A nutritional composition for improving retinopathy of prematurity

Abstract

The present invention relates to a nutritional composition for improving retinopathy of prematurity. The nutritional composition for improving retinopathy of prematurity comprises the following components in parts by mass: 0.05 to 0.3 part of casein phosphopeptide, 0.05 to 0.3 part of casein glycomacropeptide, 1.5 to 3 parts of polydextrose, and 0.1 to 3 parts of fructooligosaccharide. The nutritional composition provided by the present invention can increase the abundance of the beneficial bacterium Bifidobacterium in the intestine, reduce the abundance of Enterobacter, and at the same time reduce the area of neovascularization of the pathological retina caused by hyperoxia and reduce the area of avascular region; inhibit or prevent the oxidation process through the bidirectional regulation of the retina-intestine axis, thereby protecting retinal cells from oxidative stress damage, and has good market application value.

Description

Technical Field

[0001] The present invention relates to a nutritional composition for improving retinopathy of prematurity. Background Art

[0002] Retinopathy of prematurity (ROP) is a serious complication in premature infants, caused by the loss of normal blood vessels in the immature retina and the appearance of new pathological blood vessels and fibroplasia, which is more common in low birth weight infants and small gestational age premature infants. In recent years, thanks to the continuous improvement of the rescue, treatment and nursing levels of critically ill premature infants, the survival rates of extremely low birth weight infants and ultra-low birth weight infants have shown a straight upward trend. Among them, there are many extremely premature infants and ultra-low birth weight infants between 22 weeks and 24 weeks who have been successfully treated. However, with the increase in their survival rates, the incidence of retinopathy of prematurity is also increasing year by year.

[0003] The treatment of retinopathy of prematurity is mostly surgical treatment and non-surgical treatment. Surgical treatments include laser photocoagulation, scleral buckling, vitrectomy, etc. The most commonly used non-surgical treatment is anti-VEGF (vascular endothelial growth factor) drugs. At present, in the field of foods for premature infants and low birth weight infants, such as formula foods for special medical purposes, infant formula foods, complementary foods and nutritional supplements, etc., there is no existing technology that can inhibit or prevent the oxidation process through bidirectional regulation of the retina-intestine axis and thereby protect retinal cells from oxidative stress damage. Summary of the Invention

[0004] Aiming at the problem of how to improve retinopathy of prematurity, the present invention provides a nutritional composition for improving retinopathy of prematurity. By controlling the components and their dosages of the nutritional composition, the retinopathy problem of premature infants and low birth weight infants can be significantly improved.

[0005] To solve the above technical problems, the technical solution provided by the present invention is:

[0006] In the first aspect, the present invention provides a nutritional composition for improving retinopathy of prematurity, comprising the following components in parts by mass: casein phosphopeptide 0.05 part to 0.3 part, casein glycomacropeptide 0.05 part to 0.3 part, polydextrose 1.5 parts to 3 parts, and fructooligosaccharide 0.1 part to 3 parts.

[0007] Compared with the prior art, the nutritional composition for improving retinopathy of prematurity provided by the present invention can inhibit or prevent the oxidation process through the bidirectional regulation of the retina-gut axis, thereby protecting retinal cells from oxidative stress damage. Experimental data show that caseinomacropeptide can improve the intestinal flora and weaken oxidative stress, and polydextrose can improve the intestinal flora. The components work synergistically to improve the retinopathy caused by the retinal damage of prematurity induced by oxidative stress; the nutritional composition provided by the present invention can increase the abundance of the beneficial bacterium Bifidobacterium in the intestine, decrease the abundance of Enterobacter, and at the same time reduce the area of neovascularization in the pathological retina caused by hyperoxia and reduce the area of avascular region.

[0008] Preferably, the nutritional composition comprises the following components in parts by mass: 0.1 part to 0.3 part of casein phosphopeptide, 0.1 part to 0.2 part of caseinomacropeptide, 1.8 parts to 2.5 parts of polydextrose, and 0.1 part to 1.2 parts of fructooligosaccharide.

[0009] Preferably, the nutritional composition further comprises: protein, fat, and carbohydrate.

[0010] More preferably, the nutritional composition further comprises the following components in parts by mass: 10 parts to 21 parts of protein, 22 parts to 41 parts of fat, and 47 parts to 70 parts of carbohydrate.

[0011] Even more preferably, the nutritional composition further comprises the following components in parts by mass: 12 parts to 17 parts of protein, 25 parts to 30 parts of fat, and 50 parts to 58 parts of carbohydrate.

[0012] More preferably, the protein comprises at least one of skim milk powder, whey protein isolate, whey protein concentrate, or milk protein concentrate.

[0013] More preferably, the fat comprises at least one of soybean oil, sunflower oil, coconut oil, rapeseed oil, corn oil, or medium-chain triglycerides.

[0014] More preferably, the carbohydrate comprises at least one of demineralized whey powder or lactose. Brief Description of the Drawings

[0015] Figure 1 It is a graph showing the sequence proportion of Enterobacter in the feces of the young rats in each group of the present invention;

[0016] Figure 2 It is a graph showing the sequence proportion of Bifidobacterium in the feces of the young rats in each group of the present invention;

[0017] Figure 3 It is a graph showing the proportion of the area of avascular region in the retina of the young rats in each group of the present invention;

[0018] Figure 4It is a graph showing the proportion of the area of the neovascular region in the retinas of the young rats in each group of the present invention;

[0019] Figure 5 It is the SOD activity in the retinal tissues of the young rats in each group of the present invention;

[0020] Figure 6 It is the MDA content in the retinal tissues of the young rats in each group of the present invention;

[0021] In the figure, there is no significant difference between the same letters, and there is a significant difference between different letters (P < 0.05). Detailed implementation manners

[0022] In order to make the objectives, technical solutions and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0023] In the present invention, the materials without special instructions are all commercially available products.

[0024] Embodiment 1

[0025] This embodiment provides a nutritional composition for improving premature retinopathy, including the following components by mass: 0.05 g of casein phosphopeptide, 0.05 g of casein glycomacropeptide, 1.5 g of polydextrose, 0.1 g of fructooligosaccharide, 14 g of protein (provided by equal mass of concentrated whey protein and concentrated milk protein), 30 g of fat (provided by equal mass of soybean oil and sunflower oil), and 50 g of carbohydrate (provided by equal mass of demineralized whey powder and lactose).

[0026] Adopt the dry mixing process to mix the components of the above-mentioned nutritional composition for improving premature retinopathy evenly.

[0027] Embodiment 2

[0028] This embodiment provides a nutritional composition for improving premature retinopathy, including the following components by mass: 0.2 g of casein phosphopeptide, 0.2 g of casein glycomacropeptide, 2.1 g of polydextrose, 0.7 g of fructooligosaccharide, 14 g of protein (provided by equal mass of skim milk powder and separated whey protein), 27 g of fat (provided by equal mass of coconut oil, corn oil and medium-chain triglycerides), and 53 g of carbohydrate (provided by demineralized whey powder and lactose with a mass ratio of 5:1). The total energy of protein, fat and carbohydrate is about 2138 kJ.

[0029] Adopt the dry mixing process to mix the components of the above-mentioned nutritional composition for improving premature retinopathy evenly.

[0030] Embodiment 3

[0031] This embodiment provides a nutritional composition for improving retinopathy of prematurity, comprising the following components by mass: 0.3 g of casein phosphopeptide, 0.1 g of caseinomacropeptide, 3 g of polydextrose, 3 g of fructooligosaccharide, 17 g of protein (provided by skim milk powder and concentrated milk protein in equal mass), 25 g of fat (soybean oil, coconut oil and rapeseed oil in equal mass), and 52 g of carbohydrate (provided by demineralized whey powder).

[0032] Adopt the dry mixing process, and mix the components of the above nutritional composition for improving retinopathy of prematurity evenly.

[0033] Comparative Example 1

[0034] This comparative example provides a nutritional composition, which is similar to Example 2, except that: the casein phosphopeptide is replaced with the same mass of caseinomacropeptide, that is, the dosage of caseinomacropeptide is 0.4 g. The remaining conditions are the same as those in Example 2 and will not be repeated.

[0035] Comparative Example 2

[0036] This comparative example provides a nutritional composition, which is similar to Example 2, except that: the caseinomacropeptide is replaced with the same mass of casein phosphopeptide, that is, the dosage of casein phosphopeptide is 0.4 g. The remaining conditions are the same as those in Example 2 and will not be repeated.

[0037] Comparative Example 3

[0038] This comparative example provides a nutritional composition, which is similar to Example 2, except that: the polydextrose is replaced with the same mass of fructooligosaccharide, that is, the dosage of fructooligosaccharide is 2.8 g. The remaining conditions are the same as those in Example 2 and will not be repeated.

[0039] Comparative Example 4

[0040] This comparative example provides a nutritional composition, which is similar to Example 2, except that: the fructooligosaccharide is replaced with the same mass of polydextrose, that is, the dosage of polydextrose is 2.8 g. The remaining conditions are the same as those in Example 2 and will not be repeated.

[0041] Comparative Example 5

[0042] This comparative example provides a nutritional composition, which is similar to Example 2, except that the addition of casein phosphopeptide, casein glycomacropeptide, polydextrose and fructooligosaccharide is omitted simultaneously. The other conditions are the same as those in Example 2, that is, the nutritional composition includes the following components by mass: protein (provided by skim milk powder and whey protein isolate in equal mass) 14 g, fat (provided by coconut oil, corn oil and medium-chain triglycerides in equal mass) 27 g, and carbohydrate (provided by desalted whey powder and lactose in a mass ratio of 5:1) 53 g. The total energy of protein, fat and carbohydrate is about 2138 kJ.

[0043] Application Examples 1 to 3

[0044] Application Examples 1 to 3 respectively provide a nutritional product, which consists of a nutritional composition for improving retinopathy of prematurity and vitamins and minerals. The nutritional compositions respectively adopt the nutritional compositions of Examples 1 to 3 for improving retinopathy of prematurity. The vitamins and minerals include the following components: vitamin A 530 μg RE / 100 g, vitamin D3 16 μg / 100 g, vitamin E 12 mg α-TE / 100 g, vitamin K1 35 μg / 100 g, vitamin B1 700 μg / 100 g, vitamin B2 1064 μg / 100 g, vitamin B6 630 μg / 100 g, vitamin B 12 1.9 μg / 100 g, niacin 7.5 mg / 100 g, folic acid 150 μg / 100 g, pantothenic acid 5 mg / 100 g, vitamin C 110 mg / 100 g, biotin 18 μg / 100 g, inositol 88 mg / 100 g, sodium 290 mg / 100 g, potassium 501 mg / 100 g, copper 430 μg / 100 g, magnesium 50 mg / 100 g, iron 8.5 mg / 100 g, zinc 5.5 mg / 100 g, manganese 53 μg / 100 g, calcium 589 mg / 100 g, phosphorus 342 mg / 100 g, iodine 93 μg / 100 g, chlorine 440 mg / 100 g, selenium 12.8 μg / 100 g, choline 83 mg / 100 g, taurine 33 mg / 100 g, L-carnitine 10.8 mg / 100 g and nucleotides 21 mg / 100 g.

[0045] Adopt the dry mixing process to mix the components of the above nutritional product evenly.

[0046] Application Comparative Examples 1 to 5

[0047] Application Comparative Examples 1 to 5 respectively provide a nutritional product, which consists of a nutritional composition and vitamins and minerals. The nutritional compositions respectively adopt the nutritional compositions of Application Comparative Examples 1 to 5. The components and contents of the vitamins and minerals are the same as those in Application Example 1, and will not be repeated.

[0048] Adopt the dry mixing process and mix the components of the above nutritional products evenly.

[0049] Efficacy test

[0050] (1)Establishment and grouping of animal models

[0051] Healthy adult pregnant SD rats were selected for the experiment, provided by Spf (Suzhou) Biotechnology Co., Ltd. (China), with the certificate number SCXK (Su) 2022-0006, gestational age of 12d to 14d, and body weight of 450g to 550g. Before the experiment, each pregnant rat was placed in a separate mouse cage to adapt to the environment for 3d to 5d, with free access to water and food. The experimental environmental conditions included: light / dark cycle of 12h / 12h, background noise of 40db ± 10db, and room temperature of 20°C to 24°C. The gestation period of normal rats is 19d to 23d, and the young rats born before the 20th day of pregnancy were defined as premature young rats.

[0052] Lipopolysaccharide LPS was used to prepare a premature birth model in pregnant rats to induce premature birth in pregnant rats. On the 16th day of pregnancy of pregnant rats, the mass of pregnant rats was accurately weighed, and LPS (purchased from Thermo Fisher Scientific) was intraperitoneally injected at a dose of 350 μg / kg for 2 consecutive days.

[0053] 10 to 12 premature young rats per litter were included in the subsequent experiment, with a total of 60 to 70 rats, randomly divided into 6 groups, 10 rats in each group, and each group was fed for 30 days. The feed formulas for each group were the nutritional products of Application Examples 1 to 3 and Application Comparative Examples 1 to 5 respectively.

[0054] Within 6 hours after the premature young rats were born, the newborn premature young rats were placed in a sealed oxygen chamber with an oxygen concentration of 80% (high oxygen) in the oxygen chamber, which was alternated every 24 hours for 14 days. At 15 days, the young rats were placed in normal air and continued to be raised until 30 days after birth. A digital oxygen meter was used to monitor the oxygen concentration in the oxygen chamber, and it was measured at least 5 times a day, with an oxygen flow rate of 2L / min to 3L / min and a nitrogen flow rate of 0.5L / min to 0.8L / min, so that the oxygen concentration in the oxygen chamber during the high-oxygen period was maintained at 80% ± 2%.

[0055] (2)16S rRNA fecal intestinal flora sequencing of feces

[0056] On the 30th day of feeding, feces of the six groups of young rats were collected respectively. Before each fecal collection, each young rat was placed alone in a sterilized animal cage box and allowed to move freely for 30 min. About 0.2 g of feces from each young rat was collected, and bacterial DNA in the feces of each group of young rats was extracted according to the instructions of the QIAGEN fecal genomic DNA extraction kit. Then, PCR amplification was performed in the highly variable region V3-V4 of 16S rRNA using the universal primers 341F and 805R. After quantitative detection of the amplification products by 2% agarose gel electrophoresis, high-throughput sequencing of the 16S rRNA gene was carried out to obtain the proportion of Escherichia coli and Bifidobacterium sequences. The test results are as Figures 1 - 2 shown in Table 1.

[0057] Table 1 Proportion results of Escherichia coli and Bifidobacterium sequences in the feces of each group of young rats

[0058]

[0059] In each column of Table 1, there is no significant difference between the same letters, and there is a significant difference between different letters (P < 0.05).

[0060] As can be seen from Figure 1 Table 1, the proportion of Escherichia coli sequences in the application of Comparative Examples 1-4 groups is significantly lower than that in the application of Comparative Example 5 group, indicating that adding casein phosphopeptide, casein glycomacropeptide, polydextrose or fructooligosaccharide to the nutritional product can regulate the number of Escherichia coli; the proportion of Escherichia coli sequences in the application of Comparative Example 2 group is higher than that in the application of Comparative Examples 1, 3-4 groups, indicating that the reduction of casein glycomacropeptide weakens the regulation effect on the intestinal flora; the proportion of Escherichia coli sequences in the application of Comparative Examples 3-4 groups is lower than that in the application of Comparative Examples 1-2 groups, indicating that fructooligosaccharide and polydextrose have a greater impact on Escherichia coli; the proportion of Escherichia coli sequences in the Application Examples 1-3 groups is significantly lower than that in the application of Comparative Examples 1-5 groups, indicating that the nutritional product provided by the present invention can effectively regulate the number of Escherichia coli.

[0061] As can be seen from Figure 2 Table 1, the proportion of Bifidobacterium sequences in the application of Comparative Examples 1-4 groups is significantly higher than that in the application of Comparative Example 5 group, indicating that adding casein phosphopeptide, casein glycomacropeptide, polydextrose or fructooligosaccharide to the nutritional product can regulate the number of Bifidobacterium; among the application of Comparative Examples 1-4 groups, the proportion of Bifidobacterium sequences in the application of Comparative Example 2 group is the lowest, indicating that the reduction of casein glycomacropeptide weakens the regulation effect on the intestinal flora; the proportion of Bifidobacterium sequences in the Application Examples 1-3 groups is significantly higher than that in the application of Comparative Examples 1-5 groups, indicating that the nutritional product provided by the present invention can effectively regulate the number of Bifidobacterium.

[0062] The nutritional product provided by the present invention can reduce the number of Escherichia coli and increase the number of Bifidobacterium, thereby effectively improving the retinopathy of prematurity through microbial regulation.

[0063] (3)Area sizes of avascular area and neovascular area in the retinas of young rats

[0064] After 30 days of feeding, the young rats in each group were sacrificed. One eye was randomly selected from each group, and the residues in the retina and the residual vitreous vessels were removed to make retinal spreads. The retinal spreads were placed on polylysine-coated glass slides with the inner layer of the retina facing up, sealed with glycerol at a concentration of 50%, covered with coverslips, and stored in the dark at 4°C.

[0065] The blood vessels were stained, and the stained spreads were observed and photographed under a fluorescence microscope. The retinal blood vessels showed green fluorescence, while the avascular area was a black area where the peripheral green fluorescence terminated. The area of the avascular area and the percentage of the neovascular area in the total retinal area were measured and calculated using Adobe Photoshop CS3 image analysis software. The test results are as Figures 3 - 4 shown in Table 2.

[0066] Table 2 Proportion results of avascular area and neovascular area in the retinas of young rats in each group

[0067]

[0068] In each column of Table 2, there was no significant difference between the same letters, while there was a significant difference between different letters (P < 0.05).

[0069] From Figure 3 and Table 2, it can be seen that the proportion of the avascular area in Application Examples 1 - 3 groups was smaller than that in Application Comparative Examples 1 - 5 groups. The proportion in Application Comparative Example 5 group was the highest, and there was a significant difference compared with Application Comparative Examples 1 - 4 and Application Examples 1 - 3 groups. From Figure 4 and Table 2, it can be seen that the proportion of the neovascular area in Application Examples 1 - 3 groups was smaller than that in Application Comparative Examples 1 - 5 groups. The proportion in Application Comparative Example 5 group was the highest. This indicates that the nutritional product provided by the present invention can effectively reduce the avascular area of the immature retina and reduce the pathological neovascular proliferation caused by hyperoxia.

[0070] In addition, the retinal blood vessels in Application Comparative Examples 3 - 4 groups showed better performance than those in Application Comparative Examples 1 - 2 groups, but the difference was not significant. The difference lies in that Application Comparative Examples 3 - 4 groups contain caseinomacropeptide and casein phosphopeptide and are at the optimal content. This shows that these two components, caseinomacropeptide and casein phosphopeptide, and their contents in the nutritional product may play more functions.

[0071] (4)Determination of oxidative stress markers

[0072] The retinal blood vessels of premature infants are not fully developed. High oxygen can cause vasoconstriction of immature retinas, leading to retinal hypoxia-ischemia, vascular occlusion, and then the production of a large number of vascular growth factors. Driven by the retinal VEGF signaling pathway, pathological retinal neovascularization and fibrous tissue hyperplasia form, which can lead to retinal detachment and blindness. Oxidative stress is one of the important injury mechanisms of ROP. The oxidative stress level of premature infants can be used as a predictor of ROP, and the degree of ROP lesions worsens with the increase in the degree of oxidative stress. Superoxide dismutase (SOD) and malondialdehyde (MDA) are very sensitive oxidative stress markers. Kits were used to measure the activity of SOD and the content of MDA. The SOD kit used a total superoxide dismutase (T-SOD) test kit (hydroxylamine method), and the MDA kit used a malondialdehyde (MDA) assay kit (TBA method), both of which were purchased from Nanjing Jiancheng Bioengineering Institute.

[0073] Under the microscope, the retinas of the other eyes of the young rats in each group were quickly separated. According to the mass-volume ratio of retina to phosphate buffer (PBS buffer) of 1 g:9 mL, PBS buffer was added, and it was ground thoroughly to make a retinal homogenate. It was centrifuged at 3500 r / min for 10 min in a 4 °C low-temperature centrifuge, and the supernatant was taken and stored at -80 °C. The activity of SOD and the content of MDA in the retinal tissues of the young rats in each group were measured strictly according to the instructions of the SOD and MDA kits. The test results are as Figures 5 - 6 shown in Table 3.

[0074] Table 3 Test results of the activity of SOD and the content of MDA in the retinal tissues of the young rats in each group

[0075]

[0076] In each column of Table 3, there is no significant difference between the same letters, and there is a significant difference between different letters (P < 0.05).

[0077] As Figure 5 shown in Table 3, the SOD activity of Application Examples 1-3 groups was higher than that of Application Comparative Examples 1-5 groups. The SOD activity of Application Comparative Example 5 group was the lowest, and there was a significant difference from other groups. As Figure 6 shown in Table 3, the MDA content of Application Examples 1-3 groups was significantly lower than that of Application Comparative Examples 1-5 groups. The MDA content of Application Comparative Example 5 group was the highest, and there was a significant difference from other groups. This shows that the intake of the nutritional product provided by the present invention can effectively regulate the degree of oxidative stress. In addition, the SOD value of Application Comparative Examples 1-2 groups was significantly lower than that of Application Comparative Examples 3-4 groups, and the MDA content of Application Comparative Examples 1-2 groups was significantly higher than that of Application Comparative Examples 3-4 groups. This shows that compared with polydextrose or fructooligosaccharide, the absence of casein phosphopeptide or casein glycomacropeptide weakens the antioxidant stress effect.

[0078] The comprehensive results show that the nutritional product provided by the present invention with a nutritional composition as the main active component can inhibit or prevent the oxidation process through the bidirectional regulation of the retina-gut axis, thereby protecting retinal cells from oxidative stress damage.

[0079] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, or improvements made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A nutritional composition for improving retinopathy of prematurity, characterized in that, Comprising the following components in parts by mass: 0.05 to 0.3 parts of casein phosphopeptide, 0.05 to 0.3 parts of casein glycomacropeptide, 1.5 to 3 parts of polydextrose, and 0.1 to 3 parts of fructo-oligosaccharide.

2. The nutritional composition for improving retinopathy of prematurity according to claim 1, characterized in that, The nutritional composition comprises the following components in parts by mass: 0.1 to 0.3 parts of casein phosphopeptide, 0.1 to 0.2 parts of casein glycomacropeptide, 1.8 to 2.5 parts of polydextrose, and 0.1 to 1.2 parts of fructo-oligosaccharide.

3. The nutritional composition for improving retinopathy of prematurity according to claim 1, wherein The nutritional composition further comprises: protein, fat, and carbohydrate; The carbohydrate comprises at least one of demineralized whey powder or lactose.

4. The nutritional composition for improving retinopathy of prematurity according to claim 1, wherein The nutritional composition further comprises the following components in parts by mass: 10 to 21 parts of protein, 22 to 41 parts of fat, and 47 to 70 parts of carbohydrate; The carbohydrate comprises at least one of demineralized whey powder or lactose.

5. The nutritional composition for improving retinopathy of prematurity according to claim 1, characterized in that, The nutritional composition further comprises the following components in parts by mass: 12 to 17 parts of protein, 25 to 30 parts of fat, and 50 to 58 parts of carbohydrate; The carbohydrate comprises at least one of demineralized whey powder or lactose.

6. The nutritional composition for improving retinopathy of prematurity according to any one of claims 3 to 5, wherein, The protein comprises at least one of skim milk powder, whey protein isolate, whey protein concentrate, or milk protein concentrate.

7. The nutritional composition for improving retinopathy of prematurity according to any one of claims 3 to 5, characterized in that, The fat comprises at least one of soybean oil, sunflower seed oil, coconut oil, rapeseed oil, corn oil, or medium-chain triglycerides.

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