Subcutaneous Injection Of Sodium Selenite Research Articles

Protective Effects of Safranal Against Selenite-Induced Cataract in Rats

Cataract, which is the opacity inside clear ocular lens of eye, result in the scattering of visible light as it passes via the lens and consequently deterioration in optical image. The aim of the present study was to investigate whether safranal, an active constituent of Crocus sativus L. stigmas, has a protective effect on the cataract in the rat's pups. The animals were randomly divided into five groups, each of which consisted of 7 rat pups. Group I served as normal control (vehicle administration). For testing cataract induction, animals of Groups II, III, and IV were administered a single subcutaneous injection of sodium selenite on postpartum day 12. After sodium selenite intoxication, Group II served as control selenite, Groups III-IV received intraperitoneal safranal at doses of 200, and 300 mg/kg, respectively from the 11th day through the 17th day, while group V receive only safranal (300 mg/kg). On postpartum day 30, the rat pups were examined for cataract formation, and the lenses were isolated for further analysis. This study found that selenite caused significant (p < 0.05) cataract formation. Through the effects of selenite on the level of lipid peroxidation (MDA) which was upregulated. Furthermore, the antioxidant enzymes levels GSH levels and NRF2 protein were downregulated. In contrast, treatment with safranal could significantly (p < 0.05) ameliorate cataract formation and oxidative damage in the lens. Moreover, safranl administration significantly increased the protein expressions of Nrf2 and the GSH level, in addition to reducing the level both the MDA and the level soluble proteins in the lens. Taken together, safranal is a prospective anti-cataract agent that probably delays the onset and progression of cataracts induced by sodium selenite.

Protective Effects of Rosmarinic Acid against Selenite-Induced Cataract and Oxidative Damage in Rats.

Cataracts are the major cause of blindness and are associated with oxidative damage of the lens. In the present study, the aim was to evaluate the protective effects of rosmarinic acid on selenite-induced cataractogenesis in Sprague-Dawley rat pups. The animals were randomly divided into five groups, each of which consisted of 10 rat pups. Group I served as normal control (vehicle administration). For testing cataract induction, animals of Groups II, III, IV, and V were administered a single subcutaneous injection of sodium selenite (2.46 mg/kg body weight) on postpartum day 12. After sodium selenite intoxication, Group II served as control selenite. From the 11th day through the 17th day, Groups III-V received rosmarinic acid intraperitoneally at doses of 5, 10, and 50 mg/kg, respectively. On postpartum day 24, the rat pups were examined for cataract formation, and the lenses were isolated for further analysis of proteins and oxidative damage indicators. Selenite caused significant (p < 0.05) cataract formation. Through the effects of selenite, the protein expressions of filensin and calpain 2 were reduced, and the calcium concentrations, the level of lipid peroxidation (TBARS), and inflammation indicators (iNOS, COX-2, and NFκB) were upregulated. Furthermore, the protein expression of the antioxidant status (Nrf2, SOD, HO-1, and NQO1), the antioxidant enzymes activities (GSH-Px, GSH-Rd, and catalase), and the GSH levels were downregulated. In contrast, treatment with rosmarinic acid could significantly (p < 0.05) ameliorate cataract formation and oxidative damage in the lens. Moreover, rosmarinic acid administration significantly increased the protein expressions of filensin, calpain 2, Nrf2, SOD, HO-1, and NQO1, the antioxidant enzymes activities, and the GSH level, in addition to reducing the calcium, lipid peroxidation, and inflammation indicators in the lens. Taken together, rosmarinic acid is a prospective anti-cataract agent that probably delays the onset and progression of cataracts induced by sodium selenite.

Interaction of Coenzyme Q10 with Liposomes and its Impact on Suppression of Selenite - Induced Experimental Cataract.

Purpose: To stress the influence of Coenzyme Q10 (CoQ10) on the structural properties of liposomes as model membranes and to investigate the possible role of CoQ10 or CoQ10 doped in liposomes when topically instilled as eye drops, in preventing cataract.Methods: The molecular interaction between liposomes and Coenzyme Q10 was examined using differential scanning calorimetry (DSC) and Fourier transform infrared spectroscopy (FTIR). Rat pups were randomly divided into six groups comprising 15 pups. Group (1), control group. Group (2), untreated model of cataract, received a single subcutaneous injection of sodium selenite. Instillation of pure CoQ10 (Group 3), CoQ10 encapsulated into neutral (Group 4), positive (Group 5) and negative (Group 6) Dipalmitoyl phosphatidylcholine (DPPC) liposomes on the opacification of lenses in rat pups after sodium selenite injection was topically received.Results: The incorporated CoQ10 is probably associated with lipid bilayers where it interacts to a large extent and perturbs them. This results in strong broadening and shift to lower temperature (94°C) of the major characteristic endothermic peak of pure DPPC at 105°C. FTIR showed that the incorporation of CoQ10 into DPPC induces a conformational change in the polar region of DPPC. Ophthalmological and Biochemical studies revealed that CoQ10 alone followed by negatively charged liposomes doped with CoQ10 are more effective in reducing the progress of cataract as well as improving the lens soluble proteins levels and total antioxidant capacity.Conclusion: The interactions of CoQ10 with membrane systems may contribute to a better understanding of CoQ10 physiological properties and the development of therapeutically advanced systems.

Protective effects of Cassia tora leaves in experimental cataract by modulating intracellular communication, membrane co-transporters, energy metabolism and the ubiquitin-proteasome pathway

Context: Cataract is the clouding of eye lens which causes impairment in vision and accounts for the leading factor of global blindness. Functional food-based prevention of cataract finds application in vision research because of its availability and easy access to all classes of the society. Cassia tora Linn. (Caesalpinaceae) is an edible plant mentioned in the traditional systems of medicine for whole body health, especially to the eyes.Objective: The present study evaluates the potential of ethyl acetate fraction of Cassia tora leaves (ECT) on experimental cataract.Materials and methods: Cataract was induced by a single subcutaneous injection of sodium selenite (4 μg/g body weight) on 10th day. ECT was supplemented orally from 8th day up to 12th day at a concentration of 5 μg/g body weight and marker parameters were evaluated after 30 days.Results: The production of MPO and the activation of calpain were reduced 52.17% and 36.67% by ECT in lens tissue, respectively. It modulated the energy status by significantly increasing the activity of CCO 1 (55.56%) and ATP production (41.88%). ECT maintained the ionic balance in the lens by reducing the level of sodium (50%) and increasing the level of potassium (42.5%). It also reduced cell junction modifications and preserved a functional ubiquitin-proteasome pathway.Discussion and conclusion: The results reinforce the growing attention on wild plant food resources for preventive protection against cataract. The data suggest the value of Cassia tora leaves as a functional food for ameliorating cataract pathology.

Lupeol, a pentacyclic triterpenoid isolated from Vernonia cinerea attenuate selenite induced cataract formation in Sprague Dawley rat pups

This study investigated the inhibitory effects of active component isolated from flavonoid fraction of Vernonia cinerea (FVC), lupeol on selenite induced cataract formation. Previous reports suggest that phytochemicals or natural plant products retard the process of cataractogenesis by scavenging free oxygen radicals. Hence, the present study sought to assess the potential of lupeol on in vivo selenite induced cataract models. Lupeol, a pentacyclic triterpenoid, was isolated from the ethyl acetate fraction of methanolic extract of Vernonia cinerea, follows standard chromatographic techniques. Structural elucidation of the compound was carried out using 1H NMR, 13C NMR, Mass spectrometry together with other complementary techniques (UV and IR). From these, the isolated compound was identified as Lupeol (3′-hydroxylup-20(29)-ene). The antioxidant activity was comparatively studied using DPPH radical scavenging and FRAP assay. Lupeol exhibited higher DPPH radical scavenging activity as well as reducing power assay. In this study, cataract was induced by a single subcutaneous injection of sodium selenite (4 μg/g body weight) on rat pups. Lupeol was administered orally from 8th day upto 21st day at a concentration 25 μg/g body weight. Cataract was visualized on 16th day with the help of an ophthalmoscope and later on with the naked eye. On the 30th day, rats were euthanized by sodium pentothal injection, lenses were excised and the biochemical parameters such as activity of superoxide dismutase (SOD), catalase (CAT), Glutathione peroxidase (GPx), Glutathione reductase (GR), Glutathione-S-transferase (GST), Ca2+ ATPase, glutathione content (GSH), reactive oxygen species (ROS), lipid peroxidation products (malondialdehyde) were estimated and found effective in the treatment of cataract by lupeol.

Cassia tora leaves modulates selenite cataract by enhancing antioxidant status and preventing cytoskeletal protein loss in lenses of Sprague Dawley rat pups

Cataract is the clouding or opacity that develops in the eye's lens and is considered to be an unavoidable consequence of aging due to irreversible lens damage. Free radicals and oxidant species are reported to be the major factor responsible for the onset and pathology of cataract. No pharmacological measures are formulated to treat cataract blindness and surgical removal of the opaque lens is the only remedy till date. Boosting of antioxidant potential of the lens is proved to prevent cataract and many indigenous plants have been screened for anticataractogenic potential in the last decades. The objective of the present study was to determine whether Cassia tora leaves; the plant employed in traditional medicine for eye rejuvenation and ailments, can prevent cataract in neonatal rats. Cataract was induced by a single subcutaneous injection of sodium selenite at a dose of 4 μg/g body weight on the 10th day and Cassia tora leaves was administered orally from 8th day upto 12th day at a concentration of 5 μg/g body weight. After 30 days; lens morphology, oxidant-antioxidant equilibrium, glutathione metabolism, cytoskeletal protein/gene expressions were monitored. Lens morphology, biochemical analysis and expression studies supported the anticataractogenic effect of Cassia tora leaves. In summary, it can be suggested that the consumption of these leaves afford protection to the lens with its antioxidant action and seems to be a new therapeutic approach against cataract by preventive protection.

Luteolin Supplementation Prevents Selenite-Induced Cataractogenesis in Sprague Dawley Rat Pups.

Luteolin, a flavonoid present in leaves and stems of many plants finds mention in literature for beneficial effects on eyes. Presently, no reports are available on the in vivo anticataractogenic effect of luteolin. The current study was designed to evaluate the efficacy of luteolin on selenite-induced cataract models in vivo. The study consisted of three groups of Sprague Dawley rat pups 8-10 d old (Group I (Normal), Group II (Cataract induced), and Group III (Treatment)). Cataract was induced in Group II and Group III by a subcutaneous injection of sodium selenite (4 μg/g body weight) on the 10th day. Luteolin was administered orally from 8th day up to 12th day at a concentration of 1 μg/g body weight in Group III. After 30 d, lenses of treated animals showed normal morphology. Activities of antioxidant enzymes were increased and levels of reactive oxygen species were decreased in the luteolin-treated group when compared to the cataract-induced group. Increased Ca(2+) ATPase activity and lowered calcium level, caspase 3 activity and down-regulation of caspase 3 expression were seen in the treatment group when compared to the selenite group. Luteolin enhances the antioxidant potential and thereby lowers the oxidative damages to the lens. It also stabilizes the membrane integrity of the lens and maintains the ionic balance.

Prevention of Selenite-Induced Cataratogenesis by Ginkgo biloba Extract (Egb761) in Wistar Rats

Objective: To investigate whether Ginkgo biloba extract retards selenite-induced cataractogenesis in Wistar rat pups.Methods: On postpartum day eight, Group I rat pups (n = 12) received an intraperitoneal injection of physiological saline. Groups II and III rat pups (n = 12) received a subcutaneous injection of sodium selenite. Group III also received an intraperitoneal injection of G. biloba extract once daily on postpartum days 9–14. Both eyes of each pup were examined from postpartum day 16 up to day 30. After sacrifice, encapsulated pup lens were analyzed for mean activities of catalase, superoxide dismutase, glutathione peroxidase (GPx), glutathione S-transferase and glutathione reductase. In addition, the mean concentrations of reduced glutathione (GSH) and malondialdehyde were analyzed in samples of lens and serum.Results: Dense lenticular opacification occurs 100% in Group II, but only minimal opacification occurs in three pups of Group III (25%), no opacification in 75% of Group III, none in Group I. Compared with Groups I and III, Group II rat showed lower lenticular antioxidant enzyme activity, lower level of GSH, and higher level of malondidehyde (mean ± standard deviation SD, p < 0.05 respectively).Conclusions: The treatment with G. biloba extract is effective against oxidative stress – a crucial factor of cataractogenesis in rat pups, possibly by preventing depletion of antioxidant enzymes and by inhibiting lipid peroxidation.

Ameliorative effect of acetyl-l-carnitine and/or nifedipine against selenite-induced cataractogenesis in young albino rats

Free radical toxicity and calcium ion overload have been identified as the major two players in the causation of cataract. The current study was carried out to investigate the anti-cataractogenic effect of single and combined treatment with acetyl-l-carnitine and nifedipine in sodium selenite-induced cataract. Rat pups were divided into 5 groups; 1st group received intraperitoneal injection (i.p.) of saline and served as normal control, 2nd group received single subcutaneous injection of sodium selenite 30nmol/g body weight on p10 (postpartum day 10), 3rd and 4th groups received either acetyl-l-carnitine (200mg/kg, i.p.) or nifedipine (0.1mg/kg, i.p.) on p9, respectively, before the administration of sodium selenite, and the treatment continued till p14. Last group received the combined treatments of acetyl-l-carnitine and nifedipine in the same regimen. All animals were examined using a slit lamp and retroillumination then sacrificed on p30. Lenses were removed and processed for biochemical analyses, histopathological and electron microscopic examination. Selenite-treated groups showed significantly (P≤0.05) lower values of redox system components (glutathione and glutathione reductase activity) and anti-oxidant enzymes׳ activities (superoxide dismutase and catalase) along with increased lipid peroxidation that was accompanied by 100% opacified crystalline lenses (mature cataract) with abnormal structure as detected by electron microscopy. It is concluded that acetyl-l-carnitine or nifedipine was able to partially protect against selenite-induced abnormalities. While, combined treatment with acetyl-l-carnitine and nifedipine was superior to individual treatments in slowing down the development of cataract by restoring the anti-oxidant defense and mitigating lipid peroxidation in the lens and hence represents an attractive anti-cataractogenic remedy.

In vivo analysis of protein quality control in response to aging using transgenic mice

AbstractPurpose It has been suggested that aging affects the cells ability to protect protein integrity, which is essential for cellular homeostasis. On cell aging there is impaired protein homeostasis, which can lead to protein insolubilization that is toxic to cells. The carboxyl terminus of Hsp70‐interacting protein (CHIP) is an important ubiquitin ligase for protein quality control, and particularly in selective degradation of aggregation prone proteins. The objective of this study was to characterize the transgenic mice that overexpress CHIP/Ubc5 in lens epithelial cells and investigate the role of these proteins in the process that maintains proteins in solution and therefore participates in a number of age‐related diseases, including cataract.Methods To overexpress CHIP, the α‐crystallin promoter was used to drive the expression of CHIP‐Ubc5 in lens epithelial cells of transgenic mice. The CHIP levels of transgenic mice (Tg) were determined with western blotting analysis. Cataract was induced by single subcutaneous injection of sodium selenite and the lens transparency was assessed by slit lamp examination. The soluble and insoluble protein content in all groups was evaluated.Results The data show that the transgenic mice only overexpress the transgene in lens epithelial cells.The slit lamp analysis showed that transgenic mice appears developed less cataract compared with wild type, with concomitantly increase in ratio of the soluble and insoluble proteins.Conclusion Together the data suggest that overexpression of CHIP/Ubc5 in lens epithelial cells could prevent cataract formation by promoting reduction of protein insolubilization.

Rutin ameliorates free radical mediated cataract by enhancing the chaperone activity of α-crystallin

Cataract, the leading cause of blindness, is associated with oxidative damage and protein modification in the lens. The present study was carried out to assess the efficacy of rutin on rat-lens crystallins in selenite-induced in-vivo cataract models. Eight-day-old Sprague-Dawley rat pups were grouped as control (G I), experimental (G II) and rutin-treated (G III). The rat pups in G II, and G III received a single subcutaneous injection of sodium selenite (4μg/g body weight) and G I received a single subcutaneous injection of sterile water on the 10th day. The treatment groups (G III) were administered with rutin (1μg/g body weight) respectively from the 8th to 15th day. Cataract was visualized from the 16th day. Lens crystallins (α, β, and γ) were isolated by size exclusion chromatography. Chaperone activity of isolated crystallins was measured by heat, DTT, and oxidation-induced aggregation and refolding assays. Concentration of total protein (soluble and insoluble) and SDS-PAGE analysis of soluble proteins were also done. Treatment with rutin prevented the loss of α crystallin chaperone property, and protein insolubilization prevailed during selenite-induced cataract. These results suggest the therapeutic potential of rutin, a bioflavonoid, against selenite-induced cataract, which has been reported in this paper for the first time. The work assumes significance, as this is a novel approach in modulating the chaperone activity of lens crystallins in selenite-induced cataract by a natural product.

C-Phycocyanin Modulates Selenite-Induced Cataractogenesis in Rats

The present investigation is aimed to evaluate the anticataractogenic potential of C-phycocyanin (C-PC), extracted and purified from Spirulina platensis. Enucleated rat lenses were maintained in vitro in Dulbecco's modified Eagle medium (DMEM). Group I contained DMEM, Group II and Group III contained 100μM of sodium selenite, Group III was subdivided into three viz IIIa, IIIb, IIIc supplemented with 100, 150, 200μg of C-PC respectively. In the in vivo study, on tenth day post partum: Group I rat pups received an intraperitoneal injection of saline, Group II, IIIa, IIIb, and IIIc rat pups received a subcutaneous injection of sodium selenite (19μmol/kg bodyweight) Group IIIa, IIIb, IIIc also received an intraperitoneal injection of 100, 150, 200mg/kg body weight of C-PC, respectively, from postpartum days 9-14. On termination of the experiment, the lenses from both in vitro and in vivo studies were subjected to morphological examination and subsequently processed to estimate the activities of antioxidant enzymes namely superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, levels of reduced glutathione and lipid peroxidation products. Sodium selenite-exposed, C-PC-treated rat lenses (Group IIIc), showed significant restoration of antioxidant enzyme activity (p < 0.05) when compared to their counterpart Group II. Group IIIc conserved the levels of GSH and lipid peroxidation products at near to normal levels as compared with Group II. Results conclude the possible role of C-PC in modulating the antioxidant enzyme status, thereby retarding sodium selenite-induced cataract incidence both in vitro and in vivo.

Effect of Crocus sativus stigmas (saffron) extract on sodium selenite induced cataract formation

AbstractPurpose To study the effect of Crocus sativus (CS) stigmas (saffron) extract on sodium selenite induced cataract formation in vivo.Methods Twenty‐four Wistar rat pups were randomized into 3 groups (n=8/group). Control group received subcutaneous injection of normal saline on postnatal day 10. Selenite and CS treated group received a subcutaneous injection of sodium selenite (20 µmol/kg body weight) on postnatal day 10. CS group also received intraperitoneal injections of CS extract (60 mg/kg body weight) on postnatal days 11 and 12. On postpartum day 21, rats were sacrificed and the lenses were isolated and examined for cataract formation. Activities of superoxide dismutase (SOD) and catalase (CAT) and levels of reduced glutathione (GSH) and glutathione disulfide (GSSG) in the isolated lenses were measured. SDS‐PAGE of lens water soluble protein fraction (WSF) was done.Results Selenite resulted in significant cataract formation compared to control group, p&lt;0.001. In the CS group, cataract formation was significantly prevented compared to selenite group, p&lt; 0.0001. The mean activities of SOD and CAT were significantly increased in the CS group compared to the selenite group (p=0.025 and 0.0009 respectively). The GSSG/GSH ratio was significantly decreased in the CS group compared to selenite group (p=0.001). CS prevented selenite induced proteolysis of the lens WSF.Conclusion CS extracts prevented selenite induced cataract formation in Wistar rats possibly by enhancement of antioxidant status and inhibition of proteolysis of the lens WSF.

Inhibitory effects of grape seed proanthocyanidin extract on selenite-induced cataract formation and possible mechanism

This study investigated the inhibitory effect of grape seed proanthocyanidin extract (GSPE) on selenite-induced cataract formation in rats and the possible mechanism. Eighty 8-day-old Sprague-Dawley rats were divided randomly into 5 groups: control group, model group, three GSPE groups (low dose, medium dose and high dose). Control group received subcutaneous injection of physiological saline. Model group was given subcutaneous injection of sodium selenite (20 μmol/kg body weight) on the postpartum day 10, and once every other day for consecutive three times thereafter. GSPE treated groups were respectively administered GSPE at doses of 50, 100, and 200 mg/kg body weight intragastrically 2 days prior to the selenite injection (that was, on the postpartum day 8), and once daily for fourteen consecutive days thereafter. The opacity of lenses was observed, graded and photographed under the slit lamp microscopy and the maximal diameter of the nuclear cataract plaques was measured. The lenses were analyzed for superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-PX), malondialdehyde (MDA), calcium (Ca(2+)), nitric oxide (NO) and anti-hydroxyl radical ability (anti-OH(-)). The histomorphology of lenses was observed with HE staining under a light microscope. The levels of calpainII, and iNOS protein and mRNA expression in lenses were detected by using immunohistochemistry and real-time quantitative RT-PCR. The results showed subcutaneous injection of sodium selenite led to severe nuclear cataract in model group, and the achievement ratio of model group was 100%. As compared with model group, the degree of lenses opacity and the maximal diameter of nuclear cataract plaques were significantly reduced in GSPE-treated groups. Moreover, we observed selenite treatment caused a significant decrease in the activities of antioxidative enzymes (SOD, CAT, GSH-PX) and anti-OH(-) ability, accompanied by a significant increase in the levels of MDA, NO, Ca(2+) as well as iNOS, and calpainII protein and mRNA expression. Administration of GSPE could dose-dependently preserve the activities of these antioxidative enzymes and anti-OH(-) ability, accompanied by a significant reduction in the levels of MDA, NO, Ca(2+) as well as iNOS, and calpainII protein and mRNA expression. These results suggested that GSPE markedly prevented selenite-induced cataract formation probably by suppressing the generation of lipid peroxidation and free radicals as well as the activation of iNOS, and calpainII in the lenses.

Prevention of Selenite-Induced Cataractogenesis by an Ethanolic Extract of Cineraria maritima: An Experimental Evaluation of the Traditional Eye Medication

In the present study, the antioxidant potential of an ethanolic extract of Cineraria maritima and its efficacy in preventing selenite-induced cataractogenesis were assessed in vitro and in vivo. In the in vitro phase of the study, lenses dissected out from the eyes of Wistar rats were incubated for 24 h at 37 °C in Dulbecco's modified Eagle medium (DMEM) alone (group I), in DMEM containing 100 μM of selenite only (group II), or in DMEM containing 100 μM of selenite and 300 μg/ml C. maritima extract added at the same time (group III). Gross morphological examination of the lenses revealed dense opacification in group II, minimal opacification in group III, and no opacification in group I lenses. The mean activities of the antioxidant enzymes catalase, glutathione peroxidase, and superoxide dismutase were significantly lower in group II than in group I or group III lenses, while malondialdehyde concentration was significantly higher in group II lenses than in group I and group III lenses. In the in vivo phase of the study, dense opacification of lenses was noted in all rat pups (100%) that had received a single subcutaneous injection of sodium selenite alone (19 μM/kg body weight) on postpartum day 10, whereas cataract formation occurred in only 33.3% of rat pups that had received selenite as well as an intraperitoneal injection of the extract of C. maritima (350 mg/kg body weight) for five consecutive days. These observations suggest that the ethanolic extract of C. maritima may prevent experimental selenite-induced cataractogenesis.

Moringa oleiferaPrevents Selenite-Induced Cataractogenesis in Rat Pups

The aim of this study was to investigate the protective effects of the flavonoid fraction of Moringa oleifera leaves (FMO) on selenite cataract in vivo. Rat pups of Sprague-Dawley strain initially weighing 10–12 g on day 8 were used for the study and grouped as control (G I), selenite induced (G II), and FMO treated (G III). The rat pups in G II and G III received a single subcutaneous injection of sodium selenite (4 µg/g body weight) on day 10 and G III was administered with FMO (2.5 µg/g body weight) from day 8 to 15. Cataract was visualized from day 16. The development of cataract was assessed and rat lenses were analyzed for the activities of antioxidant enzymes (superoxide dismutase and catalase), generation of reactive oxygen species, reduced glutathione, protein oxidation, and lipid peroxidation. FMO was subjected to in vitro antioxidant assays (2,2-diphenyl-picrylhydrazyl and superoxide scavenging assays). The total phenolic content of FMO was 4.4 mg of catechin equivalent/g dried plant material. The extract showed remarkable activity on 2,2-diphenyl-picrylhydrazyl (IC50 36 µg/mL) and in superoxide radical (IC50 33.81µg/mL) scavenging assays. FMO effectively prevented the morphological changes and oxidative damage in lens. FMO maintained the activities of antioxidant enzymes and sulfhydryl content and prevented reactive oxygen species generation and lipid peroxidation. FMO was effective in preventing cataractogenesis in selenite model by enhancing the activities of antioxidant enzyme, reducing the intensity of lipid peroxidation, and inhibiting free radical generation.

Vitex negundo Modulates Selenite-Induced Opacification and Cataractogensis in Rat Pups

Recently, much interest has been generated in the search for phytochemical therapeutics, as they are largely free from adverse side effects and economical. The goal of this study was to determine the efficacy of Vitex negundo in modulating the selenite-induced oxidative stress in vivo model. Sprague-Dawley rat pups of 8 days old were used for the study and divided into control (G I), selenite induced (G II), and selenite + V. negundo treated (G III). Cataract was induced by the single subcutaneous injection of sodium selenite (4 mg/kg body weight) on the tenth day and V. negundo (2.5 mg/Kg body weight) administered intraperitoneally from eighth to 15th day. Morphological examination of the rat lenses revealed no opacification in G I and mild opacification in G III whereas dense opacification in G II (stages 4-6). Levels of selenium in G II and G III showed no significant changes. The activities of superoxide dismutase, catalase, and Ca(2+)ATPase were significantly increased in G III compared to G II (p < 0.05), while lower level of reactive oxygen species, Ca(2+), and thiobarbituric acid reactive substances were observed in G III compared G II (p < 0.05). These results indicate the therapeutic potential of methanolic extract of V. negundo on modulating biochemical parameters against selenite-induced cataract, which have been reported in this paper for the first time.

Evaluation of anticataract potential of Triphala in selenite-induced cataract: In vitro and in vivo studies

Triphala (TP) is composed of Emblica officinalis, Terminalia chebula, and Terminalia belerica. The present study was undertaken to evaluate its anticataract potential in vitro and in vivo in a selenite-induced experimental model of cataract. In vitro enucleated rat lenses were maintained in organ culture containing Dulbecco’s Modified Eagles Medium alone or with the addition of 100μM selenite. These served as the normal and control groups, respectively. In the test group, the medium was supplemented with selenite and different concentrations of TP aqueous extract. The lenses were incubated for 24 h at 37°C. After incubation, the lenses were processed to estimate reduced glutathione (GSH), lipid peroxidation product, and antioxidant enzymes. In vivo selenite cataract was induced in 9-day-old rat pups by subcutaneous injection of sodium selenite (25 μmole/kg body weight). The test groups received 25, 50, and 75 mg/kg of TP intraperitoneally 4 h before the selenite challenge. At the end of the study period, the rats’ eyes were examined by slit-lamp. TP significantly (P < 0.01) restored GSH and decreased malondialdehyde levels. A significant restoration in the activities of antioxidant enzymes such as superoxide dismutase (P < 0.05), catalase (P < 0.05), glutathione peroxidase (P < 0.05), and glutathione-s-transferase (P < 0.005) was observed in the TP-supplemented group compared to controls. In vivo TF 25mg/kg developed only 20% nuclear cataract as compared to 100% in control. TP prevents or retards experimental selenite-induced cataract. This effect may be due to antioxidant activity. Further studies are warranted to explore its role in human cataract.

Trigonella foenum-graecum (Fenugreek) Protects Against Selenite-Induced Oxidative Stress in Experimental Cataractogenesis

Cataract is the opacification in eye lens and leads to 50% of blindness worldwide. The present study was undertaken to evaluate the anticataract potential of Trigonella foenum-graecum Linn seeds (fenugreek) in selenite-induced in vitro and in vivo cataract. In vitro enucleated rat lenses were maintained in organ culture containing Dulbecco's modified Eagles medium (DMEM) alone or in addition with 100 microM selenite and served as the normal and control groups, respectively. For the test group, the medium was supplemented with selenite and T. foenum-graecum aqueous extract. The lenses were incubated for 24 h at 37 degrees C. After incubation, the lenses were processed for the estimation of reduced glutathione (GSH), lipid peroxidation product (malondialdehyde), and the antioxidant enzymes. In vivo selenite cataract was induced in 9-day-old rats by subcutaneous injection of sodium selenite (25 micromol/kg body weight). Animals in the test group were injected with different doses of aqueous extract of T. foenum-graecum 4 h before the selenite challenge. A fall in GSH and a rise in malondialdehyde levels were observed in control as compared to normal lenses. T. foenum-graecum significantly (P < 0.01) restored glutathione and decreased malondialdehyde levels. A significant restoration in the activities of antioxidant enzymes such as superoxide dismutase (P < 0.01), catalase, (P < 0.01), glutathione peroxidase (P < 0.01), and glutathione-S-transferase (P < 0.01) was observed in the T. foenum-graecum supplemented group as compared to control. In vivo, none of the eyes was found with nuclear cataract in treated group as opposed to 72.5% in the control group. T. foenum-graecum protects against experimental cataract by virtue of its antioxidant properties. Further studies are warranted to explore its role in human cataract.

Vitex negundo attenuates calpain activation and cataractogenesis in selenite models

Recent investigations have shown that phytochemical antioxidants can scavenge free radicals and prevent various diseases. Cataract is the leading cause of blindness and is associated with oxidative damage of the lens. Selenite-induced cataract in rat pups is an excellent mimic of oxidative stress-induced cataract. Selenite cataract is associated with oxidative stress, loss of calcium homeostasis, calpain activation and protein insolubilization in the lens. Our present study focuses on the isolation of flavonoids from Vitex negundo and to assess its efficacy in preventing these changes in the lens of selenite-induced cataract models. Eight-day-old Sprague-Dawley rat pups were used for the study and divided into four groups: Control (G I), Sodium selenite-induced (G II), Sodium selenite+quercetin treated (G III), Sodium selenite+flavonoids from Vitex negundo (FVN) (G IV). Cataract was induced by a single subcutaneous injection of Sodium selenite (4mg/Kg body weight) on the 10th day. Treatment groups received quercetin (1.0mg/Kg body weight) and FVN (1.0mg/Kg body weight) intraperitoneally from 8th to 15th day. Cataract was visualized from the 16th day. Morphological examination of the rat lenses revealed no opacification in G I and mild opacification in G III and G IV (stage 1) whereas dense opacification in G II (stage 4–6). The activities of superoxide dismutase (SOD), catalase, Ca2+ATPase, concentration of reduced glutathione (GSH) and protein sulfhydryl content were significantly increased in G III and G IV compared to G II, while decreased activities of calpains, lower concentration of calcium and thiobarbituric acid reactive substances (TBARS) were observed in G III and IV as compared to G II. Lens protein profile of water soluble proteins showed normal levels of expression in treated groups compared to that of selenite-induced rats. These results indicate good antioxidant and therapeutic potential of FVN in modulating biochemical parameters against selenite-induced cataract, which have been reported in this paper for the first time.