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1.
Braz. j. biol ; 83: e246451, 2023. graf
Article in English | MEDLINE, LILACS, VETINDEX | ID: biblio-1339402

ABSTRACT

Abstract Dipteryx alata Vogel is a tree species widely found in Cerrado, settling preferentially in well drained soils. Studies related to ecophysiology of D. alata may contribute to the decision making about using seedlings of this species in projects aimed at the recovery of degraded areas where seasonal flooding happens. This study aimed to assess the effects of flooding on photosynthetic and antioxidant metabolism and quality of D. alata seedlings cultivated or not under flooding during four assessment periods (0, 20, 40, and 60 days), followed by 100 days after the end of each assessment period (0+100, 20+100, 40+100, and 60+100 days), allowing verifying the potential for post-flooding recovery. Flooded plants showed lower photosynthetic efficiency than non-flooded plants, regardless of the periods of exposure. However, this efficiency was recovered in the post-flooding, with values similar to that of the non-flooded seedlings. Moreover, the damage to FV/FM was evidenced by an increase in the period of exposure to flooding, but recovery was also observed at this stage of the photosynthetic metabolism. Seedling quality decreased under flooding, not varying between periods of exposure, but remained lower although the increase observed in the post-flooding period, with no recovery after flooding. The occurrence of hypertrophied lenticels associated with physiological changes and an efficient antioxidant enzyme system might have contributed to the survival and recovery of these seedlings. Thus, this species is sensitive to flooding stress but capable of adjusting and recovering metabolic characteristics at 100 days after the suspension of the water stress, but with no recovery in seedling quality. Thus, we suggested plasticity under the cultivation condition and determined that the time of 100 days is not enough for the complete resumption of growth.


Resumo Dipteryx alata Vogel é uma arbórea de ampla ocorrência no Cerrado, se estabelecendo preferencialmente em solos bem drenados. Estudos referentes à ecofisiologia de D. alata em podem contribuir para a tomada de decisão sobre o uso de mudas dessa espécie em programas de recuperação de áreas degradadas sujeitas a alagamento temporário. Objetivamos com essa pesquisa avaliar os efeitos do alagamento no metabolismo fotossintético e antioxidante, além da qualidade de mudas dessa espécie, cultivadas ou não sob alagamento durante quatro períodos de avaliação (0, 20, 40 e 60 dias) seguidos de 100 dias após o término de cada período (0+100, 20+100, 40+100, 60+100 dias), possibilitando verificar o potencial de recuperação pós-alagamento. Observamos que as plantas alagadas apresentaram menor eficiência fotossintética e danos em FV/FM entretanto houve recuperação dessas características no pós alagamento. A qualidade das mudas reduziu sob alagamento não variando entre os períodos de exposição e embora tenha aumentado no pós-alagamento manteve-se menor não se recuperando. A ocorrência de lenticelas hipertrofiadas associadas a alterações fisiológicas e um eficiente sistema enzimático antioxidante devem ter contribuído para a sobrevivência e recuperação metabólica dessas mudas. Diante disso, sugerimos que a espécie é sensível ao estresse por alagamento, mas capaz de se ajustar e recuperar as características metabólicas 100 dias após a suspensão deste estresse hídrico, no entanto a qualidade da mudas não apresentou recuperação, assim, sugerimos plasticidade diante da condição de cultivo e ressaltamos que o tempo de 100 dias não é suficiente para a completa retomada do crescimento.


Subject(s)
Seedlings , Dipteryx , Photosynthesis , Floods , Antioxidants
2.
Braz. j. biol ; 83: e251198, 2023. tab, graf
Article in English | MEDLINE, LILACS, VETINDEX | ID: biblio-1339350

ABSTRACT

Abstract The present study was designed to investigate the effects of Gundelia tournefortii L. plant extract on different tissues in terms of DNA damage, biochemical and antioxidant parameter values in rats with high-calorie diets. With this aim, Wistar albino male rats were divided into 4 groups containing 6 rats each and the study was completed over 12 weeks duration. At the end of the implementation process over the 12 weeks, rats were sacrificed and blood and tissue samples were obtained. Analyses were performed on blood and tissue samples. According to results for DNA damage (8-OHdG), in brain tissue the OG2 group was significantly reduced compared to the NC group. For MDA results in liver tissue, OG1 and OG2 groups were determined to increase by a significant degree compared to the control group, while the OG2 group was also increased significantly compared to the obese group. In terms of the other parameters, comparison between the groups linked to consumption of a high calorie diet (HCD) and administration of Gundelia tournefortii L. in terms of antioxidant activities and serum samples obtained statistically significant results. Gundelia tournefortii L. plant extracts had effects that may be counted as positive on antioxidant parameter activity and were especially identified to improve DNA damage and MDA levels in brain tissues. Additionally, consumption of Gundelia tournefortii L. plant extract in the diet may have antiobesity effects; thus, it should be evaluated for use as an effective weight-loss method and as a new therapeutic agent targeting obesity.


Resumo O presente estudo foi desenhado para investigar os efeitos do extrato da planta Gundelia tournefortii L. em diferentes tecidos em termos de danos ao DNA, valores de parâmetros bioquímicos e antioxidantes em ratos com dietas hipercalóricas. Com esse objetivo, ratos Wistar albinos machos foram divididos em 4 grupos contendo 6 ratos cada e o estudo foi concluído ao longo de 12 semanas de duração. No final desse processo de implementação, os ratos foram sacrificados e amostras de sangue e tecido foram obtidas. As análises foram realizadas em amostras de sangue e tecido. De acordo com os resultados para danos ao DNA (8-OHdG), no tecido cerebral o grupo OG2 foi significativamente reduzido em comparação com o grupo NC. Para os resultados de MDA no tecido hepático, os grupos OG1 e OG2 aumentaram significativamente em comparação ao grupo controle, enquanto o grupo OG2 também aumentou significativamente em comparação ao grupo obeso. Quanto aos demais parâmetros, a comparação entre os grupos ligados ao consumo de dieta hipercalórica (DC) e à administração de Gundelia tournefortii L. em termos de atividades antioxidantes e amostras de soro obteve resultados estatisticamente significativos. Os extratos de plantas de Gundelia tournefortii L. tiveram efeitos que podem ser considerados positivos na atividade dos parâmetros antioxidantes e foram especialmente identificados para melhorar os danos ao DNA e os níveis de MDA nos tecidos cerebrais. Além disso, o consumo de extrato vegetal de Gundelia tournefortii L. na dieta pode ter efeitos antiobesidade; portanto, deve ser avaliado para uso como um método eficaz de perda de peso e como um novo agente terapêutico voltado para a obesidade.


Subject(s)
Animals , Rats , Asteraceae , Antioxidants , DNA Damage , Plant Extracts/pharmacology , Rats, Wistar , Obesity/drug therapy
3.
Int. arch. otorhinolaryngol. (Impr.) ; 26(1): 119-124, Jan.-Mar. 2022. tab, graf
Article in English | LILACS-Express | LILACS | ID: biblio-1364911

ABSTRACT

Abstract Introduction Aminoglycoside, as an antimicrobial medication, also has side-effects on the inner ears, bringing about hearing disorders. Curcumin has been proven to be a strong scavenger against various reactive oxygen species (ROS), and the increase in ROS production is considered to play an important role in the process of hearing disorder. Objective To prove that curcumin is an effective antioxidant to prevent cochlear damage based on malondialdehyde (MDA) expression. Methods The present research used 32 Rattus norvegicus, of the Wistar lineage, randomly divided into 8 groups: negative control, ototoxic control (a single dose of 40 mg/ml of gentamicin via intratympanic injection), 2 groups submitted to ototoxic control + curcumin treatment (100 mg/kg, 200 mg/kg), 2 groups who iunderwent ototoxic control + curcumin treatment for 7 days, and two groups submitted to curcumin treatment as prevention for 3 days + ototoxic induction. Results The results showed that the lowest dosage of curcumin (100 mg/kg) could decrease MDA expression on the cochlear fibroblastic wall of the ototoxic model; however using greater doses of curcumin (200 mg/kg) for 7 days would provide a better effect. Curcumin could also significantly decrease MDA expression when it was administered during the preototoxic exposure. Conclusion Curcumin can be used as a therapy for ototoxic prevention based on the decrease in MDA expression.

4.
Braz. j. med. biol. res ; 55: e11891, 2022. graf
Article in English | LILACS-Express | LILACS | ID: biblio-1364558

ABSTRACT

The non-enzymatic antioxidant system protects blood components from oxidative damage and/or injury. Herein, plasma non-enzymatic antioxidant capacity after acute strenuous swimming exercise (Exe) and exercise until exhaustion (Exh) was measured in rats. The experiments were carried out in never exposed (Nex) and pre-exposed (Pex) groups. The Nex group did not undergo any previous training before the acute strenuous swimming test and the Pex group was submitted to daily swimming for 10 min in the first week and 15 min per day in the second week before testing. Plasma glucose, lactate, and pyruvate were measured and plasma total protein sulfhydryl groups (thiol), trolox equivalent antioxidant capacity (TEAC), ferric reducing ability of plasma (FRAP), and total radical-trapping antioxidant parameter (TRAP) levels were evaluated. There were marked increases in plasma lactate concentrations (Nex-Control 1.31±0.20 vs NexExe 4.16±0.39 vs NexExh 7.19±0.67) and in thiol (Nex-Control 271.9±5.6 vs NexExh 314.7±5.7), TEAC (Nex-Control 786.4±60.2 vs NexExh 1027.7±58.2), FRAP (Nex-Control 309.2±17.7 vs NexExh 413.4±24.3), and TRAP (Nex-Control 0.50±0.15 vs NexExh 2.6±0.32) levels after acute swimming and/or exhaustion. Also, there were increased plasma lactate concentrations (Pex-Control 1.39±0.15 vs PexExe 5.22±0.91 vs PexExh 10.07±0.49), thiol (Pex-Control 252.9±8.2 vs PexExh 284.6±6.7), FRAP (Pex-Control 296.5±15.4 vs PexExh 445.7±45.6), and TRAP (Pex-Control 1.8±0.1 vs PexExh 4.6±0.2) levels after acute swimming and/or exhaustion. Lactate showed the highest percent of elevation in the Nex and Pex groups. In conclusion, plasma lactate may contribute to plasma antioxidant defenses, and the TRAP assay is the most sensitive assay for assessing plasma non-antioxidant capacity after strenuous exercise.

5.
Organ Transplantation ; (6): 126-2022.
Article in Chinese | WPRIM | ID: wpr-907043

ABSTRACT

Common marginal donor liver mainly consists of fatty donor liver, elderly donor liver, small volume donor liver and liver graft from donation after cardiac death (DCD), etc. The application of marginal donor liver may resolve the severe shortage of donor liver to certain extent. Nevertheless, marginal donor liver yields a higher risk of ischemia-reperfusion injury (IRI) and causes more severe IRI than normal donor liver, which is a main cause for the failure of transplantation. In addition, oxidative stress is a major risk factor causing IRI of marginal donor liver. Therefore, how to mitigate oxidative stress and alleviate IRI of marginal donor liver has become a hot spot in clinical practice. Reactive oxygen species (ROS)-mediated oxidative stress occurs throughout the whole process of IRI. In this article, the role of oxidative stress in IRI of marginal donor liver transplantation and the ROS-targeted prevention and treatment were reviewed, aiming to provide reference for clinical practice.

6.
Article in English | WPRIM | ID: wpr-906679

ABSTRACT

@#BACKGROUND: Individuals who survive a cardiac arrest often sustain cognitive impairments due to ischemia-reperfusion injury. Mesenchymal stem cell (MSC) transplantation is used to reduce tissue damage, but exosomes are more stable and highly conserved than MSCs. This study was conducted to investigate the therapeutic effects of MSC-derived exosomes (MSC-Exo) on cerebral ischemia-reperfusion injury in an in vitro model of oxygen-glucose deprivation/reperfusion (OGD/R), and to explore the underlying mechanisms. METHODS: Primary hippocampal neurons obtained from 18-day Sprague-Dawley rat embryos were subjected to OGD/R treatment, with or without MSC-Exo treatment. Exosomal integration, cell viability, mitochondrial membrane potential, and generation of reactive oxygen species (ROS) were examined. Terminal deoxynucleotidyl transferase-mediated 2’-deoxyuridine 5’-triphosphate nick-end labeling (TUNEL) staining was performed to detect neuronal apoptosis. Moreover, mitochondrial function-associated gene expression, Nrf2 translocation, and expression of downstream antioxidant proteins were determined. RESULTS: MSC-Exo attenuated OGD/R-induced neuronal apoptosis and decreased ROS generation (P<0.05). The exosomes reduced OGD/R-induced Nrf2 translocation into the nucleus (2.14±0.65 vs. 5.48±1.09, P<0.01) and increased the intracellular expression of antioxidative proteins, including superoxide dismutase and glutathione peroxidase (17.18±0.97 vs. 14.40±0.62, and 20.65±2.23 vs. 16.44±2.05, respectively; P<0.05 for both). OGD/R significantly impaired the mitochondrial membrane potential and modulated the expression of mitochondrial function-associated genes, such as PINK, DJ1, LRRK2, Mfn-1, Mfn-2, and OPA1. The abovementioned changes were partially reversed by exosomal treatment of the hippocampal neurons. CONCLUSIONS: MSC-Exo treatment can alleviate OGD/R-induced oxidative stress and dysregulation of mitochondrial function-associated genes in hippocampal neurons. Therefore, MSC-Exo might be a potential therapeutic strategy to prevent OGD/R-induced neuronal injury.

7.
Acta Pharmaceutica Sinica ; (12): 296-302, 2022.
Article in Chinese | WPRIM | ID: wpr-922938

ABSTRACT

Reactive oxygen species (ROS) is defined as the electron reduction product of oxygen with high reactivity which can maintain normal physiological functions and redox homeostasis. The tumor microenvironment is in a state of oxidative stress. ROS can affect multiple processes of tumor immune response by modulating the phenotype and functions of tumor cells and immune cells. With the rapid development of immunology, ROS-based tumor immunomodulation has been widely concerned and studied. In this review, the mechanism of ROS participating in tumor immune response is elaborated. Meanwhile, the research process and application of ROS in tumor immunomodulation in recent years are reviewed and analyzed.

8.
Journal of Clinical Hepatology ; (12): 372-380, 2022.
Article in Chinese | WPRIM | ID: wpr-920887

ABSTRACT

Objective Drug resistance is the main cause of chemotherapy failure in hepatocellular carcinoma (HCC), and thioredoxin reductase 1 (TXNRD1), as a major influencing factor for reactive oxygen species (ROS) metabolism, has been proven to be associated with the poor prognosis of patients with HCC. This study aims to explore the role of TXNRD1 in the mechanism of multidrug resistance in HCC. Methods BEL/FU cells in BEL-7402 cell line were selected as the multidrug-resistant cell line. The siRNA was used for the intervention of TXNRD1 expression; quantitative real-time PCR and Western blotting were used to measure the expression of TXNRD1; CCK-8 assay and flow cytometry were used to evaluate the effect of TXNRD1 on hepatocyte ROS accumulation, resistance to 5-fluorouracil (5-Fu) and doxorubicin (DOX), and apoptosis in vitro; a xenograft tumor model was established to investigate the effect of auranofin (AUR) on drug resistance in vivo. The two-independent-samples t test was used for comparison of continuous data between two groups. Results As a multidrug-resistant HCC cell line, BEL/Fu showed high mRNA and protein expression levels of TXNRD1 (both P < 0.05). Compared with 5-Fu or DOX treatment alone, the TXNRD1 inhibitor AUR combined with 5-Fu or DOX had had a significant reduction in the number of colony formation ( P < 0.01) and a significant increase in apoptosis ratio ( P < 0.001). The ROS scavenger N-acetylcysteine (NAC) significantly weakened the effect of TXNRD1 knockdown by siRNA on the drug resistance of BEL/Fu cells, and the application of NAC effectively reduced the apoptosis ratio of cells after siRNA interference ( P < 0.001). Animal experiments also confirmed that compared with the nude mice treated with 5-Fu alone, the nude mice treated with 5-Fu and AUR had a significantly lower tumor mass ( P < 0.001) and a significantly smaller tumor volume ( P < 0.001). Conclusion TXNRD1 plays an important role in the drug resistance of HCC, and inhibition of its level in cells can effectively improve drug resistance. As a TXNRD1 inhibitor, AUR has great application prospects in the multimodality therapy for HCC.

9.
Braz. j. biol ; 82: e241081, 2022. tab, graf
Article in English | MEDLINE, LILACS, VETINDEX | ID: biblio-1285584

ABSTRACT

Abstract This study investigated the use of melatonin to arrest the effects of apoptosis in vitrified zebrafish (D. rerio) embryos. Dechorionated embryos at 22-24 somite-stage were divided (n = 60/treatment) into a non-vitrified (Control Group, 0 M melatonin) and vitrified treatments with 0 M (T1), 1 µM (T2) and 1 mM of melatonin (T3). For vitrified treatments, a solution methanol/propylene glycol based was used and the embryos stored in -196 °C for a week. After thaw, survival rate, scanning electron microscopy, expression of anti (bcl-2) and pro-apoptotic (bax/caspase-3) genes, reactive oxygen species (ROS) formation and DNA fragmentation analyses were performed. No live embryos were obtained from vitrified treatments, observing a rapid degeneration immediately after thawing, with the vitelline layer rupture and leakage of its content, followed by breakdown of epithelial cells and melanisation of the tissue. Regarding the apoptotic process, T3 had the highest relative gene expression, for the three genes (P < 0.05) furthermore, T2 had similar expression of pro-apoptotic genes to CG (P < 0.05). ROS formation revealed that CG presented lower percentage of embryo surface area affected (3.80 ± 0.40%) (P < 0.05), in contrast, no differences were found among the other groups. T1 was most significantly (P < 0.05) damaged by DNA fragmentation. The vitrified groups with melatonin had similar damage levels of CG (P > 0.05). The inclusion of 1 µM of melatonin in the vitrifying solution, countered the effects of apoptotic process in post-thaw embryos, suggesting its utility in cryopreserving fish embryos.


Resumo Este estudo investigou o uso da melatonina para conter os efeitos da apoptose em embriões vitrificados de zebrafish (D. rerio). Embriões descorionados no estágio de 22-24 somitos foram divididos (n = 60 / tratamento) em tratamento não vitrificado (Grupo Controle, melatonina 0 M) e tratamentos vitrificados com 0 M (T1), 1 µM (T2) e 1 mM de melatonina (T3). Para os tratamentos vitrificados, utilizou-se uma solução à base de metanol/propilenoglicol e os embriões foram armazenados em -196 °C por uma semana. Após o descongelamento, foram realizadas análises de taxa de sobrevivência, microscopia eletrônica de varredura, expressão dos genes anti (bcl-2) e pró-apoptóticos (bax/caspase-3), formação de espécies reativas de oxigênio (EROS) e análises de fragmentação de DNA. Não foram obtidos embriões vivos a partir dos tratamentos vitrificados, observando uma rápida degeneração imediatamente após o descongelamento, com ruptura da camada vitelina e vazamento de seu conteúdo, seguida de quebra das células epiteliais e melanização do tecido. Em relação ao processo apoptótico. T3 apresentou expressão gênica relativa alta para os três genes (P <0,05), além disso, T2 apresentou expressão semelhante as dos genes pró-apoptóticos de GC (P <0,05). A formação de EROS revelou que GC apresentou menor percentual de área de superfície embrionária afetada (3,80 ± 0,40%) (P <0,05), ao contrário, não foram encontradas diferenças entre os outros grupos. T1 foi mais significativamente (P <0,05) danificado pela fragmentação do DNA. Os grupos vitrificados com melatonina apresentaram níveis de dano semelhantes ao do GC (P> 0,05). A inclusão de 1 µM de melatonina na solução de vitrificação, contrariou os efeitos do processo apoptótico em embriões pós-descongelamento, sugerindo sua utilidade na criopreservação de embriões de peixes.


Subject(s)
Animals , Zebrafish , Melatonin/pharmacology , Cryopreservation , Apoptosis
10.
Ciênc. rural (Online) ; 52(1): e20200955, 2022. tab, graf
Article in English | LILACS-Express | LILACSEXPRESS, LILACS, VETINDEX | ID: biblio-1286036

ABSTRACT

ABSTRACT: This study identified physiological and biochemical changes in 'Fuyu' persimmon buds during dormancy. Branches were collected between March and August 2015. Dormancy was evaluated by biological testing of isolated node cuttings at 25 °C and a photoperiod of 16 h. The variables analyzed were water content; reducing sugar content; respiratory activity; activity of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (POD) and polyphenol oxidase (PPO) enzymes; hydrogen peroxide content (H2O2) and lipid peroxidation. At the end of March 2015, the plants were already dormant, and the leaves and fruits present indicated a paradormancy effect. Induction of endodormancy may have occurred in June 2015, when chilling hours (CH) below 7.2 °C and higher CH below 12 °C began to accumulate, which coincided with the period in which there was a decrease in water content and respiratory activity, an increase in reducing sugars, a decrease in SOD, CAT, APX and PPO and an increase in H2O2. After an accumulation of 553 CH below 12 °C, the budburst capacity increased, and the buds presented increased water content, decreased reducing sugars content, increased respiratory activity, low activity in SOD, CAT, APX and POD and high levels of H2O2.


RESUMO: O objetivo deste trabalho foi identificar alterações fisiológicas e bioquímicas em gemas de caquizeiro 'Fuyu' durante a dormência. Ramos foram coletados entre março e agosto de 2015. A dormência foi avaliada pelo teste biológico de estacas de nós isolados, a 25 °C e fotoperíodo de 16 h. As variáveis analisadas foram umidade ponderal, teor de açúcares solúveis, atividade respiratória, atividade das enzimas superóxido dismutase (SOD), catalase (CAT), ascorbato peroxidase (APX), guaiacol peroxidase (POD) e polifenoloxidase (PPO), teor de peróxido de hidrogênio (H2O2) e peroxidação lipídica. No final de março de 2015 as plantas já estavam dormentes, as folhas e frutos presentes indicam efeito de paradormência. A indução da endodormência pode ter ocorrido em junho de 2015, quando iniciou acúmulo de horas de frio (HF) abaixo de 7,2 °C e maiores HF abaixo de 12 °C, que coincidiu com o período que houve diminuição da umidade e da atividade respiratória, aumento dos açúcares redutores, diminuição da atividade da SOD, CAT, APX e PPO e aumento de H2O2. Após acúmulo de 553 HF abaixo de 12 °C, a capacidade de brotação aumentou e as gemas apresentaram aumento da umidade, diminuição do teor de açúcares redutores, aumento da atividade respiratória, baixa atividade da SOD, CAT, APX e POD e elevados teores de H2O2.

11.
Electron. j. biotechnol ; 52: 76-84, July. 2021. graf, ilus
Article in English | LILACS | ID: biblio-1283597

ABSTRACT

BACKGROUND: Butyrate is a histone deacetylase inhibitor that induces apoptosis and inhibits cell proliferation of colorectal cancer cells. To improve its anticancer activity, butyrate has been evaluated mixed with drugs and different molecules. Plant antimicrobial peptides are attractive anticancer alternative molecules because they show selective cytotoxic activity against different cancer cell lines. In this work, we explore if the plant defensin c-thionin (Capsicum chinense) can improve butyrate activity on Caco-2 cell line and we also determined the mechanism of death activated. RESULTS: The combined treatment of c-thionin (3.5 mM) and butyrate (50 mM) showed higher cytotoxicity on Caco-2 cells with respect to single treatments. Also, the combined treatment reduced cell proliferation and exhibited a higher rate of apoptosis than single treatments. Combined treatment induced caspases 8 and 9 activation to an extent comparable with that of butyrate while c-thionin did not activate caspases. Additionally, reactive oxygen species generation preceded the onset of apoptosis, and superoxide anion production was higher in cells treated with the combined treatment. CONCLUSIONS: The c-thionin from Habanero chili pepper improved the butyrate cytotoxicity on Caco-2 cells. This effect occurred through apoptosis induction associated with reactive oxygen species production. Therefore, the combination of butyrate with cytotoxic antimicrobial peptides could be an attractive strategy for cancer therapy.


Subject(s)
Humans , Butyrates , Capsicum/chemistry , Adenocarcinoma , Colonic Neoplasms , Cell Cycle , Reactive Oxygen Species , Apoptosis , Caco-2 Cells , Defensins , Thionins
12.
Braz. arch. biol. technol ; 64: e21200733, 2021. tab, graf
Article in English | LILACS-Express | LILACS | ID: biblio-1360191

ABSTRACT

Abstract The post-harvest resting of the fruits can improve seed physiological quality ,once it allows the seed to complete the maturation process, so it has been a common practice in vegetable seed companies, however, there are a few studies of this technique in sweet pepper. The objective of this research was to evaluate physiological quality, and biochemical response of sweet's peppers in regarding on the stage of maturation and the post-harvest rest of the fruits. The experimental was conducted in a 4x2 factorial, being the first factor comprised four maturation stages (35, 50, 65 and 80 days after anthesis) and, the second the post-harvest management of fruits, with and without a temporary storage of seven days. Seeds were evaluated for water content, weight of thousand seeds, germination, vigor, superoxide dismutase, catalase and peroxidase activities, lipid peroxidation and hydrogen peroxide content. Fruit harvest time indicated is 80 days after anthesis (fruits 100% yellow) when seeds showed maximum germination and vigor. The post-harvest resting of the fruits was beneficial to seed physiological quality, weight of one thousand seeds and to reduce hydrogen peroxide content. Seeds of higher physiological quality showed lower superoxide dismutase and catalase enzyme activity, so they can be used as a marker of physiological quality in sweet pepper seeds.

13.
Braz. j. med. biol. res ; 54(10): e10730, 2021. graf
Article in English | LILACS | ID: biblio-1285651

ABSTRACT

Chondroitin sulfate (CS) is a type of glycosaminoglycan described as an antioxidant molecule that has been found in animal species such as fish. Tilapia (Oreochromis niloticus) represents an eco-friendly source of this compound, since its economical processing generates usable waste, reducing the negative environmental impact. This waste was used for CS extraction, purification, characterization by enzymatic degradation, and evaluation of its antioxidant effect. CS obtained from tilapia presented sulfation mainly at carbon 4 of galactosamine, and it was not cytotoxic at concentrations up to 200 µg/mL. Furthermore, 100 µg/mL of CS from tilapia reduced the levels of reactive oxygen species to 47% of the total intracellular reactive oxygen species level. The ability of CS to chelate metal ions in vitro also suggested an ability to react with other pathways that generate oxidative radicals, such as the Haber-Weiss reaction, acting intracellularly in more than one way. Although the role of CS from tilapia remains unclear, the pharmacological effects described herein indicate that CS is a potential molecule for further study of the relationship between the structures and functions of chondroitin sulfates as antioxidants.


Subject(s)
Animals , Chondroitin Sulfates , Antioxidants/pharmacology , Reactive Oxygen Species , Fishes , Glycosaminoglycans
14.
Chinese Journal of Anesthesiology ; (12): 1212-1217, 2021.
Article in Chinese | WPRIM | ID: wpr-911344

ABSTRACT

Objective:To evaluate the role of reactive oxygen species (ROS)-mediated mitochondrial pathway of apoptosis in long-term cognitive impairment induced by multiple exposures to sevoflurane in the neonatal rats.Methods:Sixty SPF healthy neonatal Sprague-Dawley rats, weighing 12-20 g, were divided into 3 groups ( n=20 each) using a random number table method: control group (group C), multiple exposures to sevoflurane for anesthesia group (group S) and ROS inhibitor group (group A). Group S and group A inhaled 3% sevoflurane for 2 h starting from 6, 7 and 8 days after birth, while group C inhaled air.In group A, ROS inhibitor N-acetylcysteine (NAC) 150 mg/kg was intraperitoneally injected before each anesthesia with sevoflurane.The spontaneous activity was evaluated by open field test on day 35 after birth.The cognitive function was determined by Morris water maze test on day 36 after birth.The rats were sacrificed after the end of Morris water maze test, and the hippocampal tissues were obtained for determination of the apoptosis rate of hippocampal neurons, reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) (by flow cytometry) and levels of Cyt c and cleaved caspase-9 and caspase-3 (by Western blot). The expression of Bcl-2 and Bax mRNA was detected by real-time polymerase chain reaction.The ultrastructure of mitochondria in hippocampal neurons was observed with a transmission electron microscope. Results:Compared with group C, the escape latency was significantly prolonged, the number of crossing the original platform was reduced, the apoptosis rate of hippocampal neurons and levels of ROS and MMP were increased, the expression of Cyt c, cleaved caspase-9, cleaved caspase-3 and Bax mRNA was up-regulated, the expression of Bcl-2 mRNA was down-regulated, the ratio of Bax/Bcl-2 was increased ( P<0.05), mitochondria were swollen, and mitochondrial cristae structure was broken in group S. Compared with group S, the escape latency was significantly shortened, the number of crossing the original platform was increased, the apoptosis rate of hippocampal neurons and levels of ROS and MMP were decreased, the expression of Cyt c, cleaved caspase-9, cleaved caspase-3 and Bax mRNA was down-regulated, the expression of Bcl-2 mRNA was up-regulated, the ratio of Bax/Bcl-2 was decreased ( P<0.05), and the mitochondrial swelling and rupture of cristae structure were improved in group A. Conclusion:The mechanism by which multiple exposures to sevoflurane induce long-term cognitive impairment may be related to activating the ROS-mediated mitochondrial pathway of apoptosis in neonatal rats.

15.
Article in Chinese | WPRIM | ID: wpr-910438

ABSTRACT

Objective:To investigate the mechanism underlying the inhibiting effect of low-glucose combined with palmitic acid on human colon cancer cells and its influence on the radiosensitivity.Methods:Under the treatment of low-glucose, palmitic acid and low-glucose combined with palmitic acid, the treatment condition that significantly inhibited the proliferation of SW480 was screened by CCK-8 assay. The reactive oxygen species (ROS) level, mitochondrial membrane potential and apoptosis rate were detected by flow cytometry. The changes in the radiosensitivity were detected by immunofluorescence-based γ-H 2AX quantification and colony formation assay. The protein expression level was detected by Western blot. Results:Compared with the control group, the condition of low-glucose combined with 120μmol/L palmitic acid significantly inhibited the proliferation of SW480 cells ( P<0.01). The expression levels of CPT1a, PFKFB3 and PKM were significantly up-regulated, the expression levels of NDUFV1, NDUFV2 and NDUFS1 were remarkably down-regulated, the ROS level was significantly increased and the ATP level was considerably reduced in the cells under metabolic stress (all P<0.01). After irradiation, the number of γ-H 2AX foci was significantly increased ( P<0.05), and the D 0 value was significantly reduced ( P<0.01), the ROS level was considerably increased ( P<0.001), the apoptosis rate was significantly increased ( P<0.001) and the expression level of γ-H 2AX protein was remarkably up-regulated ( P<0.01) in the low-glucose combined with 120μmol/L palmitic acid group. Pretreatment with NAC could reverse the changes of ROS, apoptosis and γ-H 2AX protein expression. Conclusions:The combination of low-glucose and palmitic acid can induce metabolic stress in SW480 cells, inhibit tumor proliferation and increase the radiosensitization when combined with radiotherapy by inducing the generation of ROS and DNA damage.

16.
Article in Chinese | WPRIM | ID: wpr-907564

ABSTRACT

Objective:To study the effect of thioredoxin domain containing protein 5 (TXNDC5)-peroxiredoxin 2 (Prx2) on the drug resistance of prostate cancer cells.Methods:Prostate cancer PC3 cells were cultured in vitro, treated with the chemotherapy drug cyclophosphamide (5, 10, 15 μmol/L) for 24 hours, and PC3 cells without any treatment was served as the control group. The expression levels of TXNDC5 in PC3 cells were detected by real-time fluorescent quantitative PCR (RT-qPCR) and Western blotting. PC3 cells with TXNDC5 knocking down were exposed by cyclophosphamide and CCK-8 was used to detect the cell viability of siTXNDC5 group and siNC group. The content of reactive oxygen free radicals was determined by reactive oxygen detection kit. PC3 cells and its parental cyclophosphamide-resistant ones with TXNDC5 knocking down were treated by 10 μmol/L cyclophosphamide and subjected for CCK8 assay. The expression of Prx2 in PC3 cells was detected by Western blotting after TXNDC5 was silenced. Prx2 expression was silenced in PC3 cells overexpressing TXNDC5, and cell viability and reactive oxygen free radical content were detected in Vec-Ctrl group, pcTXNDC5 group, siNC group, siPrx2 group and pcTXNDC5+ siPrx2 group. Results:Compared with the control group, cyclophosphamide treatment significantly increased the expression of TXNDC5 at mRNA and protein levels in PC3 cells. After PC3 cells were treated with cyclophosphamide (10, 15 μmol/L) for 12 h, compared with the siNC group, the cell viability in the siTXNDC5 group was significantly suppressed (0.44±0.08 vs. 0.74±0.10, t=3.647, P=0.031; 0.30±0.04 vs. 0.53±0.06, t=6.115, P=0.006). When PC3 cells were treated with 10 μmol/L cyclophosphamide for 6 and 12 h, compared with the siNC group, the production of reactive oxygen free radicals in the siTXNDC5 group was significantly increased (2.68±0.19 vs. 1.58±0.26, t=-6.027, P=0.005; 4.56±0.37 vs. 2.73±0.26, t=-6.995, P=0.003). When PC3 cells and its cyclophosphamide-resistant ones were treated with 10 μmol/L cyclophosphamide for 12 h, compared with the siNC group, the cell viability was significantly inhibited in the siTXNDC5 group. Western blotting analysis showed that the expression of Prx2 was significantly reduced when TXNDC5 was silenced. Silencing Prx2 could significantly attenuate the increase of cell viability and the decrease of reactive oxygen content resulting from TXNDC5 overexpression. PC3 cells were treated with 10 μmol/L cyclophosphamide for 12 h, and the cell viabilities of the Vec-Ctrl group, pcTXNDC5 group, siNC group, siPrx2 group and pcTXNDC5+ siPrx2 group were 0.52±0.07, 0.69±0.03, 0.56±0.05, 0.43±0.05, 0.58±0.07, respectively, and there was a statistically significant difference ( F=8.868, P=0.003). Furthermore, the cell viability in the pcTXNDC5+ siPrx2 group decreased significantly when compared to that of the pcTXNDC5 group ( P=0.045). The contents of reactive oxygen free radicals in the above 5 groups were 3.26±0.46, 2.09±0.49, 3.16±0.38, 4.62±0.26, 2.87±0.36, respectively, and there was a statistically significant difference ( F=16.037, P<0.001). The content of reactive oxygen radicals in the pcTXNDC5+ siPrx2 group was higher than that of the pcTXNDC5 group ( P=0.036). Conclusion:TXNDC5 can reduce the level of reactive oxygen free radicals in prostate cancer cells by regulating the expression of Prx2, so as to promote the drug resistance of prostate cancer cells.

17.
Article in Chinese | WPRIM | ID: wpr-906389

ABSTRACT

Objective:To explore the inhibitory effect of dihydroartemisinin (DHA) on the proliferation of HepG2 cells, elucidate the mechanism from the perspectives of oxidative damage and energy metabolism, and discuss the possibility of combined use of DHA with sorafenib (Sora). Method:Cell counting kit-8 (CCK-8) assay was used to obtain the 50% inhibitory concentration (IC<sub>50</sub>) of DHA and Sora on HepG2 and SW480 cells and Chou-Talalay method was used to obtain the combination index (CI) of DHA and Sora. HepG2 cells were classified into the control group, DHA group (10 µmol·L<sup>-1</sup>), Sora group (5 µmol·L<sup>-1</sup>), and DHA + Sora group (DHA 10 µmol·L<sup>-1</sup>, Sora 5 µmol·L<sup>-1</sup>) and then incubated with corresponding drugs for 8-12 h. Seahorse XF glycolytic rate assay kit and cell mito stress test kit were employed to respectively detect the glycolysis function of cells and oxidative phosphorylation function of mitochondria. DCFH-DA and lipid peroxidation MDA assay kit were separately used to analyze the intracellular levels of reactive oxygen species (ROS) and malondialdehyde (MDA). Western blot was applied to determine the intracellular levels of heme oxygenase-1 (HO-1) and glutamate-cysteine ligase catalytic subunit (GCLC). Result:Compared with the control group, DHA alone inhibited the ATP synthesis in mitochondrial oxidative phosphorylation and glycolysis (<italic>P</italic><0.01), increased the levels of intracellular ROS and MDA (<italic>P<</italic>0.05), and decreased the levels of HO-1 and GCLC (<italic>P<</italic>0.05) in HepG2 cells. DHA and Sora had synergistic inhibitory effect on proliferation of HepG2 and SW480 cells, with CI < 0.90. The DHA + Sora group showed stronger suppression of ATP synthesis in mitochondrial oxidative phosphorylation and glycolysis (<italic>P</italic><0.01), higher levels of intracellular ROS and MDA (<italic>P<</italic>0.01), and lower levels of intracellular antioxidation-related proteins HO-1 and GCLC in HepG2 cells (<italic>P<</italic>0.01) than the DHA group. Conclusion:DHA may increase the level of MDA by reducing HO-1 and GCLC and increasing ROS in HepG2 cells, which results in mitochondria oxidative damage, restricts cell glycolysis and mitochondrial oxidative phosphorylation, and thus finally inhibits the proliferation of HepG2 cells. DHA and Sora have synergistic inhibitory effect on the proliferation of HepG2 and SW480 cells, and the mechanism may be related to the synergistic oxidative damage that affects the mitochondrial electron transport chain and suppresses cell energy metabolism.

18.
Article in Chinese | WPRIM | ID: wpr-906174

ABSTRACT

Objective:To investigate the effect of Ginseng Radix et Rhizoma-Notoginseng Radix et Rhizoma-Chuanxiong Rhizoma extract on endothelial microparticles (EMPs)-induced vascular endothelial cell senescence, and explore the possible mechanism. Method:Human umbilical vein endothelial cells (HUVECs) were used as the research objects, and the aged model was established with 10-12 passages of replicative senescence cells. The experimental cells were divided into young group (2-4 passage cells), aged group (10-12 passage cells), only EMPs intervention group (extract EMPs produced by aged cells to intervene young cells) and low dose, middle dose and high dose drug intervention groups (200, 300, 400 mg·L<sup>-1</sup>). Senescence related <italic>β</italic>-galactosidase (SA-<italic>β</italic>-gal) staining and cell cycle propidium iodide (PI) staining were used to determine cell senescence. Cell counting kit-8 (CCK-8) assay was used to screen the drug concentration. EMPs were extracted by two-step centrifugation, EMPs labeled with phycoerythrin (PE) anti-human CD31 antibody or fluorescein isothiocyanate (FITC) annexin V were detected by flow cytometry, intracellular reactive oxygen species (ROS) were detected by 2',7'- dichlorofluorescein diacetate (DCFDA) staining. Result:After treatment with the drug, SA-<italic>β</italic>-gal activity of the aged cells significantly decreased (<italic>P</italic><0.01), the S phase arrest was restored (<italic>P</italic><0.01), and the number of CD31<sup>+</sup> EMPs and annexin V<sup>+</sup> EMPs secreted by aged cells decreased (<italic>P</italic><0.05). Compared with the young group, only EMPs intervention group could induce increased SA-<italic>β</italic>-gal activity and S phase arrest in young cells (<italic>P</italic><0.05,<italic>P</italic><0.01). However, after intervention of EMPs and the drug, EMPs-mediated increase of SA-<italic>β</italic>-gal activity was significantly inhibited and S phase arrest was restored (<italic>P</italic><0.05). The increase of intracellular ROS induced by EMPs was also significantly inhibited by the drug (<italic>P</italic><0.05,<italic>P</italic><0.01). Conclusion:Ginseng Radix et Rhizoma-Notoginseng Radix et Rhizoma-Chuanxiong Rhizoma extract can delay the senescence of vascular endothelial cells by influencing EMPs, and the mechanism may be related to the inhibition of increased intracellular ROS induced by EMPs.

19.
Article in Chinese | WPRIM | ID: wpr-906086

ABSTRACT

Objective:To investigate the effect of licochalcone A (LCA) on apoptosis in human breast cancer MDA-MB-231 cells, and to explore its possible mechanism. Method:MDA-MB-231 cells were treated with LCA of different concentrations, and<italic> </italic>cell counting kit-8 (CCK-8) assay was used to detect the cell viability. The cells were treated with LCA (10, 20, and 40 μmol·L<sup>-1</sup>) for 24 h, and apoptosis was detected by Annexin V staining with fluorescein isothiocyanate (FITC) and propidium iodide (PI) (Annexin V-FITC/PI). The level of intracellular reactive oxygen species (ROS) was detected by 2′,7′-dichlorodihydrofluorescein diacetate (DCFA-DA) fluorescent probe. Mitochondrial membrane potential (MMP) was detected by 5, 5′, 6, 6′-tetrachloro-1, 1′, 3, 3′-tetraethyl-imidacarbocyanine (JC-1) fluorescence probe. Western blot was used to detect the expression of cell apoptosis-related proteins, such as B-cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax), and endoplasmic reticulum (ER) stress-related proteins, such as C/EBP homologous protein (CHOP), activating transcription factor 4 (ATF4), protein kinase R-like ER kinase (PERK), p-PERK, eukaryotic translation initiation factor 2 alpha (eIF2<italic>α</italic>), and p-eIF2<italic>α</italic>. Result:With the increase in the drug concentration (starting from 5 μmol·L<sup>-1</sup>), the cell viability decreased (<italic>P<</italic>0.05) with IC<sub>50 </sub>of 19.05 μmol·L<sup>-1</sup> as compared with the normal group. Additionally, the apoptosis rates of the LCA groups (10, 20, 40 μmol·L<sup>-1</sup>) significantly increased (<italic>P</italic><0.05), which reached 30.2% (<italic>P</italic><0.05) at LCA concentration of 40 μmol·L<sup>-1</sup>. LCA (10, 20, and 40 μmol·L<sup>-1</sup>) decreased the expression of Bcl-2 (<italic>P<</italic>0.05) and increased Bax expression (<italic>P<</italic>0.05) in a dose-dependent manner. Besides, the intracellular ROS level was elevated (<italic>P<</italic>0.05) and mitochondrial MMP was reduced (<italic>P<</italic>0.05) after LCA (10, 20, and 40 μmol·L<sup>-1</sup>) treatment in a dose-dependent manner, leading to mitochondrial dysfunction. LCA (10, 20, and 40 μmol·L<sup>-1</sup>) induced ER stress to up-regulate the expression of CHOP, ATF4, p-PERK, and p-eIF2<italic>α</italic> (<italic>P<</italic>0.05) in a dose-dependent manner. Conclusion:LCA can induce MDA-MB-231 cell apoptosis by increasing intracellular ROS level and reducing MMP to trigger mitochondrial dysfunction and ER stress.

20.
Article in Chinese | WPRIM | ID: wpr-905907

ABSTRACT

Objective:To investigate the biological essence of the content variation of differential primary and secondary metabolites in fresh<italic> </italic>roots of <italic>Scutellaria baicalensis </italic>under drought stress. Method:The changes of metabolites were analyzed by ultra-high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass/mass (UHPLC-ESI-Q-TOF-MS/MS). Result:A total of 11 differential compounds were identified from the roots of <italic>S. baicalensis</italic> (VIP≥2). Under drought stress, citric acid content increased and shikimic acid content decreased, indicating that the drought stress weakened the primary metabolism but strengthened secondary metabolism. Drought stress raised the content and regulated the proportion of various secondary metabolites by modulating the biosynthesis and biotransformation of them. To be specific, the content of free flavonoids with many phenolic hydroxyl groups and high biological activity and pharmacological activity, such as baicalin, wogonoside, baicalein, wogonin, chrysin, eriodictyol, 5,2',6'-trihydroxy-7,8-dimethoxyflavone, 5,8-dihydroxy-6,7-dimethoxyflavone, and 3,5,7,2',6'-pentahydroxyflavanone, was significantly increased. The massive compounds, like an intricate buffer, maintain metabolism stable as quickly and accurately as possible through biosynthesis and biotransformation, thus responding to the changing environment, which reveals how the quality of genuine regional drugs is influenced and why compounds in herbal medicine are complex. Conclusion:Secondary metabolites with low content but high activity are important influencing factors of medicinal material quality and metabolites with high content and high activity are evaluation indicators of genuine regional drug quality.

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