METHOD FOR TREATMENT OF VISUAL STRESS CONDITIONS AND COMPOSITIONS USED THEREIN

Abstract

The methods relate to preventing photoreceptor damage caused by blue light by oral administration of a macular carotenoid composition comprising of lutein and zeaxanthin isomers. The photoreceptor damage is prevented through the upregulation of genes and protein synthesis manifested by an increase in the levels gene expression and proteins synthesis of Gnat1, Rhodopsin, NCAM, Rod arrestin, GAP-43, Nrf2, Ho-1 and decrease in the GFAP, NFkB (oxidative stress markers) in rat eye models exposed to visible blue light for various time periods and simultaneously treated with macular carotenoid compositions comprising of lutein zeaxanthin and their isomers. Methods described herein also relate to treatment of visual stress conditions by administering macular carotenoid compositions for treatment headache, blurred vision, photoreceptor degeneration, oxidative stress, endoplasmic reticulum stress and the like, which are caused due to prolonged exposure to blue light source. The methods described herein help to treat visual stress disorders by reducing headache and headache frequency, slowing down photoreceptor degeneration, improvement in oxidative stress and protection of retinal cells from light damage. The compositions herein are safe for humans and other animals for consumption and can be used for treatment of visual stress disorders in the subject in need thereof.

Claims

1. Method of preventing photoreceptor damage caused by blue light by oral administration of effective amount of macular carotenoid composition, comprising lutein, zeaxanthin and meso-zeaxanthin, through up-regulation of GNAT1 and NCAM.

2. Method of preventing photoreceptor damage of claim 1, wherein GNAT 1, a photoreceptor gene responsible for biosynthesis of alpha-transducin 1, is upregulated about 1.2 to 3.5 times in subjects administered with macular carotenoid composition as compared to subjects on the vehicle control group.

3. Method of preventing photoreceptor damage of claim 1, wherein NCAM a gene responsible for neurite growth is upregulated about 1.2 to 2.0 times in subjects administered with macular carotenoid composition as compared to subjects on the vehicle control group.

4. Method of preventing photoreceptor damage of claim 1, wherein blue light comes from the digital devices like television sets, computers, laptops, smart phones, tablets, electronic devices, fluorescent lighting and LED lighting.

5. Method of preventing photoreceptor damage of claim 1, wherein the macular carotenoid composition is comprised of lutein and zeaxanthin isomers such as meso-zeaxanthin and R,R zeaxanthin.

6. Method of preventing photoreceptor damage of claim 5, wherein effective amount of the macular carotenoid composition comprises of lutein, meso-zeaxanthin and R,R zeaxanthin ranges from 1 mg to 100 mg/kg body weight.

7. Method of preventing photoreceptor damage as claimed in claim 5, wherein the lutein and zeaxanthin isomers are present in the ratio of 5:1

Description

DETAILED DESCRIPTION OF FIGURES

[0049] FIG. 1 relates to the graphical representation of the rhodopsin gene expression measured in rat eyes in the treatment and vehicle control groups in terms of fold change of vehicle control group exposed to different light conditions i.e. 24 hour LED light exposure (24 h L), 16 hour light and 6 hour dark adaptation (16 h L/6 h D) and 12 hour light and 12 hour dark adaptation (12 h L/12 h D).

[0050] FIG. 2 relates to the graphical representation of the rod arrestin (Sag) gene concentration measured in rat eyes in the treatment and vehicle control groups in terms of fold change of vehicle control group exposed to different light conditions i.e. 24 hour LED light exposure (24 h L), 16 hour light and 6 hour dark adaptation (16 h L/6 h D) and 12 hour light and 12 hour dark adaptation (12 h L/12 h D).

[0051] FIG. 3 relates to the graphical representation of the graphical representation of the G protein subunit alpha transducin 1 (GNAT-1)gene expression measured in rat eyes in the treatment and vehicle control groups in terms of fold change of vehicle control group exposed to different light conditions i.e. 24 hour LED light exposure (24 h L), 16 hour light and 6 hour dark adaptation (16 h L/6 h D) and 12 hour light and 12 hour dark adaptation (12 h L/12 h D).

[0052] FIG. 4 relates to the graphical representation of the graphical representation of the nuclear factor-kappa B (NF-B) protein concentration measured in rat eyes in the treatment and vehicle control groups in terms of fold change of vehicle control group exposed to different light condition i.e. 24 hour LED light exposure (24 h L), 16 hour light and 6 hour dark adaptation (16 h L/6 h D) and 12 hour light and 12 hour dark adaptation (12 h L/12 h D).

[0053] FIG. 5 relates to the graphical representation of the graphical representation of neural cell adhesion molecule (NCAM) gene expression measured in rat eyes in the treatment and vehicle control groups in terms of fold change of vehicle control group exposed to different light conditions i.e. 24 hour LED light exposure (24 h L), 16 hour light and 6 hour dark adaptation (16 h L/6 h D) and 12 hour light and 12 hour dark adaptation (12 h L/12 h D).

[0054] FIG. 6 relates to the graphical representation of the graphical representation of the growth-associated protein-43 (GAP-43) gene expression measured in rat eyes in the treatment and vehicle control groups in terms of fold change of vehicle control group exposed to different light conditions i.e. 24 hour LED light exposure (24 h L), 16 hour light and 6 hour dark adaptation (16 h L/6 h D)and 12 hour light and 12 hour dark adaptation (12 h L/12 h D).

[0055] FIG. 7 relates to the graphical representation of the glial fibrillary acidic protein (GFAP) protein concentration measured in rat eyes in the treatment and vehicle control groups in terms of fold change of vehicle control group exposed to different light conditions i.e. 24 hour LED light exposure (24 h L), 16 hour light and 6 hour dark adaptation (16 h L/6 h D) and 12 hour light and 12 hour dark adaptation (12 h L/12 h D).

[0056] FIG. 8 relates to the graphical representation of the nuclear factor (erythroid-derived 2)-like 2 (Nrf2) protein concentration measured in rat eyes in the treatment and vehicle control groups in terms of fold change of vehicle control group exposed to different light conditions i.e. 24 hour LED light exposure (24 h L), 16 hour light and 6 hour dark adaptation (16 h L/6 h D) and 12 hour light and 12 hour dark adaptation (12 h L/12 h D).

[0057] FIG. 9 relates to the graphical representation of the graphical representation of the Haemeoxygenase I (HO-1) protein concentration measured in rat eyes in the treatment and vehicle control groups in terms of fold change of vehicle control group exposed to different light conditions i.e. 24 hour LED light exposure (24 h L), 16 hour light and 6 hour dark adaptation (16 h L/6 h D) and 12 hour light and 12 hour dark adaptation (12 h L/12 h D).

DETAILED DESCRIPTION

[0058] Retinal photoreceptor cells are uniquely adapted to function over a wide range of ambient light conditions. However, in most species prolonged intense visible light exposure can lead to photoreceptor cell damage. In nocturnal animals, the light intensity required for visual cell damage need be only 2-3 times above normal room lighting (Noell 1965). Visual cell damage can then progress to cell death and loss of vision, or retinal light damage may regress with recovery of function. Intense light exposure has been used as an environmental stressor in testing genetic animal models of photoreceptor cell degeneration. There is growing evidence that disease mechanisms in age related macular degeneration (AMD) involve oxidative stress and inflammation. Hence it becomes important to understand eye stress disorders caused due to prolonged exposure to light sources, to understand effect on retinal and brain health. As the disorders caused are not reversible, the damaging effects on eye health need to be identified in time and corrective measures need to be taken to avoid further damage to eye.

[0059] The methods described herein and the compositions used are directed to treating eye stress disorders by administering a macular carotenoid composition in an effective amount(s) to a subject in need thereof. The methods and compositions herein can treat conditions associated with eye stress, so as to protect and treat eye and improve coordination with brain, when administered for example to a subject who is exposed to various light emitting devices, including but not limited to blue light, sun rays, and other lightwaves of various intensities for a prolonged time period.

[0060] The active ingredients of macular carotenoid compositions herein are obtained by natural resources and the compositions overall are safe for administration, and thus the compositions are useful as nutraceutical compositions and/or formulations.

[0061] Constant exposure to blue light poses hazard through a modern life style and work conditions and may cause suffering of visual health to a great extent. Such people experience symptoms such as frequent and intense headache, affected vision due to photoreceptor degeneration and oxidative stress and visual stress which may have an overall impact on work performance. One of the causative factors is use of computers and laptops and exposure to blue light for extended periods of time such as for example, but not limited to 4 to 10 hours.

[0062] The term blue light used herein refers to the light in the visible spectrum in the range of 400 to 490 nm which is emitted from displays of digital devices such as computers, television laptops, smart phones and tablets, electronic devices fluorescent and LED lighting etc.

[0063] The term fold change of the vehicle control group means representation of values of the treatment groups in terms of multiples of the values for the vehicle control group exposed to the same LED blue light conditions.

[0064] The terminology subject refers to a human individual or a mammal which may be undergoing testing, which is being treated with the macular carotenoid compositions herein.

[0065] The terminology subject in need thereof can include specific individuals or mammals, who may get exposed to, or may have been exposed to blue light for a prolonged period of time such as for example but not limited to 4 to 10 hours. This causes damage to retinal and brain health and damages the photoreceptors and affects retinal cell viability.

[0066] The terminology eye stress disorders can include various eye disorders caused due to prolonged exposure to light sources, the disorders include intense and frequent headache, affected vision due to photoreceptor degeneration, increased oxidative stress, thus affecting overall retinal and brain health.

[0067] In one aspect the methods relate to preventing photoreceptor damage caused by blue light by oral administration of a carotenoid composition comprising of lutein and zeaxanthin isomers through up-regulation of GNAT1 which is a photoreceptor gene responsible for biosynthesis of alpha-transducin 1. The method of preventing photoreceptor damage caused by blue light is obtained by administration of an effective amount of multicarotenoid composition comprising lutein, zeaxanthin or isomers thereof in the dose range of 0.1-100 mg/kg body weight in an embodiment. In an embodiment, the dose range is 1-100 mg/kg body weight, more preferably in the range of 10-80 mg/kg body weight and most preferably in the range of 20-50 mg/kg body weight. In one embodiment the effective amount of the macular carotenoid composition comprises of lutein, meso-zeaxanthin and R,R zeaxanthin in the range from 0.1 mg to 100 mg/kg body weight. Further lutein and zeaxanthin isomers may be present in the ratio of 5:1 in such dose.

[0068] In another aspect the methods relate to preventing photoreceptor damage caused by blue light coming by oral administration of an effective amount of carotenoid composition comprising of lutein and zeaxanthin isomers through up-regulation of NCAM gene responsible for the neurite growth. The method of preventing photoreceptor damage caused by blue light is obtained by administration of a multicarotenoid composition comprising lutein, zeaxanthin or isomers thereof in the dose range of 0.1-100 mg/kg body weight in an embodiment. In an embodiment, the dose range is 1-100 mg/kg body weight, more preferably in the range of 10-80 mg/kg body weight and most preferably in the range of 20-50 mg/kg body weight. In one embodiment the effective amount of the macular carotenoid composition comprises of lutein, meso-zeaxanthin and R,R zeaxanthin in the range from 0.1 mg to 100 mg/kg body weight. Further lutein and zeaxanthin isomers may be present in the ratio of 5:1 in such dose.

[0069] In one more aspect the methods relate to preventing photoreceptor damage caused by blue light by oral administration of carotenoid composition comprising of lutein and zeaxanthin isomers reduction in the level of oxidative stress markers such as GFAP, NFkB. The method of preventing photoreceptor damage is obtained by administration of a multicarotenoid composition comprising lutein, zeaxanthin or isomers thereof in the dose range of 0.1-100 mg/kg body weight in an embodiment. In an embodiment, the dose range is 1-100 mg/kg body weight, more preferably in the range of 10-80 mg/kg body weight and most preferably in the range of 20-50 mg/kg body weight. In one embodiment the effective amount of the macular carotenoid composition comprises of lutein, meso-zeaxanthin and R,R zeaxanthin in the range from 0.1 mg to 100 mg/kg body weight. Further lutein and zeaxanthin isomers may be present in the ratio of 5:1 in such dose.

[0070] In one more aspect the methods relate to preventing photoreceptor damage caused by blue light by oral administration of carotenoid composition comprising of lutein and zeaxanthin isomers through upregulation of protein nuclear transcription factor Nrf2, Haemeoxygenase HO-1.The method of preventing photoreceptor damage is obtained by administration of administration of an effective amount of a multicarotenoid composition comprising lutein, zeaxanthin or isomers thereof in the dose range of 0.1-100 mg/kg body weight in an embodiment. In an embodiment, the dose range is 1-100 mg/kg body weight, more preferably in the range of 10-80 mg/kg body weight and most preferably in the range of 20-50 mg/kg body weight. In one embodiment the effective amount of the macular carotenoid composition comprises of lutein, meso-zeaxanthin and R,R zeaxanthin in the range from 0.1 mg to 100 mg/kg body weight. Further lutein and zeaxanthin isomers may be present in the ratio of 5:1 in such dose.

[0071] In yet another aspect the methods relate to preventing photoreceptor damage caused by blue light by oral administration of carotenoid composition comprising of lutein and zeaxanthin isomers through upregulation of rhodopsin (Rho) gene, Rod arrestin gene (Sag) gene, Growth associated protein 43 (GAP-43) genes. The method of preventing photoreceptor damage is obtained by administration of an effective amount of multicarotenoid composition comprising lutein, zeaxanthin or isomers thereof in the dose range 0.1-100 mg/kg body weight in an embodiment. In an embodiment, the dose range is 1-100 mg/kg body weight, more preferably in the range of 10-80 mg/kg body weight and most preferably in the range of 20-50 mg/kg body weight. In one embodiment the effective amount of the macular carotenoid composition comprises of lutein, meso-zeaxanthin and R,R zeaxanthin in the range from 0.1 mg to 100 mg/kg body weight. Further lutein and zeaxanthin isomers may be present in the ratio of 5:1 in such dose.

[0072] The method of preventing photoreceptor damage caused by blue light by oral administration of a macular carotenoid composition gives a multi fold increase in the expression of photoreceptor genes as compared to the control vehicle group in groups exposed to different duration of LED blue light. The treatment group treated with the multicarotenoid composition results in upregulation of 1.2 to 4.5 fold increase in the expression of photoreceptor genes, retinal proteins as compared to subjects on the vehicle control group. The methods herein result in the decrease in the expression of oxidative stress markers. The oxidative stress markers show a 0.5 to 3 fold decrease in the treatment group as compared to the vehicle control group. The advantageous effect for the group treated with the multicarotenoid composition is quantified in terms of increase in the expression of the retinal photoreceptor gene expression and protein synthesis as multiple of the value obtained for the vehicle control group and decrease in the expression of the oxidative stress markers as multiples of the value obtained for the vehicle control group.

[0073] In another aspect methods of preventing photoreceptor damage caused by blue light involve upregulation of GNAT 1, a photoreceptor gene responsible for biosynthesis of alpha-transducin 1, by about 1.2 to 4.5 times in subjects administered with macular carotenoid composition as compared to subjects on the vehicle control group.

[0074] In another aspect method of preventing photoreceptor damage caused by blue light involve upregulation of NCAM a gene responsible for neurite growth by about 1.2 to 4.5 times in subjects administered with macular carotenoid composition as compared to subjects on the vehicle control group.

[0075] In an embodiment, methods are described herein for treatment of eye stress disorders by administering a macular carotenoid composition to a subject in need thereof, and evaluating one or more parameters, such as for example but not limited to, improvement in intense and frequent headache, improvement in vision due to improvement in photoreceptor degeneration, reduction in oxidative and endoplasmic reticulum stress to assess improvement in overall retinal and brain health.

[0076] In one embodiment, macular carotenoid compositions herein and methods of treatment herein may include administering to a subject the macular carotenoid composition, which can include additional nutrients such as for example but not limited to fatty acids, and at least one food grade excipient selected from the group of, but not limited to, an antioxidant, oil, a surfactant, a solubilize, an emulsifier, and the like, or a combination(s) thereof.

[0077] In one embodiment, macular carotenoid compositions and methods of treatment using the composition, may comprise one or more carotenoids in various ratios, where the carotenoid(s) are lutein and zeaxanthin, either alone or in combination, and selected from the group of, but not limited to free lutein, including lutein esters and/or isomers of lutein, zeaxanthin including one or more of its isomers, such as (R,R) zeaxanthin and/or meso-zeaxanthin, derivatives thereof.

[0078] In an embodiment, a macular carotenoid composition herein is administered in a daily dose of at or about 0.1 to 100 mg/kg body weight.

[0079] In an embodiment, a macular carotenoid composition herein is administered in a daily dose of about 1 mg to about 100 mg/kg body weight.

[0080] In some embodiments, macular carotenoid compositions herein and methods of treatment using the composition are directed to administering the composition, for example in a daily dose of about 10 mg to about 80 mg/kg body weight of macular carotenoids, to the subject in need thereof.

[0081] Further the composition may be administered such that, macular carotenoids contained therein are provided in a daily dose of about 20-50 mg/kg body weight over a certain amount of treatment to treat visual fatigue.

[0082] The methods described herein relate to treatment of eye stress disorders in subjects in need thereof, such as for example mammals including a human, who are exposed to light emitting devices of various wavelengths and intensities, for a prolonged time period, by administering an effective amount of macular carotenoid composition and evaluating one or more parameters such as effect on headache recovery, retinal photoreceptor gene expression analysis, retinal protein synthesis analysis, level of oxidative stress markers, effect on oxidative stress, effect on photoreceptor degeneration and effect on endoplasmic reticulum stress, to check beneficial effects of macular carotenoids for treatment of subjects with various eye stress disorders.

[0083] The method of preventing photoreceptor damage caused by blue light coming from digital devices requires the administration of a multicarotenoid composition. The multicarotenoid composition includes combination of lutein and zeaxanthin isomers such as meso-zeaxanthin and R,R zeaxanthin. Thus the composition is comprised of lutein, meso-zeaxanthin and R,R zeaxanthin. The macular pigments may be derived for example from a plant extract and/or oleoresin containing xanthophylls and/or xanthophylls esters, and are useful for nutrition and health applications. The macular pigments may be derived from the plant extract and/or oleoresin containing xanthophylls and/or xanthophylls esters, which is safe for human consumption. In one embodiment the ratio of lutein to zeaxanthin isomers is 5:1.

[0084] In an embodiment the isomers of zeaxanthin are present in the form of meso-zeaxanthin and RR-zeaxanthin in the ratio of 3:1. A multicarotenoid composition, for example in a test composition, may contain 1 mg trans-lutein and 0.2 mg of zeaxanthin. The zeaxanthin in this embodiment may be present as 0.15 mg of meso-zeaxanthin and 0.05 mg of R,R-zeaxanthin. The ratio of the isomers of zeaxanthin (e.g. meso-zeaxanthin and R,R-zeaxanthin) is in the range of at or about 80:20 to at or about 20:80 (e.g. 4:1 to 1:4), more preferably it is present in the range of 75:25. The composition is administered to a subject in need thereof suffering from various eye stress disorders such as intense and frequent headache, affected retinal and brain health.

[0085] In an embodiment, a test composition can be scaled to the doses described above. For example, about 0.1 mg/kg body weight of test composition (e.g. based on the amount of lutein) can include 0.1 mg of lutein and 0.02 mg of zeaxanthin isomers. In another example such as described above, about 1 mg of test composition can include 1 mg lutein and 0.2 mg zeaxanthin isomers. In another example, 50 mg of test composition can include 50 mg lutein and 10 mg zeaxanthin isomers. In another example and 100 mg test composition can include 100 mg lutein and 20 mg zeaxanthin isomers. It will be appreciated that in an embodiment, the ratio of lutein and zeaxanthin isomers is at or about 5:1 in the test composition. Accordingly about .sup.th the amount of zeaxanthin isomer content is present in the composition with an amount of lutein.

[0086] In one embodiment, macular carotenoid compositions herein and methods of treatment and prevention of photoreceptor damage caused by blue light herein using the multicarotenoid compositions, may be administered to a subject in need thereof, in a form of a nutraceutical carrier, food supplement, beverage, medical food, while employing dosage forms such as granules, powders, sachets, beadlets, capsules, soft gel capsules, tablets, solutions, suspensions, and the like. The active carotenoid(s) of the compositions may be prepared by an extraction process and formulated into a composition, such as together with one or more other food grade excipient(s) and/or materials to obtain the desired form.

[0087] In one embodiment, methods of treatment/prevention of photoreceptor damage herein may comprise administering an effective amount of macular carotenoids to subjects in need thereof, to treat eye stress disorders such as intense and frequent headache, visual stress due to photoreceptor degeneration, oxidative stress and endoplasmic reticulum stress, occurring as a result from exposure to blue light, sun rays, and/or lights of varying wavelengths and/or intensities.

[0088] In one embodiment, methods of treatment/prevention of photoreceptor damage herein may comprise administering an effective amount of macular carotenoids to subjects in need thereof for treatment of eye stress disorder such as headache, and evaluating one or more parameters, such as effect on visual strain, reduction in photoreceptor degeneration, markers of endoplasmic reticulum stress, protein expressions for stress, oxidative stress markers, antioxidant enzymes and checking the effect(s) on improvement of retinal health.

[0089] Compositions and methods herein can treat various eye stress disorders, photoreceptor damage caused by blue light when symptoms are evident in subjects exposed to light sources of varying wavelengths and intensities, selected from the group of, but not limited to, electronic devices emitting blue light, UV light, flickering light, traffic signals, electronic equipment, and the like, and/or combinations thereof. More particularly the methods relates to prevention of photoreceptor damage caused by blue light exposure coming from digital devices like television sets, computers, laptops, smart phones, tablets, electronic devices, fluorescent lighting and LED lighting.

[0090] Compositions and methods herein can treat eye stress disorders, when subjects exposed for a prolonged period of time to light sources of varying wavelengths and intensities are suffering from intense and frequent headache, visual discomfort due to oxidative stress and photoreceptor degeneration and increased eye strain.

[0091] In an embodiment, subjects in need of treatment of eye stress disorders are identified and administered with an effective amount of a macular carotenoid composition, such that a daily dose of lutein alone, or zeaxanthin alone, or isomers thereof, or combinations thereof is provided to improve retinal health.

[0092] In some embodiments, methods for treatment of eye stress disorders herein and the compositions used herein, are comprised of evaluating one or more retinal health parameters such as gene expression analysis and effect on antioxidant markers.

[0093] Methods and compositions as used herein are also evaluated for in animal models (mouse) by intraperitoneal injection and eyes/retinas are then harvested for future anatomical and protein studies. Effect of compositions is checked on photoreceptor degeneration by Western blot and immunofluorescent staining on a major marker of endoplasmic reticulum stress marker. The effect of composition is also checked in animal model to check efficiency on Photo-Oxidative Retinal Damage. Effect is checked on antioxidant enzymes, photoreceptor damage caused by blue light, vision and brain health markers.

[0094] The method of preventing photoreceptor damage caused by blue light is evaluated in rat models by administration of daily oral dose of lutein zeaxanthin composition at a dose of 100 mg/kg body weight into 3 groups each exposed to different light conditions i.e. 24 hour LED light exposure, 16 hour LED light and 6 hour dark adaptation and 12 hour LED light exposure and 12 hour dark adaptation and each having treatment group and control vehicle group for a period of 60 months. At the end of 60 days the animals were sacrificed, eyes were excised and blood collected. The vision health parameters such as retinal photoreceptor gene expression retinal protein levels and levels of oxidative stress markers were measured. The gene expression of photoreceptor genes like Rhodopsin (Rho) gene, protein transcription factors nuclear factor (erythroid-derived 2)-like 2 (Nrf2),G protein subunit alpha transducin 1 gene (GNAT-1), nuclear factor-kappa B (NF-B) glial fibrillary acid protein (GFAP), nuclear factor (NF)-B, neural cell adhesion molecule (NCAM) gene, growth-associated protein-43 (GAP-43) gene, haemeoxygenase 1 protein expression were determined. While the compositions and methods herein have been described in terms of specific illustrative embodiments, any modifications and equivalents that would be apparent to those skilled in the art are intended to be included within the scope of the methods and compositions herein. The details of the methods and compositions herein, its objects, and advantages are explained hereunder in greater detail in relation to non-limiting exemplary illustrations.

EXAMPLE 1

[0095] Male wistar rats (age: 8 weeks, weight: 18020 g) were housed in a controlled environment with a 12:12-h light-dark cycle at 22 C. and provided with rat chow and water ad libitum. All experiments were conducted under the National Institutes of Health's Guidelines for the Care and Use of Laboratory Animals and approved by the Ethics Committee of the Inonu University. Rats were randomly divided into groups each containing 7 animals exposed to different light conditions based on experimental design as follows:

TABLE-US-00001 TABLE 1 Details of the experimental design Vehicle control Groups based on light exposure Test composition group (A) group (B) Group I - Intense LED light for a 24 hour light Lutein zeaxanthin at a dose of 100 mg/kg Vehicle control for cycle (24 h L) body weight for a period of 60 a period of 60 days. days. Group II - Intense LED light for a 16 hour light Lutein zeaxanthin at a dose of 100 mg/kg Vehicle control for and 6 hour dark adaptation(16 h L/6 h D) body weight for a period of 60 a period of 60 days. days. Group III - Intense LED light for a 12 hour light Lutein zeaxanthin at a dose of 100 mg/kg Vehicle control for and 12 hour dark cycle (Light/Dark; 12 h L/12 h body weight for a period of 60 a period of 60 days. D) days.

[0096] The test composition was administered in the form of oil suspension and vehicle control was only oil, without addition of test composition in it.

[0097] At the end of 60 days animals were sacrificed at the same time each day immediately following light and dark exposure. Eyes were excised and blood collected by cardiac puncture under dim red illumination. The vision health parameters such as retinal photoreceptor gene expression retinal protein levels and levels of oxidative stress markers were measured. The gene expression of Rhodopsin (Rho) gene, protein transcription factors nuclear factor (erythroid-derived 2)-like 2 (Nrf2), G protein subunit alpha transducin 1 gene (GNAT-1), nuclear factor-kappa B (NF-B) glial fibrillary acid protein (GFAP), nuclear factor (NF)-B, neural cell adhesion molecule (NCAM), growth-associated protein-43 gene (GAP-43), Haemeoxygenase 1 protein expression were determined. The data was analyzed using the GLM procedure of SAS (SAS Institute: SAS User's Guide). The treatments were compared using ANOVA and student's unpaired t test; P<0.05 was considered statistically significant.

[0098] The result for retinal gene expression analysis and retinal protein synthesis are as follows:

TABLE-US-00002 TABLE 2 Details of the composition administered on retinal gene expression and retinal protein synthesis Intense Lights (LED) Light/Dark 24 h L 16 h L/6 h D 12 h L/12 h D Test Test Test Parameters Vehicle Compo- Vehicle Compo- Vehicle Compo- tested Control sition Control sition Control sition GNAT1 0.17 0.54 0.41 0.69 1.00 1.48 NCAM 0.37 0.62 0.64 0.81 1.00 1.22 Rhodopsin 0.17 0.58 0.56 0.78 1.00 1.63 (Rho) Rod arrestin 0.21 0.61 0.55 0.69 1.00 1.42 (Sag) NFkB 2.06 1.43 1.32 1.03 1.00 0.82 GAP43 0.31 0.57 0.65 0.79 1.00 1.17 GFAP 1.85 1.26 1.42 1.09 1.00 0.81 Nrf2 0.21 0.56 0.54 0.78 1.00 1.31 HO-1 0.28 0.60 0.60 0.81 1.00 1.32

[0099] The results displayed a decreased level of retinal gene expression and retinal proteins synthesis like rhodopsin (Rho) gene, Rod arrestin (Sag)gene , GNAT-1 gene, NCAM gene, GAP-43 gene, proteins Nrf2 and HO-1 in retinal tissues and increased level of markers of oxidative stress nuclear factor kappa B(NFkB) and glial fibrillary acidic protein (GFAP) in the vehicle control group B of all the 3 groups due to damaging exposure of rat retinas to visible blue light and no treatment intervention. The treatment with test composition at a dose of 100 mg/kg body weight alleviated these effects and showed increase in the levels of gene expression and protein synthesis for Gnat1 gene, Rhodopsin gene, NCAM gene, Rod arrestin (Sag) gene, Growth associated proteins gene (GAP-43), proteins Nrf2 and Ho-1 and decrease in the oxidative stress markers GFAP, NFkB in all the three treatment groups A receiving test composition at the dose of 100 mg/kg body weight dose. As can be seen from the table the treatment groups for all the three light conditions group IA, IIA and IIIA showed an increase in expression of retinal photoreceptor gene proteins, i.e. NCAM and GNAT-1 as compared to the vehicle control.

[0100] The group I A (24 hour light group) showed almost 3 fold increase in the gene expression of GNAT-1 from 0.17 to 0.54 and a two fold increase in the level of NCAM gene from 0.37 to 0.62 as compared to the control vehicle group. Similarly the group IIIA indicated a 1.48 fold increase in the GNAT level and a 1.22 fold increase in the level of NCAM gene as compared to the control group illustrating the beneficial effect of the method of prevention of photoreceptor damage caused by blue light by administration of the multicarotenoid composition. The graphical analysis of the results is as shown in the FIGS. 1 to 9.