Composition for preventing or treating retinal diseases comprising Centella asiatica extract

Abstract

The present invention relates to a composition for preventing, ameliorating, or treating glaucoma and macular degeneration, comprising a Centella asiatica extract. Specifically, the composition increases the efficiency of cellular glucose metabolism, thereby increasing the survival rate of retinal neurons and pigment epithelial cells, and protecting cells from oxidative damage caused by A2E, and is therefore useful as a composition for preventing, ameliorating, or treating glaucoma and macular degeneration for the sake of eye health.

Description

[Technical field]

[0001] The present specification relates to novel uses of Centella asiatica extract for protecting retinal neuronal cells from oxidative stress, improving the viability of retinal neuronal cells or retinal pigment epithelial cells by increasing sugar utilization, inhibiting apoptosis of retinal neuronal cells, or preventing, ameliorating or treating glaucoma or macular degeneration. [Background technology]

[0002] Glaucoma is a disease caused by damage to the optic nerve due to increased intraocular pressure or abnormal blood circulation in the optic nerve. It is characterized by temporary or permanent visual field defects and reduced vision, and if not treated, it can lead to permanent blindness. The usual cause of the disease is increased intraocular pressure due to impaired circulation of aqueous humor. In people over 40 years old, the outlet for aqueous humor gradually narrows, and the amount of aqueous humor produced increases more than the amount of it drained, causing intraocular pressure to rise, which leads to compression and damage to the optic nerve. Glaucoma can also develop even when the intraocular pressure is normal. In particular, in Korea and Japan, there are relatively many glaucoma patients with normal intraocular pressure (about 70-80% of glaucoma patients), and the cause is known to be impaired blood circulation in the optic nerve. In fact, it is known that patients with normal intraocular pressure often show symptoms of peripheral blood circulation disorders due to metabolic diseases. Ultimately, glaucoma leads to the death of optic nerve cells, such as retinal ganglion cells (Almasieh et al., 2012), and substances that can prevent or treat the death of optic nerve cells could be used as foods and treatments to inhibit the onset and progression of glaucoma.

[0003] The macula is the part of the retina where photoreceptors are densely packed and where light is most clearly and accurately received, and plays a very important role in vision. However, the degeneration of the macula due to various causes causes vision impairment, and is called macular degeneration. Macular degeneration is a disease caused by gradual degeneration of retinal epithelial cells, retina, and choriocapillary layer. In the early stages of macular degeneration, abnormal retinal epithelial cells with extracellular deposits called drusen are observed between Bruch's membrane and retinal epithelial cells. In the later stages, it is largely divided into exudative macular degeneration and atrophic macular degeneration. In both of these two types of macular degeneration, the photoreceptors in the macula are gradually destroyed, causing a decline in the function of the macula over time and a decline in central vision. Oxidative stress and inflammation are considered to be the main causes of macular degeneration, and in particular, A2E molecules, which are by-products of intracellular metabolism, gradually accumulate in retinal pigment epithelial cells without being excreted, and then undergo an oxidative stress process, ultimately leading to cell death (Sparrow and Cai, 2001). Therefore, it is known that blocking the accumulation of the causative substance, A2E molecules, can prevent oxidation in retinal cells and reduce the risk of macular degeneration itself.

[0004] Glaucoma and macular degeneration, along with diabetic retinopathy, are one of the three major causes of blindness, and the main cause is the death of optic nerve cells or retinal nerve cells. It is known that the number of patients with these two diseases is rapidly increasing not only among the elderly but also among people in their 30s to 50s due to the recent increase in the prevalence of metabolic diseases.

[0005] In recent years, there has been active development of pharmaceuticals and functional health foods for maintaining eye health, such as preventing oxidative damage to the eyes. In particular, energy-improving agents have been developed whose main ingredient is anthocyanoside derived from wild blueberries. Herbal medicines such as Cassia berries and Wolfberry berries have long been used to maintain and improve eyesight. However, there has been no research on methods of using Centella asiatica to prevent or treat glaucoma and retinal macular degeneration, which are related to eye health. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] Korean Patent No. 10-1700903 [Patent Document 2] Korean Patent Publication No. 10-2018-0080584 Summary of the Invention [Problem to be solved by the invention]

[0007] Therefore, the present inventors recognized the importance of prevention or treatment of eye health such as glaucoma and macular degeneration, and selected natural resources that can promote proliferation of retinal nerve cells and pigment epithelial cells and protect retinal cells from oxidative damage using various natural materials. As a result, Centella asiatica extract showed excellent proliferation efficacy in optic nerve cells and retinal cells, and in particular inhibited oxidative damage caused by A2E in retinal cells. In addition, it significantly increased sugar availability in retinal cells, which can be inferred to have excellent recovery and protection effects from cell damage, so the Centella asiatica extract of the present invention has a high possibility of being commercialized as a composition for preventing, improving, or treating glaucoma and macular degeneration.

[0008] Therefore, an object of the present invention is to provide a pharmaceutical composition comprising a Centella asiatica extract for preventing, ameliorating or treating glaucoma or macular degeneration.

[0009] Another object of the present invention is to provide a health functional food composition containing Centella asiatica extract for preventing and improving glaucoma and macular degeneration.

[0010] The object of the present invention is not limited to the above-mentioned object. The object of the present invention will become clearer from the following description and can be realized by the means and combinations thereof described in the claims. [Means for solving the problem]

[0011] One aspect of the present invention provides a food composition for protecting retinal nerve cells, which comprises an extract of Centella asiatica as an active ingredient, for protecting retinal nerve cells from oxidative stress.

[0012] In one aspect of the present invention, the Centella asiatica extract provides a food composition that improves the survival rate of retinal nerve cells or retinal pigment epithelial cells by increasing sugar utilization.

[0013] In one aspect of the present invention, there is provided a food composition, wherein the Centella asiatica extract suppresses apoptosis of retinal nerve cells.

[0014] In one aspect of the present invention, the Centella asiatica extract provides a food composition that prevents death of retinal nerve cells from oxidative stress caused by A2E.

[0015] In one aspect of the present invention, the food composition provides a food composition that prevents or ameliorates retinal neuronal cell damage diseases caused by oxidative stress.

[0016] Another aspect of the present invention provides a pharmaceutical composition for preventing, ameliorating or treating retinal neuronal cell damage caused by oxidative stress, comprising an extract of Centella asiatica as an active ingredient.

[0017] In another aspect of the present invention, there is provided a pharmaceutical composition, wherein the extract of Centella asiatica improves the survival rate of retinal nerve cells or retinal pigment epithelial cells by increasing sugar utilization.

[0018] In another aspect of the present invention, there is provided a pharmaceutical composition, wherein the Centella asiatica extract inhibits apoptosis of retinal nerve cells.

[0019] In another aspect of the present invention, there is provided a pharmaceutical composition, wherein the Centella asiatica extract prevents death of retinal nerve cells from oxidative stress caused by A2E.

[0020] In another aspect of the present invention, there is provided a pharmaceutical composition, wherein the disease is macular degeneration or glaucoma.

[0021] In one or another aspect of the present invention, there is provided a composition, wherein the Centella asiatica extract is an extract extracted with water, a C1-6 lower alcohol, or a mixed solvent thereof.

[0022] In one or another aspect of the present invention, there is provided a composition, wherein the Centella asiatica extract is an extract from a 30% to 90% aqueous ethanol solution.

[0023] In one or another aspect of the present invention, there is provided a composition, wherein the Centella asiatica extract is administered at a dosage of 0.004 to 40 mg / kg / day.

[0024] In another aspect of the invention, a composition is provided which is a formulation for topical administration.

[0025] In another aspect of the invention, there is provided a composition which is an ophthalmic formulation.

[0026] In one or another aspect of the present invention, there is provided a composition, wherein the Centella asiatica extract is contained in the composition at a concentration of 5 to 500 μg / ml based on the total weight of the composition. Effect of the Invention

[0027] The Centella asiatica extract of the present invention can increase glucose metabolism, effectively proliferate retinal nerve cells and pigment epithelial cells, and protect retinal cells from oxidative damage caused by A2E, and therefore can improve, prevent, or treat eye diseases caused by damage to retinal nerve cells and pigment epithelial cells, namely, glaucoma and macular degeneration.

[0028] The effects of the present invention are not limited to those mentioned above, and should be understood to include all effects that can be inferred from the following description. [Brief description of the drawings]

[0029] [Figure 1] FIG. 1 is a diagram showing the cell growth efficacy of Centella asiatica extracts according to different extraction solvents of the present invention in retinal nerve cells. [Diagram 2] FIG. 1 shows the cell viability of retinal neurons and pigment epithelial cells treated with the composition of the present invention. [Diagram 3] FIG. 1 shows the glucose metabolic efficacy of treatment with the composition of the present invention in retinal pigment epithelial cells. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0030] Unless otherwise specified, all numbers, values ​​and / or expressions expressing ingredients, reaction conditions, and amounts of ingredients used in this specification should be understood to be modified in each case by the term "about" because those numerical values ​​are approximations that reflect various uncertainties in measurement that arise in obtaining such values ​​from inherently different things. Also, when numerical ranges are disclosed in this description, such ranges are continuous and include all values ​​from the minimum value to the maximum value inclusive, unless otherwise specified. Furthermore, when such ranges refer to integers, all integers from the minimum value to the maximum value inclusive, unless otherwise specified.

[0031] When a range is recited herein for a variable, the variable should be understood to include all values ​​within the stated range, including the endpoints recited in the range. For example, the range "5 to 10" will be understood to include the values ​​5, 6, 7, 8, 9, and 10, as well as any subranges such as 6 to 10, 7 to 10, 6 to 9, 7 to 9, and any value between the integers appropriate within the stated range, such as 5.5, 6.5, 7.5, 5.5 to 8.5, and 6.5 to 9. Also, for example, the range "10%-30%" will be understood to include all integers up to and including 30%, including values ​​such as 10%, 11%, 12%, 13%, etc., as well as any subranges such as 10%-15%, 12%-18%, 20%-30%, and any value between reasonable integers within the stated range, such as 10.5%, 15.5%, 25.5%, etc.

[0032] The present invention will be described in detail below.

[0033] The present invention relates to a composition for preventing, improving or treating glaucoma and macular degeneration, comprising a Centella asiatica extract. Specifically, the composition can be usefully used in a composition for preventing, improving or treating glaucoma and macular degeneration for eye health by increasing glucose metabolism, inducing proliferation of retinal nerve cells and pigment epithelial cells, and protecting cells from oxidative stress caused by A2E.

[0034] Centella asiatica is a perennial creeping herb belonging to the family Umbelliferae, which grows wild mainly in hot and humid regions and has been used in traditional Chinese medicine for skin diseases, antipyretics, hemoptysis, diuretics, tonics, impotence, leucorrhea, arthritis, etc. (Lee, Young-no, 2006). The main known components of Centella are triterpenic acid sugar esters, and asiaticoside, madecassoside, brahmoside, brahminoside, etc. have been reported, and it is known that asiatic acid, madecassic acid, thanksunic acid, isothankunic acid, etc. are produced when hydrolyzed. Asiaticoside and madecassoside, which are pentacyclic triterpene glycosides belonging to the α-amyrin-ursolic acid group as the main components of Centella asiatica, have been used for treating skin wounds and chronic ulcers since ancient times, and have been reported to have therapeutic and antibacterial effects for leprosy, wounds, gastric ulcers, various skin diseases, mental diseases, tuberculosis, venous diseases, dementia, etc. Due to these effects, Centella asiatica is used for various purposes such as food, skin treatment agent, wound treatment agent, memory enhancer, and tonic, and is used in the fields of medicine, cosmetics, functional foods, etc. in Korea, but no published literature has revealed that Centella asiatica extract and its main components are effective in preventing, improving, or treating glaucoma and macular degeneration.

[0035] The present invention provides a pharmaceutical composition for preventing or treating glaucoma and macular degeneration, comprising an extract of Centella asiatica, said composition being in the form of a liquid extract or a lyophilized powder of said liquid extract.

[0036] The present invention also provides a health functional food composition for preventing and improving glaucoma and macular degeneration, comprising Centella asiatica extract, which may be in the form of a liquid extract or a freeze-dried powder of the liquid extract.

[0037] Various aspects of the invention are described below.

[0038] One aspect of the present invention provides a food composition for protecting retinal nerve cells, which comprises an extract of Centella asiatica as an active ingredient, for protecting retinal nerve cells from oxidative stress.

[0039] The functional health food composition according to the present invention contains Centella asiatica extract as an active ingredient and can be ingested for the purpose of improving eyesight by inhibiting glaucoma and macular degeneration. The active ingredient can be ingested as a food manufactured as a tablet, capsule, powder, granule, pill, liquid, suspension, etc., or can be added to a general food. Unlike general medicines, the functional health food uses food as a raw material, and therefore has the advantage of being free of side effects that may occur when taking medicines for a long period of time. The content of the active ingredient can be suitably determined depending on the purpose of use (prevention or improvement). In general, the functional health food composition can contain 0.1 to 90% by weight of the active ingredient. However, for long-term intake for the purpose of health and hygiene or health regulation, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be more than the above range.

[0040] The type of the food is not particularly limited. Examples of the food to which the active ingredient can be added include drinks, meat, sausages, bread, biscuits, mochi, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, drinking water, alcoholic beverages, vitamin complexes, dairy products and dairy products, etc., including any health functional food in the ordinary sense. The nature of the food is also not particularly limited, and may be any of solid, semi-solid, gel, liquid, and powder.

[0041] Beverages can be made using the health functional food composition of the present invention. There is no particular restriction on the selection of ingredients other than the active ingredient as ingredients contained in the beverage, and various flavoring agents or natural carbohydrates can be contained as additional ingredients, similar to ordinary beverages. Examples of the above-mentioned natural carbohydrates include monosaccharides, such as glucose, fructose, etc.; disaccharides, such as maltose, sucrose, etc.; and polysaccharides, such as ordinary sugars such as dextrin, cyclodextrin, etc., and sugar alcohols such as xylitol, sorbitol, erythritol, etc. As flavoring agents other than those mentioned above, natural flavoring agents (thaumatin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The ratio of the natural carbohydrates is usually about 1 to 20 g, preferably about 5 to 12 g, per 100 g of the composition of the present invention.

[0042] In addition to the active ingredients, the health functional food composition of the present invention may contain various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and enhancers (cheese, chocolate, etc.), pectinic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohol, carbonation agents used in carbonated drinks, etc. as additives. In addition, fruit pulp for the production of natural fruit juice, fruit juice beverages, and vegetable beverages may be contained. These ingredients may be used independently or in combination. The ratio of such additives is not very important, but is generally selected in the range of 0.1 to 20% by weight in the health functional food composition of the present invention.

[0043] In one aspect of the present invention, the Centella asiatica extract provides a food composition that improves the survival rate of retinal nerve cells or retinal pigment epithelial cells by increasing sugar utilization.

[0044] In one aspect of the present invention, there is provided a food composition, wherein the Centella asiatica extract suppresses apoptosis of retinal nerve cells.

[0045] In one aspect of the present invention, the Centella asiatica extract provides a food composition that prevents death of retinal nerve cells from oxidative stress caused by A2E.

[0046] In one aspect of the present invention, the food composition provides a food composition that prevents or ameliorates retinal neuronal cell damage diseases caused by oxidative stress.

[0047] Another aspect of the present invention provides a pharmaceutical composition for preventing, ameliorating or treating retinal neuronal cell damage caused by oxidative stress, comprising an extract of Centella asiatica as an active ingredient.

[0048] In another aspect of the present invention, there is provided a pharmaceutical composition, wherein the extract of Centella asiatica improves the survival rate of retinal nerve cells or retinal pigment epithelial cells by increasing sugar utilization.

[0049] In another aspect of the present invention, there is provided a pharmaceutical composition, wherein the Centella asiatica extract inhibits apoptosis of retinal nerve cells.

[0050] In another aspect of the present invention, there is provided a pharmaceutical composition, wherein the Centella asiatica extract prevents death of retinal nerve cells from oxidative stress caused by A2E.

[0051] In another aspect of the present invention, there is provided a pharmaceutical composition, wherein the disease is macular degeneration or glaucoma.

[0052] When the pharmaceutical composition is used clinically, it can be formulated with a carrier commonly used in the pharmaceutical field to be formulated into various preparations such as oral preparations such as tablets, capsules, powders, granules, pills, liquids and suspensions; injectable preparations in the form of solutions or suspensions for injection, or ready-to-use dry powders for injection that can be prepared with distilled water for injection at the time of injection; or ointments. Pharmaceutical preparations made commercially using commonly used carriers can be administered orally or parenterally, for example, intravenously, subcutaneously, intraperitoneally or topically. Therefore, the pharmaceutical composition of the present invention can further contain suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceuticals.

[0053] Carriers, excipients and diluents that may be included in the pharmaceutical composition of the present invention may include one or more selected from lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, hydroxymethylcellulose, uncrystalline cellulose, silicified uncrystalline cellulose, povidone, crospovidone, croscarmellose sodium, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, neusilin, colloidal silicon dioxide, lactose, talc, magnesium stearate, colloidal magnesium stearyl, mineral oil, and the like.

[0054] When the composition is formulated, it is prepared using a diluent or excipient such as a filler, extender, binder, wetting agent, disintegrant, or surfactant. Solid preparations for oral administration include tablets, pills, powders, granules, troches, lozenges, capsules, etc., and such solid preparations are prepared by mixing the composition of the present invention with at least one excipient, such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose, calcium carbonate, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, tolerant solutions, emulsions, elixirs, syrups, etc., and may contain various excipients such as wetting agents, sweeteners, flavorings, preservatives, etc. in addition to water and liquid paraffin, which are commonly used simple diluents. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspensions may be used, such as propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases may be used, such as witepsol, macrogol, tween 61, cacao butter, laurin butter, and glycerol gelatin. Parenteral administration may generally be by subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection.

[0055] The preferred dosage of the pharmaceutical composition of the present invention varies depending on the age, weight, severity of disease, drug form, administration route and period of the patient, but can be appropriately selected by those skilled in the art. However, for a desirable effect, the pharmaceutical composition of the present invention may be administered at 0.01 mg / kg to 10 g / kg, preferably 1 mg / kg to 1 g / kg per day. The dosage can be administered several times a day at regular intervals, preferably once or six times in divided doses, according to the judgment of a doctor or pharmacist.

[0056] In one embodiment, the composition is a natural product that is routinely ingested and taken and can be safely used to prevent, ameliorate or treat glaucoma and macular degeneration.

[0057] The Centella asiatica extract may be one extracted by a conventional extraction method, and may be an extract that has been subjected to reduced pressure drying and freeze-drying to be in a powder form.

[0058] In one or another aspect of the present invention, there is provided a composition, wherein the Centella asiatica extract is an extract extracted with water, a C1-6 lower alcohol, or a mixed solvent thereof.

[0059] The composition for preventing, improving or treating glaucoma and macular degeneration having the above-mentioned composition of the present invention may be prepared by repeating the above-mentioned extraction process as necessary, and an extract obtained by repeating the extraction 2 to 5 times may be used. Also, the extract may be freeze-dried to prepare a powdered dry extract, which may be used in the composition for preventing, improving or treating glaucoma and macular degeneration.

[0060] The content of the alcohol in the extraction solvent is preferably maintained at a concentration of 0 to 95% by weight, more preferably 30 to 95% by weight, and more preferably 50% by weight, from the viewpoint of extraction efficiency. Any alcohol commonly used in the art can be used as the alcohol, and preferably an alcohol having 1 to 5 carbon atoms, and the alcohol may be one or more selected from methanol, ethanol, isopropanol, propanol, and butanol. More preferably, ethanol, spirits, etc. can be used as the alcohol.

[0061] In one or another aspect of the present invention, there is provided a composition, wherein the Centella asiatica extract is an extract from a 30% to 90% aqueous ethanol solution.

[0062] The extraction method is a method commonly known in the art, and may be, for example, a method using an extraction device such as supercritical extraction, subcritical extraction, high temperature extraction, high pressure extraction or ultrasonic extraction, or a method using an adsorption resin including XAD and HP-20.

[0063] The extract is concentrated under reduced pressure using a vacuum rotary evaporator or the like to obtain an extract. The obtained extract may be dried under reduced pressure, vacuum dried, boiled dried, spray dried, dried at room temperature, or freeze-dried, if necessary. In particular, the freeze-drying method has the advantage of reducing the loss of volatile organic substances in the extract.

[0064] In one aspect of the present invention, there is provided a composition, wherein the Centella asiatica extract is an extract from a 40% to 60% aqueous ethanol solution.

[0065] In one aspect of the present invention, the composition is characterized by promoting the proliferation of optic nerve cells and retinal cells in ocular health or protecting cells from oxidative stress.

[0066] In one or another aspect of the present invention, there is provided a composition, wherein the Centella asiatica extract is administered at a dosage of 0.01 mg to 10 g / kg / day.

[0067] In another aspect of the invention, a composition is provided, wherein the composition is a formulation for topical administration.

[0068] In another aspect of the invention, there is provided a composition, wherein said composition is an ophthalmic formulation.

[0069] In one or another aspect of the present invention, there is provided a composition, wherein the Centella asiatica extract is contained in the composition at a concentration of 5 to 500 μg / ml based on the total weight of the composition.

[0070] The composition effectively proliferates retinal nerve cells and pigment epithelial cells and protects the cells from oxidative stress. The composition is a natural product that is ingested or taken on a daily basis and can be safely used to prevent, improve or treat glaucoma and macular degeneration.

[0071] The Centella asiatica extract may be one extracted by a conventional extraction method, and may be an extract that has been subjected to reduced pressure drying and freeze-drying to be in a powder form.

[0072] In one aspect of the present invention, there is provided a composition, wherein the extract is a water extract, a C1 to C5 alcohol extract, or a C1 to C5 aqueous alcohol extract.

[0073] The composition for preventing, improving or treating glaucoma and macular degeneration according to the present invention may be prepared by repeating the extraction process as necessary, and an extract obtained by repeating the extraction 2 to 5 times may be used. The extract may also be freeze-dried to prepare a powdered dry extract, which may be used in the composition for preventing, improving or treating glaucoma and macular degeneration.

[0074] The content of the alcohol in the extraction solvent is preferably maintained at a concentration of 0 to 95% by weight, more preferably 30 to 95% by weight, and more preferably 50% by weight, from the viewpoint of extraction efficiency. Any alcohol commonly used in the field can be used as the alcohol, and preferably an alcohol having 1 to 5 carbon atoms, and the alcohol can be one or more selected from methanol, ethanol, isopropanol, propanol, and butanol. More preferably, the alcohol can be ethanol, spirit, or the like.

[0075] The extraction method may be a method commonly used in the art, such as a method using an extraction device for supercritical extraction, subcritical extraction, high temperature extraction, high pressure extraction or ultrasonic extraction, or a method using an adsorption resin including XAD and HP-20.

[0076] The extract is concentrated under reduced pressure using a vacuum rotary evaporator or the like to obtain an extract. The obtained extract can be dried under reduced pressure, vacuum dried, boiled dried, spray dried, dried at room temperature, or freeze-dried, if necessary. In particular, the freeze-drying method has the advantage of reducing the loss of volatile organic substances in the extract.

[0077] The Centella asiatica extract obtained by the above-mentioned method increases cellular glucose metabolism, efficiently increases the proliferation of optic nerve cells and retinal cells, and effectively inhibits cell death due to oxidative stress caused by A2E.

[0078] In one aspect of the present invention, there is provided a composition, wherein the Centella asiatica extract is an extract from a 40% to 60% aqueous ethanol solution.

[0079] [Mode for carrying out the invention] The present invention will be described in more detail below with reference to specific examples. The following examples are merely illustrative to aid in understanding the present invention, and are not intended to limit the scope of the present invention.

[0080] [Examples and Comparative Examples]

[0081] Manufacturing example: Manufacturing of Centella asiatica extract

[0082] Centella asiatica was purchased from a farmer in Hobucheon, Gyeongsangnam-do, Korea, dried in the shade, and the whole plant was crushed. 1 kg of the crushed Centella was extracted with 50% alcohol at 80℃ for 5 hours, then concentrated and freeze-dried to obtain 135 g of Centella extract (yield 13.5%).

[0083] Comparative Examples 1 to 4

[0084] As comparative examples, the hot water extract (Comparative Example 1), 30% (Comparative Example 2), 70% (Comparative Example 3) and 95% (Comparative Example 4) alcohol extracts of Centella asiatica were used.

[0085] [Experimental Example]

[0086] Experimental Example 1: Measurement of proliferation effect of retinal nerve cells and retinal pigment epithelium cells

[0087] The Centella asiatica extracts obtained in the above Preparation Examples and the Comparative Examples were measured for their proliferation effects on retinal nerve cells in order to select the most effective extract.

[0088] RGC-5 retinal nerve cells were cultured in a DMEM medium containing 10% serum and antibiotics in an incubator maintained at 37°C with 5% carbon dioxide by volume. GH3 cells were dispensed into a 96-well plate at a concentration of 5x103cells / well and cultured for 24 hours. To measure the retinal nerve cell proliferation effect of Centella asiatica extract, the medium was replaced with DMEM medium containing 10% charcoaled FBS, and the Centella asiatica extracts prepared in Preparation Example and Comparative Examples 1 to 4 were treated at 100μg / mL. After 48 hours, the proliferation rate of RGC-5 cells was measured at an absorbance of 550nm using MTT reagent, and the absorbance of each test solution was measured based on the control group to show the cell proliferation rate (%).

[0089] The retinal neuronal cell proliferation rate of Centella asiatica extract is shown in FIG.

[0090] As shown in FIG. 1, the 50% Centella asiatica alcohol extract, which is the Production Example, showed the most excellent retinal nerve cell proliferation efficacy, and significant proliferation efficacy was also observed from Comparative Examples 3 and 4.

[0091] Meanwhile, in order to measure the protective effect of the 50% alcohol extract of Centella asiatica, which has the best proliferation effect, on retinal nerve cells and retinal pigment epithelial cells, the proliferation effect of the retinal cells was measured.

[0092] RGC-5 retinal nerve cells were cultured in the same manner as described above, and ARPE-19 retinal pigment epithelial cells were cultured in DMEM / F12 medium containing 10% serum and antibiotics in an incubator maintained at 37°C with 5% carbon dioxide by volume. RGC-5 cells and ARPE-19 cells were distributed into a 96-well plate at a concentration of 5x103 cells / well and cultured for 24 hours. To measure the proliferation effect of Centella asiatica extract on retinal nerve cells and retinal pigment epithelial cells, the medium was replaced with DMEM medium and DMEM / F12 medium containing 10% charcoaled FBS, and then Centella asiatica 50% alcohol extract was treated at 50μg / mL, 100μg / mL, and 200μg / mL, respectively. After 48 hours, the cell proliferation rate of each was measured at 550nm using MTT reagent, and the absorbance of each test solution was measured based on the control group to show the cell proliferation rate (%).

[0093] The proliferation rates of retinal nerve cells and retinal pigment epithelium cells of the 50% alcohol extract of Centella asiatica are shown in FIG.

[0094] As shown in FIG. 2, the 50% Centella asiatica alcohol extract, which is a production example, showed very excellent proliferation efficacy at a concentration of 100 μg / mL or more.

[0095] Experimental Example 2: Measurement of glucose metabolism efficiency in retinal pigment epithelial cells

[0096] In order to study the glucose uptake efficacy of the 50% alcohol extract of Centella asiatica, which is the above-mentioned Preparation Example, in retinal pigment epithelial cells, a 2-NBDG uptake experiment was carried out.

[0097] The test cells used were retinal pigment epithelial cells, ARPE-19, and the medium, medium composition, and experimental conditions were the same as those in Experimental Example 1.

[0098] ARPE-19 cells were plated in a 96-well plate at a concentration of 1×104 cells / well and cultured for 24 hours. To measure glucose metabolism efficacy, the cells were treated with 50% Centella asiatica alcohol extract at 25μg / mL, 50μg / mL, and 100μg / mL, cultured for 24 hours, and then treated with serum-free DMEM / F12 medium and 100μM 2-NBDG for 30 minutes. After washing twice with cold phosphate buffered saline (PBS), glucose uptake was measured using a fluorometer (excitation 485nm; emission 525nm).

[0099] The glucose uptake efficacy of the disease-causing herb extract is shown in FIG.

[0100] As shown in FIG. 3, the Centella asiatica extract showed a very excellent glucose metabolism effect at a concentration of 100 μg / mL, which can be attributed to the retinal cell proliferation effect of the Centella asiatica extract.

[0101] Experimental Example 3. Cytoprotective effect of A2E

[0102] As part of the macular degeneration inhibitory effect, the inhibitory effect of A2E on cell death was measured.

[0103] The test cells used were retinal pigment epithelial cells, ARPE-19, and the medium, medium composition, and experimental conditions were the same as those in Experimental Example 1.

[0104] ARPE-19 cells were plated in a 24-well plate at a concentration of 3x104cells / well and cultured for 24 hours, after which A2E was added at a concentration of 3μM for 3 days. To measure the cell protection effect against oxidative stress caused by A2E, the cells were treated with 50% Centella asiatica alcohol extract at 25μg / m, 50μg / m, and 100μg / mL, and as a positive control, lutein, known to have an inhibitory effect on macular degeneration, was treated at 30μM and cultured for 48 hours. After 48 hours, the cells were irradiated with UV light for 5 minutes using a UV lamp. The cell growth rate was measured at an absorbance of 570nm using MTT reagent, and the absorbance of each test solution was measured and compared to that of a control group not treated with A2E.

[0105] The cell protective effect of the Centella asiatica extract against oxidative stress induced by A2E is shown in Table 1.

[0106] [Table 1] TIFF0007648529000001.tif41143

[0107] As shown in Table 1, 100 μg / mL of the 50% Centella asiatica alcohol extract showed about 16% cell survival effect compared to the control group, and protected cells from oxidative damage caused by A2E to the same level as lutein, which is a positive control group. Therefore, the present invention can provide a composition for preventing, improving or treating glaucoma and macular degeneration for eye health, which contains Centella asiatica extract.

[0108] Although the embodiments of the present invention have been described above with reference to the accompanying drawings, those skilled in the art will understand that the present invention can be embodied in other specific forms without changing the technical concept or essential features of the present invention. Therefore, the above-described embodiments should be understood to be illustrative and not restrictive in any respect.

Claims

1. A food composition for protecting retinal nerve cells from oxidative stress, comprising an extract of Centella asiatica as an active ingredient, wherein the extract of Centella asiatica is an extract of 50% to 60% aqueous ethanol solution.

2. The food composition according to claim 1, wherein the Centella asiatica extract improves the viability of retinal nerve cells or retinal pigment epithelial cells by increasing sugar utilization.

3. The food composition according to claim 1, wherein the extract of Centella asiatica inhibits apoptosis of retinal nerve cells.

4. The food composition according to claim 1, wherein the Centella asiatica extract prevents death of retinal nerve cells from oxidative stress caused by A2E.

5. The food composition according to claim 1 , which prevents or ameliorates retinal neuronal cell damage diseases caused by oxidative stress.

6. A pharmaceutical composition for preventing, improving or treating retinal nerve cell damage caused by oxidative stress, comprising an extract of Centella asiatica as an active ingredient, wherein the extract of Centella asiatica is an extract of 50% to 60% aqueous ethanol solution.

7. The pharmaceutical composition according to claim 6, wherein the Centella asiatica extract improves the survival rate of retinal nerve cells or retinal pigment epithelial cells by increasing sugar utilization.

8. The pharmaceutical composition according to claim 6, wherein the extract of Centella asiatica inhibits apoptosis of retinal nerve cells.

9. The pharmaceutical composition according to claim 6, wherein the Centella asiatica extract prevents death of retinal nerve cells from oxidative stress caused by A2E.

10. The pharmaceutical composition of claim 6, wherein the disease is macular degeneration or glaucoma.

11. The composition according to any one of claims 1 to 10, wherein the Centella asiatica extract is administered at a dosage of 0.01 mg / kg / day to 10 g / kg / day.

12. The pharmaceutical composition according to any one of claims 6 to 10, which is a preparation for topical administration.

13. The pharmaceutical composition of claim 12 which is an ophthalmic formulation.

14. The composition according to any one of claims 1 to 10, wherein the Centella asiatica extract is contained in the composition at a concentration of 5 to 500 µg / ml based on the total weight of the composition.

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