Reduction Of ROS Research Articles

Piperonal protects neuron-like and retinalpigment epithelial (RPE) cells from oxidative stress and apoptosis through inhibition of α-synuclein aggregation

Protein fibrillation is a crucial process in the onset of several neurodegenerative and retinal disorders due to the formation of cytotoxic species. Because of their capacity to prevent protein aggregation, small molecules have the potential to be appointed as therapeutic agents. Here, we examined the inhibitory impacts of the piperonal as a carbaldehyde-derived compound on the fibrillation process of α-synuclein and underlying cytotoxicity against neuron-like (PC12) and human retinal pigment epithelial (RPE) (ARPE-19) cells. The results showed that the values of kapp and lag time of α-synuclein were modulated with piperonal. Moreover, ANS fluorescence intensity analysis indicated that piperonal can inhibit the formation of a molten global (misfolded) state of α-synuclein, which is a necessary step in the formation of protein amyloid fibrils. Congo red absorption and circular dichroism spectroscopy also verified the inhibition of β-sheet structure formation after treatment of α-synuclein with piperonal. Furthermore, theoretical studies displayed that piperonal interacts with VAL40:HN, GLU35:O, VAL40, and LYS43 amino acid residues and forms a complex. In addition, cytotoxicity assays demonstrated that piperonal as a safe small molecule could mitigate the induced cytotoxicity by α-synuclein amyloids in PC12 and ARPE-19 cells through reduction of ROS and Bax/Bcl2 mRNA overexpression. Taken together, these outcomes showed that piperonal as a natural aldehyde compound can inhibit α-synuclein fibrillation and underlying cytotoxicity which may be developed for potential therapeutic applications in vivo.

Withanicandrin Isolated from Datura Ferox Promotes Antinociception by Modulating the Asics and TRPS Channels and Anti-Inflammation in Adult Zebrafish.

This is the first study to analyze the anti-inflammatory and antinociceptive effect of withanicandrin, isolated from Datura Ferox leaves, and the possible mechanism of action involved in adult zebrafish (ZFa). To this end, the animals were treated intraperitoneally (i. p.) with withanicandrin (4; 20 and 40 mg/kg; 20 μL) and subjected to locomotor activity and acute toxicity. Nociception tests were also carried out with chemical agents, in addition to tests to evaluate inflammatory processes induced by κ-Carrageenan 1.5 % and a Molecular Docking study. As a result, withanicandrin reduced nociceptive behavior by capsaicin at a dose of 40 mg/kg and by acid saline at doses of 4 and 40 mg/kg, through neuromodulation of TRPV1 channels and ASICs, identified through blocking the antinociceptive effect of withanicandrin by the antagonists capsazepine and naloxone. Furthermore, withanicandrin caused an anti-inflammatory effect through the reduction of abdominal edema, absence of leukocyte infiltrate in the liver tissue and reduction of ROS in thel liver tissue and presented better affinity energy compared to control morphine (TRPV1) and ibuprofen (COX-1 and COX-2).

Vitamin E alleviates chlorpyrifos induced glutathione depletion, lipid peroxidation and iron accumulation to inhibit ferroptosis in hepatocytes and mitigate toxicity in zebrafish

Organophosphates, a widely used group of pesticides, can cause severe toxicity in human beings and other non-target organisms. Liver, being the primary site for xenobiotic metabolism, is extremely vulnerable to xenobiotic-induced toxicity. Considering the numerous vital functions performed by the liver, including xenobiotic detoxification, protecting this organ from the ubiquitous pesticides in our food and environment is essential for maintaining homeostasis.In this study, we have investigated the impact of the organophosphate pesticide, Chlorpyrifos (CPF), on zebrafish liver at a concentration (300 μg/L) which is environmentally realistic. We have also demonstrated the role of dietary supplementation of α-tocopherol or Vitamin E (Vit E) (500 mg/kg feed) in mitigating pesticide-induced liver toxicity. Mechanistically, we showed that Vit E resulted in significant elevation of the Nrf2 and its downstream antioxidant enzyme activities and gene expressions, especially that of GST and GPx, resulting in reduction of CPF-induced intracellular lipid ROS and hepatic LPO. Further interrogation, such as analysis of GSH: GSSG ratio, intracellular iron concentration, iron metabolizing genes, mitochondrial dysfunction etc. revealed that CPF induces ferroptosis which can be reversed by Vit E supplementation. Ultimately, reduced concentration of CPF in zebrafish serum and flesh highlighted the role of Vit E in ameliorating CPF toxicity.

Ethyl acetate fraction of Sargassum pallidum extract attenuates particulate matterinduced oxidative stress and inflammation in keratinocytes and zebrafish

Objective: To evaluate the effect of the ethyl acetate fraction derived from Sargassum pallidum extract against particulate matter (PM)-induced oxidative stress and inflammation in HaCaT cells and zebrafish. Methods: HaCaT cells and zebrafish were used to evaluate the protective effects of the ethyl acetate fraction of Sargassum pallidum extract against PM-induced oxidative stress and inflammation. The production of nitric oxide (NO), intracellular ROS, prostaglandin E2 (PGE2), and pro-inflammatory cytokines, and the expression levels of COX-2, iNOS, and NF-κB were evaluated in PM-induced HaCaT cells. Furthermore, the levels of ROS, NO, and lipid peroxidation were assessed in the PM-exposed zebrafish model. Results: The ethyl acetate fraction of Sargassum pallidum extract significantly decreased the production of NO, intracellular ROS, and PGE2 in PM-induced HaCaT cells. In addition, the fraction markedly suppressed the levels of pro-inflammatory cytokines and inhibited the expression levels of COX-2, iNOS, and NF-κB. Furthermore, it displayed remarkable protective effects against PM-induced inflammatory response and oxidative stress, represented by the reduction of NO, ROS, and lipid peroxidation in zebrafish. Conclusions: The ethyl acetate fraction of Sargassum pallidum extract exhibits a protective effect against PM-induced oxidative stress and inflammation both in vitro and in vivo and has the potential as a candidate for the development of pharmaceutical and cosmeceutical products.

Functional features of the exopolysaccharide extracts produced by Lactiplantibacillus strains isolated from table olives.

This study evaluates the functional characteristics of the exopolysaccharide (EPS) extracts produced by various strains of Lactiplantibacillus pentosus (LPG1, 119, 13B4, and Lp13) and Lactiplantibacillus plantarum (Lp15) isolated from table olives. None of the EPS crude extracts showed cytotoxicity when administered to THP-1 human macrophage cells at dosages ranging from 6.25 to 50 μg mL-1. Many exhibited anti-inflammatory properties (reduction of pro-inflammatory cytokines TNF-α and IL-6 production) and antioxidant activity (reduction of ROS%) when macrophages were stimulated with Escherichia coli lipopolysaccharide. Notably, the EPS extract produced by the L. pentosus LPG1 strain had the best results corroborated by western blot immune analysis for differential expression of COX-2, Nrf-2, and HO-1 proteins, with the most significant antioxidant and anti-inflammatory response observed at a dosage of 50 μg mL-1. Chemical analysis revealed that the EPS extract produced by this strain contains a heteropolymer composed of mannose (35.45%), glucose (32.99%), arabinose (17.93%), xylose (7.48%), galactose (4.03%), rhamnose (1.34%), and fucose (0.77%). Finally, we conducted response surface methodology to model the EPS extract production by L. pentosus LPG1 considering pH (3.48-8.52), temperature (16.59-33.41 °C) and salt concentration (0.03-8.77% NaCl) as independent variables. The model identified linear effects of salt and pH and quadratic effects of salt as significant terms. The maximum EPS extract production (566 mg L-1) in a synthetic culture medium (MRS) was achieved at pH 7.5, salt 7.0%, and a temperature of 20 °C. These findings suggest the potential for novel applications for the EPS produced by L. pentosus LPG1 as nutraceutical candidates for use in human diets.

The protective effects of betanin against experimental gastric ulcer by reduction of ROS and suppression of inflammatory genes via NF-κB, iNOS, COX-2 and TNF-α pathways

Betanin, a prominent compound found in beetroot, however, the potential protective effects of this compound against gastric damage and ulceration have yet to be explored. This study aims to investigate the impact of purified beetroot betanin (Bet) on the activities of inflammatory enzymes and gene expressions associated with inflammation in gastric tissues. Gastric ulcer rats were induced by ethanol at dose 5ml/kg body weight. Ulcerated rats were treated by purified betanin or omeprazole; from which some inflammation-related genes and gastric-inflammation enzymes were investigated using RT-PCR and biochemical analysis, respectively. Bet effectively suppressed the activities of key enzymes involved in gastric mucosal inflammation, including 5-Lipoxygenase (5-LO), Hyaluronidase (HAase), and myeloperoxidase (MPO), by 58%, 50%, and 45%, respectively. Docking studies showed Bet binding to catalytic sites or key regions for enzymes’ activities, thus explaining the inhibitory capacity. Bet ingestion decreased the expression levels of inflammation-related genes, factor-kappa B (NF-ϰB) by 57%. This down-regulation subsequently led to a reduction in the expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and tumour necrosis factor-α (TNF-α). Bet ingestion supressed thiobarbituric acid reactive substance (TBARS) rates by 59%, augmented nitric oxide (NO) level by 107% and protected gastric tissues from damage and ulceration. Bet revealed gastro-protective effects as confirmed by a reduction in the ulceration surface area and the acidity by 75.9% and 40%, respectively, and a significant increase in the weight of mucus and the curative index by 117% and 74.5%, respectively. In conclusion, these outcomes of physiological and molecular-based investigations are indicative of potential use of Bet as a functional food additive for the prevention of gastric ulcer.

Relation between Strawberry Fruit Redness and Bioactivity: Deciphering the Role of Anthocyanins as Health Promoting Compounds.

The red colour of most berries is often associated to fruit healthiness, since it has been linked to enrichment in anthocyanins (polyphenol with antioxidative properties). However, recent studies suggest that anthocyanins could not be the major contributors to bioactivity leading to uncertainty about their role as important molecules in the generation of health-promoting properties. To shed light on this issue, spectrophotometric and HPLC techniques were used for characterizing the content of phenolic compounds, including anthocyanins, in fruits of red (Fragaria x ananassa, cv. Fortuna) and white strawberry (Fragaria vesca spp. XXVIII) species (distinguishing receptacle from achene). In addition, the effect of these extracts on the reduction of intracellular ROS was tested, as well as on the activity of antioxidant enzymes and the quantification of cell oxidation markers. The results showed that white receptacle extracts (deprived of anthocyanins) were able to protect cells from oxidative damage to a greater extent than red fruits. This could be due per se to their high antioxidant capacity, greater than that shown in red fruits, or to the ability of antioxidants to modulate the activity of antioxidant enzymes, thus questioning the positive effect of anthocyanins on the wholesomeness of strawberry fruits. The results shed light on the relevance of anthocyanins in the prevention of health-associated oxidative damage.

Shexiang Tongxin dropping pills protect against ischemic stroke-induced cerebral microvascular dysfunction via suppressing TXNIP/NLRP3 signaling pathway

Ethnopharmacological relevancePatients with ischemic stroke (IS) often continue to exhibit cerebral microcirculatory dysfunction even after receiving thrombolytic therapy. Enhancing the function of cerebral microvascular endothelia represents a pivotal advancement in the therapeutic strategy for ischemic microcirculatory disturbances. A traditional Chinese medicinal formulation named Shexiang Tongxin Dropping Pills (STDP), has been clinically employed to ameliorate microcirculatory abnormalities. Existing literature attests to the beneficial role of STDP on endothelial cells (ECs). Nevertheless, specific impacts and underlying mechanisms of STDP in rectifying IS-induced cerebral microvascular dysfunction warrant further exploration. Aim of the studyThis investigation seeks to delineate the effects of STDP on cerebral microvascular endothelial damage induced by ischemic stroke and to elucidate the underlying mechanism involved. Materials and methodsMiddle cerebral artery occlusion and reperfusion (MCAO/R) technique was employed to established ischemic stroke model in mice. The therapeutic efficacy of STDP on cerebral microvascular function was assessed through laser speckle contrast imaging, behavioral assays, and histological evaluations. Biochemical markers in the brain tissue, including GSH, SOD, MDA, and ROS, were quantified using specific assay kits. In vitro study, oxygen-glucose deprivation and reperfusion (OGD/R) was performed in bEnd.3 cells. The cytoprotective potential of STDP was then evaluated by measuring cell viability, LDH activity, endothelial permeability, and oxidative stress parameters. Important targets in critical pathway were verified by immunoblotting and immunofluorescence both in mice brain slices and bEnd.3 cells. ResultsSTDP decrease brain infarct size, repaired microvascular cerebral blood flow and attenuated neurological deficiency in MCAO/R mice. Moreover, STDP abolished MCAO/R-induced oxidative stress which was reflected by rescuing GSH content, restoration of SOD activity and T-AOC, reduction of MDA and ROS. Ex vivo, STDP increased cerebral microvascular endothelial cells viability, abolished oxidative stress and decreased their permeability after ODG/R. Mechanistically, STDP significantly suppressed endothelial ROS-TXNIP mediated the activation of NLRP3 inflammasome in vivo and in vitro. ConclusionSTDP improves ischemic stroke-induced cerebral microcirculatory deficits by regulating cerebral microvascular endothelial ROS/TXNIP/NLRP3 signaling pathway.

Anti-inflammatory, antioxidant and photoprotective activity of standardised Gaultheria procumbens L. leaf, stem, and fruit extracts in UVA-irradiated human dermal fibroblasts

Ethnopharmacological relevanceGaultheria procumbens L. is a polyphenolic-rich medicinal and food plant. Its leaves, stems, and fruits are traditional anti-inflammatory, antipyretic, antioxidant, and antimicrobial herbal medicines used to treat internal and external inflammation-related ailments, including rheumatic diseases, influenza, the common cold, fever, and skin and periodontal problems. Moreover, G. procumbens leaf extract is used for skin care as an anti-ageing and anti-wrinkle ingredient. Aim of the studyVarious environmental factors, especially solar ultraviolet radiation, accelerate skin ageing by promoting oxidative stress and inflammation. Despite the dermoprotective and anti-ageing applications, the impact of G. procumbens on human dermal fibroblasts is unknown. Therefore, the study aimed to evaluate the anti-inflammatory, antioxidant, and photoprotective activity of G. procumbens standardised leaf, stem, and fruit extracts in cellular models, including human dermal fibroblasts (Hs68 cells) under UVA-irradiation, the primary pro-ageing skin stressor. Materials and methodsHs68 fibroblasts were pre-treated (24h) with G. procumbens extracts (0.5–100 μg/mL) or reference compounds followed by UVA-irradiation (8 J/cm2). Cell viability and metabolic activity were measured by CCK-8 and MTT assays in human Hs68 and mouse L929 fibroblasts, respectively. The ROS level, SOD, and GST activities were estimated by fluorescence and spectrophotometric techniques. The pro-inflammatory potential (NF-κB transcription factor activation) was checked using THP1-Blue™ NF-κB cells, and the anti-inflammatory activity was studied by measuring IL-8, ICAM-1, and NF-κB levels and phosphorylation of Erk kinase in LPS-stimulated Hs68 cells by spectrophotometry and confocal microscopy. The UVA-induced DNA damage and cell migration were evaluated by comet and scratch assays, respectively. ResultsThe extracts did not affect the metabolic activity of mouse L929 fibroblasts and the viability of unirradiated human Hs68 cells. Additionally, the extracts noticeably enhanced the viability of UVA-irradiated Hs68 cells up to 115–120% (p < 0.001) for stem and leaf extract at 25 μg/mL. All extracts in a wide concentration range (0.5–100 μg/mL) did not activate monocytes or induce the NF-κB transcription factor in LPS-stimulated Hs68 fibroblasts. On the other hand, the extracts (5–25 μg/mL) restored the activity of endogenous antioxidant enzymes, i.e., SOD and GST, up to 120–140% (p < 0.001) in the UVA-irradiated Hs68 cells. Moreover, a statistically significant reduction of ROS, IL-8, ICAM-1, and NF-κB levels by up to 48%, 88%, 43%, and 39%, respectively (p < 0.001) and strong suppression of Erk kinase activation was observed for the extracts (25–50 μg/mL) in LPS-stimulated human fibroblasts. The total DNA damage (% tail DNA) in irradiated Hs68 cells was also strongly decreased by up to 66–69% (p < 0.001) at 50 μg/mL. However, the treatment with the extracts did not relevantly enhance the cell migration of Hs68 fibroblasts. ConclusionsThe results suggest that G. procumbens may effectively protect human skin fibroblast from UVA irradiation. The leaf and stem extracts were the most potent antioxidants, while fruit and stem extracts revealed the strongest anti-inflammatory activity. The observed effects support the traditional use of aerial plant parts (leaves, stems, and fruits) in treating inflammation-related skin disorders cross-linked with oxidative stress and the topical application of Gaultheria extracts as anti-ageing agents intended for skin care.

In-silico prediction and in vitro validation of antioxidant, antibacterial and antifungal potential of Black Turmeric (Curcuma caesia Roxb.) essential oils and its constituents

The growing interest in the health and pharmaceutical industries has led to a significant increase in demand for essential oils. Curcuma caesia, as a member of the Zingiberaceae family, has been used for its high-quality essential oils (EOs) with remarkable therapeutic properties. The current study aimed at predicting the in-silico interactions of C. caesia volatile constituents against the protein responsible for oxidative and microbial stress in order to investigate their potential for industrial and pharmaceutical applications. The accuracy of the in-silico predictions was confirmed by carrying out the in vitro validation of the antioxidant, antibacterial, and antifungal activity of the EOs derived from the rhizome and leaves, as well as its volatile constituents. EOs from the rhizome and leaf samples of C. caesia were extracted through the hydro distillation method and yielded a percentage (%) of 1.80 ± 0.02 and 0.95 ± 0.02 (fresh weight basis, v/w), respectively. A total of 38 and 29 volatile constituents were identified in the rhizome and leaf EOs by gas-chromatography mass spectrometry analysis, representing 92.55% and 90.75% of the total peak area, respectively. The rhizome and leaf EOs showed oxygenated sesquiterpene (39.91% and 27.69%) as the predominate fractions followed by sesquiterpene hydrocarbons (13.36% and 18.44%). Curzerenone (18.09%), epicurzerenone (12.47%), eucalyptol (11.57%), camphor (5.94%), and germacrone (4.47%) were the major constituents in the rhizome EO, whereas, the leaf EO was dominated by eucalyptol (12.53%), camphor (7.23%), curzerenone (5.24%), germacrone (4.81%) and curzerene (4.9%). The in vitro biological activities such as antioxidant, antibacterial, and antifungal potential of C. caesia EOs were evaluated. The antioxidant evaluation using enzymatic and non-enzymatic assays, showed excellent potential of C. caesia EOs as compared to standard drugs. Specifically, in the enzymatic assay, leaf EO of C. caesia was resulted into the reduction of ROS and an increase in the activity of superoxide dismutase (SOD), catalase (CAT), and glutathione S-transferase (GST) in OZ-stimulated PMNs. The EOs also displayed a remarkable antibacterial activity against Staphylococcus aureus, elucidating it as the most susceptible strain. Further, the in-silico prediction of the constituents was conducted using Swiss-ADME, and molecular docking analysis. Swiss-ADME revealed the drug-likeness and safety properties of constituents present in C. caesia EOs. The molecular docking study was carried out to interpret the possible interactions of phytoconstituents qualified in the Swiss-ADME study with two bacterial proteins (Tyrosyl-tRNA synthetase from Staphylococcus aureus (PDB ID: 1JIJ) and Glucosamine 6-phosphate synthase from Escherichia coli (PDB ID: 1XFF)), one fungal protein (N-myristoyl transferase from Candida albicans (PDB ID: 1IYL)) and one oxidative protein (Human peroxiredoxin 5 (PDB ID: 1HD2)). Furthermore, to validate the result of molecular docking, the biological activity of higher affinity constituents was also evaluated. As a result, the in vitro findings supported the in-silico prediction. Therefore, C. caesia or its constituents like curzerenone, camphor, eucalyptol, epicurzerenone, germacrone, viridifloral, muurola-4,10(14)-dien-1-ol, and globulol might be a valuable source for developing antioxidant and antimicrobial drugs. To the best of present author’s knowledge, this is the first scientific report on the in-silico prediction of C. caesia constituents against bacterial, fungal and oxidative proteins through molecular docking, which may contribute to the development of various alternatives against multi-drug resistance.

A Review on Understanding the Effects and Mechanisms of Salinity Tolerance in Rice (Oryza sativa L.)

Salinity, along with drought, is one of the key abiotic stressors that has posed a danger to the advancement and evolution of cereal crops like rice and wheat. Water shortage and a lack of irrigation water availability are the main causes of salty soil formation. Rice is salt sensitive and glycophyte, wheat is moderately salt tolerant. Wild tolerant cultivars like Oryza coarctata and Oryza alta are more tolerant than traditional cultivars such as Pokkali and Nona Bokra in rice. Salt stress affects crop plants’ processes like ionic imbalance, osmotic and oxidative stress. Na+ should be low in the shoots of the plant which is restricted by various transporters in the cell membrane of the roots in soil. High K+ &amp; Na+/K+ homeostasis should be maintained. Many RILs and NILs have been developed which acts as a donor for salinity tolerant genes. FL478 is a recombinant inbred line in which candidate genes are situated in the Saltol region of chromosome 1 region which is obtained by a cross between Pokkali x IR29. Increase in world’s population, rice output must be increased by at least 25% by 2030 and 50% by 2050.Salinity stress is a polygenic character which involves several genes works in harmony. For evolution of salinity tolerant cultivars, we need to access the physiological, biochemical genetic responses of the crop plant which helps in transfer of candidate genes from donor parents to elite high yielding salt sensitive cultivars. Especially in rice salt tolerant mechanisms like, Ion equilibrium regulation, Adjustment of osmotic potential, Reduction of ROS, Nutrient disequilibrium, and Regulation of PGRs. Conventional, MABC, MAS and direct gene transfer by transgenic methods. This review paper's main objective is to understand the mechanisms of the crop plants to salinity effects and development of salt tolerant cultivars by modern approaches which fulfill the food scarcity of staple food crops with increasing population.

Protective effects of tanshinone IIA on Porphyromonas gingivalis-induced atherosclerosis via the downregulation of the NOX2/NOX4-ROS mediation of NF-κB signaling pathway

Tanshinone IIA (TSA), an active component isolated from Danshen, possess high medicinal values against atherosclerosis by reducing vascular oxidative stress, inhibiting platelet aggregation, and protecting the endothelium from damage. The periodontal pathogen Porphyromonas gingivalis (P. gingivalis) has been proven to accelerate the development of atherosclerosis. We aim to determine the effects of TSA on P. gingivalis-induced atherosclerosis in ApoE-knockout (ApoE−/−) mice. After feeding with a high-lipid diet and infected with P. gingivalis three times per week for four weeks, TSA-treated (60 mg/kg/d) mice greatly inhibited atherosclerotic lesions both morphologically and biochemically and exhibited significantly reduction ROS, 8-OHdG, and ox-LDL levels in serum compared with P. gingivalis-infected mice. Additionally, TSA-treated mice were observed a marked reduction of ROS, 8-OHdG and ox-LDL in the serum, mRNA levels of COX-2, LOX-1, NOX2 and NOX4 in the aorta, as well as the levels of NOX2, NOX4, and NF-κB. These results suggest that TSA attenuates oxidative stress by decreasing NOX2 and NOX4 and downregulating NF-κB signaling pathway, which might be contributed to the amelioration of atherosclerosis.

Anticancer effect of antioxidant-rich methanolic extract of Rauvolfia serpentina (L.) Benth. ex Kurz leaves in HepG2 and HeLa cells: A mechanistic insight

Rauvolfia serpentina (L.) Benth. ex Kurz is a pharmaceutically, and agriculturally important medicinal plant traditionally used against snake and insect bites, dysentery, abdominal pain, fever, and insanity. However, no detailed study has been done on anticancer potency of crude leaf extract of Rauvolfia serpentina. Therefore, we investigated the anticancer activity of methanolic extract of Rauvolfia serpentina leaf (MERSL) in human hepatocarcinoma HepG2 and cervical cancer HeLa cells along with its antioxidant capacity. GC-MS analysis revealed the presence of many anticancer phytochemicals including campesterol, stigmasterol, beta-sitosterol, alpha-amyrin, lupeol, phytol, and D-pinitol. The antioxidant activity of MERSL was confirmed by DPPH (IC50 = 106.16 μg/ml), ABTS (IC50 = 146.67 μg/ml), FRAP (IC50 = 1.4 mg/ml), and total antioxidant capacity assay invitro, and reduction of H2O2-induced ROS in noncancer fibroblast F111 cells. Crystal violet assay showed that MERSL reduced cell viability in HepG2 (IC50 = 274.12 μg/ml) and HeLa (IC50 = 231.33 μg/ml) at 24 h, but not in F111 cells at these IC50 concentrations. Further, wound healing and clonogenic assays confirmed anticancer activity, which resulted in decreased cell migration and revival capacity in HepG2 and HeLa cells. Apoptotic changes were visualized by acridine orange-ethidium bromide and DAPI staining using fluorescence and confocal microscopy. Also, comet assay indicated DNA damage. An increase in intracellular ROS, calcium influx, reduction of mitochondrial mass and membrane potential, and cell cycle arrest, established by flow cytometry and fluorimetry, highlighted the possible underlying mechanism of cell death by MERSL in HepG2 and HeLa cells. Therefore, this study evidenced that Rauvolfia serpentina leaves could be an encouraging pharmacological application for developing anticancer strategies.

Metformin promotes in vitro maturation of oocytes from aged mice by attenuating mitochondrial oxidative stress via SIRT3-dependent SOD2ac.

Human female fecundity decreases irreversibly as chronological age rises, adversely affecting oocyte quality, consequently worsening pregnancy outcomes and increasing the extent of birth defects. The first-line type 2 diabetes treatment metformin has been associated with delayed aging and reduction of oxidative stress; yet it remains unclear if metformin confers any benefits for oocytes from aged mice, particularly in the context of the assisted human reproductive technology (ART) known as in vitro maturation (IVM). Here, we found that adding metformin into the M16 culture medium of oocytes from aged mice significantly improved both oocyte maturation and early embryonic development. This study showed that metformin reduced the extent of meiotic defects and maintained a normal distribution of cortical granules (CGs). RNA-seq analysis of metformin-treated oocytes revealed genes apparently involved in the reduction of mitochondrial ROS. Further, the results supported that the metformin improved mitochondrial function, reduced apoptosis, increased the extent of autophagy, and reduced mitochondrial ROS via SIRT3-mediated acetylation status of SOD2K68 in oocytes from aged mice. Thus, this finding demonstrated a protective effect for metformin against the decreased quality of oocytes from aged mice to potentially improve ART success rates and illustrated a potential strategy to prevent or delay reproductive aging.

Leptin and prolactin reduce cryodamage in normozoospermic human semen samples during cryopreservation

ObjectivesCryopreservation has destructive effects on the function and structure of spermatozoa. It is known that leptin and prolactin play an active role in decreasing the rates of reactive oxygen species and DNA fragmentation, as well as enhancing sperm motility. Hence, this experiment aimed to investigate the effects of leptin and prolactin as pro-survival factors on the normozoospermic human semen samples during cryopreservation. Material and methodsSemen samples were collected from 15 healthy, fertile men ranging from 25 to 40 years. Cryopreservation of the samples was performed in liquid nitrogen over a period of two weeks, using five varying concentrations of leptin/prolactin, 0, 10, 100, 500, and 1000ng/ml respectively. Sperm motility, total caspase activity, and mitochondrial and cytosolic ROS were measured by flowcytometry, TUNEL, and other appropriate tests after thawing of the samples. ResultsBoth hormones were observed to have positive effects on the motility of the samples post-cryopreservation, the highest improvement being in the 100ng/ml concentration leptin and prolactin in comparison to the control group (P=0.01 and P=0.041, respectively). A significant reduction of mitochondrial ROS was also observed in 100 and 1000ng/ml of leptin (P=0.042), and there was a considerable decrease in the cytosolic ROS in the 100ng/ml of prolactin in comparison to the control group (P=0.048). Total caspase activity was also highly reduced in the 100, 500, and 1000ng/ml of leptin compared to the control group (P=0.039). Interestingly, both hormones also significantly decreased DNA fragmentation in 1000ng/ml compared to the control group (P=0.042). ConclusionIt can be concluded that leptin and prolactin act as protective agents against cryodamage to spermatozoa during cryopreservation.

Targeting on Nrf2/Sesn2 Signaling to Rescue Cardiac Dysfunction during High-Fat Diet-Induced Obesity.

Obesity is of concern to the population because it is known to cause inflammation and oxidative stress throughout the body, leading to patient predisposition for health conditions such as diabetes, hypertension, and some cancers. However, some proteins that are activated in times of oxidative stress may provide cytoprotective properties. In this study, we aim to gain further understanding of the interconnection between Nrf2 and Sesn2 during obesity-related stress and how this relationship can play a role in cardio-protection. Cardiomyocyte-specific Sesn2 knockout (cSesn2−/−) and Sesn2 overexpressed (tTa-tet-Sesn2) mice and their wildtype littermates (Sesn2flox/flox and tet-Sesn2, respectively) were assigned to either a normal chow (NC) or a high-fat (HF) diet to induce obesity. After 16 weeks of dietary intervention, heart function was evaluated via echocardiography and cardiac tissue was collected for analysis. Immunoblotting, histology, and ROS staining were completed. Human heart samples were obtained via the LifeLink Foundation and were also subjected to analysis. Overall, these results indicated that the overexpression of Sesn2 appears to have cardio-protective effects on the obese heart through the reduction of ROS and fibrosis present in the tissues and in cardiac function. These results were consistent for both mouse and human heart samples. In human samples, there was an increase in Sesn2 and Nrf2 expression in the obese patients’ LV tissue. However, there was no observable pattern of Sesn2/Nrf2 expression in mouse LV tissue samples. Further investigation into the link between the Sesn2/Nrf2 pathway and obesity-related oxidative stress is needed.

Sulindac acetohydrazide derivative attenuates against cisplatin induced organ damage by modulation of antioxidant and inflammatory signaling pathways

This study aimed to explore the mechanisms of action of a sulindac acetohydrazide derivative, N'-(4-dimethylaminobenzylidene)-2-1-(4-(methylsulfinyl) benzylidene)-5-fluoro-2-methyl-1H-inden-3-yl) acetohydrazide, against anticancer drug cisplatin induced organ damage. Using a rodent model, various markers of organ function and signaling pathways were examined and validated by molecular docking studies. The study involves five groups of animals: control, DMSO, CDDP, CDDP + DMFM, and DMFM. Biochemical enzyme activity, histopathology, tissue antioxidant, and oxidative stress markers were examined. RT-PCR and western blot analyses were conducted for the expression of inducible cyclooxygenase enzyme (COX-2), nuclear factor kappa beta (NF-κB), p65, IL-1, TNF-α, and inducible nitric oxide synthase (iNOS). Flow cytometry analysis of CD4 + TNF-α, CD4 + COX-2, and CD4 + STAT-3 cells in whole blood was performed. Structural and dynamic behavior of DMFM upon binding with receptor molecule molecular docking and dynamic simulations were performed using bioinformatics tools and software. Treatment with DMFM reversed cisplatin-induced malondialdehyde (MDA) and nitric oxide (NO) induction, whereas the activity of glutathione peroxidase (GPx), and superoxide dismutase (SOD) in the kidney, heart, liver, and brain tissues were increased. DMFM administration normalized plasma levels of biochemical enzymes. We observed a marked decline in CD4 + STAT3, TNF-α, and COX2 cell populations in whole blood after treatment with DMFM. DMFM downregulated the expression factors related to inflammation at the mRNA and protein levels, i.e., IL-1, TNF-α, iNOS, NF-κB, STAT-3, and COX-2. Dynamic simulations and in silico docking data supports the experimental findings. Our experimental and in silico results illustrated that DMFM may affect protective action against cisplatin-induced brain, heart, liver, and kidney damage via reduction of inflammation and ROS.

Biochemical characterization and structure-based in silico screening of potent inhibitor molecules against the 1 cys peroxiredoxin of bacterioferritin comigratory protein family from Candidatus Liberibacter asiaticus

Bacterioferritin comigratory protein family 1 Cys peroxiredoxin from Candidatus Liberibacter asiaticus (CLaBCP) is an important antioxidant defense protein that participates in the reduction of ROS, free radicals, and peroxides. In the present study, we report the biochemical studies and in silico screening of potent antibacterial molecules against CLaBCP. The CLaBCP showed enzymatic activity with the Km value 54.43, 94.34, 120.6 µM, and Vmax of 59.37, 69.37, 70.0 µM min−1 for H2O2, TBHP, CHP respectively. The residual peroxidase activity of CLaBCP was analyzed at different ranges of pH and temperatures. The CLaBCP showed structural changes and unfolding in the presence of its substrates and guanidinium chloride by CD and fluorescence. The structure-based drug design method was employed to screen and identify the more efficient molecule against CLaBCP. The validated CLaBCP model was used for the virtual screening of potent antibacterial molecules. The docking was performed at CLaBCP active site to evaluate the binding energy of the top five molecules (LAS 34150849, BDE 33184869, LAS 51497689, BDE 33672484, and LAS 34150966). All identified molecule has a higher binding affinity than adenanthin analyzed by molecular docking. Molecular dynamics studies such as RMSD, Rg, SASA, and PCA results showed that the CLaBCP inhibitor(s) complex is more stable than the CLaBCP-adenanthin complex. MMPBSA results suggested that the identified molecule could form a lower energy CLaBCP-inhibiter(s) complex than the CLaBCP-adenanthin complex. The screened molecules may pave the route for the development of potent antibacterial molecules against CLa. Communicated by Ramaswamy H. Sarma

Selective blue-filtering spectacle lens protected primary porcine RPE cells against light emitting diode-induced cell damage.

This study aimed to investigate whether use of a selective-blue-filtering (S-BF) lens can protect cultured primary porcine RPE cells against photo-irradiation. Transmittance of S-BF and UV-filtering (UVF) lenses was characterised spectrophotometrically. RPE cells were exposed to 1700 lux of white (peak λ at 443 and 533 nm; 0.44 mW/cm2) or blue (peak λ at 448 and 523 nm; 0.85 mW/cm2) LED light for 16 h to evaluate the influence of light source on the culture. The effect of the S-BF and UVF ophthalmic lenses on RPE cell cultures under blue light irradiation was then investigated. Cell viability was compared using trypan blue and MTT assays. Intracellular ROS production was detected by a fluorescein probe CM-H2DCFDA. Expression levels of catalase and Prdx3 were analysed by western blot. Trypan blue staining showed blue light caused more cell death than no light (p = 0.001) or white light (p = 0.005). MTT assay supported the hypothesis that exposure to blue light damaged RPE cells more severely than no light (p = 0.002) or white light (p = 0.014). Under blue light, use of the S-BF lens, which blocked 17% more blue light than the UVF lens, resulted in higher cellular viability (S-BF: 93.4±1.4% vs UVF: 90.6±1.4%; p = 0.022; MTT: 1.2-fold; p = 0.029). Blue and white light both significantly increased ROS production. The S-BF lens protected cells, resulting in lower levels of ROS and higher expression of catalase and Prdx3. To conclude, blue LED light exposure resulted in significant cytotoxicity to RPE cells. Partial blockage of blue light by an S-BF lens led to protective effects against retinal phototoxicity, which were mediated by reduction of ROS and increased levels of antioxidant enzymes.

Therapeutic Potential of Isorhamnetin following Acetaminophen-Induced Hepatotoxicity through Targeting NLRP3/NF-κB/Nrf2.

Acetaminophen (APAP)-induced acute liver injury (ALI) is the principal cause of acute liver failure (ALF) in some countries including the United States and with few available treatments. Isorhamnetin is a bioflavonoid that is found in medicinal plants like Hippophae rhamnoides L. and Ginkgo biloba L. with promising potential to regulate inflammatory responses. In this study, we evaluated the possible effect of isorhamnetin in prevention of APAP-induced ALI and analyzed further the involvement of oxidative stress and inflammation-associated factors. Male C57BL/6 mice were given isorhamnetin (25 or 100 mg/kg b.w., p.o.) three times at 48, 24, and 1 h before APAP administration (300 mg/kg b.w., i.p.). Functional indicators of liver injury were measured as well as analysis of oxidative stress- and inflammation-associated indices and liver histopathology was also conducted. Isorhamnetin at the higher dose of 100 mg/kg significantly lowered serum levels of ALT, ALP, and AST in addition to reduction of ROS, TBARS, IL-6, TNFα, NF-kB, NLRP3, caspase 1, and MPO and significantly prevented reduction of GSH, SOD activity, sirtuin 1, and Nrf2. Additionally, isorhamnetin alleviated pathological changes of the liver tissue and suitably reversed NF-kB and Nrf2 immunoreactivity. These findings show protective effect of isorhamnetin against acetaminophen-induced liver injury through reducing oxidative stress, inflammation, and pyroptosis which is attributed to its regulation of NF-kB, Nrf2, NLRP3, and sirtuin 1.