Blue light damage resistant composition and preparation method and application thereof

Abstract

The invention belongs to the field of health-care food, and particularly relates to a blue light damage resistant composition as well as a preparation method and application thereof. The formula of the composition comprises the following raw material components: a blueberry extract, a Chinese wolfberry fruit extract, a chrysanthemum flower extract, a dendrobium officinale extract, a cassia seed extract, a mulberry leaf extract, a rhizoma polygonati extract and a radix asparagi extract. The composition provided by the invention is reasonable in formula, high in safety and free of discomfort after oral administration, and the composition which is good in taste, convenient to take and capable of effectively preventing, relieving or assisting in treating eye injuries or eye diseases caused by blue light is provided for people who are in contact with the blue light for a long time.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to a composition for preventing and treating eye damage caused by blue light, and a preparation method and application thereof. Background Art

[0002] In recent years, with the popularity of electronic devices such as computers and smart phones, people are increasingly exposed to artificial light sources, and long-term light exposure has become a major challenge affecting the public's visual health. The phototoxicity of visible light or artificial light from screens such as televisions, computers, and mobile phones, especially blue light, has been widely studied. Studies have shown that short-wave blue light between 400-460nm is the most harmful and can induce symptoms such as eye fatigue, dryness, pain, blurred vision, headaches, and insomnia. Blue light not only damages most tissues in the eye, but also, compared with other colors of light, it can penetrate the cornea and lens and directly reach the retina, causing irreversible photochemical damage. Therefore, blue light is particularly harmful to the eyes, which is called blue light hazard or blue light damage.

[0003] Blue light damage to the eye is a high risk factor for a variety of eye diseases such as age-related macular degeneration (AMD), retinitis pigmentosa (RP) and myopia. Epidemiological studies have shown that AMD is a human retinal degenerative disease that affects people over 55 years old in industrialized countries and is the leading cause of blindness.

[0004] As teenagers are exposed to blue light such as mobile phone screens for longer and longer periods of time, the trend of myopia is becoming more and more obvious. Currently, blue light has become the main eye disease that threatens the vision of teenagers around the world. According to statistics, the global prevalence of myopia has exceeded 28%, and it is estimated that by 2050, about half of the world's population (nearly 5 billion people) will suffer from myopia to some extent. The composition of the present invention has a good therapeutic effect on eye diseases caused by blue light damage, and is particularly suitable for teenagers who are exposed to electronic products for a long time.

[0005] Currently, there are not many Chinese medicine eye protection health products on the market, and there is a lack of guidance from Chinese medicine theory, so the efficacy and quality are difficult to guarantee. Developing an effective health product that can combat blue light damage and protect vision has become the most urgent task at present.

[0006] There is no relevant record of blue light damage in ancient Chinese medicine books, but according to the theory of traditional Chinese medicine, after the syndrome differentiation of eye diseases caused by blue light, it is known that eye diseases caused by blue light are a mixture of deficiency and excess. The initial symptoms are mainly external evil damaging the body fluid, and wind and fire attack the eyes. Over time, the main symptom is the loss of yin fluid. External evil damaging the body fluid causes the loss of yin fluid, so it is necessary to use Chinese medicine for clearing heat, nourishing yin, and promoting body fluid.

[0007] There are also some traditional Chinese medicine products in the prior art that can resist blue light damage and protect eyesight, such as the formula of CN114081111A "A functional plant beverage and its preparation method" includes cassia seed, honeysuckle, mulberry leaf, chrysanthemum, selfheal, apple, raspberry, blueberry, black grape, and wolfberry. The functional plant beverage can relieve eye fatigue and protect the eyes, but it focuses more on clearing heat and fails to solve the problem of eye fluid loss caused by blue light irradiation. In addition to adding heat-clearing and yin-nourishing drugs such as blueberry, chrysanthemum, cassia seed, wolfberry, and mulberry leaf, the raw materials of the present invention also add dendrobium officinale, asparagus, and polygonatum to take advantage of their fluid-generating effect, so that the whole prescription combines the effects of clearing heat, nourishing yin, and promoting fluid. It has been verified by the effect test that the present invention has good efficacy on oxidative stress, inflammatory response, etc. caused by blue light, especially for the body's reduced tear secretion. It shows a stronger therapeutic effect, has a good effect of promoting fluid and moisturizing dryness, and can be used to relieve blue light damage, prevent, relieve and assist in the treatment of eye diseases such as dry eyes. Summary of the invention

[0008] In view of the many limitations of current drugs on the market, the present invention aims to provide a composition for preventing or treating eye diseases caused by blue light damage, helping people who are exposed to blue light such as artificial light sources for a long time to avoid eye diseases such as retinopathy. The drug of the present invention uses Chinese herbal medicine extracts as the main raw materials, has a simple preparation process, significant efficacy, and high safety, and is suitable for the prevention or treatment of eye diseases caused by blue light damage.

[0009] The raw materials used in the present invention have rich nutritional and health-care values ​​and can effectively alleviate blue light damage to the eyes through reasonable compounding.

[0010] Blueberry: A deciduous shrub of the Ericaceae family and the Vaccinium genus. The pulp is rich in nutrients and has the effects of enhancing memory, preventing brain nerve aging, protecting eyesight, anti-cancer, softening blood vessels, enhancing human immunity, anti-oxidation and slowing down aging.

[0011] Chrysanthemum: Dried inflorescence of Chrysanthemum morifolium Ramat., a plant of the Asteraceae family. It tastes spicy, sweet, bitter, and slightly cold. It enters the lung and liver meridians. It has the effects of dispersing wind-heat, calming liver yang, clearing the liver and improving eyesight, and clearing away heat and detoxification.

[0012] Lycium barbarum: This product is the dried mature fruit of Lycium barbarum L., a plant of the Solanaceae family. The fruit is harvested in summer and autumn when it turns red, dried with hot air, and the fruit stalks are removed, or dried in the sun after the skin is wrinkled, and the fruit stalks are removed. It tastes sweet and neutral. It enters the liver and kidney meridians. It has the effect of nourishing the liver and kidneys, improving intelligence and eyesight.

[0013] Cassia seed: This product is the dried mature seeds of Cassia obtusifolia L. or Cassia tora L. of the leguminous plant. The mature fruits are harvested in autumn, dried in the sun, the seeds are beaten, and impurities are removed. It tastes sweet, bitter, salty, and slightly cold. It enters the liver and large intestine meridians. It has the effects of clearing away heat, improving eyesight, and moistening the intestines and relieving constipation.

[0014] Dendrobium officinale: This product is the dried stem of Dendrobium officinale Kimura et Migo, a plant of the orchid family. It is harvested from November to March of the following year, and the impurities are removed. Some fibrous roots are cut off, and the stems are twisted into spiral or spring shapes while being heated and dried; or cut into sections, dried or dried at low temperature. The former is commonly known as "Tiepi Fengdou" (earring dendrobium); the latter is commonly known as "Tiepi Shihu". It tastes sweet and slightly cold. It enters the stomach and kidney meridians. It has the effects of nourishing the stomach, promoting fluid production, nourishing yin and clearing away heat.

[0015] Mulberry leaf: This product is the dried leaves of Morus alba L., a plant of the Moraceae family. It is harvested after the first frost, impurities are removed, and it is dried in the sun. It tastes sweet, bitter, and cold. It enters the lung and liver meridians. It has the effects of dispersing wind-heat, clearing the lungs and moistening dryness, and clearing the liver and improving eyesight.

[0016] Polygonatum sibiricum: This product is the dried rhizome of Polygonatum kingianum Coll.et Hemsl., Polygonatum sibiricum Red. or Polygonatum cyrtonema Hua, all of the Liliaceae family. It is commonly known as "big polygonatum", "chicken-headed polygonatum" or "ginger-shaped polygonatum" according to its shape. It is collected in spring and autumn, the fibrous roots are removed, washed, and slightly blanched in boiling water or steamed until thoroughly cooked, and then dried. It tastes sweet and neutral. It enters the spleen, lung and kidney meridians. It has the effects of replenishing qi and nourishing yin, strengthening the spleen, moistening the lungs and benefiting the kidneys.

[0017] Asparagus cochinchinensis (Lour.) Merr. is a dried root of Asparagus cochinchinensis (Lour.) Merr., a plant of the Liliaceae family. It is collected in autumn and winter, washed, the stem base and fibrous roots are removed, and the root is boiled or steamed in boiling water until thoroughly cooked. The outer skin is removed while hot, washed, and dried. It tastes sweet, bitter, and cold. It enters the lung and kidney meridians. It has the effects of nourishing yin and moistening dryness, clearing the lungs and promoting the production of body fluids.

[0018] The extract used in the present invention can be prepared into an extract from the above-mentioned raw materials blueberry, wolfberry, chrysanthemum, dendrobium officinale, cassia seed, mulberry leaf, polygonatum, and asparagus by conventional extraction methods or compounded with respective commercially available extracts. Conventional extraction methods usually use water extraction, for example, the raw material is added with 10 times the amount of water and refluxed for 1 hour, filtered, and the filtrate is sprayed dry to obtain the corresponding extract. The inventor has conducted quality control research on the commercially available extracts of the above-mentioned raw materials and found that the effective ingredients of different commercially available extracts are similar, and all can achieve the anti-blue light damage effect of the present invention.

[0019] The raw materials of the composition of the present invention are composed of blueberry, chrysanthemum, wolfberry, dendrobium officinale, cassia seed, mulberry leaf, polygonatum, asparagus and the like. Calculated by weight, the composition includes: 0.01-0.2 parts of blueberry extract, 0.5-1.2 parts of wolfberry extract, 0.1-1.0 parts of chrysanthemum extract, 0.5-1.2 parts of dendrobium officinale extract, 0.5-1.2 parts of cassia seed extract, 0.1-1.0 parts of mulberry leaf extract, 0.5-1.5 parts of polygonatum extract and 0.6-1.2 parts of asparagus extract. Preferably, the composition comprises, by weight: 0.08-0.12 parts of blueberry extract, 0.2-0.5 parts of wolfberry extract, 0.2-0.5 parts of chrysanthemum extract, 0.2-0.5 parts of dendrobium officinale extract, 0.2-0.5 parts of cassia seed extract, 0.5-0.9 parts of mulberry leaf extract, 0.8-1.0 parts of polygonatum extract, and 0.8-1.0 parts of asparagus extract.

[0020] Most preferably, the composition is as follows, in parts by weight: 0.1 parts of blueberry extract, 0.38 parts of wolfberry extract, 0.38 parts of chrysanthemum extract, 0.38 parts of dendrobium officinale extract, 0.38 parts of cassia seed extract, 0.76 parts of mulberry leaf extract, 1 part of polygonatum extract, and 1 part of asparagus extract.

[0021] The product is prepared from the above extracts; wherein the auxiliary materials include the following ingredients: corn oil, soybean oil, olive oil, gelatin, pectin, carrageenan, agar, starch, glycerin, brilliant blue, titanium dioxide, sugar, food additives, and distilled water. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] The in vitro effect evaluation of the prescription of this application includes Figure 1 Results of reactive oxygen species determination in cells (a. control group; b. model group; c. lutein group; d. prescription group); Figure 2 Quantitative results of intracellular reactive oxygen species; Figure 3 The results of LDH release assay in cell supernatant; Figure 4 is the result of determination of intracellular MD content; Figure 5 It is the result of determination of intracellular SOD activity; Figure 6 It is the result of determination of intracellular GSH level; Figure 7The results of intracellular GSH-PX activity determination; Figure 8 The results are for the determination of cellular IL-6 levels; Fig. 9 The results are for the determination of cellular IL-1β levels; Fig.10 The results are for the determination of TNF-α levels in cells; Fig.11 The results of the cell VEGF level determination. The in vivo pharmacodynamic evaluation of the present application prescription, wherein Fig.12 Comparison of eyeball appearance in different groups of rats (a. control group; b. model group; c. lutein group; d. patent CN114081111A group; e. low-dose prescription group; f. high-dose prescription group; g. Dendrobium officinale, Polygonatum sibiricum and Asparagus cochinchinensis group); Fig.13 Comparison of tear secretion volume among rats in different groups; Fig.14 Comparison of retinal morphology in rats of different groups (a. control group; b. model group; c. lutein group; d. patent CN114081111A group; e. low-dose prescription group; f. high-dose prescription group; g. Dendrobium officinale, Polygonatum sibiricum and Asparagus cochinchinensis group); Fig.15 The results of MDA determination in the serum of rats in different groups; Fig.16 The results of SOD determination in the serum of rats in different groups; Fig.17 The results of CAT determination in the serum of rats in different groups; Fig.18 The results of IL-6 determination in the serum of rats in different groups; Fig.19 The results of IL-1β determination in the serum of rats in different groups; Fig. 20 The results of TNF-α determination in the serum of rats in different groups; Fig.21 The results of VEGF determination in the serum of rats in different groups. DETAILED DESCRIPTION

[0023] According to the above prescription, the composition product of the present application is prepared. The following is an example of a representative preparation embodiment.

[0024] Embodiment 1 Composition liquid beverage (prescription best embodiment)

[0025] Prescription: 0.1g blueberry extract, 0.38g wolfberry extract, 0.38g chrysanthemum extract, 0.38g dendrobium candidum extract, 0.38g cassia seed extract, 0.76g mulberry leaf extract, 1g polygonatum extract, 1g asparagus extract; weigh the prescribed extracts, add 450ml distilled water, mix well, sterilize at 120℃ for 30min and then fill.

[0026] Embodiment 2 Composition solid beverage (prescription best embodiment)

[0027] Prescription: 0.1g blueberry extract, 0.38g wolfberry extract, 0.38g chrysanthemum extract, 0.38g dendrobium candidum extract, 0.38g cassia seed extract, 0.76g mulberry leaf extract, 1g polygonatum extract, 1g asparagus extract; weigh the prescribed extracts, add 0.1% sucralose and 39.9% maltodextrin, granulate according to the conventional granule preparation method, dry and package.

[0028] Example 3 Composition solid beverage

[0029] Prescription: 0.1g blueberry extract, 0.38g wolfberry extract, 0.38g chrysanthemum extract, 0.38g dendrobium candidum extract, 0.38g cassia seed extract, 0.76g mulberry leaf extract, 1g polygonatum extract, 1g asparagus extract; weigh the prescribed extracts, add 0.1% sucralose, mix well and sterilize and package.

[0030] Example 4 Composition Tableted Candy (Prescription Best Embodiment)

[0031] Prescription: 0.1g blueberry extract, 0.38g wolfberry extract, 0.38g chrysanthemum extract, 0.38g dendrobium candidum extract, 0.38g cassia seed extract, 0.76g mulberry leaf extract, 1g polygonatum extract, 1g asparagus extract; weigh the prescribed extracts, add 10% sodium carboxymethyl starch, mix well, press into tablets, sterilize and package.

[0032] Example 5 Composition Gel Candy (Prescription Best Embodiment)

[0033] Prescription: 0.1g blueberry extract, 0.38g wolfberry extract, 0.38g chrysanthemum extract, 0.38g dendrobium officinale extract, 0.38g cassia seed extract, 0.76g mulberry leaf extract, 1g polygonatum extract, 1g asparagus extract; weigh the extracts in the prescription, add corn oil suspension according to the ratio of extract to corn oil of 3:7, use a mold to prepare the shape, sterilize, and package. Experimental Example 1: Evaluation of the in vitro effect of the composition prescription of the present invention

[0034] 1.1 Instruments and reagents

[0035] Milli-Q ultrapure water system (Millipore, USA); ultra-low temperature storage box (Qingdao Haier Special Electric Co., Ltd., China); SPARK multifunctional microplate reader (TECAN, Switzerland); biological safety cabinet (THERMO FISHER, USA); Heracell VIOS160i carbon dioxide incubator (THERMO FISHER, USA); CKX31SF inverted microscope (OLYMPUS CORPORATION, Japan); AX224ZH one-tenth balance (Ohaus Instrument (Changzhou) Co., Ltd.); one-millionth balance (Xinghui (Tianjin) Biotechnology Co., Ltd.); digital illuminometer (Taishi Electronic Industry Co., Ltd.); PL-LED100F LED light source (Beijing Princes Technology Co., Ltd.)

[0036] The composition was prepared according to the proportions in Example 1, wherein wolfberry extract (Lanzhou Waterless Biotechnology Co., Ltd.); blueberry extract, chrysanthemum extract, dendrobium officinale extract, cassia seed extract, asparagus extract (Shanghai Yiji Industrial Co., Ltd.); mulberry leaf extract, polygonatum sibiricum extract (Xi'an Tianben Bioengineering Co., Ltd.); phosphate buffered saline (PBS, Beijing Solebow Technology Co., Ltd.); thiazolyl blue (MTT, Beijing Solebow Technology Co., Ltd., 54231222003); DMEM culture medium (Gibco, USA, 6124009); fetal bovine serum (Biological Industries, Gibco, USA, 04-001-1ACS; penicillin / streptomycin (double antibody) (PS, Gibco, USA, 15140122); dimethyl sulfoxide (DMSO, Sigma-Aldrich, 829R031); trypsin-EDTA (Gibco, USA, 2026-04-3C); ARPE-19 cells (Shanghai Zhongqiao Xinzhou Biotechnology Co., Ltd.); lactate dehydrogenase (LDH), malondialdehyde (MDA), glutathione (GSH), glutathione peroxidase (GSH-Px), catalase (CAT), superoxide dismutase (superoxide SOD) detection kits were purchased from Nanjing Jiancheng Bioengineering Institute with batch numbers 20240418, 20240419, 20240422, 20240410, 20240420, and 20240419; ROS detection kit (batch number 011124240403) and BCA protein quantification kit (batch number 060723240430) were purchased from Biotech Co., Ltd.

[0037] 1.2 Grouping and Dosing

[0038] The cells were seeded in 6-well plates and the cell density was adjusted to 1×10 5 / ml, 2ml was added to each well, and cultured for 24h, and then divided into a blank group (no blue light, no drug), a model group (blue light, no drug) and a drug group (blue light, drug). The drug groups were divided into a lutein group, a prescription group, a heat-clearing group (chrysanthemum, mulberry leaf, cassia seed, blueberry), and a yin-nourishing and body-excretion-promoting group (wolfberry, dendrobium officinale, polygonatum, asparagus). After 24h of drug administration, blue light irradiation (100W / m 2, 120min), the blank group was kept away from light, the supernatant was collected after the modeling was completed, and the supernatant was centrifuged (1000rpm, 10min, 4℃), and the LDH activity and IL-6, IL-β, TNF-α, and VEGF levels were determined by taking the supernatant; the cells were rinsed twice with pre-cooled PBS, 1ml of trypsin was added for digestion for 5min, 2ml of full culture was added to terminate the digestion, the bottom of the well plate was repeatedly blown and hit until all the cells fell off, the cell suspension was transferred to a pre-cooled EP tube, centrifuged (1000rpm, 10min, 4℃), the supernatant was discarded, 1mlPBS was added for centrifugation again, the supernatant was discarded, 500μlPBS was added, ultrasonic disruption (300W, ultrasonic 5s, interval 30s, disruption 5 times), centrifuged (3000rpm, 10min, 4℃), and the supernatant was taken to determine the MDA, GSH levels and SOD, GSH-PX activities.

[0039] 1.3LDH determination

[0040] The cell supernatant collected under "1.2" was aspirated and the LDH release in the cell supernatant was measured according to the instructions of the kit to evaluate the degree of cell damage.

[0041] 1.4 Determination of MDA, GSH levels and SOD, GSH-PX activities in cells

[0042] Take the cell supernatant or suspension obtained under "1.2" and determine the malondialdehyde (MDA), glutathione (GSH) levels and superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities in the cells according to the kit instructions. 1.5 Determination of IL-6, IL-β, TNF-α, and VEGF in cells

[0043] The cell supernatant collected under "1.2" was aspirated and the levels of IL-6, IL-β, TNF-α, and VEGF in the cell supernatant were determined according to the instructions of the kit.

[0044] 1.6 ROS determination

[0045] The cells were seeded in a 12-well plate and the cell density was adjusted to 1×10 4 / ml, 0.7ml was added to each well, and cultured for 24h. For grouping and dosing, refer to "1.2". After the model was established, the supernatant was discarded, and the cells were washed twice with PBS. The DCFH-DA fluorescent probe was added, and the cells were stained at 37℃ for 30min, then washed with PBS to remove the fluorescent probe that did not enter the cells. The intracellular ROS was semi-quantitatively analyzed by laser confocal microscopy, and the images were imported into Image J software for analysis.

[0046] 1.7 Data Processing

[0047] The experimental data were analyzed by one-way ANOVA and plotted using Graphpad prism software.

[0048] 1.8 In vitro pharmacodynamic evaluation results

[0049] See attached for the results Figure 1-11 .in Figure 1 Results of reactive oxygen species determination in cells (a. control group; b. model group; c. lutein group; d. prescription group); Figure 2 It is the quantitative result of intracellular reactive oxygen species; Figure 3 The results of LDH release assay in cell supernatant; Figure 4 is the result of determination of intracellular MD content; Figure 5 It is the result of determination of intracellular SOD activity; Figure 6 It is the result of determination of intracellular GSH level; Figure 7 The results of intracellular GSH-PX activity determination; Figure 8 The results are for the determination of cellular IL-6 levels; Fig. 9 The results are for the determination of cellular IL-1β levels; Fig.10 The results are for the determination of TNF-α levels in cells; Fig.11 The results are for the determination of cellular VEGF levels.

[0050] The degree of cell damage was evaluated by measuring lactate dehydrogenase in the cell supernatant. After blue light irradiation, the LDH activity in the cell supernatant increased significantly, indicating that blue light irradiation for two hours can cause cell damage. After the drug was administered for pre-protection, the LDH activity in the cells decreased significantly, indicating that the prescription can protect cells from eye diseases caused by blue light damage (see the results). Figure 3 ).

[0051] The ability of the prescription to resist oxidative stress was evaluated by measuring the oxidative stress product MDA, non-enzymatic antioxidant GSH, and antioxidant enzymes SOD and GSH in cells. According to the results, the oxidative stress response of cells was significantly reduced after pre-protection with medication. The oxidative stress levels of the prescription group and the lutein group were similar, indicating that the prescription had a strong ability to resist oxidative stress. The anti-oxidative stress abilities of the heat-clearing group and the body fluid-generating group were not as good as those of the prescription group, indicating that the combination of the two could exert a better antioxidant ability (see the results). Figure 4-7 ), and ROS was significantly reduced after administration (see Figure 1-2 ).

[0052] The anti-inflammatory ability of the prescription was investigated by measuring the inflammatory factors in the cells. After blue light irradiation, the levels of IL-6, IL-β, TNF-α, and VEGF in the cell supernatant increased significantly, indicating that blue light irradiation can cause inflammatory reactions. After pre-protection with the drug, the levels of inflammatory factors decreased significantly, indicating that the prescription has good anti-inflammatory ability (see the results). Figure 8-11 ).

[0053] Experimental Example 2: In vivo pharmacodynamic evaluation of the composition of the present invention

[0054] 1.1 Instruments and reagents

[0055] DKB-501A water bath (Shanghai Jinghong Experimental Equipment Co., Ltd.); 1 / 10,000 electronic analytical balance (Ohaus Instrument (Changzhou) Co., Ltd., model: AX2242H); 1 / 100,000 electronic analytical balance (METTLE-TOLEDO, Switzerland, model: AX205); 1 / 100,000 electronic analytical balance (METTLE-TOLEDO, Switzerland, model: XPR6UD5); ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd., model: KO-400B); Milli-Q ultrapure water system (Millpore, USA); high-speed refrigerated centrifuge (Thermo Fisher Scientific, USA); Fisher, model: ST16R); microcentrifuge (Eppendorf, Germany); vortexer (Labnet, USA, model: S0220-230V-EU); SPARK multifunctional microplate reader (TECAN, Switzerland); handheld digital slit lamp microscope (Suzhou Kangjie Medical Co., Ltd., model: KJ5S3 / KJ5S4); blue light board (Henan Qinlan Medical Equipment Co., Ltd., model: GBH-N-02)

[0056] Lycium barbarum extract (Lanzhou Waterless Biotechnology Co., Ltd.); blueberry extract, chrysanthemum extract, dendrobium officinale extract, cassia seed extract, asparagus extract (Shanghai Yiji Industrial Co., Ltd.); mulberry leaf extract, polygonatum sibiricum extract (Xi'an Tianben Bioengineering Co., Ltd.); malondialdehyde (MDA), catalase (CAT), superoxide dismutase (SOD) detection kits (Nanjing Jiancheng Bioengineering Institute, batch numbers 20240914, 20240828, 20240419 respectively); rat interleukin-6 (IL-6), rat interleukin-1β (IL-1β), rat tumor necrosis factor α (TNF-α), rat vascular endothelial growth factor (VEGF) ELISA kits (Jiangsu Enzyme Immunity Industrial Co., Ltd., MM-0190R2, MM-0047R2, MM-018 0R2, MM-0807R1); phenol red cotton thread for tear detection (Tianjin Jingming New Technology Development Co., Ltd., 20100040); lutein (Sichuan Weikeqi Biotechnology Co., Ltd.; WP24051706); Tween-80 (Aladdin, S110840); compound tobekaan (Bausch and Lomb Co., Ltd., H20023088); trichloroacetaldehyde hydrate (Aladdin, 302-17-0); FAS eyeball fixative (Wuhan Sewell Biotechnology Co., Ltd., CR2402057)

[0057] 1.2 Experimental animals

[0058] 56 male SD rats.

[0059] 1.3 Grouping and Dosing

[0060] Preparation of lutein solution: Accurately weigh 125 mg of lutein into a brown volumetric flask, add 10% Tween 80-normal saline solution to make up to 25 ml.

[0061] Preparation of patent CN114081111A solution: select medicinal materials according to "Example 1 of Chinese patent application CN114081111A", namely 3.3g of apple, 1.7g of fresh raspberry, 1.2g of blueberry, 1.2g of black grape, 1.2g of wolfberry, 4.8g of honeysuckle, 10.4g of cassia seed, 1.7g of mulberry leaf, 4.8g of chrysanthemum, and 1.7g of selfheal. Mix them evenly and add 50ml of pure water. Extract at 90℃ for 40min, filter, add 50ml of pure water to the residue, extract at 90℃ for 40min, filter, combine the two filtrates, mix them evenly and then administer.

[0062] Prescription group: Preparation of prescription high-dose solution: adopt the prescription ratio of Example 1, use normal saline as solvent to prepare a liquid composition, dilute for use, prescription high-dose group solution: 45 mg of blueberry extract, 170 mg of wolfberry extract, chrysanthemum extract, cassia seed extract, and dendrobium candidum extract, 340 mg of mulberry leaf extract, 450 mg of polygonatum extract and asparagus extract, add normal saline to make up to 50 ml.

[0063] Preparation of low-dose solution: dilute the above high-dose solution twice.

[0064] Preparation of Dendrobium officinale, Polygonatum sibiricum and Asparagus solution: only Dendrobium officinale, Polygonatum sibiricum and Asparagus are added, according to the same ratio as the prescription in Example 1, and according to the above method, a mixed solution is prepared with normal saline as solvent. Rats are randomly divided into a blank group, a model group, a lutein group, a patent CN114081111A group, a prescription low-dose group, a prescription high-dose group, and a Dendrobium officinale, Polygonatum sibiricum and Asparagus group, 8 rats in each group, and rats are fed and watered normally during the experiment. Adaptive feeding starts to be administered after one week, and the blank group and the model group are given normal saline, and the administration volume is 10ml / kg. Modeling starts one week after administration, and drug treatment is given at the same time during modeling. The blank group is placed in the dark, and after modeling is completed, a slit lamp is used to observe the state of the rat eyeball, and the tear secretion volume is determined by the phenol red cotton thread method. The rat serum is taken to determine the oxidative stress index and inflammatory factors, and a part of the eyeball is removed and stored in the eyeball fixative for pathological tissue sections to observe the retinal morphological changes, and the other part is quickly peeled off the retina on ice for subsequent experiments.

[0065] 1.4 Observation of eyeball appearance

[0066] Rats were anesthetized by intraperitoneal injection of 10% chloral hydrate. After anesthesia, the rats were dilated by instillation of 1% compound tropicamide eye drops. The rats were fixed in front of the slit lamp, the angle and distance of the slit lamp were adjusted appropriately, the fundus lesions of the rats were observed, and images were collected.

[0067] 1.5 H&E staining

[0068] The fixed eyeball specimens were dehydrated with gradient ethanol, transparentized with xylene, embedded in paraffin and cut into 7 μm sections. After baking, the sections were dewaxed with xylene and rehydrated with gradient ethanol, followed by HE staining. After sealing with neutral gum, the pathological changes of retinal tissue were observed under a microscope.

[0069] 1.6 Tear secretion volume measurement

[0070] After the modeling was completed, the same person tested the tear secretion of each group of mice. The mice were anesthetized by intraperitoneal injection of 10% chloral hydrate. After the corneal reflex disappeared, the lower eyelid of the mouse was gently pulled down, and one end of the phenol red cotton thread was placed into the conjunctival sac with toothed forceps. The position was 1 / 3 of the middle and outer canthus between the inner and outer canthus. Be careful not to let the forceps and phenol red cotton thread touch the cornea to prevent stimulating the lacrimal glands, causing a large amount of tear secretion, and affecting the measurement results. After placing the phenol red cotton thread, start timing, take out the phenol red cotton thread after 30 seconds, and use a ruler to measure the distance of the red part of the phenol red cotton thread (in mm).

[0071] 1.7 Determination of serum MDA level and CAT and SOD activity

[0072] Rats were anesthetized by intraperitoneal injection of 10% chloral hydrate. After the rats were anesthetized, they were fixed on a mouse board, and an incision was made in the abdomen of the rats using surgical scissors. After the internal organs were exposed, the internal organs were spread apart using absorbent cotton to fully expose the abdominal aorta. The injection needle was slowly inserted and gently pulled back to extract blood. The collected blood was transferred to a 10 ml centrifuge tube and placed at 4°C overnight. After the serum was fully precipitated, the tube was centrifuged (4000 rpm, 10 min, 4°C), the supernatant was collected, and stored in a -80°C refrigerator and thawed before measurement.

[0073] The serum MDA level and CAT and SOD activities were determined according to the kit instructions.

[0074] 1.8 Determination of IL-6, IL-β, TNF-α, and VEGF in serum

[0075] The levels of IL-6, IL-β, TNF-α, and VEGF in serum were determined according to the kit instructions.

[0076] 1.9 Data Processing

[0077] The experimental data were analyzed by one-way ANOVA and plotted using Graphpad prism software.

[0078] 1.10 In vivo pharmacodynamic evaluation results

[0079] See attached for the results Figure 12-21 .in Fig.12 Comparison of eyeball appearance in different groups of rats (a. control group; b. model group; c. lutein group; d. patent CN114081111A group; e. low-dose prescription group; f. high-dose prescription group; g. Dendrobium officinale, Polygonatum sibiricum and Asparagus cochinchinensis group); Fig.13 Comparison of tear secretion volume among rats in different groups; Fig.14Comparison of retinal morphology in rats of different groups (a. control group; b. model group; c. lutein group; d. patent CN114081111A group; e. low-dose prescription group; f. high-dose prescription group; g. Dendrobium officinale, Polygonatum sibiricum and Asparagus cochinchinensis group); Fig.15 The results of MDA determination in the serum of rats in different groups; Fig.16 The results of SOD determination in the serum of rats in different groups; Fig.17 The results of CAT determination in the serum of rats in different groups; Fig.18 The results of IL-6 determination in the serum of rats in different groups; Fig.19 The results of IL-1β determination in the serum of rats in different groups; Fig. 20 The results of TNF-α determination in the serum of rats in different groups; Fig.21 The results of VEGF determination in the serum of rats in different groups.

[0080] Compared with the blank group, the model group had obvious ocular neovascularization, and it tended to spread to the center of the eyeball, indicating that blue light exposure can lead to an increase in ocular neovascularization. Compared with the model group, the ocular neovascularization of rats in different drug groups decreased to varying degrees, among which the lutein group and the high-dose prescription group had the most obvious reduction in neovascularization (see the results). Fig.12 ).

[0081] Because blue light damage can cause yin deficiency and hyperactivity of fire in the human body, which in turn causes loss of body fluid in the eyes, the effect of the prescription on nourishing yin and promoting body fluid was investigated by measuring the tear secretion volume in the eyes. Compared with the blank group, the tear secretion volume in the model group was significantly reduced, indicating that blue light irradiation can cause loss of eye fluid. After two weeks of drug pre-protection and use, the tear secretion volume increased to varying degrees compared with the model group. The increase in the lutein group and the patent CN114081111A group was not obvious. Both the low-dose and high-dose prescription groups could increase the tear secretion volume of rats in a dose-dependent manner. The tear secretion volume of the prescription group increased significantly compared with the lutein group and the patent CN114081111A group (compared with the lutein group, P < 0.05; compared with the patent CN114081111A group, P < 0.01). After the single administration of Dendrobium officinale, Polygonatum sibiricum, and Asparagus cochinchinensis, the tear secretion volume increased, but the effect was not obvious. This shows that when Dendrobium officinale, Polygonatum sibiricum, and Asparagus cochinchinensis are used in combination with blueberry, wolfberry, chrysanthemum, cassia seed, and mulberry leaf, the effect of promoting body fluid and moistening dryness is better (see the results). Fig.13 ).

[0082] Compared with the control group, the retinal morphology of the model group changed significantly, especially the outer nuclear layer became thinner and the edges were blurred, which was consistent with the symptoms of retinopathy caused by blue light damage, indicating that the blue light damage model was successfully established. Compared with the model group, the outer nuclear layer of the retina in each drug-treated group thickened to varying degrees, indicating that the drug has a certain therapeutic effect on blue light damage. Among them, the lutein group recovered most significantly, and the thickness of the outer nuclear layer was almost the same as that of the blank group. The thickness of the retinal outer nuclear layer of the patent CN114081111A group and the Dendrobium officinale, Polygonatum sibiricum, and Asparagus cochinchinensis group increased slightly compared with the model group, but there was still a difference compared with the control group, indicating that the two were slightly insufficient in their anti-blue light damage efficacy. The thickness of the retinal outer nuclear layer in the high and low dose groups of the prescription increased, and was dose-dependent. Among them, the thickness of the retinal outer nuclear layer in the high dose group of the prescription was not significantly different from that in the lutein group and the control group, indicating that its anti-blue light damage efficacy was equivalent to that of lutein (results are shown in Fig.14 ).

[0083] After blue light irradiation, the level of oxidative stress product MDA in the serum of the model group increased significantly, and the activities of SOD and CAT decreased significantly, indicating that blue light irradiation can cause oxidative stress in the body. The levels of MDA in different drug administration groups decreased, and the activities of SOD and CAT increased significantly. Compared with the lutein group and the patent CN114081111A group, the degree of oxidative stress in the prescription group was comparable, indicating that the prescription had comparable antioxidant stress capacity compared with lutein and patent CN114081111A (results in Figure 15-17 ).

[0084] Compared with the blank group, the levels of inflammatory factors in the serum of the model group increased, indicating that blue light irradiation can cause inflammatory reactions in the body. After administration, the levels of inflammatory factors decreased to varying degrees. The levels of inflammatory factors in the prescription group, lutein group, and patent CN114081111A group were not much different, indicating that the anti-inflammatory ability of the prescription, lutein, and patent CN114081111A was comparable (see the results). Figure 18-21 ).

[0085] The results showed that the blue light damage model was successfully established, and it was well reflected in oxidative stress and inflammatory response. In terms of anti-inflammatory and antioxidant, the positive drug lutein group had the best efficacy among all treatment groups. In comparison, there was no significant difference in the oxidative stress and inflammatory factor levels of the low- and high-dose groups of the prescription corresponding to this patent, and both were able to significantly inhibit the oxidative stress and inflammatory response of the body caused by blue light irradiation; in terms of tear secretion volume, this patent had the best efficacy, which was significantly different from the positive drug group, indicating that this patent was superior to the positive drug lutein in nourishing yin and promoting fluid production. At the same time, compared with the prescription provided by patent CN114081111A and the combined extracts of Dendrobium officinale, Polygonatum sibiricum, and Asparagus cochinchinensis, this patent has a stronger preventive and therapeutic effect on blue light damage.

Claims

1. A composition for preventing, alleviating or assisting in the treatment of eye damage or eye diseases caused by blue light damage, characterized in that: The invention comprises multiple extracts as effective substances. The composition is composed of the following extracts in parts by weight: 0.01-0.2 parts of blueberry extract, 0.5-1.2g parts of wolfberry extract, 0.1-1.0 parts of chrysanthemum extract, 0.5-1.2 parts of dendrobium officinale extract, 0.5-1.2 parts of cassia seed extract, 0.1-1.0 parts of mulberry leaf extract, 0.5-1.5 parts of polygonatum extract and 0.6-1.2 parts of asparagus extract.

2. The composition according to claim 1, characterized in that: The composition consists of the following extracts in parts by weight: 0.08-0.12 parts of blueberry extract, 0.2-0.5 parts of wolfberry extract, 0.2-0.5 parts of chrysanthemum extract, 0.2-0.5 parts of dendrobium officinale extract, 0.2-0.5 parts of cassia seed extract, 0.5-0.9 parts of mulberry leaf extract, 0.8-1.0 parts of polygonatum extract, and 0.8-1.0 parts of asparagus extract.

3. The composition according to claim 2, characterized in that: The composition consists of the following extracts in parts by weight: 0.1 part of blueberry extract, 0.38 part of wolfberry extract, 0.38 part of chrysanthemum extract, 0.38 part of dendrobium officinale extract, 0.38 part of cassia seed extract, 0.76 part of mulberry leaf extract, 1 part of polygonatum extract, and 1 part of asparagus extract.

4. Use of the composition according to any one of claims 1 to 3 in the preparation of an anti-blue light damage product for preventing, alleviating or assisting in the treatment of eye damage or eye diseases caused by blue light damage.

5. The use according to claim 4, characterized in that: The composition is prepared in the form of liquid beverage, solid beverage, compressed candy, and jelly candy.

6. The use according to claim 5, characterized in that: The product against blue light damage includes the following auxiliary materials: corn oil, soybean oil, olive oil, gelatin, pectin, carrageenan, agar, starch, glycerin, brilliant blue, titanium dioxide, sugar, food additives, and distilled water.

7. The use according to claim 5 or 6, characterized in that: The composition according to any one of claims 1 to 3 is used together with auxiliary materials to prepare solid beverages, liquid beverages, compressed candies or jelly candies.

8. The use according to claim 7, characterized in that: The method for preparing a solid beverage comprises the following steps: taking the composition according to any one of claims 1 to 3 and the auxiliary material according to claim 6, granulating, sterilizing, and packaging to obtain a solid beverage.

9. The use according to claim 7, characterized in that: The method for preparing a liquid beverage comprises the following steps: taking the composition according to any one of claims 1 to 3 and the auxiliary material according to claim 6, and preparing a liquid beverage according to a beverage preparation process.

10. The use according to claim 7, characterized in that: The method for preparing compressed candy comprises the following steps: taking the composition according to any one of claims 1 to 3 and the auxiliary materials according to claim 6, compressing, sterilizing, and packaging to obtain compressed candy.

11. The use according to claim 7, characterized in that: The method for preparing gel candy comprises the following steps: taking the composition according to any one of claims 1 to 3 and the auxiliary materials according to claim 6, preparing according to the gel candy preparation process, sterilizing, and packaging to obtain the gel candy.

12. Use of the composition according to any one of claims 1 to 3 in the preparation of a blue light-resistant health-care drug or health-care food for preventing, alleviating or assisting in the treatment of eye damage or eye diseases caused by blue light damage.

13. The use according to claim 12, wherein the eye injury or eye disease is selected from retinopathy, dry eyes, and decreased vision.

14. The use according to claim 12 or 13, wherein the health-care medicine or health-care food is prepared in the form of a liquid beverage, a solid beverage, a compressed candy, or a jelly candy.

15. A blue light damage-resistant liquid beverage for preventing, alleviating or assisting in the treatment of eye damage or eye diseases caused by blue light damage, prepared by the following steps: 1) Weigh the extracts according to the following composition: 0.1 g of blueberry extract, 0.38 g of wolfberry extract, 0.38 g of chrysanthemum extract, 0.38 g of dendrobium officinale extract, 0.38 g of cassia seed extract, 0.76 g of mulberry leaf extract, 1 g of polygonatum extract, and 1 g of asparagus extract; 2) Add 450 ml of distilled water, mix well, sterilize at 120℃ for 30 minutes and then fill.

16. A blue light damage-resistant solid beverage for preventing, alleviating or assisting in the treatment of eye damage or eye diseases caused by blue light damage, prepared by the following steps: 1) Weigh the extracts according to the following composition: 0.1 g of blueberry extract, 0.38 g of wolfberry extract, 0.38 g of chrysanthemum extract, 0.38 g of dendrobium officinale extract, 0.38 g of cassia seed extract, 0.76 g of mulberry leaf extract, 1 g of polygonatum extract, and 1 g of asparagus extract; 2) Add 0.1% sucralose and 39.9% maltodextrin, granulate according to the conventional granule preparation method, dry and package.

17. A blue light damage-resistant solid beverage for preventing, alleviating or assisting in the treatment of eye damage or eye diseases caused by blue light damage, prepared by the following steps: 1) Weigh the extracts according to the following composition: 0.1 g of blueberry extract, 0.38 g of wolfberry extract, 0.38 g of chrysanthemum extract, 0.38 g of dendrobium officinale extract, 0.38 g of cassia seed extract, 0.76 g of mulberry leaf extract, 1 g of polygonatum extract, and 1 g of asparagus extract; 2) Add 0.1% sucralose, mix well and sterilize and package.

18. A compressed candy for preventing, alleviating or assisting in the treatment of eye damage or eye diseases caused by blue light damage, which is prepared by the following steps: 1) Weigh the extracts according to the following composition: 0.1 g of blueberry extract, 0.38 g of wolfberry extract, 0.38 g of chrysanthemum extract, 0.38 g of dendrobium officinale extract, 0.38 g of cassia seed extract, 0.76 g of mulberry leaf extract, 1 g of polygonatum extract, and 1 g of asparagus extract; 2) Add 10% sodium carboxymethyl starch, mix well, compress into tablets, sterilize and package.

19. A gel candy that prevents, alleviates or assists in treating eye damage or eye diseases caused by blue light damage, prepared by the following steps: 1) Weigh the extracts according to the following composition: 0.1 g blueberry extract, 0.38 g wolfberry extract, 0.38 g chrysanthemum extract, and 0.5 g iron stone. Dendrobium officinale extract 0.38g, Cassia seed extract 0.38g, Mulberry leaf extract 0.76g, Polygonatum sibiricum extract 1g, Asparagus cochinchinensis extract 1g; 2) Add corn oil suspension according to the ratio of extract to corn oil of 3:7, prepare the suspension using a mold, sterilize, and package.

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