Compound polysaccharide for delaying blue light induced eye aging and preparation method thereof

Abstract

The invention relates to the technical field of polysaccharide extraction, in particular to a compound polysaccharide for delaying blue light induced eye aging and a preparation method thereof, and the polysaccharide compound is prepared by compounding tremella polysaccharide, sparassis crispa polysaccharide and pleurotus geesteranus polysaccharide according to a specific proportion. The preparation method comprises the following steps: respectively carrying out steam explosion pretreatment to extract three edible fungus polysaccharides, preparing a monosaccharide solution through a hot water extraction-alcohol precipitation method and impurity removal, compounding according to the mass ratio of tremella polysaccharide to sparassis crispa polysaccharide to pleurotus geesteranus polysaccharide of (1-3): (1-2): (0.5-2), uniformly stirring under mild conditions, and freeze-drying to obtain a compounded product. The compound polysaccharide has the beneficial effects that the compound polysaccharide shows a remarkable anti-aging effect in a fruit fly blue light irradiation model, the H2O2 level and ROS fluorescence intensity of eyes can be reduced, and eye pigment accumulation is reduced, so that eye aging induced by blue light is delayed.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of polysaccharide extraction, and particularly relates to a compound polysaccharide for delaying blue light-induced eye aging and a preparation method thereof. BACKGROUND

[0002] In recent years, people's lifestyle has undergone tremendous changes, and with the rapid development of semiconductor lighting technology, people's use of electronic products has increased year by year, and it is inevitable to receive blue light released from various electronic screens and indoor lighting devices. Blue light is the light with the shortest wavelength (380-500 nm) in the visible spectrum, which belongs to high-energy short-wave light and has relatively high energy. Its harm has attracted increasing attention, and it can penetrate the cornea and directly act on the retina, accelerating the aging process of the eye. Excessive exposure to blue light can promote the production of reactive oxygen species (ROS). Studies have shown that blue light can trigger oxidative stress in eye cells, leading to a significant increase in intracellular ROS levels. In the eye, especially in the retina and cornea, the influence of blue light can break the redox balance in cells, leading to the accumulation of ROS, and ultimately leading to the occurrence of diseases such as cataracts, glaucoma, and age-related macular degeneration. H2O2 is a form of ROS, and research has found that long-term exposure to blue light can increase the production of H2O2. The accumulation of H2O2 in cells can also cause oxidative stress and cell death, and excessive H2O2 can damage cell structure and affect normal cell function. Seeking a method to delay blue light-induced eye aging has attracted widespread attention. Dietary regulation is considered an effective measure for the body to adapt to the blue light environment, and is one of the beneficial, environmentally friendly and cost-effective methods to improve the damage of blue light to the eye. It is of great significance to find active ingredients in natural products that can effectively delay blue light-induced eye aging.

[0003] It has been confirmed that tremella polysaccharide, hydrangea polysaccharide and pholiota nameko polysaccharide have potential antioxidant activity and anti-aging ability when used alone, but reports of their synergistic effect are relatively rare. Single polysaccharide has limited effect in protecting eye tissue induced by blue light. Therefore, it is of great significance to study the extraction and compounding method of the three polysaccharides and develop a stable, effective and synergistically active natural active combination. SUMMARY

[0004] The technical problem to be solved by the present application is to provide a compound polysaccharide for delaying blue light-induced eye aging and a preparation method thereof. The compound polysaccharide is compounded by tremella polysaccharide, hydrangea polysaccharide and pholiota nameko polysaccharide in a specific ratio, and combined with steam explosion pretreatment technology, which significantly enhances the synergistic antioxidant activity and effectively alleviates the oxidative damage and aging of the eye caused by blue light.

[0005] In order to solve the above technical problems, the technical scheme adopted by the present application is to provide a preparation method of a compound polysaccharide for delaying blue light-induced eye aging, comprising the following steps: (1) polysaccharides are extracted from tremella, pholiota nameko and hypsizygus marmoreus, wherein the tremella is pretreated by steam explosion, extracted by hot water extraction-alcohol precipitation method, and purified by membrane separation to obtain tremella polysaccharide; The pholiota nameko and hypsizygus marmoreus are pretreated by steam explosion, extracted by hot water extraction-alcohol precipitation method, and deproteinized to obtain pholiota nameko polysaccharide and hypsizygus marmoreus polysaccharide; (2) the tremella polysaccharide, pholiota nameko polysaccharide and hypsizygus marmoreus polysaccharide are dissolved in deionized water respectively to form tremella polysaccharide solution, pholiota nameko polysaccharide solution and hypsizygus marmoreus polysaccharide solution with a mass concentration of 1-5% w / v; (3) the mixed solution is obtained by compounding the tremella polysaccharide, pholiota nameko polysaccharide and hypsizygus marmoreus polysaccharide according to the mass ratio of 1-3:1-2:0.5-2, and stirring uniformly at 40-50 DEG C; (4) the mixed solution is freeze-dried to obtain the compound polysaccharide.

[0006] The present application has the following beneficial effects: the steam explosion pretreatment breaks the cell wall structure of tremella, pholiota nameko and hypsizygus marmoreus by instant pressure release of high-temperature and high-pressure steam, so that the polysaccharide is more easily dissolved out, and the polysaccharide molecular conformation may be changed to expose more active groups. The high mannose content of tremella polysaccharide enhances the adhesion and moisturizing properties, the high glucose content of pholiota nameko polysaccharide contributes to the immune regulation, and the free radical scavenging ability of hypsizygus marmoreus polysaccharide cooperates with the above two to jointly alleviate the oxidative stress induced by blue light. The compound polysaccharide after compounding reduces the levels of ROS and H2O2 in the eyes, reduces the accumulation of pigments, and thus delays the aging of the eyes. BRIEF DESCRIPTION OF DRAWINGS

[0007] Figure 1 Fig. 2 is the influence of different experimental group polysaccharides of example 2 of the present application on the H2O2 content in the eyes of fruit flies under blue light radiation (different lowercase letters represent significant differences, P<0.05); Figure 2 Fig. 3 is the influence of different experimental group polysaccharides of example 2 of the present application on the ROS fluorescence intensity in the eyes of fruit flies under blue light radiation (different lowercase letters represent significant differences, P<0.05); Figure 3 Fig. 4 is the influence of different experimental group polysaccharides of example 2 of the present application on the compound eyes of fruit flies under blue light radiation. DETAILED DESCRIPTION

[0008] In order to explain the technical content, the purposes and effects of the present application in detail, the following will be described in combination with the embodiments and the accompanying drawings.

[0009] The key idea of the present application is: Firstly, the cell wall structure of tremella, pholiota and hohenbuehelia was effectively destroyed by steam explosion pretreatment technology, which made polysaccharide more fully dissolved and significantly improved the polysaccharide yield. Compared with the traditional hot water extraction method, the polysaccharide extraction rate was increased by more than 30%, and the biological activity structure of polysaccharide was better maintained.

[0010] Secondly, different purification processes were adopted according to the characteristics of different raw materials: membrane separation purification was used for tremella polysaccharide, which could effectively remove small molecular impurities and improve the product purity to more than 90%; deproteinization was used for pholiota and hohenbuehelia polysaccharide, which reduced the production cost while ensuring the purity. This differential treatment not only ensured the product quality, but also took into account the economy of the process.

[0011] Thirdly, through the compounding of specific mass ratio (1-3):(1-2):(0.5-2), the three polysaccharides produced significant synergistic effect. Experiments proved that the antioxidant activity of the compound was increased by more than 40%, which was significantly higher than that of single polysaccharide or simple combination of any two polysaccharides.

[0012] Fourthly, the compounding was carried out at a mild temperature of 40-50℃, which ensured the full mixing of each component and avoided the damage to the active structure of polysaccharide caused by high temperature. The final freeze-drying process effectively maintained the physical stability and biological activity of the compound polysaccharide, and prolonged the shelf life of the product.

[0013] The preparation method has clear process route and simple operation, which is suitable for industrial production. The prepared compound polysaccharide shows excellent effect in delaying blue light-induced eye aging, providing high-quality raw materials for the development of related functional foods and health products.

[0014] The present application provides a preparation method of compound polysaccharide for delaying blue light-induced eye aging, comprising the following steps: (1) extracting polysaccharide from tremella, pholiota and hohenbuehelia, wherein the tremella is pretreated by steam explosion, and the polysaccharide is extracted by hot water extraction-alcohol precipitation method and combined with membrane separation purification to obtain tremella polysaccharide; The pholiota and hohenbuehelia are pretreated by steam explosion, and the polysaccharide is extracted by hot water extraction-alcohol precipitation method and deproteinized to obtain pholiota polysaccharide and hohenbuehelia polysaccharide; (2) dissolving the tremella polysaccharide, pholiota polysaccharide and hohenbuehelia polysaccharide in deionized water to form tremella polysaccharide solution, pholiota polysaccharide solution and hohenbuehelia polysaccharide solution with a mass concentration of 1-5% w / v; (3) compounding the tremella polysaccharide, pholiota polysaccharide and hohenbuehelia polysaccharide according to the mass ratio of (1-3):(1-2):(0.5-2) to obtain a mixed solution, and stirring uniformly at 40-50℃; (4) freeze-drying the obtained mixed solution to obtain compound polysaccharide.

[0015] From the above description, in the above preparation method, the steam explosion pretreatment breaks the cell wall structure of tremella, gloioprella and hohenbuehelia by instant pressure release of high temperature and high pressure steam, making polysaccharide more easily dissolved out, and possibly changing the polysaccharide molecular conformation to expose more active groups. The high mannose content of tremella polysaccharide enhances adhesion and moisturizing properties, the high glucose content of gloioprella polysaccharide contributes to immune regulation, and the free radical scavenging ability of hohenbuehelia polysaccharide cooperates with the two to jointly alleviate blue light-induced oxidative stress. The compounded polysaccharide delays eye aging by reducing eye ROS and H2O2 levels and reducing pigment accumulation.

[0016] Preferably, in the above preparation method, the steam explosion pretreatment in step (1) has the following conditions: pressure 0.5-1.5 MPa, pressure maintaining time 60-120 s.

[0017] From the above description, the above limited optimization of steam explosion parameters balances the cell wall breaking effect and the integrity of polysaccharide structure, and avoids excessive degradation.

[0018] Preferably, in the above preparation method, the steam explosion pretreatment of tremella in step (1) has the following conditions: pressure 1.0 MPa, pressure maintaining time 60 s.

[0019] From the above description, the above limited conditions can maximize the maintenance of the active structure of tremella polysaccharide and improve the extraction rate.

[0020] Preferably, in the above preparation method, the steam explosion pretreatment of gloioprella and hohenbuehelia in step (1) has the following conditions: pressure 1.2 MPa, pressure maintaining time 60 s.

[0021] From the above description, the above limited conditions are optimized for the cell structure characteristics of gloioprella and hohenbuehelia to improve the yield of polysaccharide.

[0022] Preferably, in the above preparation method, the membrane separation and purification in step (1) uses an ultrafiltration membrane with a molecular weight cut-off of 8-12 kDa.

[0023] From the above description, the above limitation effectively removes small molecular impurities and improves the purity and biological activity of tremella polysaccharide.

[0024] Preferably, in the above preparation method, the mass ratio of tremella polysaccharide: gloioprella polysaccharide: hohenbuehelia polysaccharide in step (3) is 2:1.5:1.

[0025] From the above description, the above limited preferred ratio is verified by a fruit fly model to have the best effect of reducing the content of eye ROS and H2O2.

[0026] Preferably, in the above preparation method, the freeze-drying conditions in step (4) are: a vacuum degree less than 20 Pa, and a cold trap temperature lower than -50℃.

[0027] From the above description, the above definition ensures the physical stability and biological activity retention of the compounded polysaccharide.

[0028] The present application also provides the compounded polysaccharide prepared by the above preparation method for delaying blue light-induced eye aging.

[0029] Example 1 A preparation method of a compounded polysaccharide for delaying blue light-induced eye aging, comprising the following steps: (1) Extraction of polysaccharide: Tremella polysaccharide: Take 500 g of fresh Tremella, soak in water for 3 min, and then perform steam explosion pretreatment (pressure 1.0 MPa, time 60 s). After pretreatment, the Tremella is dried at 60℃ to constant weight, and then crushed to pass through an 80-mesh sieve. Take 100 g of the Tremella powder, add 40 times the weight of distilled water, and extract in a water bath at 90℃ for 3 hours. Centrifuge to obtain the supernatant, concentrate, and then add 3 times the volume of pre-cooled anhydrous ethanol for precipitation. Collect the precipitate, remove the protein by the Sevag method, purify with a 10 kDa ultrafiltration membrane, and freeze-dry to obtain the Tremella polysaccharide.

[0030] Clavaria polysaccharide: Take 500 g of Clavaria, perform steam explosion pretreatment (pressure 1.2 MPa, time 60 s) in the same way, dry and crush, add distilled water at a solid-liquid ratio of 1:30, extract at 90℃ for 3 hours, and then freeze-dry after alcohol precipitation and protein removal to obtain the Clavaria polysaccharide.

[0031] Hypsizygus marmoreus polysaccharide: The preparation method is the same as that of the Clavaria polysaccharide.

[0032] (2) Preparation of compounded polysaccharide: Take 2 g, 1.5 g, and 1 g of Tremella polysaccharide, Clavaria polysaccharide, and Hypsizygus marmoreus polysaccharide, respectively, dissolve them in 100 mL of deionized water to form a mixed solution with a mass concentration of 4.5%.

[0033] (3) Stir the mixed solution at 45℃ for 30 min to ensure thorough mixing.

[0034] (4) Freeze-dry the mixed solution under the conditions of a vacuum degree of 10 Pa and a cold trap temperature of -60℃ to obtain a compounded polysaccharide powder.

[0035] Example 2 1. Preparation of conventional polysaccharide (1) Extraction of Tremella polysaccharide: The Tremella was soaked in water for 3 min and then dried at 60°C. The dried Tremella was ground and sieved to 80 mesh. 100 g of the ground Tremella was pretreated with anhydrous ethanol at 65°C for 3 h to remove part of the pigments and fats. The Tremella powder was dissolved in distilled water (2 L) at 90°C for 2 times (3 h each time) and centrifuged at 4000 rpm / min for 10 min. The supernatant was concentrated and precipitated with pre-cooled anhydrous ethanol at 4°C for 24 h. The precipitate was centrifuged at 8000 rpm / min for 10 min at 4°C and then deproteinized with Sevag reagent for 4 times. Anhydrous ethanol was added and the mixture was left at 4°C overnight. The precipitate was centrifuged and then freeze-dried to obtain the crude Tremella polysaccharide. The crude polysaccharide was dissolved in distilled water and prepared into a solution with a certain concentration using an ultrafiltration membrane (10 KD) with a suitable pore size. The solution was filtered using a membrane separation system with the operating pressure controlled at 0.5 MPa and the temperature controlled at 25°C. The permeate and the retentate were collected and freeze-dried to obtain the purified Tremella polysaccharide sample.

[0036] (2) Extraction of Sparassis and Hypsizygus polysaccharide: The Sparassis and Hypsizygus were soaked in water for 3 min and then dried at 60°C. The dried Sparassis and Hypsizygus were ground and sieved to 80 mesh. The ground powder was treated with anhydrous ethanol to remove pigments and fat impurities for 3 h and then dried at 60°C. The dried Sparassis powder and Hypsizygus powder were dissolved in distilled water (1:30 = w:v) at 90°C for 3 h with continuous stirring. The mixture was centrifuged at 4500 r / min for 10 min and the supernatant was concentrated to 1 / 5 of the original volume in a rotary evaporator. Then, 3 times the volume of anhydrous ethanol was added to the concentrate and the mixture was left at 4°C overnight. The polysaccharide was deproteinized using the Sevag method and then dialyzed in distilled water for 72 h. The freeze-dried Sparassis and Hypsizygus crude polysaccharide was obtained.

[0037] (3) Compounding: The three polysaccharides were dissolved separately to prepare solutions with a mass concentration of 5 mg / mL. The Sparassis polysaccharide, the Hypsizygus polysaccharide and the Tremella polysaccharide were mixed in a ratio of 2:1.5:1 and freeze-dried to obtain the compound polysaccharide product without steam explosion pretreatment.

[0038] 2. Preparation of compound polysaccharide for delaying blue light-induced eye aging (1) Extraction of Tremella polysaccharide: The Tremella was soaked in water for 3 min and then subjected to steam explosion. The sample was subjected to steam explosion at a pressure of 1.0 MPa for 60 s. The Tremella polysaccharide was extracted using the conventional polysaccharide preparation method to obtain the steam explosion pretreated and purified Tremella polysaccharide sample.

[0039] (2) Extraction of Polysaccharides from Sparassis and Hypsizygus marmoreus: Sparassis and Hypsizygus marmoreus were soaked in water for 3 min, and then were subjected to steam explosion. The samples were subjected to steam explosion at a pressure of 1.2 MPa for 60 s. The polysaccharides from Sparassis and Hypsizygus marmoreus were extracted by using a conventional polysaccharide preparation method, to obtain crude polysaccharides from Sparassis and Hypsizygus marmoreus after steam explosion pretreatment.

[0040] (3) Compounding: three kinds of polysaccharides were dissolved respectively to prepare solutions with a mass concentration of 5 mg / mL, and were mixed and stirred according to Tremella polysaccharide:Sparassis polysaccharide:Hypsizygus marmoreus polysaccharide = 2:1.5:1, and were freeze-dried, the freeze-drying conditions being: vacuum degree less than 20 Pa, and cold trap temperature lower than -50°C. The steam explosion pretreated compounding polysaccharide product was obtained.

[0041] 3. Structure analysis of polysaccharide components Experimental method: (1) Molecular weight: the molecular weight of the polysaccharide sample was determined by using a gel permeation chromatograph. The chromatographic column was Viscotek A600M (300 mm × 7.8 mm), and the mobile phase was 0.01 M NaNO3 aqueous solution. After the sample was dissolved and filtered through a 0.22 μm filter membrane, 50 μL of the sample was injected, and elution analysis was performed at a flow rate of 0.5 mL / min.

[0042] (2) Monosaccharide composition: after the polysaccharide component sample was hydrolyzed by using 2 M trifluoroacetic acid, the hydrolyzate was subjected to 1-phenyl-3-methyl-5-pyrazolone derivatization treatment. Subsequently, the derivatized sample was subjected to high performance liquid chromatography analysis. The standard samples including mannose, glucose, galactose and fucose were also subjected to the same derivatization treatment. The derivatized monosaccharides were separated on a Waters SunFire C18 chromatographic column (4.6 mm × 250 mm).

[0043] Experimental results: The molecular weights of Tremella polysaccharide, Sparassis polysaccharide and Hypsizygus marmoreus polysaccharide without steam explosion pretreatment were 4.70 × 10 3 , 6.69 × 10 2 , 5.24 × 10 5 respectively. After steam explosion pretreatment, the molecular weights of the polysaccharides were reduced, and the molecular weights of Tremella polysaccharide, Sparassis polysaccharide and Hypsizygus marmoreus polysaccharide were 4.50 × 10 3 , 2.60 × 10 2 , 4.21 × 10 5 respectively. The molecular weights of Tremella polysaccharide, Sparassis polysaccharide and Hypsizygus marmoreus polysaccharide are shown in Table 1.

[0044] Table 1 Tremella polysaccharide, Sparassis polysaccharide and H. sinuatus polysaccharide are composed of fucose, galactose, glucose and mannose. Tremella polysaccharide is mainly composed of mannose, and Sparassis polysaccharide and H. sinuatus polysaccharide are mainly composed of glucose. After steam explosion pretreatment, the molar percentage of monosaccharides of polysaccharides changes. The content of mannose in Tremella polysaccharide increases, the contents of glucose and mannose in Sparassis polysaccharide increase, and the contents of fucose, galactose and mannose in H. sinuatus polysaccharide increase. The monosaccharide composition of Tremella polysaccharide, Sparassis polysaccharide and H. sinuatus polysaccharide is shown in Table 2.

[0045] Table 2 4. Blue light-induced Drosophila eye aging model evaluation (1) Drosophila culture: 3-day-old standard type w1118 Drosophila melanogaster normally grown under natural light were selected for subsequent experiments. The formula of the basic culture medium is: 10 g of agar, 6.0 g of yeast, 52.0 g of sucrose, 68.0 g of corn powder and 3.2 mL of propionic acid are added per 330 mL of culture medium. The experimental culture medium of the polysaccharide dose group is based on the basic culture medium, and 0.156 mg / mL of different polysaccharide components are added. Blue light irradiated Drosophila were incubated in an incubator with a temperature control of 25±1℃ and a humidity of 55%, with a 12 h blue light / dark cycle. Blue light was emitted by a seawater tank LED lighting lamp, and the peak value of blue light was measured using a spectral illuminometer. The results showed that the peak wavelength of blue light was 457.3 nm, the main wavelength was 462.5 nm, the average illuminance was 2500 lx, and the EB of blue light hazard small light source was 2.39 W / m 2 . Drosophila were fed in the basic culture medium group (control group) and the polysaccharide dose group (Tremella polysaccharide group, Sparassis polysaccharide group, H. sinuatus polysaccharide group, and complex polysaccharide group), and the fresh food was replaced every 3-5 days.

[0046] The specific composition of each group is as follows: Control group: no polysaccharide sample was added to the basic culture medium; Tremella polysaccharide group: polysaccharide sample after steam explosion pretreatment and purification obtained in Example 2; Sparassis polysaccharide group: crude polysaccharide obtained after steam explosion pretreatment and freeze-drying in Example 2; H. sinuatus polysaccharide group: crude polysaccharide obtained after steam explosion pretreatment and freeze-drying in Example 2; Non-steam explosion-complex polysaccharide group: non-steam explosion pretreatment complex polysaccharide product obtained in Example 2; Steam explosion-complex polysaccharide group: steam explosion pretreatment complex polysaccharide product obtained in Example 2; (2) Eye observation: 40 d old female fruit flies were transferred to EP tubes after anesthesia with anhydrous ether, and were frozen and fixed at -80°C for 3-4 h. Then, the eyes of fruit flies were observed and photographed using an ultra-depth-of-field digital microscope.

[0047] (3) Measurement of ROS fluorescence intensity in the eyes: 40 d old female fruit flies were starved for 2 h, then anesthetized with anhydrous ether, and the eyes were dissected. ROS in the eyes of fruit flies was detected using DCFH-DA reactive oxygen ROS fluorescent probe.

[0048] (4) Measurement of H2O2 content in the eyes: 40 d old female fruit flies were starved for 2 h, then anesthetized with anhydrous ether, and the eyes were dissected. The H2O2 content in the eyes of fruit flies was measured according to the H2O2 content detection kit instructions.

[0049] Experimental results The H2O2 content in the eyes of fruit flies was measured according to the H2O2 content detection kit instructions. The results are shown in Figure 1 , and the results show that, relative to the control group, the H2O2 content in the eyes of fruit flies in the tremella polysaccharide group, the clavaria polysaccharide group, the hymenopellus polysaccharide group, the non-steam explosion-compound polysaccharide group, and the steam explosion-compound polysaccharide group was significantly reduced by 0.32 times ( P <0.05), 0.32 times ( P <0.05), 0.22 times ( P <0.05), 0.31 times ( P <0.05), and 0.75 times ( P <0.05), respectively; relative to the non-steam explosion-compound polysaccharide group, the H2O2 content in the eyes of fruit flies in the steam explosion-compound polysaccharide group was significantly reduced by 0.30 times ( P <0.05), and the H2O2 content in the steam explosion-compound polysaccharide group was the lowest. The results show that compound polysaccharide can significantly reduce the H2O2 content in the eyes of fruit flies under blue light irradiation, and the steam explosion pretreated compound polysaccharide has the best effect.

[0050] ROS in the eyes of fruit flies was detected using DCFH-DA reactive oxygen ROS fluorescent probe. The results are shown in Figure 2 , and the results show that, for the fluorescence intensity of ROS in the eyes of fruit flies, relative to the control group, the fluorescence intensity of ROS in the eyes of fruit flies in the tremella polysaccharide group, the clavaria polysaccharide group, the hymenopellus polysaccharide group, the non-steam explosion-compound polysaccharide group, and the steam explosion-compound polysaccharide group was reduced by 0.25 times ( P <0.05), 0.24 times ( P <0.05), 0.21 times ( P <0.05), 0.22 times ( P <0.05), and 0.39 times ( P<0.05); Compared with the non-steam-explosion-polysaccharide group, the fluorescence intensity of ROS in the eyes of fruit flies in the steam-explosion-polysaccharide group was significantly reduced by 0.10 times ( P The ROS fluorescence intensity was lowest in the steam explosion-compound polysaccharide group (<0.05). These results indicate that the compound polysaccharide can reduce the ROS fluorescence intensity in the eyes of fruit flies under blue light irradiation, and the steam explosion pretreatment of the compound polysaccharide showed the best effect.

[0051] like Figure 3 As shown, when fruit flies were exposed to blue light, the pigment color of their eyes was duller than that of the Tremella fuciformis polysaccharide group, Hydrangea macrophylla polysaccharide group, Pleurotus ostreatus polysaccharide group, the non-steam-explosion-compound polysaccharide group, and the steam-explosion-compound polysaccharide group, and the pigment accumulation was more severe. The eye pigment in the steam-explosion-compound polysaccharide group was the brightest red. The results indicate that compound polysaccharides can reduce pigment accumulation in the eyes of fruit flies under blue light irradiation, and the steam-explosion pretreated compound polysaccharide group showed the best effect.

[0052] The above description is merely an embodiment of the present invention and does not limit the patent scope of the present invention. Any equivalent modifications made based on the content of the present invention's specification and drawings, or direct or indirect applications in related technical fields, are similarly included within the patent protection scope of the present invention.

Claims

1. A method for the preparation of a complex of polysaccharides for delaying blue light induced eye aging, characterized in that, Comprise the following steps: (1) extracting polysaccharides from Tremella fuciformis, Sparassis crispa and Hypsizygus marmoreus, wherein, the Tremella fuciformis is pretreated by steam explosion, extracted by hot water extraction-alcohol precipitation method and purified by membrane separation to obtain Tremella fuciformis polysaccharide; Sparassis crispa and Hypsizygus marmoreus are pretreated by steam explosion, extracted by hot water extraction-alcohol precipitation method, and deproteinized to obtain Sparassis crispa polysaccharide and Hypsizygus marmoreus polysaccharide; (2) dissolving Tremella fuciformis polysaccharide, Sparassis crispa polysaccharide and Hypsizygus marmoreus polysaccharide in deionized water respectively to form Tremella fuciformis polysaccharide solution, Sparassis crispa polysaccharide solution and Hypsizygus marmoreus polysaccharide solution with a mass concentration of 1-5% w / v; (3) compounding the polysaccharides according to the mass ratio of Tremella fuciformis polysaccharide:Sparassis crispa polysaccharide:Hypsizygus marmoreus polysaccharide=(1-3):(1-2):(0.5-2) to obtain a mixed solution, and stirring uniformly at 40-50℃; (4) freeze-drying the mixed solution to obtain a compounded polysaccharide.

2. A process for the preparation of a complex polysaccharide for delaying blue light induced eye aging as claimed in claim 1, wherein, The steam explosion pretreatment conditions in step (1) are: pressure 0.5-1.5 MPa, pressure maintaining time 60-120 s.

3. A process for the preparation of a complex polysaccharide for retarding blue light induced eye aging as claimed in claim 1, wherein, The steam explosion pretreatment conditions of Tremella fuciformis in step (1) are: pressure 1.0 MPa, pressure maintaining time 60 s.

4. The process as claimed in claim 1, wherein the process for the preparation of the complex polysaccharide for retarding blue light induced eye aging, wherein the process comprises of the steps of: The steam explosion pretreatment conditions of Sparassis crispa and Hypsizygus marmoreus in step (1) are: pressure 1.2 MPa, pressure maintaining time 60 s.

5. The process as claimed in claim 1, wherein the process for the preparation of the complex polysaccharide for retarding blue light induced eye aging, wherein the process comprises the steps of: The membrane separation purification in step (1) uses an ultrafiltration membrane with a molecular weight cut-off of 8-12 kDa.

6. A process for the preparation of a complex polysaccharide for retarding blue light induced eye aging as claimed in claim 1, wherein, The mass ratio of Tremella fuciformis polysaccharide:Sparassis crispa polysaccharide:Hypsizygus marmoreus polysaccharide in step (3) is 2:1.5:

1.

7. The preparation method according to claim 1, characterized in that, The freeze-drying conditions in step (4) are: vacuum degree less than 20 Pa, cold trap temperature lower than-50℃.

8. A compounded polysaccharide for delaying blue light-induced eye aging prepared by the preparation method of any one of claims 1-7.

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