Comparison of acute oxidative stress on rat lung induced by nano and fine-scale,soluble and insoluble metal oxide particles: NiO and TiO2

The aim of the present study is to understand the association between metal ion release from nickel oxide (NiO) nanoparticles and induction of oxidative stress in the lung. NiO nanoparticles have cytotoxic activity through nickel ion release and subsequent oxidative stress. However, the interaction of oxidative stress and nickel ion release in vivo is still unclear. In the present study, we examined the effect of metal ion release on oxidative stress induced by NiO nanoparticles. Additionally, nano and fine TiO₂ particles as insoluble particles were also examined. Rat lung was exposed to NiO and TiO₂ nanoparticles by intratracheal instillation. The NiO nanoparticles released Ni²⁺ in dispersion. Bronchoalveolar lavage fluid (BALF) was collected at 1, 24, 72 h and 1 week after instillation. The lactate dehydrogenase (LDH) and HO-1 levels were elevated at 24 and 72 h after instillation in the animals exposed to the NiO nanoparticles. On the other hand, total hydroxyoctadecadienoic acid (tHODE), which is an oxidative product of linoleic acid, as well as SP-D and α-tochopherol levels were increased at 72 h and 1 week after instillation. Fine NiO particles, and nano and fine TiO₂ particles did not show lung injury or oxidative stress from 1 h to 1 week after instillation. These results suggest that Ni²⁺ release is involved in the induction of oxidative stress by NiO nanoparticles in the lung. Ni²⁺ release from NiO nanoparticles is an important factor inoxidative stress-related toxicity, not only in vitro but also in vivo.     

Profiling of rutin‐mediated alleviation of cadmium‐induced oxidative stress in Zygophyllum fabago

Zygophyllum fabago grows in arid, saline soil, or disturbed sites, such as former industrial or mining areas. This species is able to grow in coarse mineral substrates contaminated with heavy metals. To investigate the effects of the flavonoid rutin (Rtn) on certain heavy metal stress responses such as antioxidant defense systems and water status, seedlings were subjected to 100 and 200 μM CdCl₂ treatment without or with 0.25 and 1 mM Rtn for 7 and 14 d (days). Cd stress decreased growth (RGR), water content (RWC), leaf osmotic potential (Ψ_Π), and chlorophyll fluorescence, all of which could be partly alleviated by addition of Rtn. Activities of superoxide dismutase, peroxidase (POX), ascorbate peroxidase, and glutathione reductase increased within the first 7 d after exposure to Cd. However, failure of antioxidant defense in the scavenging of reactive oxygen species (ROS) was evidenced by an abnormal rise in superoxide anion radical ( ${\mathrm{O}}_2^{??}$) and hydrogen peroxide contents and a decline in hydroxyl radical (OH^?) scavenging activity, resulting in enhancement of lipid peroxidation (TBARS) as a marker of Cd‐induced oxidative stress. However, exogenously applied Rtn considerably improved the stress tolerance of plants via a reduction in Cd accumulation, modulation of POX activity, increase of proline (Pro) content, decrease in TBARS and ROS content and consequent lowering of oxidative damage of membrane. Overall, 0.25 and 1 mM Rtn could protect Z. fabago from the harmful effects of 100 μM Cd‐induced oxidative stress throughout the experiment. © 2014 Wiley Periodicals, Inc. Environ Toxicol 30: 816–835, 2015.     

氧化应激致心室重构的研究进展

心室重构意味着血管壁中氧化应激水平的提高,当机体受到有害刺激时,氧化和抗氧化之间失衡,导致大量活性氧簇（ ROS）的产生,加速心室重构的进展。近年来,新型抗氧化剂成为治疗或预防心室重构的靶点,观察抗氧化剂治疗在心室重构患者的长期应用是当下趋势。该文就氧化应激与心室重构之间的关系及相关药物治疗进行综述。     

Effect of dietary antioxidant on mercuric chloride induced lung toxicity and oxidative stress

This study was conducted to evaluate the effect of environmental contaminant mercuric chloride on levels of trace elements and oxidative parameters in rat lungs and to investigate the efficacy of possible protection by natural antioxidant diallylsulphide (DAS) against lung injury. Twenty-four healthy male rats were randomly divided into four groups: I – control, II – DAS (200?mg/kg), III – HgCl₂ (50?mg/kg), and IV – DAS (200?mg/kg) + HgCl₂ (50?mg/kg). Mercuric chloride induced oxidative stress was indicated by a significant decrease in levels of superoxide dismutase, catalase, and glutathione peroxidase as compared to the control group (p–<0.05). Also, hydroxyproline (HYP) content in lung tissues of mercuric chloride-treated group was significantly increased (p?<?0.05). DAS markedly attenuated mercuric-induced biochemical alterations in lungs by upregulating the activities of antioxidant enzymes. These findings indicated that within the doses selected, DAS can provide significant protection against HgCl₂-induced toxicity.     

氧化应激在糖尿病视网膜病发病机制中的研究进展

糖尿病视网膜病是糖尿病最常见和严重的微血管并发症之一。近年来大量研究表明,氧化应激和糖尿病视网膜病变的发生发展有一定的相关性。高血糖引起体内氧化应激反应,氧化应激又通过损害内皮细胞、线粒体,生成异常代谢通路,促进炎性反应等加重对视网膜组织的损害,造成视网膜缺血、甚至脱离等严重后果;而抗氧化治疗在临床上也取得了一定的成效。本文主要围绕氧化应激和糖尿病视网膜病的发病关系展开综述。     

Oxidative damage mediated iNOS and UCP-2 upregulation in rat brain after sub-acute cyanide exposure: dose and time-dependent effects

Cyanide-induced chemical hypoxia is responsible for pronounced oxidative damage in the central nervous system. The disruption of mitochondrial oxidative metabolism has been associated with upregulation of uncoupling proteins (UCPs). The present study addresses the dose- and time-dependent effect of sub-acute cyanide exposure on various non-enzymatic and enzymatic oxidative stress markers and their correlation with inducible-nitric oxide synthase (iNOS) and uncoupling protein-2 (UCP-2) expression. Animals received (oral) triple distilled water (vehicle control), 0.25 LD50 potassium cyanide (KCN) or 0.50 LD50 KCN daily for 21 d. Animals were sacrificed on 7, 14 and 21 d post-exposure to measure serum cyanide and nitrite, and brain malondialdehyde (MDA), reduced glutathione (GSH), glutathione disulfide (GSSG), cytochrome c oxidase (CCO), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR) and catalase (CA) levels, together with iNOS and UCP-2 expression, and DNA damage. The study revealed that a dose- and time-dependent increase in cyanide concentration was accompanied by corresponding CCO inhibition and elevated MDA levels. Decrease in GSH levels was not followed by reciprocal change in GSSG levels. Diminution of SOD, GPx, GR and CA activity was congruent with elevated nitrite levels and upregulation of iNOS and UCP-2 expression, without any DNA damage. It was concluded that long-term cyanide exposure caused oxidative stress, accompanied by upregulation of iNOS. The upregulation of UCP-2 further sensitized the cells to cyanide and accentuated the oxidative stress, which was independent of DNA damage.     

Long-term exposure to repetitive hyperbaric oxygen results in cumulative oxidative stress in rat lung tissue

Context: Despite its known benefits, hyperbaric oxygen (HBO) is also reported to enhance the production of reactive oxygen species and can cause oxidative stress in several tissues. Previous studies had shown that HBO-induced oxidative stress is directly proportional to both its exposure pressure and duration. Nevertheless, these studies were usually performed with single-session HBO exposure but its clinical use commonly depends on long-term exposure periods.

Objective: To clarify the oxidative effect of long-term repetitive HBO in the lung tissue of rats.

Materials and methods: Male Sprague-Dawley rats were divided into six study groups exposed to consecutive HBO sessions (2.8 atm/90?min) for 5, 10, 15, 20, 30, and 40 days. Animals were sacrificed 24?h after the last HBO session. An additional control group was set to obtain normal data. Lung malondialdehyde (MDA) and carbonylated protein (PCC) levels were determined as measures of oxidative stress along with the activities of the antioxidant enzymes superoxide dismutase (SOD) and glutathione peroxidase.

Results: None of the measured parameters showed any changes among the groups exposed to 5–15 HBO sessions. However, MDA, PCC, and SOD were found to be significantly increased in the 20 to 40 session groups.

Discussion and conclusion: These results indicate that repetitive treatment with HBO may cause oxidative stress in critical tissues including the lung. Although HBO-mediated free radicals are accepted to be responsible for the benefits of this therapeutic modality, especially in cases with prolonged exposure, possible injurious effects of supranormal values of bio-oxidative products need to be considered.     

Analysis of neuroglobin mRNA expression in rat brain due to arsenite-induced oxidative stress

Arsenic (As) in drinking water is a toxicant causing several health problems including nervous system disturbance. Neuroglobin (Ngb) is a tissue globin in nervous system playing protective role against oxidative stress in many injuries. This study was to investigate how long arsenite exposure (sodium arsenite 7.5 mg/kg/day) could induce oxidative stress in blood and brain of rats and to determine whether Ngb expression in rat brain changed due to oxidative stress. Results showed that superoxide dismutase (SOD) activity and malondialdehyde (MDA) level in serum and brain homogenates and reactive oxygen species (ROS) generation in red blood cells (RBCs) did not change in the rats exposed to arsenite for 8 weeks. In the rats exposed to arsenite for 16 weeks, SOD activity decreased (serum: P < 0.05; brain homogenates: P < 0.01) and MDA level increased (P < 0.01) in serum and brain homogenates; ROS production increased (P < 0.01) in RBC. When oxidative stress occurred, Ngb mRNA expression did not change in whole brain, cerebral cortex, midbrain, and hippocampus; however, Ngb mRNA expression increased significantly (P < 0.05) in cerebellum compared to the control group. This study suggests that arsenite exposure for 16 weeks can lead to oxidative stress of blood and brain of rats. Ngb may play a protective role incerebellum when oxidative stress occurs due to arsenite exposure. © 2010 Wiley Periodicals, Inc. Environ Toxicol, 2012.     

Isoniazid-induced apoptosis in HepG2 cells: Generation of oxidative stress and Bcl-2 down-regulation

Isoniazid (INH) is a first-line antibiotic used in the treatment of infections caused by Mycobacterium tuberculosis. However it has a serious limitation of being hepatotoxic. Delineating the mechanism underlying INH-induced hepatotoxicity may be beneficial in devising ways to counteract its toxic manifestations. Studies in human hepatoma HepG2 cells have indicated that INH exposure causes induction of apoptosis. This study was aimed at identifying the key components/pathways of the INH-induced apoptotic pathway using HepG2 cells. HepG2 cells were exposed to increasing concentrations of INH (6.5, 13, 26, and 52?mM). Hydrogen peroxide (0.3?mM) served as positive control. After incubating for specific time intervals cells were harvested and evidences of cytotoxicity, oxidative stress, and apoptosis were sought. The findings indicated that INH exposure causes increased ROS generation along with alteration in levels of enzymatic antioxidants such as Superoxide dismutase, Catalase, and Glucose-6-Phosphate dehydrogenase. Altered Bcl-2/Bax content, cytochrome-c translocation, caspase activation, and DNA fragmentation emphasized involvement of apoptosis.     

Comparative study of cytotoxicity, oxidative stress and genotoxicity induced by four typical nanomaterials: the role of particle size, shape and composition

Although the biological effects of some nanomaterials have already been assessed, information on toxicity and possible mechanisms of various particle types are insufficient. Moreover, the role of particle properties in the toxic reaction remains to be fully understood. In this paper, we aimed to explore the interrelationship between particle size, shape, chemical composition and toxicological effects of four typical nanomaterials with comparable properties: carbon black (CB), single wall carbon nanotube, silicon dioxide (SiO(2)) and zinc dioxide (ZnO) nanoparticles. We investigated the cytotoxicity, genotoxicity and oxidative effects of particles on primary mouse embryo fibroblast cells. As observed in the methyl thiazolyl tetrazolium (MTT) and water-soluble tetrazolium (WST) assays, ZnO induced much greater cytotoxicity than non-metal nanoparticles. This was significantly in accordance with intracellular oxidative stress levels measured by glutathione depletion, malondialdehyde production, superoxide dismutase inhibition as well as reactive oxygen species generation. The results indicated that oxidative stress may be a key route in inducing the cytotoxicity of nanoparticles. Compared with ZnO nanoparticles, carbon nanotubes were moderately cytotoxic but induced more DNA damage determined by the comet assay. CB and SiO(2) seemed to be less effective. The comparative analysis demonstrated that particle composition probably played a primary role in the cytotoxic effects of different nanoparticles. However, the potential genotoxicity might be mostly attributed to particle shape.     

Oxidative stress and lung pathology following geogenic dust exposure

This study was designed to evaluate markers of systemic oxidative stress and lung histopathology following subacute exposure to geogenic dust with varying heavy metal content collected from a natural setting prone to wind erosion and used heavily for off‐road vehicle recreation. Adult female B6C3F1 mice were exposed to several concentrations of dust collected from seven different types of surfaces at the Nellis Dunes Recreation Area in Clark County, Nevada, designated here as CBN 1‐7. Dust representing each of the seven surface types, with an average median diameter of 4.2 μm, was selected and administered via oropharyngeal aspiration to mice at concentrations from 0.01 to 100 mg of dust kg^–1 of body weight. Exposures were given four times spaced a week apart over a 28 day period to mimic a month of weekend exposures. Lung pathology was evaluated while plasma markers of oxidative stress included levels of reactive oxygen and nitrogen species, superoxide dismutase, total antioxidant capacity and total glutathione. Overall, results of these assays to evaluate markers of oxidative stress indicate that no single CBN surface type was able to consistently induce markers of systemic oxidative stress at a particular dose or in a dose–response manner. All surface types were able to induce some level of lung inflammation, typically at the highest exposure levels. These data suggest that dust from the Nellis Dunes Recreation Area may present a potential health risk, but additional studies are necessary to characterize the full extent of health risks to humans. Copyright © 2016 John Wiley & Sons, Ltd.     

Opportunities for future therapeutic interventions for hyperoxaluria: targeting oxidative stress

Introduction: Oxalate is a toxic byproduct of metabolism and is normally produced in quantities easily removed from the body. However, under specific circumstances oxalate production is increased resulting in deposition of calcium oxalate (CaOx) crystals in the kidneys as well as other organs causing inflammation and injury. Excessive buildup of crystal deposits in the kidneys causes eventual loss of renal function requiring renal transplantation.

Areas covered: Cellular exposure to CaOx crystals induces the production of reactive oxygen species (ROS) with the involvement of renin-angiotensin aldosterone system (RAAS), mitochondria, and NADPH oxidase. Inflammasomes are activated and pro-inflammatory cytokines, such as IL-1β and IL-18 are produced. We reviewed results of experimental and clinical studies of crystal renal epithelial cell interactions with emphasis on cellular injury and ROS production.

Expert opinion: Treatment should depend upon the level of hyperoxaluria and whether it is associated with CaOx crystal deposition. Persistent low grade or intermittent hyperoxaluria can be treated with antioxidants, free radical scavengers. Hyperoxaluria associated with CaOx crystal deposition will require administration of angiotensin II receptor blockers, and NADPH oxidase or NLRP3 inflammasome inhibitors. DASH-style diet will be beneficial in both cases.     

Effect of Centella asiatica on arsenic induced oxidative stress and metal distribution in rats

Concomitant oral supplementation of Centella asiatica (100, 200 or 300 mg kg(-1), orally once daily) during arsenic exposure (20 ppm in drinking water for 4 weeks) was investigated in rats for its protective value. The animals exposed to arsenic (III) showed a significant inhibition of delta-aminolevulinic acid dehydratase (ALAD) activity, a marginal decrease in glutathione (GSH) and an increase in zinc protoporphyrin (ZPP) level in blood. Hepatic and renal glutathione (GSH) decreased, while oxidized glutathione (GSSG) and thiobarbituric acid reactive substance (TBARS) levels increased significantly in the liver, kidney and brain. The activities of brain superoxide dismutase (SOD) and catalase decreased marginally on arsenic exposure. Concomitant administration of Centella asiatica showed a significant protective action on inhibited blood ALAD activity and restored the blood GSH level, whereas most of the other blood biochemical parameters remained unchanged on Centella asiatica supplementation. Interestingly, most of the hepatic biochemical variables indicative of oxidative stress showed protection. There was, however, a significant protection observed in the altered kidney GSSG level and hepatic and brain TBARS. Only a marginal beneficial effect of Centella asiatica on blood and liver arsenic concentration was noted, particularly at the highest dose studies (300 mg kg(-1)). No effect of Centella asiatica on most of the altered renal biochemical parameters was noted. The results thus lead to the conclusion that simultaneous supplementation of Centella asiatica significantly protects against arsenic-induced oxidative stress but does not influence the arsenic concentration in these organs. It can thus be suggested that co-administration of Centella asiatica protects animals from arsenic-induced oxidative stress but exhibits no chelating property. Further studies are recommended for determining the effect of co-administration of Centella asiatica during chelation therapy with a thiol chelator.     

Evaluation of oxidative stress induction in rats following exposure to silver nanorods

The study investigated the oxidative stress induction by the 10 and 25?nm silver nanorods (SNRs) following intra-tracheal instillation in rats after 1?day, 1 week, 1 month and 3 months post instillation periods at 1 and 5?mg/kg b.w. doses. The blood was withdrawn by retro orbital plexus method after exposure periods and different oxidative stress markers were estimated. The results showed that the both sizes of SNRs induced increased levels of malondialdehyde (MDA) and depleted glutathione (GSH) levels after 1?day and 1 week post exposure periods. The 10 and 25?nm SNRs at both doses displayed that significantly reduced levels of superoxide dismutase (SOD) and catalase following 1?day and 1 week post exposure periods. Also, the results have shown that decrease in total antioxidant capacity (TAC) of both sizes of SNRs significantly following 1?day and 1 week post exposure periods, indicating the oxidative stress induction by SNRs. In spite, there were no significant changes in oxidative stress markers following 1 month and 3 months post exposure periods may be due to recovery. The increased levels of MDA and decreased levels of GSH, SOD, catalase and TAC activity are strongly associated to ROS production and lipid peroxidation, suggesting the induction of oxidative stress in rats. The 10?nm SNRs at 5?mg/kg b.w. dose exposures in rats have shown greater changes in all oxidative stress parameters, indicating the greater induction of oxidative stress when compared with the 25?nm SNRs, representing the size–dose-dependent induction of oxidative stress of SNRs.     

Methamidophos induces cytotoxicity and oxidative stress in human peripheral blood mononuclear cells

Previous studies have shown that organophosphate pesticide (OP) exposure is associated with oxidative stress. Methamidophos (MET) is an OP widely used in agriculture, which is regarded as a highly toxic pesticide and it is a potent inhibitor of acetylcholinesterase. The aim of this study was to evaluate whether MET can induce oxidative stress at low concentrations in primary cultures of human peripheral blood mononuclear cells (PBMCs). PBMCs from healthy individuals were exposed to MET (0–80 mg/L) for 0–72 h. We performed the MTT and neutral‐red assays to assess the cytotoxicity. As indicators of oxidative stress, the levels of reactive oxygen species (ROS) were assessed using flow cytometry, and the malondialdehyde (MDA) and reduced glutathione (GSH) levels were determined. MET decreased the viability of PBMCs in a dose‐dependent manner. At concentrations of 3, 10, or 20 mg/L for 24 h, MET increased the ROS production significantly compared with the vehicle control. Similarly, MET increased the levels of MDA at the same concentrations that increased ROS (10 and 20 mg/L); however, no changes in GSH levels were observed. These results suggest that MET increased the generation of oxidative stress in PBMCs. © 2015 Wiley Periodicals, Inc. Environ Toxicol 32: 147–155, 2017.     

Cytotoxicity,oxidative stress,and inflammation in human Hep G2 liver epithelial cells following exposure to gold nanorods

The gold nanorods (GNRs) are great potentials in imaging, therapy, biosensing, and many other commercial applications. However, GNRs interactions with human cells and potential health risks remain not well known. The present investigation aimed to evaluate the in vitro toxicity of 10 and 25?nm GNRs (10–50?μg/mL) following exposure for 48?h in human Hep G2 liver epithelial cells using 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), lactate dehydrogenase (LDH) leakage, glutathione (GSH) estimation, lipid peroxidation (TBARS), caspase-3 levels, and interleukin-8 (IL-8) release assays. Exposure of GNRs to cells results in decrease in cell viability and causes cell membrane damage through LDH leakage results in cytotoxicity. The IC₅₀ (concentration required to inhibit 50% of cells) values of 10?nm GNRs, 25?nm GNRs, and quartz (toxic control)-treated cells were found to be 19.9, 26.8, and 36.35?μg/mL, suggesting the higher cytotoxicity of GNRs. The GNRs exposure to liver cells found in depleted GSH levels, increased lipid peroxidation, and increased caspase-3 levels leads to induction of oxidative stress. In addition, enhanced levels of IL-8 were found, a sign of inflammation. The 10?nm GNRs have shown significant toxicity against all biochemical assays when compare to 25?nm GNRs and quartz-treated cells. Finally, the data indicate that the concentration size-dependent in vitro toxicity of GNRs toward liver Hep G2 cells. The toxicity of GNRs may be due to cell membrane damage, induction of oxidative stress, and inflammatory mediator release. Further investigations are necessitated to elucidate the in vivo toxicity of GNRs.     

咯利普兰对H_2O_2致PC12细胞氧化应激损伤的保护作用

目的探讨PDE4抑制剂咯利普兰(rolipram)对H2O2致PC12细胞氧化应激损伤的保护作用及其作用机制。方法以H2O2损伤PC12细胞为氧化应激损伤模型,MTT法检测细胞存活率;碘化丙啶(Propidium iodide,PI)染色流式细胞术检测细胞周期变化;化学比色法测定细胞清除羟自由基(·OH)和超氧阴离子自由基(O2·)的能力;以及培养上清液中的丙二醛(MDA)、谷胱甘肽(GSH)和一氧化氮(NO)含量、超氧化物歧化酶(SOD)活性。Western blot和Real-time RT-PCR检测细胞内硫氧还蛋白(Trx)和诱导型一氧化氮合酶(iNOS)的表达。结果咯利普兰能明显提高H2O2损伤的PC12细胞存活率,恢复细胞增殖;提高细胞清除·OH和O2·的能力;降低MDA和NO含量,提高GSH含量和SOD活性;使Trx蛋白和mRNA表达增加,同时下调iNOS蛋白和mRNA表达。结论咯利普兰对H2O2致PC12细胞损伤具有保护作用,其作用机制可能与提高PC12细胞的抗氧化能力相关。     

Effects of occupational exposure to trace levels of halogenated anesthetics on the liver,kidney, and oxidative stress parameters in operating room personnel

Abstract

This study was conducted to measure the concentration of halogenated anesthetics in breathing zone air of the personnel and evaluate their effects on hepatic and renal functions as well as oxidative stress parameters. Levels of these gases in air samples were lower than the national recommended exposure limits (50?ppm). Occupational exposure to anesthetic gases significantly induced oxidative stress in the operating room personnel, marked by increased plasma levels of transaminase enzymes accompanied by the marked decline of enzymatic and non-enzymatic antioxidant levels, and the rise of MDA levels (p?<?0.05); these adverse effects would increase with increasing work experience.     

Assessment of oxidative stress induced by gold nanorods following intra-tracheal instillation in rats

Gold nanorods (GNRs) are used for their wide variety of applications in various industries. There is a little availability of data related to toxicity and ecological implications of these GNRs. The study evaluated the oxidative stress induction following intra-tracheal instillation of 1 and 5?mg/kg b.w. doses of 10 and 25?nm GNRs by estimating various oxidative stress markers including lipid peroxidation (malondialdehyde; MDA), glutathione (GSH), superoxide dismutase (SOD), catalase and total antioxidant capacity (TAC) after 1?day, 1?week, 1?month, and 3?months post exposure periods. The results have shown increased MDA levels and decreased GSH levels following 1?day and 1?week post exposure periods, indicating induction of oxidative stress. Also, the SOD, catalase and TAC levels were significantly decreased following exposure of both 10 and 25?nm GNRs after 1?day and 1?week after exposures, indicating the inhibition of antioxidant defense mechanisms. Moreover, the 10?nm GNRs at 5?mg/kg dose displayed greater changes in all the estimated parameters, representing dose and size based induction of oxidative stress by GNRs. In contrast, a little change was observed during 1?month and 3?months post exposure periods, may be due to recovery. Finally, the GNRs induced dose-size-dependent oxidative stress induction by various oxidative stress markers following intra-tracheal instillation in rats.     

Anti-apoptotic and cytoprotective effect of Enicostemma littorale against oxidative stress in Islets of Langerhans

Context Oxidative stress induces apoptosis within Islets of Langerhans in diabetes mellitus (DM). Enicostemma littorale blume, herb of the Gentianaceae family is used as an anti-diabetic agent across rural India.

Objective This report demonstrates potent anti-apoptotic and cyto-protective activity of Enicostemma littorale MeOH extract (EL MeOH ext.) against 50?μM H₂O₂ in isolated rat Islets.

Materials and methods In this study, the whole plant methanolic extract of EL with doses 0.25–4?mg/mL each for the preincubation duration of 0.5–4?h against 50?μM H₂O₂ were tested for optimum protective dose and time by Trypan blue dye exclusion assay. Islet intracellular reactive oxygen species (ROS) was quantified by DCFDA staining and cell death using PS/PI & FDA/PI staining. Further, comet assay, biochemical assessment of caspase-3 and antioxidant enzyme activities along with immunoblotting of PARP-1, caspase-3, TNF-α activation and p-P38 MapK (stress kinase) induction was performed.

Results The optimized dose of EL MeOH ext. 2?mg/mL for 2?h was used throughout the study, which significantly decreased total Intracellular ROS and cell death. Further, caspase-3 activity, PARP-1 cleavage, p-P38 MapK (stress kinase) activation and TNF-α levels, which had been significantly elevated, were normalized. Antioxidant enzymes like catalase, superoxide dismutase, reduced glutathione and glutathione peroxidase, along with Comet assay, demonstrated that pretreatment with EL MeOH ext. can augment antioxidant enzyme activities and protect from DNA damage.

Discussion and conclusions Significant anti-apoptotic and cyto-protective effects were mediated by EL with Islets of Langerhans subjected to oxidative stress-induced cell death.