Application of coniferyl aldehyde in eye protection product

Abstract

The invention provides application of coniferyl aldehyde in eye protection products, and belongs to the technical field of eye health. The coniferyl aldehyde is found to have an eye protection effect for the first time, and particularly has a protection effect on retina light injury. Cell model and animal model experiments prove that coniferyl aldehyde has a protective effect on eye cell apoptosis, retina light injury and the like, so that coniferyl aldehyde can be applied to development and application of drugs for treating retina light injury diseases and functional products for relieving visual fatigue and the like.

Description

Technical Field

[0001] The invention belongs to the technical field of eye health, and specifically relates to the application of coniferyl aldehyde in eye protection products. Background Art

[0002] Eye health is an important part of national health, involving people of all ages throughout their lives. However, as electronic products gradually penetrate people's lives, excessive use of eyes has become the norm, troubling people from children to the elderly. Myopia problems in children and adolescents, high blue light exposure problems in young and middle-aged people, and macular degeneration problems in the elderly are all related to varying degrees of retinal light damage. In particular, age-related macular degeneration (AMD) is a difficult-to-treat disease that seriously damages vision and is one of the leading causes of blindness in adults over 50 years old worldwide. With the aging of the population, the incidence of AMD is also increasing year by year. It is estimated that the number of AMD patients worldwide will reach 288 million by 2040, and there is currently no clinically effective drug treatment except for anti-VEGF drugs. It can be seen that related retinal diseases and even visual impairment caused by retinal light damage have a very serious impact on the physical and mental health and quality of life of the people, and increase the burden on families and society. Therefore, preventing retinal light damage and promoting visual health have become important issues that need to be urgently addressed in the Healthy China strategy. Summary of the invention

[0003] The invention provides application of coniferyl aldehyde in eye protection products. Coniferyl aldehyde has a protective effect on retinal light damage.

[0004] The invention provides application of coniferyl aldehyde in preparing eye-protecting medicine.

[0005] Preferably, the eye protection includes preventing and / or treating eye diseases or symptoms related to retinal light damage.

[0006] Preferably, coniferyl aldehyde is used in the preparation of drugs for resisting retinal cell apoptosis.

[0007] Preferably, the eye diseases or symptoms associated with retinal light damage include at least one of the following diseases or symptoms: blurred vision, decreased vision, dryness, tearing, congestion and visual fatigue caused by retinal light damage.

[0008] Preferably, the dosage form of the drug includes an external dosage form or an oral dosage form.

[0009] The present invention also provides the use of coniferyl aldehyde in preparing a medicine for resisting oxidative stress damage caused by light.

[0010] The invention also provides an eye protection medicine, comprising coniferyl aldehyde and pharmaceutically acceptable excipients.

[0011] Preferably, the working concentration of coniferyl aldehyde is not less than 30 μM.

[0012] The present invention also provides the use of coniferyl aldehyde in the preparation of a product for relieving visual fatigue.

[0013] Preferably, the type of the product includes ordinary food, health food, external eye patches or medical device products.

[0014] The present invention also provides a functional food for relieving visual fatigue, which uses coniferyl aldehyde as an active ingredient and also includes food-acceptable excipients.

[0015] Preferably, the dosage form of the functional food includes at least one of the following: ointment, powder, liquid, tablet, gel, capsule and granule.

[0016] Beneficial effects: The present invention discovers for the first time that coniferyl aldehyde has an eye-protecting effect, especially a protective effect on retinal light damage. The present invention constructs a light stress model of human primary Müller cells derived from peripheral retinal tissue, and finds that after incubation with coniferyl aldehyde, the cell viability of Müller cells after being stimulated by strong light can be significantly restored, and the cell viability of Müller cells after strong light stress can be restored to the level under weak light irradiation conditions, indicating that coniferyl aldehyde can significantly protect Müller cells from strong light damage. The present invention also constructs a zebrafish blue light eye injury model and a mouse retinal light injury model. After treatment with coniferyl aldehyde, it is found that coniferyl aldehyde has a protective effect on ocular cell apoptosis, retinal light damage, etc. Therefore, coniferyl aldehyde can be applied to the development and application of drugs for treating retinal light damage diseases and functional products such as relieving visual fatigue. Description of the Drawings

[0017] Figure 1 It is a result diagram of the anti-light damage effect of coniferyl aldehyde on human primary Müller cells; Note: ****P<0.0001; P<0.001; * represents the statistical difference of the same group under different light intensity conditions, ns represents no statistical difference; # represents the statistical difference between the coniferyl aldehyde administration group and the control group under strong light irradiation conditions;

[0018] Figure 2 It is a typical diagram of the fluorescence intensity of apoptotic cells in the eyes of zebrafish after treatment with coniferyl aldehyde and lutein. The green fluorescent particles pointed by the red arrows in the figure are apoptotic cells;

[0019] Figure 3 It is a quantitative diagram of the fluorescence intensity of apoptotic cells in the eyes of zebrafish after treatment with coniferyl aldehyde and lutein; Note: ***P<0.001; ##P<0.01; * represents the statistical difference between the model group and the normal control group, # represents the statistical difference between different administration groups and the model group;

[0020] Figure 4It is a H&E staining map of mouse retina; in the figure, A: H&E staining map of mouse retina tissue and the thickness map of outer nuclear layer of retina, scale bar: 500μm and 20μm; B: Quantitative map of the thickness of outer nuclear layer of retina, number of mice (n = 6); Note: ****P<0.0001; #P<0.0001, * represents the statistical difference between the model group and the control group, # represents the statistical difference between different drug administration groups and the model group;

[0021] Figure 5 It is a TUNEL staining map of apoptotic cells in mouse retina; in the figure, A: TUNEL fluorescence staining map of mouse retina, DAPI blue fluorescence represents cell nuclei, TUNEL red fluorescence represents apoptotic cells, scale bar: 500μm and 200μm; B: Quantitative map of the fluorescence intensity of apoptotic cells in retina, number of mice (n = 6); Note: ****P<0.0001; ##P<0.01, P<0.001, * represents the statistical difference between the model group and the control group, # represents the statistical difference between different drug administration groups and the model group;

[0022] Figure 6 And it is a map of the content of malondialdehyde in mouse plasma, number of mice (n = 8); Note: **P<0.01; ##P<0.01, * represents the statistical difference between the model group and the control group, # represents the statistical difference between different drug administration groups and the model group, ns represents no significant difference compared with the model group;

[0023] Figure 7 It is a map of the determination results of total antioxidant capacity (T-AOC) in mouse plasma, number of mice (n = 4); in the figure: ***P<0.001; ##P<0.01; P<0.001; #P<0.001, * represents the statistical difference between the model group and the control group, # represents the statistical difference between different drug administration groups and the model group. Detailed implementation mode

[0024] The present invention provides the application of coniferaldehyde in the preparation of drugs for protecting eyes.

[0025] The coniferaldehyde in the present invention has the molecular formula of C 10 H 10 O3, with a molecular weight of 178.18, and can be a conventional commercially available reagent in the art, such as purchased from TargetMol in the examples of the present invention, product number: T2S1907.

[0026] When using coniferyl aldehyde according to the present invention, it is preferably dissolved in DMSO to prepare a corresponding solution. The present invention has been confirmed by both cell models and animal models that coniferyl aldehyde has a protective effect on retinal light damage, specifically manifested as: anti-apoptosis of retinal cells and anti-oxidative stress damage caused by light. The retinal light damage-related eye diseases or symptoms described in the present invention preferably include at least one of the following diseases or symptoms: blurred vision, decreased vision, dryness, tearing, congestion, and visual fatigue caused by retinal light damage.

[0027] The present invention provides an eye-protecting drug, comprising coniferyl aldehyde and a pharmaceutically acceptable excipient.

[0028] The drug of the present invention uses coniferyl aldehyde as the active ingredient, and the working concentration of coniferyl aldehyde is preferably not less than 30 μM.

[0029] It has been confirmed in the examples of the present invention that the coniferyl aldehyde dose-dependently improves the apoptosis of retinal cells caused by light, and has a significant eye-protecting effect against ocular light damage; the protective effect of coniferyl aldehyde on the outer nuclear layer of the retina is equivalent to that of the positive drug lutein, indicating that coniferyl aldehyde has a significant effect on improving retinal light damage; coniferyl aldehyde can significantly reduce the level of membrane lipid peroxidation, and has a good resistance and protective effect against oxidative stress damage caused by light. Therefore, an eye-protecting drug with coniferyl aldehyde as the active ingredient can be prepared.

[0030] The present invention does not particularly limit the dosage form of the drug, and preferably includes external dosage forms or oral dosage forms. Among them, the external dosage forms include paste, gel, liquid eye drops, etc., and the oral dosage forms include tablets, capsules, granules, etc.; and there is no particular limitation on the preparation method of the drug. It can be prepared by mixing based on the dosage form and pharmaceutically acceptable excipients using conventional methods in the art.

[0031] The present invention provides the use of coniferyl aldehyde in the preparation of a product for relieving visual fatigue.

[0032] The type of the product described in the present invention preferably includes ordinary food, health food, external eye patches or medical device products.

[0033] The present invention provides a functional food for relieving visual fatigue, with coniferyl aldehyde as the active ingredient.

[0034] The present invention does not particularly limit the dosage form of the functional food, and it can be, for example, paste, powder, liquid, tablet, gel, (soft) capsule, granule, etc.

[0035] In order to further illustrate the present invention, the following describes in detail the application of coniferyl aldehyde provided by the present invention in eye-protecting products in combination with examples, but they should not be construed as limiting the protection scope of the present invention.

[0036] Example 1 Cell Model Experiment

[0037] 1. Experimental method:

[0038] 1.1 Cultivation and subculture of human primary Müller cells in the retina

[0039] Human primary Müller cells in the retina were obtained from the donor's retina tissue for cultivation.

[0040] Obtaining human primary Müller cells: Use surgical scissors and forceps to separate the retina from the retinal pigment epithelium - choroid - sclera eye cup. Place a 1 cm 2 sized retinal tissue into a T25 culture flask containing 5 mL of complete DMEM culture medium. Wrap the culture flask with tin foil and then place it in an environment at 4°C overnight. Preheat the trypsin digestion solution in a 37°C water bath. Transfer the retinal tissue to a new culture flask containing 5 mL of preheated trypsin digestion solution and place it in a 37°C, CO2 incubator for 60 min. Use sterile forceps to transfer the digested retina to a cell culture dish containing 5 mL of complete DMEM culture medium and cut it into small pieces (1×1 mm) under a dissecting microscope. Transfer these small pieces of retinal tissue back to the T25 culture flask together with the complete DMEM culture medium. Use an 18G needle with a 90° bend at the top to evenly distribute these small retinal tissues at the bottom of the culture flask and press them onto the bottom of the culture flask under the microscope. Then, carefully add 2 mL of complete DMEM culture medium. Place the culture flask vertically in a cell incubator (37°C, CO2) for 15 min to allow the retinal fragments to better adhere to the bottom of the T25 culture flask, and then place the T25 culture flask horizontally and culture it in the incubator. When culturing for the 7th day, add 2 mL of complete DMEM culture medium. Minimize the interference with the culture flask during the whole process.

[0041] Cultivation of human primary Müller cells: When culturing for the 10th day, replace the old 4 mL of complete DMEM culture medium with new one, and then maintain a frequency of changing the culture medium twice a week. It takes about 2 - 3 weeks for human primary Müller cell colonies to appear from the tissue, and then another 2 - 3 weeks to reach a density of 80% - 90%.

[0042] Subculture of human primary Müller cells: The digestion time of human primary Müller cells is longer than that of general cell lines. Initially, subculture is carried out at a ratio of 1:1, and after P2, a ratio of 1:2 to 1:3 can be adopted. Usually, P3 cells can be used for cell experiments. Although it can generally be subcultured up to P10 under normal circumstances, it also depends on the situation of different donors. The specific subculture steps are as follows: Remove the culture medium and rinse with 3 mL of sterile PBS solution. Remove the PBS solution and add 2 mL of trypsin digestion solution to each T25 culture flask for digestion (37 °C, 6 - 8 min). When more than half of the cells detach from the bottom of the culture flask, add 2 mL of complete DMEM medium to terminate the digestion. Pipette to dissociate all the cells and transfer all the liquid to a 15 mL centrifuge tube. Precipitate human primary Müller cells by centrifugation (200 g, 5 min, 20 °C). Resuspend the cell pellet with 1 mL of complete DMEM medium and transfer it to a new T25 culture flask containing 3 mL of complete DMEM culture, then place it back in the incubator for culture.

[0043] Cryopreservation and recovery of human primary Müller cells: Digest and collect human primary Müller cells as described above. Resuspend the cell pellet with 1 mL of cryopreservation solution and transfer it to a cryotube. Transfer the cryotube-containing cell program cooling box to an -80 °C refrigerator for cryopreservation, and transfer the cryotube to a liquid nitrogen tank for storage the next day. When recovering the cells, thaw the cryotube in a pre-warmed 37 °C water bath. After it completely melts, add the suspension to 4 mL of pre-prepared complete DMEM culture medium, mix well, and centrifuge at 200 g for 5 min at room temperature. Then discard the supernatant, resuspend the cells with 4 mL of complete DMEM culture medium, and transfer them to a cell culture flask, and place it in a 37 °C, 5% CO2 incubator for culture.

[0044] 1.2 Photostress modeling and coniferyl aldehyde drug treatment of human retinal primary Müller cells

[0045] Seed human primary Müller cells in the logarithmic growth phase at 5,000 cells / well in a 96-well plate and culture them in a 37 °C, 5% CO2 incubator for 24 h. Aspirate the culture medium. Set 6 replicates for the coniferyl aldehyde drug solution group, and add 100 μL of coniferyl aldehyde drug solution diluted with DMEM solution in advance to each well, with a final concentration of 30 μM. The control group adds 100 μL of DMEM solution. Use a strong light irradiation system to perform photostress modeling on human retinal primary Müller cells cultured in vitro. The strong light irradiation group uses strong light of 32 k Lux for 4 h, and the weak light irradiation group uses weak light of 5 k Lux for 4 h. Then use the AlamarBlue kit to detect cell viability.

[0046] 1.3 Cell viability detection

[0047] After the cells were stimulated by light, the cell viability was detected using the AlamarBlue kit. The specific steps were as follows: Discard the cell culture supernatant, wash twice with 100 μL PBS, add 100 μL of AlamarBlue reagent diluted 1:10 with DMEM solution, and incubate in a 37°C, 5% CO2 incubator for 4 h. Read the fluorescence of each well using a microplate reader at an excitation wavelength of 544 nm and an emission wavelength of 590 nm.

[0048] 1.4 Statistical analysis of data

[0049] The results of statistical processing were expressed as mean ± SE. GraphPad Prism (Version 9.3.1) software was used for statistical analysis, and paired t-tests were used to compare the statistical differences between groups.

[0050] 2. Experimental results:

[0051] The above method was used to construct a light stress model of human primary Müller cells derived from peripheral retinal tissue to explore the effect of 30 μM coniferyl aldehyde on the viability of Müller cells after strong light irradiation. The results are as Figure 1 shown. Compared with weak light irradiation, the viability of human primary Müller cells in the control group decreased significantly after 4 h of strong light irradiation. However, after pre-incubation with 30 μM coniferyl aldehyde for 24 h, the viability of Müller cells could be significantly restored, and the viability of Müller cells after strong light stress was restored to the level under weak light irradiation conditions, indicating that coniferyl aldehyde can significantly protect Müller cells from strong light damage.

[0052] Example 2 Experiment on zebrafish blue light eye injury model

[0053] This experiment was completed by Hangzhou Huante Biotechnology Co., Ltd. (Project No.: 8254).

[0054] 1. Experimental method:

[0055] 1.1 Experimental animals

[0056] Zebrafish were all raised in fish culture water at 28°C (water quality: 200 mg of instant sea salt was added to every 1 L of reverse osmosis water, the conductivity was 450 - 550 μS / cm; pH was 6.5 - 8.5; hardness was 50 - 100 mg / L CaCO3). They were provided by the fish breeding center of Huante Biotechnology Co., Ltd. The license number for the use of experimental animals was: SYXK (Zhe) 2022 - 0004. The feeding management met the requirements of international AAALAC accreditation (accreditation number: 001458), and the IACUC ethical review number was: IACUC - 2024 - 8254 - 01.

[0057] 1.2 Instruments, Consumables and Reagents

[0058] Dissecting microscope (SZX7, OLYMPUS, Japan); CCD camera (VertA1, Shanghai Tusen Vision Technology Co., Ltd., China); Precision electronic balance (CP214, OHAUS, USA); 6-well plate (Zhejiang Beilambo Biotechnology Co., Ltd., China); Electric focusing continuous zoom fluorescence microscope (AZ100, Nikon, Japan); Blue light instrument (50w450nm, China).

[0059] Dimethyl sulfoxide (DMSO, batch number BCCD8942, Sigma, Switzerland); Pronase E (batch number G12511Y118034, Shanghai Yuanye Bio-Technology Co., Ltd., China); Acridine orange (AO, batch number C12894919, Shanghai Macklin Biochemical Co., Ltd., China); Methyl cellulose (batch number C2004046, Shanghai Aladdin Biochemical Technology Co., Ltd., China).

[0060] Positive drug lutein, yellow powder, batch number A2316085, Shanghai Aladdin Biochemical Technology Co., Ltd., solvent is DMSO, dosage for zebrafish: 62.5 μg / mL.

[0061] 1.3 Determination of the Maximum Test Concentration (MTC)

[0062] Wild-type AB strain zebrafish at 1 day post-fertilization (1 dpf) were randomly selected for blue light irradiation after hatching to establish a zebrafish blue light eye injury model. At 3 dpf, model zebrafish with good developmental status were randomly selected and distributed into 6-well plates, with 30 zebrafish in each well (experimental group). Different concentrations of coniferyl aldehyde were administered by water solution (Table 1), and a normal control group and a model group were set up simultaneously, with a volume of 3 mL in each well. After treatment at 28 °C for 1 day, the MTC of coniferyl aldehyde on model zebrafish was determined.

[0063] 1.4 Evaluation of the Anti-Blue Light and Eye Protection Efficacy

[0064] Wild-type AB strain zebrafish at 1 dpf were randomly selected after hatching and irradiated with blue light to establish a zebrafish blue light eye injury model. At 3 dpf, model zebrafish with good developmental status were randomly selected and placed in 6-well plates, with 30 zebrafish in each well (experimental group). Coniferyl aldehyde at a safe concentration (Table 2) and lutein at a concentration of 62.5 μg / mL as the positive control were dissolved in water and administered. At the same time, a normal control group and a model group were set up, with a volume of 3 mL per well. After treatment at 28°C for 1 day, the zebrafish in each experimental group were stained with AO in the dark for 30 min, washed 3 times with standard dilution water, and then 10 zebrafish were randomly selected from each experimental group and photographed under a fluorescence microscope. Image J software was used to analyze and collect data, and the fluorescence intensity of apoptotic cells in the zebrafish eyes was analyzed and statistically analyzed. The anti-blue light eye protection effect of coniferyl aldehyde was evaluated based on the statistical analysis results of the above indicators.

[0065] The results of statistical processing were expressed as mean ± SE. SPSS 26.0 software was used for statistical analysis.

[0066] 2. Experimental results:

[0067] 2.1 MTC determination results

[0068] As shown in the results of Table 1, the MTC of coniferyl aldehyde against the anti-blue light eye protection effect in zebrafish was 3.12 μg / mL. When the treatment concentration of coniferyl aldehyde was 50 μg / mL, the mortality rate of zebrafish was 100%.

[0069] Table 1 Results of the experiment on exploring the concentration of coniferyl aldehyde for anti-blue light eye protection effect (n = 30)

[0070]

[0071] 2.2 Evaluation of anti-blue light eye protection effect

[0072] Under the conditions of this example, the results are shown in Table 2 and Figure 2 and Figure 3 as follows. The zebrafish blue light eye injury model showed apoptosis of eye cells, while after treatment with coniferyl aldehyde, the apoptosis of zebrafish eye cells could be significantly reduced. Compared with the positive drug lutein, coniferyl aldehyde showed an anti-apoptotic effect equivalent to that of lutein at a lower dose of 3.12 μg / mL, indicating that coniferyl aldehyde has a significant eye protection effect against blue light-induced eye damage.

[0073] Table 2 Results of the experiment on evaluating the anti-blue light eye protection effect of coniferyl aldehyde (n = 10)

[0074]

[0075]

[0076] Example 3 Experiment on the mouse light injury model

[0077] 1. Experimental methods

[0078] 1.1 Experimental animals

[0079] 8-week-old male BALB / c mice were purchased from Zhejiang Vital River Laboratory Animal Technology Co., Ltd. Before the experiment began, all mice were adaptively fed with a 12:12 h light / dark cycle in an animal room at a certain temperature and humidity for 1 week, and were provided with normal food and drinking water every day. All experimental operations followed the guidelines for animal care and use of the Animal Research Institute of Zhejiang University.

[0080] 1.2 Construction of the mouse light damage model

[0081] A self-made light box with a length of 108 cm, a width of 50 cm, and a height of 72 cm was used. The light intensity was detected by a lux meter to be 8500 - 10000 Lux. After eye examination, 60 mice with normal eyes were randomly divided into a control group, a model group, a lutein group (100 mg / kg), a coniferyl aldehyde low-dose group (10 mg / kg), and a coniferyl aldehyde high-dose group (50 mg / kg), with 12 mice in each group. Lutein and coniferyl aldehyde were suspended in a 5% carboxymethyl cellulose sodium (CMC-Na) solution and administered by gavage once a day. Mice in the blank and model groups were gavaged with an equal volume of 5% CMC-Na. After 10 consecutive days of administration, the mice in the model group, lutein group, coniferyl aldehyde low-dose group, and high-dose group were dark-treated for 36 h, and then atropine ophthalmic gel was applied to both eyes of the mice to dilate the pupils. The mice were placed in a self-made light device with a light intensity of 8000 - 10000 Lux (the blank group was raised in a normal environment) and exposed to continuous light for 24 h. After the light exposure ended, blood was collected by enucleating the eyeballs of each group of mice, and the eyeball tissues were collected for subsequent relevant tests.

[0082] 1.3 Experimental instruments

[0083] Table 3 Experimental instruments used in Example 3

[0084]

[0085]

[0086] 1.4 Experimental consumables and reagents

[0087] Table 4 Experimental consumables and reagents required in Example 3

[0088]

[0089]

[0090] 1.5 Histopathological examination

[0091] Six mouse eyeball tissues were taken from each group for hematoxylin-eosin staining (HE) and apoptotic cell TUNEL fluorescence staining, and the histopathological changes of mouse eyeball tissues were observed using an optical microscope (completed by Wuhan Saiwei Biotechnology Co., Ltd.).

[0092] 1.5.1 The experimental procedure for H&E staining was carried out according to the hematoxylin-eosin (H&E) high-definition constant staining kit.

[0093] 1.5.2 The experimental procedure for TUNEL staining was carried out according to the Tunel kit.

[0094] 1.6 Determination of MDA and T-AOC levels in mouse plasma

[0095] Mouse blood was collected into an anticoagulant tube and centrifuged at 3000 RPM for 10 min at 4°C, and the upper plasma was collected. The contents of MDA and T-AOC were determined according to the method provided by the kit manufacturer.

[0096] 1.7 Statistical analysis

[0097] Prism 8 software was used for drawing statistical graphs and performing statistical analysis. All data were expressed as mean ± standard error. The differences between two groups were analyzed using independent sample t-tests, and P < 0.05 indicated that the differences were statistically significant.

[0098] 2. Experimental results

[0099] 2.1 H&E histopathological evaluation

[0100] The hierarchical structure of the retina includes the retinal ganglion cell layer, the inner nuclear layer of the retina, the outer nuclear layer of the retina, and the retinal pigment epithelium. The outer nuclear layer of the retina is mainly composed of photoreceptor cells, which are responsible for receiving light stimuli and converting light signals into electrical signals. The inner nuclear layer of the retina contains different types of neuron cells, which can convert electrical signals into nerve impulses. The inner / outer nuclear layer cells of the retina play an important role in the reception and conduction of visual signals. The H&E staining results of the retinal tissue are as Figure 4 shown. Light damage caused morphological changes such as loosening and reduced thickness in the outer nuclear layer of the retina in the model group mice, indicating that light caused damage to the outer nuclear layer cells of the retina ( Figure 4 A in it). However, both low-dose and high-dose coniferyl aldehyde could increase the thickness of the outer nuclear layer of the retina and better improve the morphological changes in the outer nuclear layer of the retina caused by light ( Figure 4 A and B in it), and the protective effect of coniferyl aldehyde on the outer nuclear layer of the retina was equivalent to that of the positive drug lutein, indicating that coniferyl aldehyde has a significant effect on improving retinal light damage.

[0101] 2.2 TUNEL staining of apoptotic cells in the retina

[0102] The TUNEL fluorescence staining results of apoptotic cells in retinal tissue are as follows Figure 5 shown. Light illumination induced a large number of apoptotic cells in the retinas of mice in the model group (red fluorescence, Figure 5 in A), while coniferyl aldehyde dose-dependently improved the apoptosis of retinal cells caused by light illumination, and already had an effect equivalent to that of lutein at a low dose of 10 mg / kg ( Figure 5 in A and B), indicating that coniferyl aldehyde has a protective effect on retinal light damage.

[0103] 2.3 Determination of plasma MDA level

[0104] Plasma malondialdehyde (MDA) is one of the main products of cell membrane lipid peroxidation. The content of MDA can be used as an index of lipid peroxidation to reflect the degree of cell membrane lipid peroxidation. The results of the MDA measurement level are as follows Figure 6 shown. Light illumination led to an increase in the plasma MDA level of mice in the model group, while coniferyl aldehyde at 50 mg / kg significantly reduced the MDA level, indicating that coniferyl aldehyde can significantly reduce the level of cell membrane lipid peroxidation and has a protective effect on oxidative stress damage caused by light illumination.

[0105] 2.4 Determination of plasma total antioxidant capacity (T-AOC) level

[0106] Plasma total antioxidant capacity (T-AOC) refers to the total antioxidant level composed of various antioxidant substances and antioxidant enzymes. To protect cells from oxidative stress damage caused by reactive oxygen free radicals, the total antioxidant capacity can be used to evaluate the antioxidant capacity of bioactive substances. The results of the T-AOC level determination are as follows Figure 7 shown. Light illumination led to a decrease in the total antioxidant capacity of mice in the model group, while coniferyl aldehyde at low and high doses could significantly increase the total antioxidant capacity in vivo, indicating that coniferyl aldehyde has a good resistance to oxidative stress damage caused by light illumination.

[0107] Although the above embodiments have described the present invention in detail, they are only a part of the embodiments of the present invention, rather than all embodiments. People can also obtain other embodiments according to these embodiments without creative efforts, and these embodiments all fall within the protection scope of the present invention.

Claims

1. Application of coniferyl aldehyde in preparing eye-care products.

2. The application according to claim 1, wherein The eye-care products include at least one of the following: (1) drugs for preventing and / or treating eye diseases or symptoms related to retinal light damage; (2) products for relieving visual fatigue.

3. Application of coniferyl aldehyde in preparing drugs for anti-retinal cell apoptosis.

4. Application of coniferyl aldehyde in preparing drugs for anti-oxidative stress damage caused by light.

5. A drug for protecting eyesight, characterized in that, It includes coniferyl aldehyde and pharmaceutically acceptable excipients.

6. The drug according to claim 4, characterized in that, The working concentration of the coniferyl aldehyde is not less than 30 μM.

7. Application of coniferyl aldehyde in preparing products for relieving visual fatigue.

8. The application according to claim 7, characterized in that, The types of the products include ordinary foods, health foods, external eye patches or medical device products.

9. A functional food for relieving visual fatigue, characterized in that, Taking coniferyl aldehyde as the active ingredient, it also includes food-acceptable excipients.

10. The functional food according to claim 9, characterized in that, The dosage forms of the functional foods include at least one of the following: ointments, powders, liquids, tablets, gels, capsules and granules.

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