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.

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.

Design, synthesis, and antifungal mechanism of carbaline fluorescent probes / 药学学报

Three carboline fluorescent probes F1-F3 were designed and synthesized, based on lead compound JYJ-19, an antifungal compound discovered previously by our group. The antifungal activity in vitro results showed that compound F1 had moderate antifungal activity (MIC80 = 32 μg·mL-1). The stokes shift of F1 is 70 nm. The fluorescent probe F1 has good optical properties and can be used for fluorescence imaging research. Subcellular localization experiments results showed that F1 was enriched in the mitochondria of fungal cells. The detection of intracellular reactive oxygen species levels shows that JYJ-19 enhances intracellular reactive oxygen species levels. The above results indicated that carboline compounds could exert antifungal effects by acting on fungal mitochondria.

Effect and mechanism of reactive oxygen species-responsive nanoparticles on the regulation of human gingival fibroblast function and inflammation induced by lipopolysaccharide / 口腔疾病防治

Objective@#To investigate the effects of PssL-NAC reactive oxygen species (ROS)-responsive nanoparticles on intracellular ROS production, inflammatory factor levels, collagen production, cell function and Toll-like receptor 4 (TLR4), NF-κB nuclear factor-κB (p65) pathway protein expression in human gingival fibroblasts (HGFs) induced by Porphyromonas gingivalis-lipopolysaccharide (P.g-LPS).@*Methods@#This study was reviewed and approved by the ethics committee. PssL-NAC microspheres containing oil soluble antioxidant N-acetylcysteine (NAC) were obtained by connecting the hydrophobic end of polycaprolactone (PCL) and the hydrophilic end of polyethylene glycol (PEG) via thioketal (TK) bonds in response to ROS, and self loading in the aqueous and oil phases. After preparation of the PssL-NAC microspheres and aqueous NAC solution, successful synthesis of the nanoparticles was verified by transmission electron microscopy. Then, HGFs were exposed to P.g-LPS (0, 5, or 10 μg/mL), P.g-LPS (0, 5, or 10 μg/mL)+NAC, and P.g-LPS (0, 5, or 10 μg/mL)+PssL-NAC, and the ROS levels in the different groups were observed under confocal microscopy to determine the concentration of P.g-LPS for use in subsequent experiments. The groups were as follows: control group (no treatment), P.g-LPS group (HGFs treated with P.g-LPS), NAC group (HGFs treated with P.g-LPS and NAC), and PssL-NAC group (HGFs treated with P.g-LPS and PssL-NAC). Cell counting kit-8 (CCK-8) assays verified the biosafety of PssL-NAC. The ROS levels in the different groups were detected by DCFH-DA probes and observed via confocal microscopy. Real-time qPCR (RT-qPCR) was used to monitor the gene expression levels of the intracellular inflammatory cytokines interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), collagen 1 (COL1) and collagen 3 (COL3). The effect of PssL-NAC on the migration of HGFs was observed via the scratch test. The protein expression of TLR4-NF-κB, and phosphorylated p65 (p-p65) in the TLR4-NF-κB pathway was evaluated by Western blot.@*Results@#PssL-NAC had no significant effect on HGF proliferation (P>0.05). At elevated P.g-LPS concentrations, PssL-NAC maintained intracellular ROS levels approximately twice those in the control group (P<0.001). PssL-NAC significantly decreased P.g-LPS-induced IL-6 (P<0.001) and TNF-α (P<0.001) gene expression and increased COL1 gene expression (P<0.001). After P.g-LPS stimulation, PssL-NAC restored cell migration to the control level (P>0.05) and decreased the protein expression of TLR4 (P<0.001), p65 (P = 0.006), and p-p65 (P = 0.017) in the TLR4-NF-κB pathway.@*Conclusion@#PssL-NAC maintains the appropriate intracellular ROS concentration, alleviates P.g-LPS-induced inflammation in HGFs through the TLR4-NF-κB pathway, and restores the cell functions of collagen production and migration in an inflammatory environment.

Research progress on the application of metal nanoenzymes in the prevention and treatment of plaque biofilm-associated oral diseases / 口腔疾病防治

@#Oral plaque biofilms are one of the bases for the survival and metabolism of different bacteria. With the emergence of drug-resistant bacteria due to antibiotic abuse, the prevention and treatment of plaque biofilm-associated oral diseases are becoming increasingly difficult. Although some research progress has been made in the field of biofilm formation and destruction, there is still a lack of effective clinical therapies for plaque biofilm-associated oral diseases. Metal nanoenzymes possess the physical properties of nanoparticles and exhibit catalytic activity similar to that of natural enzymes. The nanoscale size of metal nanoenzymes provides a greater specific surface area to help reactive oxygen species spread rapidly to active catalytic sites and improve the antioxidant properties of nanoenzymes. Additionally, metal nanoenzymes are easy to produce using different methods, such as electrochemical reduction, solvent thermal synthesis and microwave-assisted synthesis. Moreover, metal nanoenzymes can produce a high concentration of hydroxyl radicals, catalyze plaque biofilm degradation, lyse glucan and inhibit biofilm formation by oxidative stress reactions, as well as kill bacteria by releasing metal ions. Thus, metal nanoenzymes are expected to become a new option for the prevention and treatment of oral plaque biofilm-associated diseases. However, metal nanoenzymes can enter organisms through oral, intravenous and respiratory routes, triggering potential toxic effects such as pulmonary toxicity, hepatotoxicity and neurotoxicity. In a complex biological environment, the occurrence of metal nanoenzymes toxicity may involve multiple mechanisms, and the mechanism of action and safety need to be thoroughly investigated. In this paper, we intend to describe the research progress on metal nanoenzymes through an overview of their properties, antibacterial mechanisms, biotoxicity and applications in the prevention and treatment of oral plaque biofilm-related diseases, which may provide new ideas for the prevention and treatment of these diseases.

Tumor-targeted metabolic inhibitor prodrug labelled with cyanine dyes enhances immunoprevention of lung cancer

Recent progress in targeted metabolic therapy of cancer has been limited by the considerable toxicity associated with such drugs. To address this challenge, we developed a smart theranostic prodrug system that combines a fluorophore and an anticancer drug, specifically 6-diazo-5-oxo-l-norleucine (DON), using a thioketal linkage (TK). This system enables imaging, chemotherapy, photodynamic therapy, and on-demand drug release upon radiation exposure. The optimized prodrug, DON-TK-BM3, incorporating cyanine dyes as the fluorophore, displayed potent reactive oxygen species release and efficient tumor cell killing. Unlike the parent drug DON, DON-TK-BM3 exhibited no toxicity toward normal cells. Moreover, DON-TK-BM3 demonstrated high tumor accumulation and reduced side effects, including gastrointestinal toxicity, in mice. This study provides a practical strategy for designing prodrugs of metabolic inhibitors with significant toxicity stemming from their lack of tissue selectivity.

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.

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.

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.

Pro-oxidation status of rotator cuff tissue and expression of Beclin-1 and mTOR in patients with rotator cuff injury / 中国骨伤

OBJECTIVE@#To explore pro-oxidative state of rotator cuff tissue and expression levels of Beclin-1 and mam-malian target of rapamycin(mTOR) in patients with acute and chronic rotator cuff injury, and then analyzed relationship between rotator cuff injury and oxidative stress and autophagy.@*METHODS@#Forty patients with rotator cuff injury were seleceted from July 2019 to December 2020, and divided into male chronic injury group, male acute injury group, female chronic injury group, and female acute injury group, 10 patients in each group. All patients were performed rotator cuff repair under arthroscopy. The sample of tendon at the rotator cuff injury site of the patient was taken during operation, and total reactive oxygen species (ROS) and superoxide dismutase(SOD) were detected by detection kit;expression of Beclin-1 and mTOR mRNA were detected by reverse transcription-polymerase chain reaction (RT-PCR), and Western-blot was applied to detect protein expression of Beclin-1 and p-mTOR/mTOR.@*RESULTS@#There were no significant difference in expression of ROS, SOD, Beclin-1mRNA and mTOR mRNA between male and female chronic injury groups, and between male and female acute injury groups (P>0.05); ROS, SOD and Beclin-1mRNA in male chronic injury group were higher than those in male chronic injury group, while mTOR mRNAand protein decreased (P<0.05);ROS, SOD and Beclin-1 mRNA in female chronic injury group were up-regulated compared with female acute injury group, while mTOR mRNA was down-regulated (P<0.05).@*CONCLUSION@#Chronic rotator cuff injury is more likely to stimulate the pro-oxidation state of rotator cuff tissue than acute rotator cuff injury, which could up-regulating expression of autophagy factor Beclin-1 and down-regulating expression of mTOR. Therefore, patients with chronic rotator cuff injury may have higher levels of oxidative stress and autophagy.

Guiqi Yiyuan Ointment protects rat left lung from bystander effect of right lung injury induced by ~(12)C~(6+) beam / 中国中药杂志

This study observed the effects of Guiqi Yiyuan Ointment(GQYY) on the left lung subjecting to bystander effect of right lung injury induced by ~(12)C~(6+) beam in rats and decipher the underlying mechanism from NOD-like receptor protein 3(NLRP3)/apoptosis-associated speck-like protein containing a CARD(ASC)/cysteinyl aspartate specific proteinase-1(caspase-1) pathway. Wistar rats were randomized into 7 groups: blank, model, inhibitor [200 mg·kg~(-1), N-acetylcysteine(NAC)], western drug [140 mg·kg~(-1) amifostine(AMI)], and high-, medium-, and low-dose(4.8, 2.4, and 1.2 g·kg~(-1), respectively) GQYY groups. The model of bystander effect damage was established by 4 Gy ~(12)C~(6+) beam irradiation of the right lung(with the other part shielded by a lead plate). The pathological changes in the lung tissue, the level of reactive oxygen species(ROS) in the lung tissue, and the levels of superoxide dismutase(SOD) and malondialdehyde(MDA) in the serum were observed and measured in each group. Furthermore, the mRNA and protein levels of NLRP3, ASC, caspase-1, and phosphorylated nuclear factor-κB p65(p-NF-κB p65)/nuclear factor-κB p65(NF-κB p65) were determined. Compared with the blank group, the model group showed thickened alveolar wall, narrowed alveolar cavity, and presence of massive red blood cells and inflammatory infiltration in the alveolar wall and alveolar cavity. In addition, the model group showed elevated ROS levels in both left and right lungs, elevated MDA level, lowered SOD level, and up-regulated mRNA and protein levels of NLRP3, ASC, caspase-1, and p-NF-κB p65/NF-κB p65. Compared with the model group, the drug administration in all the groups reduced inflammatory cell infiltration in the lung tissue. The inhibitor group and the western drug group showed enlarged alveolar cavity, thinned interstitium, and reduced inflammation. There was a small amount of alveolar wall rupture in the high-and medium-dose GQYY groups and reduced inflammatory cell infiltration in the low dose GQYY group. Compared with the model group, drug administration lowered level of ROS in the left and right lungs, lowered the MDA level, elevated the SOD level, and down-regulated the mRNA and protein levels of NLRP3, ASC, caspase-1, and p-NF-κB p65/NF-κB p65. GQYY can effectively reduce the damage caused by radiation and bystander effect, which may be associated with the ROS-mediated NLRP3 inflammasome activation.

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.

Effect of a disintegrin and metalloprotease 17 deletion on production of reactive oxygen species and mitochondrial function in nasopharyngeal carcinoma cells / 中国生物制品学杂志

@#Objective To investigate the effects of a disintegrin and metalloproteinase 17(ADAM17) deletion on the production of reactive oxygen species(ROS) and mitochondrial function in nasopharyngeal carcinoma(NPC) cells.Methods Three groups of ADAM1 7 interfering plasmid ADAM17 shRNA and empty plasmid ADAM17-shRNA-NC were transfected into NPC cell line(CNE1) and detected for the interference efficiency by RT-PCR and Western blot to select shRNA with the best interference effect for the follow-up experiments.The cell proliferation was detected by CCK-8 assay,while the cell growth by clone formation test,the apoptosis and changes in mitochondrial membrane potential(MMP) by flow cytometry,the level of mitochondrial oxidative damage product ROS by fluorescence microscope,the contents of oxidative stress markers MDA and SOD by malondialdehyde(MDA) kit and superoxide dismutase(SOD) kit and the expression of mitochondrial damage markers Bax/Bcl-2,cleaved-caspase 9/caspase 9,cleaved-caspase 3/caspase 3 and c-Myc by Western blot.Results ADAM17-shRNA2 group showed the best interference effect.Compared with shRNA-NC group,the proliferation rate of cell in ADAM17-shRNA 2 group decreased significantly(t=8.964,P=0.036);the number of colonies were significantly reduced(t=10.351,P=0.014);the number of apoptosis increased significantly(t=11.25,P=0.008);the fluorescence intensity representing ROS level in cells increased obviously;the mitochondrial membrane potential decreased significantly(t=9.233,P=0.013);the SOD content decreased(t=7.233,P=0.034) and MDA content increased(t=7.415,P=0.038) significantly;the levels of Bax/Bcl-2,cleaved-caspase 9/caspase 9 and cleaved-caspase 3/caspase 3 significantly increased(t=8.985,9.021 and 7.789,P=0.023,0.011 and 0.031,respectively),while the expression of c-Myc proteins significantly decreased(t=10.352,P=0.004).Conclusion Interfering with ADAM1 7 induced SOD decrease and MDA increase by promoting oxidation,thereby alleviating oxidative damage of cell membrane,which also promoted the expression level of ROS in mitochondrion,reduced MMP,inhibited cell proliferation in vitro,and promoted apoptosis.

Mogroside regulates the oxidative stress response of retinal pigment epithelial cells induced by H 2O 2 through silent information regulator of transcription 1/nuclear factor erythroid-2-related actor 2 signaling pathway / 中华眼底病杂志

Objective:To observe and preliminarily explore the effect of mogroside on oxidative stress of retinal pigment epitheliaum (RPE) cells induced by hydrogen peroxide (H 2O 2) and its possible mechanism. Methods:A experimental study. The RPE cells were divided into control group, H 2O 2 group, silent information regulator of transcription 1 (SIRT1) inhibitor EX527 group (EX527 group), mogroside group, mogroside+EX527 group. Methyl thiazolete trazolium method was used to detect cell survival rate. Flow cytometry was used to detect cell apoptosis rate. 2' ,7'-dichlorodihydrofluorescein diacetate fluorescent probe method, xanthine method and enzyme-linked immunosorbent assay method were used to detect the level of reactive oxygen species (ROS), superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in cells respectively. Real-time quantitative polymerase chain reaction and Western blot were used to detect relative expressions of SIRT1, nuclear factor erythroid-2-related actor 2 (Nrf2), heme oxygenase-1 (HO-1) mRNA and protein in cells. One-way ANOVA was used for comparison among groups. The pairwise comparison between groups was tested by the least significant difference t test. Results:Compared with the control group, the H 2O 2 group cell survival rate decreased, the apoptosis rate increased, the ROS level in the cells increased, the SOD activity decreased, the MDA content increased, and the relative expression of SIRT1, Nrf2, HO-1 mRNA and protein decreased ( P<0.05). Compared with H 2O 2 group, the cell survival rate decreased, apoptosis rate increased, the cell ROS level increased, SOD activity decreased, MDA content increased, SIRT1, Nrf2, HO-1 mRNA and protein expression decreased in EX527 group ( P<0.05); the cell survival rate increased, apoptosis rate decreased, ROS level decreased, SOD activity increased, MDA content decreased, and the relative expression of SIRT1, Nrf2, HO-1 mRNA and protein increased in mogroside group ( P<0.05). Compared with the mogrosides group, the cell survival rate decreased, the apoptosis rate increased, the level of ROS increased, SOD activity decreased, MDA content increased, SIRT1, Nrf2, HO-1 mRNA and protein decreased in mogrosides+EX527 group ( P<0.05). Conclusions:Mogrosides can alleviate the oxidative stress response of visual RPE cells induced by H 2O 2, promote cell proliferation, and reduce cell apoptosis. Mogrosides may exert antioxidant effects by activating the SIRT1/Nrf2 signaling pathway.