that PCL particles showed no calcium mapping, while CPO/PCL particles showed obvious and uniform calcium mapping. The concentrations of O2 and H2O2 released by CPO/PCL particles were lower than those of CPO group, and the oxygen release time was longer. The expressions of Alp, Runx2, Ocn and Opn increased with the higher content of CPO/PCL particles under hypoxia in osteogenic differentiation culture and normal culture, and the induction was more obvious under osteogenic differentiation conditions (all P<0.05). HE staining results showed that the muscle tissue fibers around the injection site were scattered and disorderly distributed, with varying sizes and thicknesses at day 7 after particle injection. Significant vascular congestion, widened gaps, mild interstitial congestion, local edema, inflammatory cell infiltration, and large area vacuolization were observed in some tissues of rats. At day 14 after microparticle injection, the muscle tissue around the injection site and the tissue fibers at the microparticle implantation site were arranged neatly, and the gap size was not thickened, the vascular congestion, local inflammatory cell infiltration, and vacuolization were significantly improved compared with those at day 7. The immunohistochemical staining results showed that the expressions of CD3 and CD68 positive cells significantly increased in the surrounding muscle tissue, and were densely distributed in a large area at day 7 after particle injection. At day 14 of microparticle injection, the numbers of CD3 and CD68 positive cells in peripheral muscle tissue and tissue at the site of particle implantation were lower than those at day 7 (all P<0.01).@*CONCLUSIONS@#CPO/PCL particles have good oxygen release activity, low damage to tissue, and excellent biocompatibility.

Rats , Animals , Osteogenesis , Core Binding Factor Alpha 1 Subunit , Rats, Sprague-Dawley , Hydrogen Peroxide/pharmacology , Cell Differentiation , Oxygen , Hypoxia , Cells, Cultured
International Journal of Oral Science ; (4): 10-10, 2023.
Article in English | WPRIM | ID: wpr-971599


Xenogenic organ transplantation has been considered the most promising strategy in providing possible substitutes with the physiological function of the failing organs as well as solving the problem of insufficient donor sources. However, the xenograft, suffered from immune rejection and ischemia-reperfusion injury (IRI), causes massive reactive oxygen species (ROS) expression and the subsequent cell apoptosis, leading to the xenograft failure. Our previous study found a positive role of PPAR-γ in anti-inflammation through its immunomodulation effects, which inspires us to apply PPAR-γ agonist rosiglitazone (RSG) to address survival issue of xenograft with the potential to eliminate the excessive ROS. In this study, xenogenic bioroot was constructed by wrapping the dental follicle cells (DFC) with porcine extracellular matrix (pECM). The hydrogen peroxide (H2O2)-induced DFC was pretreated with RSG to observe its protection on the damaged biological function. Immunoflourescence staining and transmission electron microscope were used to detect the intracellular ROS level. SD rat orthotopic transplantation model and superoxide dismutase 1 (SOD1) knockout mice subcutaneous transplantation model were applied to explore the regenerative outcome of the xenograft. It showed that RSG pretreatment significantly reduced the adverse effects of H2O2 on DFC with decreased intracellular ROS expression and alleviated mitochondrial damage. In vivo results confirmed RSG administration substantially enhanced the host's antioxidant capacity with reduced osteoclasts formation and increased periodontal ligament-like tissue regeneration efficiency, maximumly maintaining the xenograft function. We considered that RSG preconditioning could preserve the biological properties of the transplanted stem cells under oxidative stress (OS) microenvironment and promote organ regeneration by attenuating the inflammatory reaction and OS injury.

Mice , Humans , Rats , Animals , Swine , PPAR gamma/pharmacology , Reactive Oxygen Species/pharmacology , Heterografts , Hydrogen Peroxide/pharmacology , Rats, Sprague-Dawley , Rosiglitazone/pharmacology , Oxidative Stress
Chinese Journal of Burns ; (6): 35-44, 2023.
Article in Chinese | WPRIM | ID: wpr-971147


Objective: To investigate the influence of reactive oxygen species (ROS) responsive self-assembled nanomicelle loaded with pyroptosis inhibitor on full-thickness skin defects in diabetic rats. Methods: Experimental research methods were employed. A nucleotide-binding oligomerization domain (NOD) 1/2 inhibitor (NOD-IN-1) was encapsulated with nanomicelle polyethylene glycol-block-polypropylene sulfide (PEG-b-PPS), and the resulting product was called PEPS@NOD-IN-1. The morphology and hydration particle size of PEG-b-PPS and PEPS@NOD-IN-1 were observed by transmission electron microscope and particle size analyzer, respectively, and the encapsulation rate and drug loading rate of PEPS@NOD-IN-1 to NOD-IN-1 and the cumulative release rate of NOD-IN-1 by PEPS@NOD-IN-1 in phosphate buffer solution (PBS) alone and hydrogen peroxide-containing PBS within 40 h were measured and calculated by microplate reader, and the sample number was 3. Twenty-four male Sprague-Dawley rats aged 6-7 weeks were injected with streptozotocin to induce type 1 diabetes mellitus. Six full-thickness skin defect wounds were made on the back of each rat. The injured rats were divided into PBS group, NOD-IN-1 group, PEG-b-PPS group, and PEPS@NOD-IN-1 group with corresponding treatment according to the random number table, with 6 rats in each group. The wound healing was observed on post injury day (PID) 3, 7, and 12, and the wound healing rate was calculated. The ROS levels in wound tissue were detected by immunofluorescence method on PID 3. On PID 7, the granulation tissue thickness in wound was assessed by hematoxylin-eosin staining, the mRNA expressions of NOD1 and NOD2 were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction, and the protein expressions of NOD1, NOD2, and GSDMD-N terminals were detected by Western blotting. Six wounds from different rats in each group were taken for detection of the above indicators. Wound tissue (3 samples per group) was taken from rats in PBS group and PEPS@NOD-IN-1 group on PID 7, and transcriptome sequencing was performed using high-throughput sequencing technology platform. Differentially expressed genes (DEGs) significantly down-regulated in PEPS@NOD-IN-1 group as compared with PBS group were screened, and the enrichment analysis of Kyoto Encyclopedia of Genes and Genomes (KEGG) was performed. The DEG heatmap of the NOD-like receptor pathway, a pyroptosis-related pathway, was made. Protein-protein interaction (PPI) analysis of DEGs in heatmap was performed through the STRING database to screen key genes of PEPS@NOD-IN-1 regulating the NOD-like receptor pathway. Data were statistically analyzed with analysis of variance for repeated measurement, one-way analysis of variance, and Tukey test. Results: PEG-b-PPS and PEPS@NOD-IN-1 were in spherical structures of uniform size, with hydration particle sizes of (134.2±3.3) and (143.1±2.3) nm, respectively. The encapsulation rate of PEPS@NOD-IN-1 to NOD-IN-1 was (60±5)%, and the drug loading rate was (15±3)%. The release of NOD-IN-1 from PEPS@NOD-IN-1 in PBS alone was slow, and the cumulative release rate at 40 h was only (12.4±2.3)%. The release of NOD-IN-1 from PEPS@NOD-IN-1 in hydrogen peroxide-containing PBS within 10 h was very rapid, and the cumulative release rate at 10 h reached (90.1±3.6)%. On PID 3 and 7, the wounds of rats in the four groups were gradually healed, and the healing in PEPS@NOD-IN-1 group was better than that in the other three groups. On PID 12, the wound scab area in PBS group was large, the wound epithelialization in NOD-IN-1 group and PEG-b-PPS group was obvious, and the wound in PEPS@NOD-IN-1 group was close to complete epithelialization. Compared with those in PBS group, NOD-IN-1 group, and PEG-b-PPS group, the wound healing rates on PID 7 and 12 in PEPS@NOD-IN-1 group were significantly increased (P<0.05), the level of ROS in wound tissue on PID 3 was significantly decreased (P<0.05), the thickness of granulation tissue in wound on PID 7 was significantly thickened (P<0.05), and the mRNA expressions of NOD1 and NOD2 and the protein expressions of NOD1, NOD2, and GSDMD-N terminals in wound tissue on PID 7 were significantly decreased (P<0.05). KEGG pathway analysis showed that DEGs significantly down-regulated in PEPS@NOD-IN-1 group as compared with PBS group were significantly enriched in NOD-like receptors, hypoxia-inducible factors, mitogen-activated protein kinases, and tumor necrosis factor (TNF) pathways. In the DEG heatmap of NOD-like receptor pathway, the genes regulating pyroptosis mainly involved NOD1, NOD2, NOD-like receptor thermoprotein domain-related protein 3, Jun, signal transduction and transcriptional activator 1 (STAT1), TNF-α-induced protein 3. The PPI results showed that NOD1, NOD2, and STAT1 were the key genes of PEPS@NOD-IN-1 regulating the NOD-like receptor pathway. Conclusions: PEPS@NOD-IN-1 can down-regulate the level of local ROS in wounds and the expression of NOD1, NOD2, and GSDMD-N terminals, the key regulators of pyroptosis, thereby promoting the repair of full-thickness skin defect wounds in diabetic rats. PEPS@NOD-IN-1 can also significantly down-regulate the pyroptosis, inflammation, and hypoxia-related pathways of wounds, and regulate NOD-like receptor pathways by down-regulating key genes NOD1, NOD2, and STAT1.

Rats , Male , Animals , Reactive Oxygen Species , Wound Healing , Rats, Sprague-Dawley , Diabetes Mellitus, Experimental , Hydrogen Peroxide , Pyroptosis , Skin Abnormalities , Soft Tissue Injuries , NLR Proteins , Hypoxia , RNA, Messenger
Chinese Journal of Burns ; (6): 15-24, 2023.
Article in Chinese | WPRIM | ID: wpr-971145


Objective: To investigate the effects and mechanism of interleukin-4-modified gold nanoparticle (IL-4-AuNP) on the wound healing of full-thickness skin defects in diabetic mice. Methods: Experimental research methods were adopted. Gold nanoparticle (AuNP) and IL-4-AuNP were synthesized by improving the methods described in published literature. The morphology of those two particles were photographed by transmission electron microscopy, and their particle sizes were calculated. The surface potential and hydration particle size of the two particles were detected by nanoparticle potentiometer and particle size analyzer, respectively. The clearance rate of IL-4-AuNP to hydrogen peroxide and superoxide anion was measured by hydrogen peroxide and superoxide anion kits, respectively. Mouse fibroblast line 3T3 cells were used and divided into the following groups by the random number table (the same below): blank control group, hydrogen peroxide alone group treated with hydrogen peroxide only, hydrogen peroxide+IL-4-AuNP group treated with IL-4-AuNP for 0.5 h and then treated with hydrogen peroxide. After 24 h of culture, the reactive oxygen species (ROS) levels of cells were detected by immunofluorescence method; cell count kit 8 was used to detect relative cell survival rate. The macrophage Raw264.7 mouse cells were then used and divided into blank control group and IL-4-AuNP group that treated with IL-4-AuNP. After 24 h of culture, the expression of arginase 1 (Arg-1) in cells was observed by immunofluorescence method. Twelve male BALB/c mice (mouse age, sex, and strain, the same below) aged 8 to 10 weeks were divided into IL-4-AuNP group and blank control group, treated accordingly. On the 16th day of treatment, whole blood samples were collected from mice for analysis of white blood cell count (WBC), red blood cell count (RBC), hemoglobin level, or platelet count and the level of aspartate aminotransferase (AST), alanine transaminase (ALT), urea, or creatinine. The inflammation, bleeding, or necrosis in the heart, liver, spleen, lung, and kidney tissue of mice were detected by hematoxylin-eosin (HE). Another 36 mice were selected to make diabetic model, and the full-thickness skin defect wounds were made on the back of these mice. The wounds were divided into blank control group, AuNP alone group, and IL-4-AuNP group, with 12 mice in each group, and treated accordingly. On the 0 (immediately), 4th, 9th, and 15th day of treatment, the wound condition was observed and the wound area was calculated. On the 9th day of treatment, HE staining was used to detect the length of neonatal epithelium and the thickness of granulation tissue in the wound. On the 15th day of treatment, immunofluorescence method was used to detect ROS level and the number of Arg-1 positive cells in the wound tissue. The number of samples was 6 in all cases. Data were statistically analyzed with independent sample t test, corrected t test, Tukey test, or Dunnett T3 test. Results: The size of prepared AuNP and IL-4-AuNP were uniform. The particle size, surface potential, and hydration particle size of AuNP and IL-4-AuNP were (13.0±2.1) and (13.9±2.5) nm, (-45.8±3.2) and (-20.3±2.2) mV, (14±3) and (16±4) nm, respectively. For IL-4-AuNP, the clearance rate to hydrogen peroxide and superoxide anion were (69±4)% and (52±5)%, respectively. After 24 h of culture, the ROS level of 3T3 in hydrogen peroxide alone group was significantly higher than that in blank control group (q=26.12, P<0.05); the ROS level of hydrogen peroxide+IL-4-AuNP group was significantly lower than that in hydrogen peroxide alone group (q=25.12, P<0.05) and close to that in blank control group (P>0.05). After 24 h of culture, the relative survival rate of 3T3 cells in hydrogen peroxide+IL-4-AuNP group was significantly higher than that in hydrogen peroxide alone group (t=51.44, P<0.05). After 24 h of culture, Arg-1 expression of Raw264.7 cells in IL-4-AuNP group was significantly higher than that in blank control group (t'=8.83, P<0.05).On the 16th day of treatment, there were no significant statistically differences in WBC, RBC, hemoglobin level, or platelet count and the level of AST, ALT, urea, or creatinine of mice between blank control group and IL-4-AuNP group (P>0.05). No obvious inflammation, bleeding or necrosis was observed in the heart, liver, spleen, lung, and kidney of important organs in IL-4-AuNP group, and no significant changes were observed compared with blank control group. On the 0 and 4th day of treatment, the wound area of diabetic mice in blank control group, AuNP alone group, and IL-4-AuNP group had no significant difference (P>0.05). On the 9th day of treatment, the wound areas both in AuNP alone group and IL-4-AuNP group were significantly smaller than that in blank control group (with q values of 9.45 and 14.87, respectively, P<0.05), the wound area in IL-4-AuNP group was significantly smaller than that in AuNP alone group (q=5.42, P<0.05). On the 15th day of treatment, the wound areas both in AuNP alone group and IL-4-AuNP group were significantly smaller than that in blank control group (with q values of 4.84 and 20.64, respectively, P<0.05), the wound area in IL-4-AuNP group was significantly smaller than that in AuNP alone group (q=15.80, P<0.05); moreover, inflammations such as redness and swelling were significantly reduced in IL-4-AuNP group compared with the other two groups. On the 9th day of treatment, compared with blank control group and AuNP alone group, the length of neonatal epithelium in the wound of diabetic mice in IL-4-AuNP group was significantly longer (all P<0.05), and the thickness of the granulation tissue in the wound was significantly increased (with q values of 11.33 and 9.65, respectively, all P<0.05). On the 15th day of treatment, compared with blank control group, ROS levels in wound tissue of diabetic mice in AuNP alone group and IL-4-AuNP group were significantly decreased (P<0.05). On the 15th day of treatment, the number of Arg-1 positive cells in the wounds of diabetic mice in IL-4-AuNP group was significantly more than that in blank control group and AuNP alone group, respectively (all P<0.05). Conclusions: IL-4-AuNP is safe in vivo, and can improve the oxidative microenvironment by removing ROS and induce macrophage polarization towards M2 phenotype, thus promote efficient diabetic wound healing and regeneration of full-thickness skin defects in diabetic mice.

Mice , Male , Animals , Interleukin-4 , Gold/pharmacology , Diabetes Mellitus, Experimental , Creatinine , Hydrogen Peroxide , Reactive Oxygen Species , Superoxides , Metal Nanoparticles , Soft Tissue Injuries , Antibodies , Inflammation , Necrosis , Hemoglobins
Journal of Experimental Hematology ; (6): 247-253, 2023.
Article in Chinese | WPRIM | ID: wpr-971132


OBJECTIVE@#To investigate the changes and roles of reactive oxygen species (ROS) and nuclear factor erythroid 2-related factor 2 (Nrf2) related antioxidases during erythroid development.@*METHODS@#Flow cytometry was used to detect the sensibility of peripheral red blood cells of wild-type mice to a strong oxidant hydrogen peroxide (H2O2). Erythroid cells from different developmental stages in bone marrow (BM) were obtained using fluorescence-activated cell sorter and the ROS levels were detected by flow cytometry. RT-qPCR was used to detect the changes of expression levels of Nrf2 and related antioxidases in erythroid cells from different developmental stages in BM. The ROS levels of the peripheral blood and BM nucleated erythrocytes in Nrf2 knockout mice were further examined. The expression level of Nrf2 in erythroid precursors isolated from 14.5 d embryonic liver of wild-type mice during differentiation and culture in vitro was detected.@*RESULTS@#In the peripheral blood of wild-type mice, the ROS level of reticulocytes and mature erythrocytes treated with H2O2 increased about 4 times and 7 times, respectively (P<0.01). In BM erythrocytes, the ROS level gradually decreased as the cells matured (r=0.85), while the expression level of Nrf2 and its related anti-oxidative genes increased (r=0.99). The ROS levels in peripheral blood erythrocytes and BM nucleated erythrocytes of Nrf2 knockout mice were significantly increased compared with wild-type mice (P<0.01). The expression of Nrf2 increased during the early erythroid development after embryonic liver cell sorting (P<0.01).@*CONCLUSION@#The expression levels of Nrf2 and its related factors vary during erythropoiesis. Nrf2 at physiological level plays an important antioxidant role during the erythroid development.

Animals , Mice , Hydrogen Peroxide , Mice, Knockout , NF-E2-Related Factor 2/metabolism , Oxidative Stress , Reactive Oxygen Species/metabolism
Chinese Journal of Natural Medicines (English Ed.) ; (6): 619-630, 2023.
Article in English | WPRIM | ID: wpr-1010975


Six new ent-abietane diterpenoids, abientaphlogatones A-F (1-6), along with two undescribed ent-abietane diterpenoid glucosides, abientaphlogasides A-B (7-8) and four known analogs were isolated from the aerial parts ofPhlogacanthus curviflorus (P. curviflorus). The structures of these compounds were determined using high-resolution electrospray ionization mass spectrometry (HR-ESI-MS), one-dimensional and two-dimensional nuclear magnetic resonance (NMR) spectroscopy, electronic circular dichroism (ECD) spectra, and quantum chemical calculations. Notably, compounds 5 and 6 represented the first reported instances of ent-norabietane diterpenoids from the genus Phlogacanthus. In the β-hematin formation inhibition assay, compounds 2, 4, 7-10, and 12 displayed antimalarial activity, with IC50 values of 12.97-65.01 μmol·L-1. Furthermore, compounds 4, 5, 8, and 10 demonstrated neuroprotective activity in PC12 cell injury models induced by H2O2 and MPP+.

Abietanes/pharmacology , Antimalarials , Hydrogen Peroxide , Biological Assay , Plant Components, Aerial
Chinese Journal of Biotechnology ; (12): 3464-3480, 2023.
Article in Chinese | WPRIM | ID: wpr-1007970


Mitophagy is a process whereby cells selectively remove mitochondria through the mechanism of autophagy, which plays an important role in maintaining cellular homeostasis. In order to explore the effect of mitophagy genes on the antioxidant activities of Saccharomyces cerevisiae, mutants with deletion or overexpression of mitophagy genes ATG8, ATG11 and ATG32 were constructed respectively. The results indicated that overexpression of ATG8 and ATG11 genes significantly reduced the intracellular reactive oxygen species (ROS) content upon H2O2 stress for 6 h, which were 61.23% and 46.35% of the initial state, respectively. Notable, overexpression of ATG8 and ATG11 genes significantly increased the mitochondrial membrane potential (MMP) and ATP content, which were helpful to improve the antioxidant activities of the strains. On the other hand, deletion of ATG8, ATG11 and ATG32 caused mitochondrial damage and significantly decreased cell vitality, and caused the imbalance of intracellular ROS. The intracellular ROS content significantly increased to 174.27%, 128.68%, 200.92% of the initial state, respectively, upon H2O2 stress for 6 h. The results showed that ATG8, ATG11 and ATG32 might be potential targets for regulating the antioxidant properties of yeast, providing a new clue for further research.

Mitophagy/genetics , Saccharomyces cerevisiae/genetics , Antioxidants , Hydrogen Peroxide/pharmacology , Reactive Oxygen Species
Braz. j. biol ; 83: 00264, 2023. tab, graf
Article in English | LILACS, VETINDEX | ID: biblio-1339364


Abstract Allium cepa L. is a commonly consumed vegetable that belongs to the Amaryllidaceae family and contains nutrients and antioxidants in ample amounts. In spite of the valuable food applications of onion bulb, its peel and outer fleshy layers are generally regarded as waste and exploration of their nutritional and therapeutic potential is still in progress with a very slow progression rate. The present study was designed with the purpose of doing a comparative analysis of the antioxidant potential of two parts of Allium cepa, i.g., bulb (edible part) and outer fleshy layers and dry peels (inedible part). Moreover, the inhibitory effect of the onion bulb and peel extracts on rat intestinal α-glucosidase and pancreatic α-amylase of porcine was also evaluated. The antioxidant potential of onion peel and bulb extracts were evaluated using 2,2-diphenyl- 1-picryl hydrazyl (DPPH), ferric-reducing antioxidant power assay (FRAP), 2,2'-azino-bis- 3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radical scavenging assay, H2O2 radical scavenging activity and Fe2+ chelating activity. Total flavonoids and phenolic content of ethanolic extract of onion peel were significantly greater as compared to that of onion bulb. Ethanolic extract of onion peel also presented better antioxidant and free-radical scavenging activity as compared to the ethanolic extract of bulb, while the aqueous extract of bulb presented weakest antioxidative potential. Onion peel extract's α-glucosidase inhibition potential was also correlated with their phenolic and flavonoid contents. The current findings presented onion peel as a possible source of antioxidative agents and phenolic compounds that might be beneficial against development of various common chronic diseases that might have an association with oxidative stress. Besides, outer dry layers and fleshy peels of onion exhibited higher phenolic content and antioxidant activities, compared to the inner bulb. The information obtained by the present study can be useful in promoting the use of vegetable parts other than the edible mesocarp for several future food applications, rather than these being wasted.

Resumo Allium cepa pertence à família Liliaceae e é rica em nutrientes e antioxidantes. Apesar das expressivas aplicações alimentares do bulbo da cebola, sua casca e outras camadas externas são geralmente consideradas resíduos, e seu potencial nutricional e terapêutico ainda é pouco explorado. O presente estudo foi delineado com o objetivo de investigar comparativamente o potencial antioxidante de duas partes de Allium cepa, por exemplo o bulbo (parte comestível) e camadas externas e cascas secas (parte não comestível). Além disso, o efeito inibitório dos extratos do bulbo de cebola e casca sobre a α-glucosidase intestinal de ratos e α-amilase pancreática suína também foi avaliado. O potencial antioxidante dos extratos da casca de cebola e bulbo foi avaliado utilizando-se 2,2-difenil-1-picrilhidrazil (DPPH), método de poder antioxidante de redução do ferro (FRAP), método 2,2'-azino-bis-3-etilbenzotiazolina-6-ácido sulfônico (ABTS) de eliminação de radicais, atividade de eliminação de radicais H2O2 e atividade quelante do Fe2+. Os flavonoides totais e os teores fenólicos do extrato de etanol da casca de cebola foram significativamente maiores quando comparados ao do bulbo. O extrato de etanol da casca de cebola também apresentou melhor atividade antioxidante e eliminação de radicais livres quando comparado ao extrato de etanol do bulbo, enquanto o extrato aquoso de bulbo apresentou menor potencial antioxidante. O potencial de inibição da α-glicosidase dos extratos de casca de cebola correlacionou-se com seus teores fenólicos e de flavonoides. Os resultados encontrados identificaram que a casca de cebola é uma possível fonte de agentes antioxidantes e compostos fenólicos que podem ser benéficos contra o desenvolvimento de várias doenças crônicas que estão associadas ao estresse oxidativo. Além disso, as camadas externas secas e as cascas da cebola exibiram maior conteúdo fenólico e atividades antioxidantes, em comparação com o bulbo interno. As informações obtidas pelo presente estudo podem promover o uso de outras partes vegetais além do mesocarpo comestível para futuras aplicações em alimentos, ao invés de serem desperdiçadas.

Animals , Rats , Onions , Antioxidants , Swine , Plant Extracts/pharmacology , alpha-Glucosidases , Hydrogen Peroxide