Composition for maintaining tear film stability and improving dry eye, and its application

Abstract

A plant-based composition characterized in that the plant-based composition comprises goji berry extract, blueberry extract and lutein esters, wherein the weight ratio of goji berry extract to blueberry extract is (10-100):1.

Description

Technical field

[0001] The present application concerns the field of foodstuffs and relates in particular to a composition for maintaining tear film stability and improving dry eyes, as well as its application. State of the art

[0002] The cornea is a transparent, thin layer at the front of the eye. Completely transparent, it lies at the front of the eyeball and has a transverse oval shape. The cornea contains highly sensitive nerve endings. When a foreign object touches the cornea, the eyelid closes involuntarily to protect the eye. To maintain its transparency, the cornea has no blood vessels and receives nutrients and oxygen from the tear film and aqueous humor. However, instability of the tear film and damage to the ocular surface, caused by abnormalities in the quality or quantity of tear fluid or by hydrodynamic abnormalities, can produce symptoms such as dry eyes and trigger dry eye syndrome. Alterations in the barrier function of the corneal epithelium are the cause of visual disturbances and eye discomfort in dry eye.According to relevant data statistics, the incidence of dry eye worldwide ranges from approximately 5.5% to 33.7%. This condition causes symptoms of eye discomfort, visual disturbances, and potentially harmful instability of the tear film on the ocular surface, accompanied by an increase in tear film osmolarity and inflammation of the ocular surface. Dry eye symptoms are influenced by a combination of factors, including tear production volume, tear evaporation rate, and the presence of inflammation. The pathological core of dry eye is often associated with damage to ocular surface tissues due to oxidative stress and inflammation, as well as with meibomian gland dysfunction. Gelatinase B, an inflammatory marker, is continuously elevated in the tear fluid of patients with dry eye.Gelatinase B levels in patients with moderate to severe dry eye correlate with clinical examination findings. Therefore, further research on gelatinase B can help us understand the degree of dry eye treatment and achieve the best possible treatment outcomes for patients. It is against this background that the present application is submitted. The invention and its advantages

[0003] One or more embodiments of the present application provide a plant-based composition and its application. The plant-based composition can improve gelatinase B levels and prevent or treat dry eye.

[0004] One or more embodiments of the present application provide a plant composition comprising goji berry extract and blueberry extract in a weight ratio of (10-100):1 as well as lutein esters.

[0005] In some implementations of the present application, the herbal composition comprises goji berry extract and blueberry extract in a weight ratio of (10-50):1 as well as lutein esters.

[0006] In some implementations of the present application, the herbal composition comprises goji berry extract and blueberry extract in a weight ratio of (30-50):1 as well as lutein esters.

[0007] One or more embodiments of the present application further provide for the use of the plant composition in the manufacture of a product for improving dry eye.

[0008] One or more embodiments of the present application further provide a product for improving dry eye, wherein the product for improving dry eye comprises the herbal composition as well as food-compatible excipients.

[0009] In some implementations of the present application, the dosage form of the product for improving dry eye includes an oral dosage form.

[0010] In some implementations of the present application, the product for improving dry eye includes at least one type of liquid drink, tablet, capsule, powder, jelly and candy.

[0011] One or more embodiments of the present application further provide for the use of the plant composition in the manufacture of a product for improving the gelatinase B level.

[0012] One or more embodiments of the present application further provide a product for improving the gelatinase B level, wherein the product for improving the gelatinase B level comprises the plant composition as well as food-compatible excipients.

[0013] In some implementations of the present application, the dosage form of the product for improving gelatinase B levels includes an oral dosage form.

[0014] In some implementations of the present application, the product for improving gelatinase B levels comprises at least one type of liquid beverage, tablet, capsule, powder, jelly and candy.

[0015] Compared to conventional technology, the present application has the following advantageous effects:

[0016] Lutein esters are a precursor form of lutein (usually extracted from plants such as marigolds) which, after oral administration, is enzymatically hydrolyzed in the human intestine, converted into bioactive free lutein, and then absorbed and selectively accumulates in ocular tissue. The applicant has stated that lutein can contribute to maintaining tear film stability and improving dry eye symptoms through two key processes: antioxidant protection against photooxidative damage and suppression of inflammatory responses.

[0017] The present application uses lutein esters, goji berry extract and blueberry extract as main active ingredients, the latter two being combined in a specific ratio to achieve the regulation of gelatinase B levels in corneal epithelial cells, thus offering a new approach to the prevention and treatment of dry eye. Brief description of the drawings

[0018] To more clearly explain the technical solutions in the embodiments of the present application and to provide a more comprehensive understanding of the present application and its advantageous effects, the drawings used in the description of the embodiments are briefly presented below. It is obvious that the drawings listed in the following description represent only some embodiments of the present application. A person skilled in the art in this field can obtain further drawings based on these drawings without any inventive step. Fig. Figure 1 shows the results of the cytotoxicity test of sodium chloride solution using the CCK-8 method; Fig.Figure 2 shows the results of the improving effect of the herbal composition on the cell model of dry eye; in the figure, “#” indicates a significant difference compared to the control group, and “*” indicates a significant difference compared to the model group. Example(s) of implementation

[0019] The present application is explained in more detail below with reference to the drawings, implementations, and embodiments. It is understood that these implementations and embodiments serve only to illustrate the present application and not to limit its scope. The purpose of providing these implementations and embodiments is to enhance and deepen the understanding of the disclosed content of the present application. It should also be understood that the present application can be realized in many different forms and is not limited to the implementations and embodiments described in this document.Experts in this field may make various changes or modifications without altering the essence of the present application, and the resulting equivalent forms shall also fall within the scope of protection of the present application. Furthermore, numerous specific details are provided in the following description to facilitate a more comprehensive understanding of the present application. It should be understood that the present application can also be implemented without one or more of these details.

[0020] Unless otherwise defined, all technical and scientific terms used in this document have the same meaning as they are generally understood by those skilled in the technical field of the present application. The terms used in the description of the present application in this document serve only to describe the implementations and embodiments and are not intended to limit the present application. terms

[0021] Unless otherwise stated or in case of contradictions, the terms or phrases used in this document have the following meanings:

[0022] The scope of the terms “and / or”, “or / and”, and “as well as / or” used in this document includes each of the two or more listed relevant points, as well as any and all combinations of the listed relevant points. The aforementioned “any” and “all” combinations include any two of the listed relevant points, any number of the listed relevant points, or the combination of all listed relevant points. It should be noted that when at least three points are connected by a combination of at least two conjunctions such as “and / or”, “or / and”, or “as well as / or”, in the present application, this technical solution naturally includes the technical solutions that are all connected by a “logical AND”, and also naturally includes the technical solutions that are all connected by a “logical OR”.For example, "A and / or B" includes the three parallel solutions A, B, and A+B. Another example: The technical solution "A, and / or, B, and / or, C, and / or, D" includes each individual point from A, B, C, D (i.e., the technical solutions all connected by "logical OR"), also includes any and all combinations of A, B, C, D, i.e., includes the combination of any two or any three points from A, B, C, D, and also includes the combination of all four points A, B, C, D (i.e., the technical solutions all connected by "logical AND").

[0023] Insofar as the present application refers to “several”, “several types”, “multiple”, “several elements”, etc., this refers, unless specifically specified, to a quantity greater than or equal to 2. For example, “one or more types” means one type or greater than or equal to two types.

[0024] The expressions used in this document, such as "their combination", "any combination", "any combination", etc., include all suitable combinations of any two or more than two of the listed items.

[0025] In this document, the word "suitable" in expressions such as "suitable combination", "suitable way", "any suitable way", etc., refers to the fact that the technical solution of the present application can be implemented, the technical problem of the present application can be solved, and the expected technical effect of the present application can be achieved.

[0026] In this document, terms such as "preferred," "better," "more advantageous," and "is suitable as" are used merely to describe implementations or embodiments with better results. It is understood that they do not constitute a limitation of the scope of protection of the present application.

[0027] In the present application, expressions such as "further", "even further", "in particular", etc. serve descriptive purposes and indicate differences in content, but should not be understood as a limitation of the scope of protection of the present application.

[0028] In the present application, "optional," "can be chosen," or "may be chosen" means that something may or may not be present; that is, it refers to the selection of one of the two parallel solutions, "present" or "not present." If several instances of "optional" appear in a technical solution, each "optional" indication is independent unless otherwise specified and there are no contradictions or mutual limitations.

[0029] In the present application, terms such as "first aspect", "second aspect", "third aspect", "fourth aspect", etc., are used for descriptive purposes only and should not be understood as indicating or suggesting a relative importance or quantity, nor should they implicitly indicate the importance or quantity of the specified technical features. Furthermore, "first", "second", "third", "fourth", etc., serve only the purpose of a non-exhaustive enumeration, and it is understood that they do not constitute a closed limitation of the quantity.

[0030] In the present application, openly described technical features include both closed technical solutions consisting of the listed features and open technical solutions containing the listed features.

[0031] In this application, unless otherwise specified, for numerical intervals (i.e., numerical ranges), the optional distributions of numbers within the said numerical interval are considered continuous, and they include the two endpoints of the numerical range (i.e., the minimum and maximum values) as well as every value between these two endpoints. Unless otherwise specified, if a numerical interval refers only to integers within that interval, it includes the two integer endpoints of the numerical range as well as every integer between these two endpoints. In this document, this is equivalent to directly listing every integer. For example, “t is an integer chosen from 1~10” means that t is any integer chosen from the group of integers consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10.Furthermore, if multiple areas are provided to describe features or properties, these areas can be combined. In other words, unless otherwise specified, the areas disclosed in this document should be understood to include all sub-areas contained therein.

[0032] Unless otherwise specified, the temperature parameters in this application permit both isothermal treatment and fluctuations within a defined temperature range. It is understood that the aforementioned isothermal treatment permits temperature fluctuations within the precision range of the instrument control. Fluctuations in ranges such as ±5°C, +4°C, ±3°C, ±2°C, and ±1°C are permitted.

[0033] In the present application, %(w / w) and wt% both denote weight percent, %(v / v) refers to volume percent, and %(w / v) refers to mass-volume percent.

[0034] All documents mentioned in the present application are cited in this application as if each document were cited individually. Unless they conflict with the purpose and / or technical solution of the present application, the documents cited in the present application are cited in full and for all purposes. When reference is made in the present application to documents, the definitions of the relevant technical features, terms, nouns, phrases, etc., in the cited documents are also cited. When reference is made in the present application to documents, examples and preferred embodiments of the cited relevant technical features may also be included in the present application as references, but only to the extent that they enable the implementation of the present application.It is understood that in the event of a conflict between the cited content and the description in the present application, the present application shall prevail or the cited content shall be adaptively corrected in accordance with the description in the present application.

[0035] The goji berry, also called "Ci" or "Gouqizi," in modern medicine usually refers to the red, dried fruit of the Solanaceae plant Lycium barbarum (Ningxia Goji). Traditional Chinese medicine considers goji berries to nourish the liver and kidneys and to clear the eyes. Using modern molecular analysis techniques, researchers have isolated various active components from goji berries, including goji berry polysaccharides, polyphenols, flavonoids, and carotenoids. Goji berry polysaccharides (Lycium barbarum polysaccharides, LBP) make up about 40% of the aqueous extract of dried goji berries and are the most important biologically active components of the goji berry. They possess various functions, such as anti-inflammatory and antioxidant effects.

[0036] Blueberries are a rich source of anthocyanins. Anthocyanins are plant polyphenolic compounds with strong antioxidant properties that can prevent chemical light damage to the retina; they promote blood flow in the microcirculation of the back of the eye, improve metabolism, and stimulate rhodopsin synthesis, thus relieving eye strain, improving vision, and protecting the retina. However, research on the effects of blueberries on dry eye is currently limited, and studies on the mechanism of action and active components of blueberries for dry eye are extremely rare.

[0037] Marigold extract: A natural dye produced from the flowers of the daisy plant Tagetes using a subcritical biotechnological process with low-temperature extraction, its main components being lutein and carotenoids. Common specifications of marigold extract contain 5%, 10%, and 20% lutein esters.

[0038] When harmful light, such as blue light from screens or ultraviolet radiation, enters the eye, it generates a large number of reactive oxygen species (ROS) on the eye's surface (cornea, conjunctiva) and in the meibomian glands. Lutein, a type of carotenoid, is a highly efficient antioxidant that can directly neutralize these free radicals. The meibomian glands are key structures for the secretion of the lipid layer (the "oil layer") of the tear film. Oxidative damage to their gland cells leads to abnormalities in the quality and quantity of lipid secretion, causing the tear fluid to evaporate too quickly. The antioxidant effect of lutein helps protect these gland cells and maintain normal lipid secretion function, thereby stabilizing the tear film and reducing evaporation.Oxidative stress activates inflammatory signaling pathways such as NF-κB on the ocular surface, leading to the release of pro-inflammatory factors (such as IL-1β, IL-6, and TNF-α). These inflammatory factors can damage goblet cells (which secrete the mucin layer of the tear film), injure the corneal epithelium, and exacerbate meibomian gland dysfunction. Studies have shown that lutein can downregulate the expression of these pro-inflammatory factors and reduce the inflammatory response on the ocular surface. A healthier ocular surface with lower levels of inflammation is beneficial for:

[0039] The normal secretion of mucin by goblet cells (to stabilize the inner layer of the tear film).

[0040] Maintaining the integrity of the corneal and conjunctival epithelium.

[0041] The normal function of the meibomian glands.

[0042] Ultimately, all these factors together promote the stability of the tear film and the moistening of the ocular surface.

[0043] The present application has established that the combination of lutein esters with goji berry extract and blueberry extract can lower gelatinase B levels in corneal epithelial cells. Based on this discovery, the present application offers a composition for maintaining tear film stability and improving dry eye symptoms and proposes the following technical solution: First aspect of the present application

[0044] The present application provides for a herbal composition comprising goji berry extract and blueberry extract in a weight ratio of (10-100):1, for example 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, 21:1, 22:1, 23:1, 24:1, 25:1, 26:1, 27:1, 28:1, 29:1, 30:1, 31:1, 32:1, 33:1, 34:1, 35:1, 36:1, 37:1, 38:1, 39:1, 40:1, 41:1, 42:1, 43:1, 44:1, 45:1, 46:1, 47:1, 48:1, 49:1, 50:1, 51:1, 52:1, 53:1, 54:1, 55:1, 56:1, 57:1, 58:1, 59:1, 60:1, 61:1, 62:1, 63:1, 64:1, 65:1, 66:1, 67:1, 68:1, 69:1, 70:1, 71:1, 72:1, 73:1, 74:1, 75:1, 76:1, 77:1, 78:1, 79:1, 80:1, 81:1, 82:1, 83:1, 84:1 85:1, 86:1, 87:1, 88:1, 89:1, 90:1, 91:1, 92:1, 93:1, 94:1, 95:1, 96:1, 97:1, 98:1, 99:1, 100:1, as well as lutein esters in a weight quantity not lower than that of the blueberry extract.

[0045] Optionally, the herbal composition includes goji berry extract and blueberry extract in a weight ratio of (10-50):1, as well as lutein esters in a weight quantity not less than that of the blueberry extract.

[0046] The herbal composition may optionally include goji berry extract and blueberry extract in a weight ratio of (30-50):1 as well as lutein esters in a weight quantity that is not lower than that of the blueberry extract. Second aspect of the present application

[0047] The present application offers the use of the plant composition in the manufacture of a product for improving dry eye.

[0048] In the present application, “improvement of dry eye” refers to the improvement of complaints such as dry eyes caused by instability of the tear film and damage to the ocular surface due to abnormalities in the quality or quantity of the tear fluid or to hydrodynamic abnormalities, as well as the improvement of abnormal changes in the barrier function of the corneal epithelium and the relief of visual abnormalities and eye discomfort.

[0049] In the present application, "test subject" refers to the consumer of the product.

[0050] In the present application, the "test subject" is an animal, preferably a mammal, more preferably a human, the test subject including, inter alia, food consumers. The test subject in the present invention is preferably a mammal. The term "mammal" refers mainly to warm-blooded, vertebrate mammals, including, but not limited to: e.g., cat, dog, hare, bear, fox, wolf, monkey, deer, rodent (such as rat, mouse), pig, cattle, sheep, horse, human, etc., preferably primates, more preferably humans. In the present application, "patient" refers to an animal, preferably a mammal, such as a human, or also a rat or mouse. Third aspect of the present application

[0051] The present application offers a product for improving dry eye, wherein the product for improving dry eye comprises the herbal composition as well as food-compatible excipients.

[0052] In the present application, "food" refers to all types of finished products and raw materials for human consumption or enjoyment, as well as to articles traditionally considered both food and Chinese medicines, but not to articles intended for therapeutic purposes, which are capable of regulating the physiological functions of an animal body (human or other animals) and often provide a beneficial effect. The area in which "food" produces a beneficial effect in the body is not particularly limited; it may be a systemic effect or an effect produced only locally. In the present application, "food" refers to a product that can be consumed directly.It is understood that the foodstuffs in the present application may contain, in addition to the aforementioned plant extract composition, any other suitable edible substances. In some embodiments, the other edible substances may be selected from those substances whose addition is permitted under food regulations, and furthermore, they may contain no substances whose addition is prohibited under food regulations. Unless otherwise specified, the food regulations refer to those in force at the time of production.

[0053] “Improvement of dry eye” refers to the definition in the second aspect.

[0054] In this application, “food-compatible additives” refers to edible raw materials, edible additives, or a combination of both. “Edible raw material” refers to an edible substance that, on its own, can provide a health-promoting function, but is not the plant extract composition; “edible additive” refers to an edible substance that, on its own, cannot provide a health-promoting function. It is understood that “food-compatible additives” may be selected from substances whose addition is permitted under the Administrative Regulations for Health-Promoting Foods, and furthermore, they do not include substances whose addition is prohibited under the Administrative Regulations for Health-Promoting Foods. Unless otherwise specified, the Administrative Regulations for Health-Promoting Foods refer to the regulations in force at the time of production.

[0055] Alternatively, the product for improving dry eye relief can be administered orally.

[0056] The product for improving dry eye conditions described in this application includes, but is not limited to, liquid drinks, tablets, capsules, powders, jelly and sweets. Fourth aspect of the present application

[0057] The present application offers the use of the plant composition in the manufacture of a product for improving gelatinase B levels.

[0058] In the present application, "improvement of gelatinase B levels" refers to a beneficial improvement in gelatinase B levels, meaning that the "improvement of gelatinase B levels" has a beneficial effect on health. In some specific implementations, "improvement of gelatinase B levels" refers to a reduction in gelatinase B levels in test subjects that is above the healthy level.

[0059] “Test subject” refers to the definition in the second aspect. Fifth aspect of the present application

[0060] The present application offers a product for improving gelatinase B levels, wherein the product for improving gelatinase B levels comprises the plant composition as well as food-compatible excipients.

[0061] Alternatively, the product for improving gelatinase B levels can be administered orally.

[0062] In the present application, the product for improving gelatinase B levels includes, but is not limited to, liquid drinks, tablets, capsules, powders, jelly and sweets.

[0063] “Food-compatible additives” refers to the definition in the third aspect.

[0064] The implementation examples of the present application are described in detail below with reference to the examples. It is understood that these examples serve only to illustrate the present application and not to limit its scope. For experimental methods in the following examples without specific conditions being stated, reference may be made primarily to the guidelines given in the present application, or alternatively, to the experimental manuals of the field, conventional conditions, manufacturer's recommended conditions, or to experimental methods known in the field.

[0065] In the following specific embodiments, unless otherwise stated, there may be slight deviations in the measurement parameters of the raw material components within the weighing accuracy range. Acceptable deviations are permitted for temperature and time parameters, caused by the measurement accuracy of the instruments or the operational accuracy.

[0066] Blueberry extract: A powdered material produced from blueberry fruit as raw material by homogenization, enzymatic degradation with pectinase and cellulase, aqueous extraction, purification, concentration, drying, etc., with a total anthocyanin content (calculated as cyanidin-3-O-glucoside chloride) ≥40%.

[0067] Goji berry extract: A powdered material produced from goji berries as raw material by crushing, extracting, filtering, concentrating, drying, with a goji berry polysaccharide content ≥10%.

[0068] Marigold extract: A natural dye produced from the flowers of the daisy plant Tagetes using a subcritical biotechnological process with low-temperature extraction, its main components being lutein and carotenoids. Common specifications of marigold extract contain 5%, 10%, and 20% lutein esters. 1 cell experiment

[0069] 1.1. Purpose of the experiment: Efficacy testing on a hyperosmotically induced dry eye cell model with corneal epithelial cells treated with lutein esters alone, as well as combined with blueberry extract (40% blueberry anthocyanins), combined with goji berry extract (10% goji berry polysaccharides) and various combined samples with different ratios.

[0070] 1.2. Experimental method: Hyperosmotically induced dry eye cell model with corneal epithelial cells.

[0071] 1.3. Experimental materials and instruments: Special medium for HCECs cells (DMEM / F12 + 15% FBS + 5 µg / mL insulin + 10 ng / mL human EGF + 1% P / S), CCK-8 test kit, sodium chloride powder, human gelatinase B Elisa kit.

[0072] 1.4. Experimental groups: Empty group, model group, lutein ester group, lutein ester combined with blueberry extract group, lutein ester combined with goji berry extract group, lutein ester combined with blueberry extract and goji berry extract group, where the goji berry extract:blueberry anthocyanin ratios were 5:1, 10:1, 30:1, 50:1, 100:1.

[0073] 1.5. Experimental instruments: cell incubator, microplate reader, thermostatically controlled drying oven, etc. 1.6. Experimental procedure:

[0074] (1) The culture of human corneal epithelial cells

[0075] Human corneal epithelial cells (HCECs) are an internationally widely used cell line for research into the pathology, physiology, and pharmacological effects of dry eye. The HCECs were cultured at 37°C in a humidified incubator with 5% CO2 in complete medium; the medium was changed every 3 days, and when the cell density reached 90%, the cells were passaged at a ratio of 1:2 to 1:3. (2) Investigation of the cytotoxicity of sodium chloride solution using the CCK-8 method

[0076] HCEC cells in the logarithmic phase were detached with trypsin, adjusted to a suitable density with complete medium, and evenly seeded into a 96-well plate. When the cell density in the wells reached 80%, the cells were randomly divided into an empty group and groups containing 0.15%, 0.3%, 0.45%, 0.48%, 0.51%, 0.54%, 0.57%, and 0.6% (wt%) sodium chloride solution (the sodium chloride solutions were prepared with medium; the corresponding osmotic pressures were 300, 350, 400, 450, 460, 470, 480, 490, and 500 mOsm / L). Each well was treated with 100 µL of medium, and the cells were treated for 24 hours. The control group received 100 µL of medium, while the experimental groups received sodium chloride solutions in medium of varying concentrations. After 12 hours, the cell viability of each group was determined using the CCK-8 method.

[0077] Specifically, the culture medium in the wells was removed and 100 µL of a 10% CCK-8 solution was added, followed by incubation at 37°C for 1–4 hours. After incubation, the absorbance of each well was measured at 450 nm. The relative cell viability of each group was calculated using the following formula. Relative cell viability % = (ODexperimental group - ODempty group) / (ODcontrol group - ODempty group) × 100%. (OD, optical density) (3) Improving effect of the samples on the dry eye cell model

[0078] Cells were seeded into a 96-well plate and randomly assigned to a control group, a model group (appropriately concentrated sodium chloride solution), and a sample-treatment group (sodium chloride solution + sample treatment). Once the cell density in the wells reached 80%, medium containing sodium chloride and the sample was added to the empty group, appropriately concentrated sodium chloride solution to the model group, and medium containing sodium chloride and the sample to the sample-treatment group. Incubation was carried out for 24 hours. The supernatant was removed, and the gelatinase B content was determined using the gelatinase B ELISA kit. 2. Test results

[0079] Investigation of the cytotoxicity of sodium chloride solution using the CCK-8 method

[0094] Table 1 Osmotic pressure (mOsm / L) Cell viability (%) 300 100,00 350 89,37 400 75,61 450 64,87 460 64,07 470 60,61 480 59,65 490 52,33 500 51,49

[0080] Combining the results from Table 1 and Fig.1. The hyperosmotic solution with 450 mOsm / L, in which cell viability remained above 50%, was selected as the modeling reagent.

[0081] (2) Improving effect of the samples on the dry eye cell model Table 2 group Gelatinase B (µg / mL) control group 0,187 Model group 0,384 1 Lutein nests 0,33 2 Lutein esters + blueberry extract 0,309 3 Lutein esters + blueberry extract + goji berry extract Blueberry extract: Goji berry extract 1: 5 0,293 1: 10 0,203 45 1: 30 0,198 6 1: 50 0,21 7 1: 100 0,254 8 Lutein esters + goji berry extract 0,261

[0082] The results from Table 2 and Fig. 2 show: The gelatinase B level in the hyperosmotically induced group, i.e. the model group, was significantly higher than in the control group, which demonstrates the successful construction of the hyperosmotically induced cellular dry eye model.

[0083] Lutein esters offer a composition for a healthier, less inflammatory ocular surface environment that promotes:

[0084] The normal secretion of mucin by goblet cells (to stabilize the inner layer of the tear film).

[0085] Maintaining the integrity of the corneal and conjunctival epithelium.

[0086] The normal function of the meibomian glands.

[0087] Ultimately, all these factors together promote the stability of the tear film and the moistening of the ocular surface.

[0088] The results from Table 2 and Fig.Figure 2 shows that the lutein ester group significantly reduced gelatinase B levels in hyperosmotically induced corneal epithelial cells. The treatment groups with the combined samples of lutein ester, blueberry extract, goji berry extract, and various mixing ratios continued to significantly reduce gelatinase B levels in hyperosmotically induced corneal epithelial cells, with ratios of 1:10, 1:30, and 1:50 showing greater efficacy. This result demonstrates that blueberry extract (containing 40% blueberry anthocyanins) and goji berry extract (containing 10% goji berry polysaccharides) can significantly downregulate hyperosmotically induced gelatinase B levels in corneal epithelial cells at ratios of 1:10, 1:30, and 1:50.

[0089] The improvement rate was calculated as: (Gelatinase B level of the model group - Gelatinase B level of the experimental group (1 to 7)) / Gelatinase B level of the model group * 100%. Based on the mathematical model for synergistic effect (improvement rate * proportion of blueberry extract / 19.53% + improvement rate * proportion of goji berry extract / 32.03%), the interaction index γ was calculated for the different ratios; the results are shown in Table 3.

[0090] The results show that blueberry extract and goji berry extract exhibit a significant synergistic effect (γ > 1) in regulating gelatinase B levels in corneal epithelial cells under ratios of 1:10, 1:30, 1:50, and 1:100. The synergistic effect of blueberry and goji berry extract was particularly pronounced under ratios of 1:10, 1:30, and 1:50. Table 3 Nr . Sample group Improvement rate γ 1 Lutein nests 14.06% / 2 Lutein esters + blueberry extract 19.53% / 3 Lutein esters + blueberry extract + goji berry extract Blueberry extract: Goji berry extract 1: 5 23.70% 0,82 4 1: 10 47.14% 1,56 5 1: 30 48.44% 1,54 6 1: 50 45.31% 1,43 7 1: 100 33.85% 1,06 8 Lutein esters + goji berry extract 32.03% /

[0091] The technical features of the implementations and embodiments described above can be combined in any suitable way. To keep the description concise, not all possible combinations of the individual technical features in the implementations and embodiments mentioned above have been described. However, as long as there are no contradictions in the combination of these technical features, they should all be considered to be covered by this description.

[0092] The exemplary embodiments described above merely express some implementation forms of the present application in order to make the technical concept of the present application concrete and understandable in detail, but must not be interpreted as limiting the scope of protection of the patent application. It should be noted that those skilled in the art in this field can make several modifications and improvements without deviating from the concept of the present application, all of which fall within the scope of protection of the present application. Furthermore, it should be understood that those skilled in the art in this field, after reading the aforementioned teaching content of the present application, can make various changes or modifications to the present application, the resulting equivalent forms also falling within the scope of protection of the present application.It is also understood that technical solutions obtained by those skilled in the art in this field through logical analysis, deduction, or limited trials based on the technical concept provided by the present application are all within the scope of protection of the attached claims of the present application. Therefore, the scope of protection of the patent in the present application should be determined by the content of the attached claims. The description and drawings may be used to clarify the content of the claims.

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