Effect of octreotide on oxidative stress in the erythrocyte and kidney tissue in adriamycin-induced experimental nephrotic syndrome model / Efeito do octreotide no estresse oxidativo em eritrócitos e no tecido renal no modelo de síndrome nefrótica experimental induzida por adriamicina

Abstract Introduction: Nephrotic syndrome (NS) is one of the reasons of end-stage kidney disease, and elucidating the pathogenesis and offer new treatment options is important. Oxidative stress might trigger pathogenesis systemically or isolated in the kidneys. Octreotide (OCT) has beneficial antioxidant effects. We aimed to investigate the source of oxidative stress and the effect of OCT on experimental NS model. Methods: Twenty-four non-uremic Wistar albino rats were divided into 3 groups. Control group, 2 mL saline intramuscular (im); NS group, adriamycin 5 mg/kg intravenous (iv); NS treatment group, adriamycin 5 mg/kg (iv) and OCT 200 mcg/kg (im) were administered at baseline (Day 0). At the end of 21 days, creatinine and protein levels were measured in 24-hour urine samples. Erythrocyte and renal catalase (CAT) and thiobarbituric acid reactive substance (TBARS) were measured. Renal histology was also evaluated. Results: There was no significant difference among the 3 groups in terms of CAT and TBARS in erythrocytes. Renal CAT level was lowest in NS group, and significantly lower than the control group. In treatment group, CAT level significantly increased compared with NS group. In terms of renal histology, tubular and interstitial evaluations were similar in all groups. Glomerular score was significantly higher in NS group compared with control group and it was significantly decreased in treatment group compared to NS group. Conclusions: Oxidative stress in NS might be due to the decrease in antioxidant protection mechanism in kidney. Octreotide improves antioxidant levels and histology in renal tissue and might be a treatment option.

Resumo Introdução: Síndrome nefrótica (SN) é uma das causas de doença renal em estágio terminal. É importante elucidar a patogênese e oferecer novas opções de tratamento. Estresse oxidativo pode desencadear a patogênese sistemicamente ou isoladamente nos rins. O octreotide (OCT) tem efeitos antioxidantes benéficos. Nosso objetivo foi investigar a fonte de estresse oxidativo e efeito do OCT no modelo experimental de SN. Métodos: Dividimos 24 ratos albinos Wistar não urêmicos em 3 grupos. Grupo controle, 2 mL de solução salina intramuscular (im); grupo SN, adriamicina 5 mg/kg intravenosa (iv); grupo tratamento SN, adriamicina 5 mg/kg (iv) e OCT 200 mcg/kg (im) foram administrados no início do estudo (Dia 0). Aos 21 dias, mediram-se os níveis de creatinina e proteína em amostras de urina de 24 horas. Mediu-se a catalase (CAT) eritrocitária e renal e a substância reativa ao ácido tiobarbitúrico (TBARS). Avaliou-se também histologia renal. Resultados: Não houve diferença significativa entre os três grupos em termos de CAT e TBARS em eritrócitos. O nível de CAT renal foi menor no grupo SN e significativamente menor que no grupo controle. No grupo tratamento, o nível de CAT aumentou significativamente em comparação com o grupo SN. Quanto à histologia renal, as avaliações tubular e intersticial foram semelhantes em todos os grupos. O escore glomerular foi significativamente maior no grupo SN em comparação com o grupo controle e diminuiu significativamente no grupo de tratamento em comparação com o grupo SN. Conclusões: Estresse oxidativo na SN pode ser devido à diminuição do mecanismo de proteção antioxidante nos rins. O octreotide melhora níveis de antioxidantes e histologia do tecido renal e pode ser uma opção de tratamento.

Down-regulation of XBP1s alleviates the senescence of renal tubular epithelial cells induced by hypoxia/reoxygenation through Sirt3/SOD2/mtROS signaling pathway / 器官移植

Objective To investigate the role and mechanism of spliced X-box binding protein 1 (XBP1s) in the senescence of primary renal tubular epithelial cells induced by hypoxia/reoxygenation (H/R). Methods Primary renal tubular epithelial cells were divided into the normal control group (NC group), H/R group, empty adenovirus negative control group (Ad-shNC group), targeted silencing XBP1s adenovirus group (Ad-shXBP1s group), empty adenovirus+H/R treatment group (Ad-shNC+H/R group) and targeted silencing XBP1s adenovirus+H/R treatment group (Ad-shXBP1s +H/R group), respectively. The expression levels of XBP1s in the NC, H/R, Ad-shNC and Ad-shXBP1s groups were measured. The number of cells stained with β-galactosidase, the expression levels of cell aging markers including p53, p21 and γH2AX, and the levels of reactive oxygen species (ROS), malondialdehyde (MDA) and superoxide dismutase (SOD) were determined in the Ad-shNC, Ad-shNC+H/R and Ad-shXBP1s+H/R groups. Chromatin immunoprecipitation was employed to verify Sirtuin 3 (Sirt3) of XBP1s transcription regulation, and the expression levels of Sirt3 and downstream SOD2 after down-regulation of XBP1s were detected. Mitochondrial reactive oxygen species (mtROS) were detected by flow cytometry. Results Compared with the NC group, the expression level of XBP1s was up-regulated in the H/R group. Compared with the Ad-shNC group, the expression level of XBP1s was down-regulated in the Ad-shXBP1s group (both P<0.001). Compared with the Ad-shNC group, the number of cells stained with β-galactosidase was increased, the expression levels of p53, p21 and γH2AX were up-regulated, the levels of ROS, MDA and mtROS were increased, the SOD activity was decreased, the expression level of Sirt3 was down-regulated, and the ratio of Ac-SOD2/SOD2 was increased in the Ad-shNC+H/R group. Compared with the Ad-shNC+H/R group, the number of cells stained with β-galactosidase was decreased, the expression levels of p53, p21 and γH2AX were down-regulated, the levels of ROS, MDA and mtROS were decreased, the SOD activity was increased, the expression level of Sirt3 was up-regulated and the ratio of Ac-SOD2/SOD2 was decreased in the Ad-shXBP1s+H/R group (all P<0.05). Conclusions Down-regulation of XBP1s may ameliorate the senescence of primary renal tubular epithelial cells induced by H/R, which probably plays a role through the Sirt3/SOD2/mtROS signaling pathway.

Research progress on association between macrophages and ischemia-reperfusion injury / 器官移植

Ischemia-reperfusion injury (IRI) is an extremely complicated pathophysiological process, which may occur during the process of myocardial infarction, stroke, organ transplantation and temporary interruption of blood flow during surgery, etc. As key molecules of immune system, macrophages play a vital role in the pathogenesis of IRI. M1 macrophages are pro-inflammatory cells and participate in the elimination of pathogens. M2 macrophages exert anti-inflammatory effect and participate in tissue repair and remodeling and extracellular matrix remodeling. The balance between macrophage phenotypes is of significance for the outcome and treatment of IRI. This article reviewed the role of macrophages in IRI, including the balance between M1/M2 macrophage phenotype, the mechanism of infiltration and recruitment into different ischemic tissues. In addition, the potential therapeutic strategies of targeting macrophages during IRI were also discussed, aiming to provide reference for alleviating IRI and promoting tissue repair.

Buzhong Yiqitang Regulates Apoptosis via Nrf2/ROS Pathway to Alleviate Cisplatin Resistance in A549/DDP Cells / 中国实验方剂学杂志

ObjectiveTo explore the mechanism of Buzhong Yiqitang-containing serum in alleviating the cisplatin resistance in human non-small cell lung cancer （A549/DDP） cells via regulating the nuclear factor E2-related factor 2 （Nrf2）/reactive oxygen species （ROS） signaling pathway. MethodThe serum containing Buzhong Yiqitang was prepared and A549/DDP cells were cultured and randomly grouped： blank （10% blank serum）， cisplatin （10% blank serum+20 mg·L-1 cisplatin）， Buzhong Yiqitang （10% Buzhong Yiqitang-containing serum+20 mg·L-1 cisplatin）， ML385 （10% blank serum+5 μmol·L-1 ML385+20 mg·L-1 cisplatin）， Buzhong Yiqitang+ML385 （10% Buzhong Yiqitang-containing serum+5 μmol·L-1 ML385+20 mg·L-1 cisplatin）， tertiary butylhydroquinone （TBHQ） （10% blank serum+5 μmol·L-1 TBHQ+20 mg·L-1 cisplatin）， and Buzhong Yiqitang+TBHQ （10% Buzhong Yiqitang-containing serum+5 μmol·L-1 TBHQ+20 mg·L-1 cisplatin）. The median inhibitory concentration （IC50） of cisplatin in each group was determined by the cell counting kit-8 （CCK-8） method and the resistance index （RI） was calculated. The apoptosis rate was detected by flow cytometry. The ROS content of each group was determined with the DCFH-DA fluorescence probe. Western blot was employed to determine the protein levels of Nrf2， cleaved cysteinyl aspartate-specific protease-3 （cleaved Caspase-3）， cytochrome C （Cyt C）， and B-cell lymphoma-2 （Bcl-2）. ResultCompared with those in the cisplatin group， the IC50 and RI of A549/DDP cells to cisplatin in Buzhong Yiqitang， ML385， and Buzhong Yiqitang+ML385 groups decreased （P˂0.05）. Compared with the blank group， the cisplatin， Buzhong Yiqitang， ML385， and Buzhong Yiqitang+ML385 groups showed increased apoptosis rate of A549/DDP cells （P˂0.05）. Compared with the blank group， cisplatin promoted the expression of Nrf2 （P˂0.05）. Compared with the cisplatin group， Buzhong Yiqitang， ML385， and Buzhong Yiqitang+ML385 inhibited the expression of Nrf2 （P<0.05）， elevated the ROS level （P˂0.05）， up-regulated the protein levels of cleaved Caspase-3 and Cyt C， and down-regulated the protein level of Bcl-2 （P<0.05）， which were the most significant in the Buzhong Yiqitang+ML385 group. Compared with the cisplatin group， the TBHQ group showed increased IC50 and RI of cisplatin （P<0.05）， decreased apoptosis rate of A549/DDP cells （P<0.05）， up-regulated protein levels of Nrf2 and Bcl-2 （P<0.05）， lowered level of ROS （P˂0.05）， and down-regulated protein levels of cleaved Caspase-3 and Cyt C （P<0.05）. Compared with the TBHQ group， Buzhong Yiqitang+TBHQ decreased the IC50 and RI of cisplatin in A549/DDP cells （P<0.05）， increased the apoptosis rate （P<0.05）， down-regulated the protein levels of Nrf2 and Bcl-2 （P<0.05）， increased ROS （P˂0.05）， and up-regulated the protein levels of cleaved Caspase-3 and Cyt C （P<0.05）. ConclusionBuzhong Yiqitang induced apoptosis by inhibiting Nrf2/ROS pathway to alleviate cisplatin resistance in A549/DDP cells.

The potential role of apigenin in the enhancement of the antioxidant adaptation to exercise by bioactive dietary compounds

Oxidative stress (OS) occurs when the antioxidant defense system is overwhelmed by the predominance of reactive oxygen species (ROS) and pro-oxidant factors. Several diseases such as hypertension, insulin resistance, type 2 diabetes mellitus and neurodegenerative diseases are characterized by chronic OS. Physical exercise constitutes an affordable tool to prevent or ameliorate these conditions. However, during physical activity, acute ROS are produced inducing an activation in peroxisome proliferator activated receptor-Gamma Coactivator-1alpha (PGC-1α), and nuclear factor erythroid-2 related factor 2 (Nrf2), PGC-1α/Nrf2 pathway. This signaling pathway facilitates interaction with antioxidant response elements (ARE), thereby initiating an upregulation in the expression of antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and mitochondrial biogenesis. In both cases, whether involving healthy animals or individuals engaged in physical exercise, supplementation with antioxidant scavengers leads to a reduction in the expression and activity of PGC-1α, SOD, CAT, and GPX across various tissues, which is not observed with indirect antioxidants. The preventive role of physical exercise against chronic OS is avoided when executed in conjunction with supplementation of scavenger antioxidants. However, similar to exercise, the indirect antioxidant apigenin can activate the PGC1-α/Nrf2 signaling pathway. Here, we summarize evidence supporting apigenin as a non-nutritional supplement that could enhance the adaptive effects of exercise, improving the endogenous antioxidant defense. Therefore, apigenin could be an interesting supplement to enhance the endogenous antioxidant adaptation induced by exercise in healthy subjects, but also to improve the effectiveness of exercise to prevent oxidative stress-associated diseases.

El estrés oxidativo (OS) ocurre cuando el sistema de defensa antioxidante es sobrepasado por el predominio de especies reactivas de oxígeno (ROS) y factores prooxidantes. Varias enfermedades como la hipertensión, la resistencia a la insulina, la diabetes mellitus tipo 2 y enfermedades neurodegenerativas se caracterizan por un OS crónico. El ejercicio físico constituye una herramienta asequible para prevenir o mejorar estas enfermedades. Sin embargo, durante la actividad física, se producen ROS agudas que inducen una activación en la vía PGC-1α/Nrf2. Esta vía de señalización facilita la interacción con los elementos de respuesta antioxidante (ARE), iniciando así una regulación que permite la expresión de enzimas antioxidantes, incluidas SOD, CAT, GPX y biogénesis mitocondrial. En ambos casos, ya sea que se trate de animales sanos o de individuos que practican ejercicio físico, la suplementación con antioxidantes "scavengers" conduce a una reducción en la expresión y actividad de PGC-1α, SOD, CAT y GPX en varios tejidos, lo que no se observa con antioxidantes "indirectos". El papel preventivo del ejercicio físico contra el OS crónico se atenúa cuando se realiza en conjunto con la suplementación de antioxidantes "scavengers". Sin embargo, de manera similar al ejercicio, la apigenina es un antioxidante "indirecto" que puede activar la vía de señalización PGC1-α/Nrf2. Aquí, resumimos la evidencia que respalda a apigenina como un suplemento no-nutricional que podría mejorar los efectos adaptativos del ejercicio, mejorando la defensa antioxidante endógena de sujetos sanos que no tienen suficiente tiempo para hacer ejercicio.

Effect of mobile phone radiofrequency electromagnetic radiations on oxidative stress and feeding behaviour in Sprague Dawley (SD) rats

Objectives: Radiofrequency electromagnetic radiation (RF-EMR) from mobile phones is known to produce a stress response because of its effect on hypothalamus. Mobile phones have become an integral part of our lives with increasing usage not only in terms of number of users but also increase in talk time. The present study aimed to study the effect of mobile phone radiofrequency electromagnetic radiations on oxidative stress and feeding behaviour assessment in Sprague Dawley (SD) rats. Materials and Methods: Twelve male SD rats of 10–12 weeks old, weighing 180–220 g, were housed and allowed to acclimatise in a room with 12:12 h light-dark cycle with ad libitum amount of food and reverse osmosis (RO) water before the start of the study. Then, rats were divided into control and RF-EMR exposed groups, and everyday feed intake and body weight were measured. At the end of the study period, blood sample was collected through retro orbital puncture for biochemical investigations. Results: The present study showed significant increase in malondialdehyde and serum corticosterone levels and decrease feeding behaviour in rats exposed to RF-EMR in rats exposed to RF-EMR. Conclusion: This study proves that mobile RF-EMR causes oxidative stress and oxidative damage leading to decreased feeding behaviour in SD rats.

Radicales libres y sistema antioxidante

Los radicales libres son compuestos caracterizados por tener un electrón desapareado en su orbital externo, condición que los torna altamente reactivos, es decir, tienen la propiedad de interactuar a través de reacciones controladas por difusión con proteínas, lípidos y ácidos nucleicos. También se les ha designado como especies reactivas de oxígeno (ERO), especies reactivas de nitrógeno (ERN) o especies reactivas de azufre (ERA). En el organismo humano se generan, principalmente, en la cadena transportadora de electrones mitocondrial, donde específicamente participan los complejos respiratorios I y III que tienen la propiedad de reducir al oxígeno y convertirlo en anión superóxido; así mismo, pueden formarse haciendo uso de una gran diversidad de reacciones enzimáticas y no enzimáticas en las que intervienen sustancias que la célula sintetiza o que se ingieren con los alimentos y algunos medicamentos. El ser humano dispone de un sistema antioxidante, que es de naturaleza enzimática y no enzimática, el cual tiene como función proteger al organismo de la acción nociva de los radicales libres; comprende enzimas -como catalasa, superóxido dismutasa, tiorredoxina, etc.- y compuestos no enzimáticos -como glutatión, ferritina, mioglobina, etc.-, pero no son lo suficientemente eficientes para protegerlo, por lo que es necesaria la ingesta de alimentos que contengan en su composición sustancias con propiedades antioxidantes cuya acción protectora dependerá de su reactividad química, así como de su concentración; estos compuestos antioxidantes se encuentran principalmente en las frutas y verduras, habiéndose identificado polifenoles, flavonoides, carotenoides, vitamina C, vitamina E, etc. Un número considerable de evidencias sugiere que la ingesta de sustancias antioxidantes protege al organismo del efecto dañino de los radicales libres, pero cuando prevalece la acción oxidante sobre la antioxidante puede conducirse al estrés oxidativo, condición que está estrechamente vinculada con una gran diversidad de enfermedades crónicas no transmisibles como cáncer, diabetes mellitus, obesidad, psoriasis, aterosclerosis, entre otras. Todo ello parece indicar que el término "estado estable redox celular" describe de manera apropiada la constante adaptación a una situación de rápido recambio químico, y sugiere que las sustancias implicadas en este proceso se designen como "especies biológicamente reactivas" en razón de la existencia de compuestos nocivos como el peróxido de hidrógeno, peroxinitrito, etc., que no son propiamente radicales libres, pero ejercen efectos dañinos a las células.

Free radicals are compounds characterized by having an unpaired electron in their outer orbit, a condition that makes them highly reactive, i.e., they interact through diffusion-controlled reactions with proteins, lipids and nucleic acids. They have also been referred to as reactive oxygen species (ROS), reactive nitrogen species (RNS) or reactive sulfur species (RSS). In the human organism, they are mainly produced in the mitochondrial electron transport chain, where respiratory complexes I and III specifically participate and reduce oxygen by converting it into superoxide anion. Likewise, they can be formed through a wide variety of enzymatic and non-enzymatic reactions involving substances that are synthesized by cells or are ingested with food and some medicines. Human beings have an antioxidant system which is both enzymatic and non-enzymatic in nature and whose function is to protect the organism from the harmful action of free radicals. This system includes enzymes-such as catalase, superoxide dismutase, thioredoxin, etc.-and non-enzymatic compounds- such as glutathione, ferritin, myoglobin, etc. However, they are not efficient enough to protect it, so it is necessary to eat foods that contain substances with antioxidant properties whose protective action will depend on their chemical reactivity and their concentration. These antioxidant compounds are mainly found in fruits and vegetables, where polyphenols, flavonoids, carotenoids, vitamin C, vitamin E, etc. have been identified. A significant amount of evidence suggests that the intake of antioxidant substances protects the body from the damaging effect of free radicals, but when the oxidative action prevails over the antioxidant action, it can lead to oxidative stress, a condition that is closely linked to a wide variety of chronic non-communicable diseases including cancer, diabetes mellitus, obesity, psoriasis, atherosclerosis, among others. All this seems to indicate that the term "cellular redox steady state" more appropriately describes the constant adaptation to a situation of rapid chemical turnover and suggests that the substances involved in this process be designated as "biologically reactive species" due to the existence of harmful compounds such as hydrogen peroxide, peroxynitrite, etc., which are not-strictly speaking-free radicals but have toxic effects on cells.

Effects of Gundelia tournefortii L. on biochemical parameters, antioxidant activities and DNA damage in a rat model of experimental obesity / Efeitos de Gundelia tournefortii L. em parâmetros bioquímicos, atividades antioxidantes e danos ao DNA em um modelo de obesidade experimental de rato

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.

Do Dipteryx alata Vogel seedlings recover the quality and the photosynthetic and antioxidant responses in the post-flooding? / Mudas de Dipteryx alata Vogel recuperam a qualidade e as respostas fotossintéticas e antioxidantes no pós-alagamento?

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.

Application progress of ROS-responsive liposomes in anti-tumor research / 药学实践杂志

Reactive oxygen species（ROS） responsive liposomes are prepared based on the high level of ROS expression in the tumor microenvironment, enabling precise drug delivery to the tumor site. With the addition of photosensitizer, the controllability of drugs in liposomes can be further enhanced.

Role of ROS/TXNIP/NLRP3 signaling pathway in high glucose-induced pyroptosis of human embryonic trophoblast cells / 中华内分泌外科杂志

Objective:To investigate the changes of ROS/TXNIP/NLRP3 signaling pathway in pyroptosis of human embryonic trophoblast cells induced by high glucose.Methods:Human embryonic trophoblast cells were cultured in vitro to establish high glucose injury model, and they were randomly divided into control group, high glucose (HG) group and HG + ROS inhibitor N-acetyl-L-cysteine (HG + NAC) group. MTT assay was used to detect the cell survival rate. The level of ROS in each group was detected by dihydroethidine ROS fluorescence probe. Expression of TXNIP and NLRP3 mRNA was detected by real-time quantitative PCR (RT-qPCR). Western blot analysis was used to detect the expression levels of TXNIP, NLRP3, Caspase-1, interleukin (IL) -1β, tumor necrosis factor-α (TNF-α) and GSDMD proteins. In addition, pyroptosis was detected by flow cytometry.Results:The optimal glucose concentration for high glucose-induced injury of human embryonic trophoblast cells was 30 mmol/L. Compared with the control group (96.27±3.10) %, the survival rate of human embryonic trophoblast cells in HG group (55.44±2.15) % was significantly lower ( P<0.05), while the fluorescence intensity (ROS level) of 7 'dichlorofluorescein (DCF), the expression levels of TXNIP and NLRP3 proteins, the number of pyroptosis, expression levels of Caspase-1, GSDMD, IL-1β and TNF-α proteins were significantly higher ( P<0.05) ; Compared with HG group, the survival rate of human embryonic trophoblast cells in HG+NAC group (84.75±2.33) % was significantly higher ( P<0.05), the fluorescence intensity (ROS level) of DCF, the expression levels of TXNIP and NLRP3 proteins, the number of pyroptosis, and expression levels of Caspase-1, GSDMD, IL-1β and TNF-α proteins were significantly lower ( P<0.05) . Conclusion:Inhibition of ROS level in human embryonic trophoblast cells induced by high glucose may promote cell proliferation and reduce the occurrence of pyroptosis by inhibiting TXNIP/NLRP3 signaling pathway.

Research progress on the role of mitochondrial dysfunction in the occurrence, progression and treatment of periodontitis / 口腔疾病防治

@#Periodontitis is a widespread disease worldwide, with the primary cause of tissue loss being an immune inflammatory response mediated by bacteria. Increasing evidence has revealed a significant correlation between mitochondrial dysfunction and the occurrence and progression of periodontitis. This paper provides a review of current research on the role of mitochondrial dysfunction in the occurrence and development of periodontitis and related therapies from the perspectives of oxidative stress, inflammatory responses, and the regulation of mitochondrial homeostasis. Mitochondria are the main source and target of cellular reactive oxygen species. Mitochondrial dysfunction can generate large amounts of reactive oxygen species, exacerbating local oxidative stress in periodontal tissues and causing cell toxicity and tissue damage. Mitochondria are also the center of cellular inflammatory responses, and the positive feedback loop of inflammation induced by mitochondrial dysfunction may explain the persistent and unresolved nature of periodontitis. Biomaterials loaded with pharmacological agents show potential in restoring mitochondrial function, controlling the development of periodontitis, and promoting periodontal tissue regeneration. However, the key sites of mitochondrial dysfunction in the occurrence and development of periodontitis are not yet fully understood, and the improvement of mitochondrial function in periodontal therapy is still in the experimental stage. Future research efforts should focus on the effect of mitochondrial dysfunction on periodontal cells and explore its specific mechanism in the occurrence and progression of periodontitis in order to provide new insights into the treatment of periodontitis.

Research progress on multifunctional natural antioxidants in oral therapies / 口腔疾病防治

@#Oxidative stress is closely associated with the development of oral diseases such as caries, periodontitis and endodontitis. The accompanying oxidative stress during inflammation could aggravate tissue damage. However, numerous studies have shown that some dental materials, such as composite resins, bleach, drugs for root canal irrigation and dental implants, can give rise to abundant free radicals, which have adverse effects on peripheral tissues. Therefore, it is essential to supplement with extra antioxidants against free radicals. Plant-derived natural antioxidants have attracted great attention in biomedicine because of their excellent biocompatibility and easy access. This paper focused on the redox imbalance in the oral cavity and the application of natural antioxidants to oral therapy and their modification of dental materials. Current research shows that by constructing polyphenol-based metal organic nanoenzymes or adding vitamins and polyphenols to bionic hydrogels, the safety and utilization rate of antioxidants can be significantly improved. However, these polymer delivery systems have problems such as poor degradability, hepatotoxicity and nephrotoxicity, and the research is still in its infancy. In terms of material modification, it is crucial to choose the type and ratio of natural antioxidants and raw materials, as well as appropriate modification methods. A strong chemical bond between the antioxidant and the raw material may lead to the failure of antioxidant release from the modified composite, lowering the antioxidant activity. At the same time, the selection of polyphenols rich in pyrogallol functional groups can retain more free phenolic hydroxyl groups after chemical modification, which is conducive to greater antioxidant activity by the implant materials. Although research on natural antioxidants in oral therapy has made progress, there is a lack of data supporting clinical trials and long-term application effects, and further research is still needed.

Berberine inhibits autophagy and promotes apoptosis of fibroblast-like synovial cells from rheumatoid arthritis patients through the ROS/mTOR signaling pathway / 南方医科大学学报

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.

In vitro inhibition of carnosic acid against influenza A virus infections / 药学学报

Carnosic acid (CA) is the main phenolic diterpenoid active ingredient in plants such as rosemary and sage, and has antiviral, antioxidant, anti-inflammatory effects and so on, however, its antiviral activity against influenza virus infections was not reported. In this study, antiviral activities against influenza A virus infections of three main bioactive ingredients from rosemary, including rosmarinic acid, CA and ursolic acid, were evaluated using virus titer titration assay, and CA showed remarkable inhibition on influenza H5N1 replication in A549 cells. The antiviral activity of CA was further confirmed and its mechanism of action was investigated using the indirect immunofluorescence assay (IFA), Western blot and real-time fluorescence quantification polymerase chain reaction (qRT-PCR). The results showed that the 50% effective concentration (EC50) of CA against influenza H5N1 in A549 cells and MDCK cells were 4.30 and 3.64 μmol·L-1, respectively. Meanwhile, CA also showed inhibition on influenza virus 2009panH1N1 (EC50: 10.1 μmol·L-1) and H3N2 (EC50: 12.8 μmol·L-1) replications in A549 cells. Mechanistic studies showed that antiviral activity of CA is related to its induction of heme oxygenase-1 (HO-1) in A549 cells and suppression on production of reactive oxygen in H5N1-infected cells.

Effect and mechanism of Bajitianwan on preventing D-galactose-induced osteoblast bone loss / 药学实践杂志

Objective To explore the effect and mechanism of Bajitianwan on preventing D-galactose (D-gal)-induced osteoblast bone loss. Methods Osteoblasts isolated from 24 h old Wistar rats were injured by D-gal and intervened with Bajitianwan extract. The osteoblastic proliferation and differentiation were determined by MTT and alkaline phosphatase (ALP), respectively. The cell reactive oxygen species (ROS) levels were detected by DCFH-DA fluorescent probes. The expression of cellular oxidation-related protein nuclear factor erythroid 2-related factor 2 (Nrf2), phosphorylated protein kinase B (p-AKT), protein kinase B (AKT), heme oxygenase-1 (HO-1), and NADPH quinone oxidoreductase 1 (NQO1) were detected by Western blotting. The intranuclear expression of Nrf2 protein was measured by immunofluorescence. Results Bajitianwan extract had significantly increased the osteoblastic proliferation and differentiation, and significantly reduced the intracellular ROS level. Bajitianwan extract had activated the PI3K/AKT pathway via activating the phosphorylation of AKT in osteoblasts, and promoted NQO1 and HO-1 expression. In addition, Bajitianwan had significantly promoted the expression of Nrf2 in the nucleus of osteoblasts, activating Nrf2 signaling pathway, and further promoted bone formation. Conclusion This study confirmed that Bajitianwan could prevent D-gal injured osteoblastic bone loss for the first time. The mechanism might be related to the regulation of oxidative stress associated PI3K/AKT and Nrf2 signaling pathway.

Effect of sodium fluoride on myocardial damage by regulating apoptosis through JNK signaling pathway / 环境与职业医学

Background It has been found that fluoride may cause cardiomyocyte damage. c-Jun N-terminal kinases (JNK) signaling pathway plays an important role in apoptosis, but its role in fluorosis-induced cardiomyocyte damage is still unknown yet. Objective To explore the toxic effect of sodium fluoride (NaF) on H9c2 cardiomyocytes of rats and whether NaF affects cardiomyocyte apoptosis through the JNK signaling pathway. Methods According to the concentrations of sodium fluoride and whether sp600125 (JNK inhibitor) was added, cardiomyocytes of rats were divided into six groups, including control group, SP600125 group (SP group), 0.24, 0.48, and 0.96 mmol·L−1 NaF groups, and 0.96 mmol·L−1 NaF+SP600125 group (NaF+SP group). Cardiomyocytes exposed to NaF for 24 h were observed using a fluorescence inverted microscope. The changes of cell viability at 24, 48, and 72 h after the treatment were detected by CCK-8 method. The levels of reactive oxygen species (ROS) at 24 h after the treatment in H9c2 cardiomyocytes were determined by fluorescent probe method. The expression levels of Bcl-2, Bax, Caspase-3, and JNK mRNA at 24 h after the treatment were detected by real-time PCR. The protein expression levels of Bcl-2, Bax, Caspase-3, and p-JNK at 24 h after the treatment were detected by Western blotting. Results Compared with the control group, after being exposed to 0.48 and 0.96 mmol·L−1 NaF for 24 h, the cell growth density decreased. With the increase of NaF concentration, rounded cells and some suspended dead cells appeared. At 24h after exposure to NaF, the cell viability of the 0.48 and 0.96 mmol·L−1 NaF groups decreased compared with the control group (P<0.05). At 48h and 72h after exposure to NaF, the cell viability levels of the NaF treated groups were significantly lower than that of the control group (P<0.05). After NaF exposure for 24 h, compared with the control group, the intracellular ROS levels were increased (P<0.05); the mRNA expression levels of Bcl-2 were decreased to varying degrees, especially in the 0.48 and 0.96 mmol·L−1 NaF groups (P<0.05); the mRNA expression levels of Bax, Caspase-3, and JNK were increased (P<0.05); the protein expression levels of Bcl-2 were reduced (P<0.05); the protein expression levels of Bax, Caspase-3, and p-JNK were elevated (P<0.05). Compared with the 0.96 mmol·L−1 NaF group, the cell viability of the NaF+SP group was increased, the intracellular ROS level was decreased, the mRNA expression levels of Bax, Caspase-3, and JNK were decreased, the protein expression level of Bcl-2 was increased, and the protein expression levels of Bax, Caspase-3, and p-JNK were decreased (P<0.05); the expression level of Bcl-2 mRNA had a rising trend but showed no statistical significance (P>0.05). Conclusion Cardiomyocyte damage after excessive fluoride exposure may result from fluoride inducing excessive ROS production in cardiomyocytes, which may activate the JNK signaling pathway and induce cardiomyocyte apoptosis.

Red light-emitting diode light mediated by the KEAP1-NRF2/HO-1 pathway induces osteogenic differentiation and attenuates the oxidative stress damage of human periodontal ligament stem cells induced by high glucose / 口腔疾病防治

Objective@# To explore the effects of red LED light mediated by the Kelch-like ECH-associated protein 1-nuclear factor erythroid 2-related factor 2/heme oxygenase-1 (KEAP1-NRF2/HO-1) pathway on osteogenic differentiation and oxidative stress damage of human periodontal ligament stem cells (hPDLSCs) induced by high glucose, which provides a basis for the application of red light-emitting diode (LED) light in cell antioxidative damage.@*Methods@#hPDLSCs were identified by flow cytometric analysis, alkaline phosphatase (ALP) staining and Alizarin red-S staining; hPDLSCs were pretreated in a high glucose environment for 48 hours and irradiated with 1, 3, or 5 J/cm2 red LED light. A CCK-8 assay was performed to choose the radiant exposure that had the strongest effect on promoting the cell proliferation rate for subsequent experiments. hPDLSCs were divided into a control group, a high glucose group and a high glucose+light exposure group. ALP staining, ALP activity, Alizarin red-S staining and quantitative calcified nodules were used to detect the osteogenic differentiation of hPDLSCs; qRT-PCR and Western blot were used to detect the gene and protein expression levels of ALP, runt-related transcription factor 2 (RUNX2) and osterix (OSX); the relative mRNA expression levels of antioxidant enzyme-related genes superoxide dismutase 2 (SOD2) and catalase (CAT) in hPDLSCs were detected by qRT-PCR; reactive oxygen species (ROS) levels were detected by fluorescence microscopy and flow cytometry; the tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) levels in cell supernatants were detected by ELISA; the NRF2-specific inhibitor ML385 was used to inhibit the NRF2 pathway; ALP staining and ALP activity were used to detect the markers of early osteogenic differentiation; qRT-PCR was used to detect the gene expression of ALP, RUNX2 and OSX; and the protein expression levels of KEAP1, NRF2 and HO-1 were detected by Western blot.@*Results @# Identified, and irradiant exposure of 5 J/cm2 was chosen for subsequent experiments. Red LED light irradiation (5 J/cm2) improved the osteogenic differentiation of hPDLSCs induced by high glucose (P<0.05), increased the mRNA and protein levels of ALP, RUNX2 and OSX (P<0.05), upregulated the mRNA expression levels of SOD2 and CAT (P<0.05), reduced the levels of ROS (P<0.05), and reduced TNF-α and IL-1β levels in the cell supernatants (P<0.05). When ML385 was added to inhibit the NRF2 pathway, the ALP activity of cells was decreased (P<0.05); the gene expression levels of ALP, RUNX2 and OSX were downregulated (P<0.05); the protein level of KEAP1 was upregulated (P<0.05); and the protein levels of NRF2 and HO-1 were downregulated (P<0.05)@*Conclusion@#Red LED light may promote the proliferation and osteoblastic differentiation of hPDLSCs induced by high glucose through the KEAP1-NRF2/HO-1 pathway and reduce the oxidative stress damage to hPDLSCs induced by high glucose.

Effect of H2O2 on Secondary Metabolism in Atractylodes chinensis and Its Mechanism / 中国实验方剂学杂志

ObjectiveTo investigate the effect of exogenous H2O2 on secondary metabolism in Atractylodes chinensis and its mechanism. MethodFresh rhizomes of A. chinensis were treated with 5.0， 1.0， 0.2， 0.04 mmol·L-1 H2O2 solution and clean water， and the relationships between the contents of reactive oxygen species， activities of antioxidant enzymes， activities of key enzymes of secondary metabolites， and contents of secondary metabolites in A. chinensis were compared. ResultUnder treatment with exogenous H2O2， the content of reactive oxygen species and malondialdehyde （MDA） in the fresh rhizomes of A. chinensis were significantly elevated on the 4th day， and returned to normal level on the 6th-8th day. The activities of superoxide dismutase （SOD）， catalase （CAT） and peroxidase （POD） were all increased first and then decreased， and reached the peak on the 4th， 4th-6th and 2th-4th day， respectively. The activities of 3-hydroxy-3-methyl glutaryl coenzyme A reductase （HMGR） and acetyl CoA carboxylase （ACC）， key enzymes of the secondary metabolites， were remarkably enhanced， and under treatments with different concentrations of H2O2， the activities of key synthetic enzymes of the secondary metabolites in 0.2 mmol·L-1 H2O2 group were increased most， with the highest biosynthesis of secondary metabolites. The contents of atractylodin， β-eudesmol， atractylone， atractylenolide Ⅱ， and atractylenolide Ⅲ on the 6th day of 0.2 mmol·L-1 H2O2 treatment were 89.5%， 108.7%， 308.8%， 64.7% and 9.3%， respectively higher than those in the control. ConclusionThe antioxidant enzymes and secondary metabolites in A. chinensis synergistically maintain the balance of reactive oxygen species， and exogenous H2O2 can improve the medicinal quality of A. chinensis remarkably.

Research progress on metabonomic studies in periodontitis / 口腔疾病防治

@#Metabolomics reflects the endogenous metabolite changes in organisms through qualitative and quantitative detection of small molecules in biological samples, revealing the metabolic changes during disease development. Metabolomic studies of periodontitis further elucidate the etiology, diagnosis and predictive markers of periodontitis at the levels of metabolites and metabolic pathways. In this paper, the concept and research methods of metabonomics were summarized, and the current status of the metabonomics of saliva and gingival crevicular fluid in the study of periodontitis was reviewed. Previous studies have shown that metabolites such as short-chain fatty acids and amino acids and metabolic pathways such as glutamic acid and pyrimidine metabolism might promote the occurrence of periodontitis, and it was suggested that lactic acid, γ-amino-butyrate, butyric acid and lysophosphatidic acid might be potential diagnostic markers of periodontitis. The metabolomics study of periodontitis still faces challenges such as high heterogeneity of results and fluctuation of metabolites. In the future, its study could be optimized through multicenter prospective studies to provide fresh approaches for the etiology and diagnosis of periodontitis.