Application of high-ferulic acid whole wheat flour food in preparation of products for prolonging life of animals

Abstract

The invention discloses application of high-ferulic acid whole wheat flour food in preparation of products for prolonging life of animals. According to the method, corn flour, whole wheat flour and refined flour are used as raw materials to prepare fruit fly culture media of materials with different ferulic acid content differences, and then different culture media are used for feeding fruit flies under oxidative damage, high-temperature stress and natural aging models. The method comprises the following steps: respectively obtaining survival curves corresponding to culture mediums with different ferulic acid contents for feeding drosophila melanogaster under different models, and analyzing to find that compared with a culture medium with low ferulic acid whole wheat flour and a culture medium with low ferulic acid fine flour, the survival curves of the culture mediums with different ferulic acid contents are different; by feeding the high-ferulic acid whole wheat flour culture medium and the corn flour culture medium, the resistance of the fruit flies to oxidative damage, high-temperature stress and natural aging is enhanced, and the median survival time and the maximum survival time of the fruit flies are remarkably prolonged. It is found for the first time that the ferulic acid content in food is related to senescence delaying, and the method has important significance in developing anti-senescence products.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, and particularly relates to application of high-ferulic acid whole-wheat flour food in preparing products for prolonging animal lifespan. Background Art

[0002] Aging refers to the irreversible functional degeneration and decreased ability to adapt to the environment that occurs during an organism's life cycle. These changes increase the likelihood of illness and death. Based on the theory that food and medicine have the same origin, in-depth research on the role of natural antioxidants in the diet in combating oxidative stress can better explore their potential in delaying aging. With the improvement of people's health awareness, the main goal of food is no longer to meet the energy required for human physiological functions. There is an increasing demand for foods with additional physiological benefits. Epidemiological studies have shown that diet plays an important role in preventing chronic diseases such as diabetes, neurodegenerative diseases and heart disease. Scientific dietary adjustments such as limiting calorie intake and supplementing foods rich in vitamins or dietary fiber can delay aging and slow the onset of age-related diseases.

[0003] Currently, most in vivo anti-aging studies are conducted using fruit flies or mice. As a mature genetic model organism, fruit flies have advantages such as rapid generation, short life cycle, low maintenance costs, and not overly stringent ethical issues. In addition, fruit flies share many conserved biological pathways with humans, and more than 70% of known human disease-causing genes are conserved. In addition, many human organs have similar functions in fruit flies, and there are many similarities in digestion, absorption, and metabolism. Fruit flies have been frequently used in the study of human disease models and are suitable as in vivo tools for high-throughput preliminary screening of potential drugs and development of disease therapies. Summary of the Invention

[0004] The technical problem to be solved by the present invention is how to delay aging.

[0005] In order to solve the above technical problems, the present invention first provides a new use of cereal flour or cereal flour products.

[0006] The present invention provides the use of cereal flour or cereal flour products in any of the following A1)-A4):

[0007] A1) preparing products for delaying animal aging;

[0008] A2) preparing products that extend animal lifespan;

[0009] A3) preparing products for enhancing the resistance of animals to high temperature stress;

[0010] A4) preparing products for enhancing the resistance of animals to oxidative damage;

[0011] The ferulic acid content in the cereal flour is at least 0.96 mg / g;

[0012] The ferulic acid content in the cereal flour product is at least 0.37 mg / g.

[0013] In the above application, the cereal flour is made from cereal grains having a ferulic acid content of at least 0.96 mg / g.

[0014] The cereals include cereals, legumes, and tuber crops. Cereals include rice (e.g., indica rice, japonica rice, and gherkin rice), wheat (e.g., wheat, barley, oats, and rye), corn, sorghum, millet, sorghum, millet, and buckwheat; legumes include soybeans, broad beans, peas, mung beans, adzuki beans, and kidney beans; and tuber crops include sweet potatoes (also known as sweet potatoes or white potatoes), potatoes, yams, taro, and cassava.

[0015] In one embodiment of the present invention, the cereal flour is corn flour.

[0016] The corn flour is corn flour made from corn kernels having a ferulic acid content of at least 0.96 mg / g. Furthermore, the corn flour is corn flour made from corn kernels having a ferulic acid content higher than or equal to that of the control variety Zhengdan 958. The corn flour can be directly obtained by commercial purchase, or it can be obtained by grinding it by itself according to conventional methods. Preferably, the corn flour is prepared according to the following method: fresh and mature kernels are selected, and the selected kernels are washed to remove impurities and soil on the surface to obtain washed kernels; then the washed kernels are soaked in water to allow them to fully absorb water to obtain soaked kernels; then the soaked kernels are subjected to a slurry treatment to obtain cornmeal; finally, the sieved cornmeal is heated and baked to obtain corn flour. In a specific embodiment of the present invention, the corn flour is corn flour made from Zhengdan 958 kernels.

[0017] In another embodiment of the present invention, the cereal flour is wheat flour.

[0018] The wheat flour includes wheat flour made from wheat grains having a ferulic acid content of at least 0.96 mg / g. Furthermore, the wheat flour is whole wheat flour. The whole wheat flour can be directly obtained commercially, or it can be obtained by grinding it by itself according to conventional methods. Preferably, the whole wheat flour is prepared as follows: select grains free of pests and diseases, remove the seed shells and impurities, and mix them evenly with water (add 5 ml of water for every 50 grams of grains), soak for 24 hours to obtain soaked wheat; then grind the soaked wheat to obtain ground wheat; finally, pass the ground wheat through a 60-mesh sieve to obtain whole wheat flour. If there are grains that are not fully ground, grind them again with a stone mill until all pass through a 60-mesh sieve. In a specific embodiment of the present invention, the whole wheat flour is whole wheat flour made from Shangmai 189 grains or whole wheat flour made from Yunong 907 grains.

[0019] In the above applications, the cereal flour products are foods or feeds processed with cereal flour as raw materials, including but not limited to food, beverages, dietary supplements, nutritional products, health products, etc.

[0020] Furthermore, the cereal flour product may also include additives such as preservatives, thickeners, colorants, sweeteners, leavening agents, moisture retainers, and coagulants.

[0021] Furthermore, the cereal flour product is animal feed, specifically culture medium.

[0022] In a specific embodiment of the present invention, the culture medium comprises the above-mentioned corn flour, agar powder, and brown sugar. Furthermore, the solutes of the culture medium are composed of the above-mentioned corn flour, agar powder, brown sugar, yeast powder, and a preservative. Still further, the culture medium is composed of the above-mentioned corn flour, agar powder, brown sugar, yeast powder, a preservative, and water. Furthermore, the culture medium (1 liter) is composed of 6250 mg of the above-mentioned corn flour, 700 mg of agar powder, 6750 mg of brown sugar, 2450 mg of yeast powder, 4 ml of propionic acid, 17.5 ml of ethanol, 1.75 g of butylparaben, and water.

[0023] In another specific embodiment of the present invention, the culture medium comprises the above-mentioned wheat flour, agar powder and brown sugar. Further, the solutes of the culture medium are composed of the above-mentioned wheat flour, agar powder, brown sugar, yeast powder and a preservative. Still further, the culture medium is composed of the above-mentioned wheat flour, agar powder, brown sugar, yeast powder, a preservative and water. Furthermore, the culture medium (1 liter) is composed of 6250 mg of the above-mentioned wheat flour, 700 mg of agar powder, 6750 mg of brown sugar, 2450 mg of yeast powder, 4 ml of propionic acid, 17.5 ml of ethanol, 1.75 g of butylparaben and water.

[0024] In the above application, the high temperature stress is high temperature stress of 29°C or above, specifically high temperature stress of 29°C.

[0025] In the above application, the oxidative damage is damage induced by hydrogen peroxide, specifically damage induced by 5% hydrogen peroxide.

[0026] In order to solve the above technical problems, the present invention also provides a cereal flour product.

[0027] The cereal flour product provided by the present invention is food or feed, comprising the cereal flour described above; the ferulic acid content in the cereal flour product is at least 0.37 mg / g; and the function of the cereal flour product is any one of the following B1) to B4):

[0028] B1) Delaying animal aging;

[0029] B2) Extend animal lifespan;

[0030] B3) Enhance the animal's resistance to high temperature stress;

[0031] B4) Enhance the animal body's resistance to oxidative damage.

[0032] Any of the following methods D1)-D4) also falls within the scope of protection of the present invention:

[0033] D1) A method for delaying aging in animals, comprising the step of causing the animals to consume or feed the cereal flour product as food or feed;

[0034] D2) A method for extending the lifespan of an animal, comprising the step of causing the animal to consume or feed the above-mentioned cereal flour product as food or feed;

[0035] D3) A method for enhancing the resistance of an animal to high temperature stress, comprising the step of causing the animal to consume or feed the cereal flour product as food or feed;

[0036] D4) A method for enhancing the resistance of an animal body to oxidative damage, comprising the step of allowing the animal to eat or feed the above-mentioned cereal flour product as food or feed.

[0037] In any of the above applications, products or methods, the delaying of animal aging or the prolonging of animal lifespan is embodied in any one of the following 1)-6):

[0038] 1) Prolong the maximum survival time of animals under high temperature stress model;

[0039] 2) Prolong the maximum survival time of animals in an oxidative damage model;

[0040] 3) Prolong the maximum survival time of animals in a natural aging model;

[0041] 4) Improve the median survival time of animals under high temperature stress model;

[0042] 5) Increase the median survival time of animals in an oxidative damage model;

[0043] 6) Improve the median survival time of animals in a natural aging model.

[0044] Any of the above-mentioned animals include mammals and insects. Among them, the mammals include humans; the insects include fruit flies. The fruit flies are specifically wild-type female fruit flies (w 118 ).

[0045] The chemical formula of any of the above-mentioned ferulic acid is C 10 H 10 O4, CAS No. 1135-24-6, and the structural formula is shown in Formula I.

[0046]

[0047] The present invention uses corn flour of Zhengdan 958, whole wheat flour of Yunong 907, Shangmai 189, Zhoumai 18 and Jimai 38, and refined flour of Jimai 22 as raw materials to prepare six fruit fly culture media with different ferulic acid content. Then, the six culture media are used to feed wild-type female fruit flies (w 118 ), and obtained wild-type female fruit flies (w 118 ) The survival curves of culture media with different ferulic acid contents were analyzed, and it was found that compared with the culture media with low ferulic acid whole wheat flour and refined flour, the culture media with high ferulic acid whole wheat flour and corn flour could enhance the survival of wild-type female fruit flies (w 118 ) and significantly improves the median and maximum survival time of fruit flies. This study is the first to demonstrate that ferulic acid content in food is associated with delayed aging, which is of great significance for the development of anti-aging products. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1Figure 1 shows the ferulic acid content, in vitro antioxidant capacity, and feeding experiment of Drosophila culture media containing materials with different ferulic acid content. A shows cornmeal, whole wheat flour, and refined flour in Drosophila culture media. B shows the ferulic acid content in Drosophila culture media (n=4). C shows the in vitro antioxidant activity of Drosophila culture media determined by the ABTS assay (n=4). D shows the in vitro antioxidant activity of Drosophila culture media determined by the DPPH assay (n=4). E shows the in vitro antioxidant activity of Drosophila culture media determined by the FRAP assay (n=4). F shows the Drosophila feeding experiment (n=10). Note: Error bars are ±SD. Different letters above the error bars in Figures BE indicate significant differences between groups at α=0.01 as tested by one-way ANOVA.

[0049] Figure 2 The wild-type female fruit flies (w 118 )Survival curve of Drosophila culture medium containing materials with different ferulic acid content.

[0050] Figure 3 The wild-type female fruit flies (w 118 )Survival curve of Drosophila culture medium containing materials with different ferulic acid content.

[0051] Figure 4 The wild-type female fruit flies (w 118 )Survival curve of Drosophila culture medium containing materials with different ferulic acid content. DETAILED DESCRIPTION

[0052] The present invention will be further described in detail below in conjunction with specific embodiments. The examples provided are only for illustrating the present invention and are not intended to limit the scope of the present invention. The examples provided below can serve as a guide for further improvements by those skilled in the art and are not intended to limit the present invention in any way.

[0053] Unless otherwise specified, the experimental methods in the following examples are conventional methods and were performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials and reagents used in the following examples, unless otherwise specified, were all commercially available.

[0054] The approval number of Shangmai 189 in the following embodiment is Guoshenmai 20230048.

[0055] Yunong 907 in the following embodiment is a new plant variety that Henan Agricultural University applied for protection registration and authorized announcement to the state on January 1, 2022. It belongs to the dry common wheat Triticum aestivum L., application number: 20221000011, announcement number: CNA030955G, variety right number: CNA20221000011.

[0056] The approval number of Jimai 38 in the following embodiments is Guoshenmai 980003.

[0057] The approval number of Zhoumai 18 in the following embodiments is Guoshenmai 2005006.

[0058] The approval number of Jimai 22 (regional trial code: 984121) in the following examples is Guoshenmai 2006018.

[0059] The approval number of Zhengdan 958 in the following embodiments is Guoshenyu20000009.

[0060] The wild type fruit flies in the following examples are w 118 strains, as recorded in the literature "Liu Bo et al., Effect of Hydrogen Peroxide on W 118 Effects of strains of Drosophila on lifespan, fertility, locomotion and antioxidant capacity, Journal of Central South Medical Sciences, Vol. 49, No. 2, 2021.

[0061] The chemical formula of ferulic acid in the following examples is C 10 H 10 O4, CAS No. 1135-24-6, and the structural formula is shown in Formula I.

[0062]

[0063] Example 1. Preparation of Drosophila Culture Medium with Different Ferulic Acid Contents and Its Phenolic Acid Content, In Vitro Antioxidant Capacity Detection, and Feeding Experiment

[0064] 1. Obtaining Materials for Preparing Drosophila Culture Medium

[0065] Corn flour of Zhengdan 958, whole wheat flour of Yunong 907 (ferulic acid content in grains of 0.96 mg / g), whole wheat flour of Shangmai 189 (ferulic acid content in grains of 0.98 mg / g), whole wheat flour of Zhoumai 18 (ferulic acid content in grains of 0.65 mg / g), whole wheat flour of Jimai 38 (ferulic acid content in grains of 0.64 mg / g), and refined flour (hereinafter referred to as refined flour) of Jimai 22 (ferulic acid content in grains of 0.821 mg / g) were prepared respectively. The specific preparation method is as follows:

[0066] Corn flour is prepared as follows: fresh, mature corn kernels are selected and cleaned to remove surface impurities and dirt to obtain cleaned kernels. The cleaned kernels are then soaked in water to allow them to fully absorb water, obtaining soaked kernels. The soaked kernels are then ground to obtain cornmeal. Finally, the sieved cornmeal is heated and toasted to obtain corn flour.

[0067] To prepare whole wheat flour, select pest-free kernels and remove the hulls and impurities. Add 5 ml of water per 50 g of kernels, mix thoroughly, place in a ziplock bag, and temper for 24 hours. Grind the kernels using a stone mill and pass through a 60-mesh sieve. If any kernels are not fully ground, grind again using a stone mill until all kernels pass through the 60-mesh sieve. Store the ground whole wheat flour in a refrigerator at -80°C.

[0068] The preparation method for refined flour is as follows: Select pest-free kernels and remove the hulls and impurities. Add 5 ml of water to every 50 g of kernels, mix thoroughly, place in a ziplock bag, and temper for 24 hours. Grind using a Brabender 180101 laboratory mill. The bran flour is the bran flour, while the flour flour is the refined flour. Store the ground flour in a freezer at -80°C.

[0069] 2. Preparation of Drosophila Culture Medium with Different Ferulic Acid Content

[0070] The corn flour of Zhengdan 958, the whole wheat flour of Yunong 907, the whole wheat flour of Shangmai 189, the whole wheat flour of Zhoumai 18, the whole wheat flour of Jimai 38 and the refined flour of Jimai 22 ( Figure 1 A) Prepare Drosophila culture medium as raw materials.

[0071] 1. Preparation of cornmeal Drosophila culture medium

[0072] 1) Prepare cornmeal Drosophila culture medium according to the following recipe: agar powder 700 mg, cornmeal (Zhengdan 958 cornmeal) 6250 mg, brown sugar 6750 mg, and dilute to 1 L with water.

[0073] 2) Place the mixture on a magnetic stirrer and stir until the brown sugar is dissolved.

[0074] 3) Heat the food in an electric stove, stirring constantly with a spoon to prevent it from burning. Bring to a boil and keep boiling for 1 minute before stopping.

[0075] 4) Stir the food with a spatula. When the food cools to 50°C, add 2450 mg of dry yeast powder (Angel Yeast Powder, production license number SC20142050300018) and stir until the food is completely dissolved.

[0076] 5) When the food has cooled to 40°C, add a preservative (4 mL of propionic acid, 17.5 mL of ethanol, and 1.75 g of butylparaben (Sangon Biotech (Shanghai) Co., Ltd., Catalog No. A501824-0250)). Dispense the food into culture tubes using a syringe, 3 mL per tube. Label the tube with the tube number and date. Place a layer of sterile gauze over the tube opening. Allow the tube to rest for 12 hours, then securely plug the tube opening with an ethanol-sterilized sponge plug. Store in a refrigerator at 4°C. The shelf life is one week.

[0077] 2. Preparation of whole wheat flour Drosophila culture medium

[0078] Prepare whole-wheat Drosophila medium according to the following recipe: 700 mg agar powder, 6250 mg whole-wheat flour (Yunong 907 whole-wheat flour, Shangmai 189 whole-wheat flour, Zhoumai 18 whole-wheat flour, or Jimai 38 whole-wheat flour), 6750 mg brown sugar, and dilute to 1 L with water.

[0079] 2) The remaining steps are the same as steps 2)-5) in "Preparation of cornmeal Drosophila culture medium".

[0080] 3. Preparation of Drosophila culture medium

[0081] 1) Prepare a Drosophila culture medium using the following formula: 700 mg agar powder, 6250 mg flour (Jimai 22 flour), 6750 mg brown sugar, and dilute to 1 L with water.

[0082] 2) The remaining steps are the same as steps 2)-5) in "Preparation of cornmeal Drosophila culture medium".

[0083] 3. Detection of ferulic acid content in Drosophila culture medium

[0084] The ferulic acid content in the fruit fly culture medium prepared in step 2 using Zhengdan 958 corn flour, Yunong 907 whole wheat flour, Shangmai 189 whole wheat flour, Zhoumai 18 whole wheat flour, Jimai 38 whole wheat flour, and Jimai 22 refined flour as raw materials was detected respectively. The determination of ferulic acid content was based on the method described in “Zhang GL, Zhou PC, Gong YL, et al. Boosting the antioxidant potential of pasta by a premature stop mutation in wheat keto-acythiolase-2[J]. Food Chemistry, 2022, 385: 132634.”

[0085] The results are as follows Figure 1As shown in Figure B, the results show that: among them, the ferulic acid content in the fruit fly culture medium prepared with Zhengdan 958 corn flour as raw material is 0.49 mg / g, the ferulic acid content in the fruit fly culture medium prepared with Yunong 907 whole wheat flour as raw material is 0.37 mg / g, the ferulic acid content in the fruit fly culture medium prepared with Shangmai 189 whole wheat flour as raw material is 0.38 mg / g, the ferulic acid content in the fruit fly culture medium prepared with Zhoumai 18 whole wheat flour as raw material is 0.22 mg / g, the ferulic acid content in the fruit fly culture medium prepared with Jimai 38 whole wheat flour as raw material is 0.27 mg / g, and the ferulic acid content in the fruit fly culture medium prepared with Jimai 22 refined flour as raw material is 0.08 mg / g. It can be seen that the ferulic acid content in the fruit fly culture medium prepared with Zhengdan 958 corn flour as raw material is significantly higher than that in the fruit fly culture medium prepared with wheat flour (whole wheat flour and refined flour) as raw material, which is consistent with the difference in ferulic acid content between the two crops themselves. In addition, the ferulic acid content in two fruit fly culture media prepared with Yunong 907 whole wheat flour and Shangmai 189 whole wheat flour as raw materials (hereinafter referred to as high ferulic acid whole wheat flour culture media) was significantly higher than that in two fruit fly culture media prepared with Zhoumai 18 whole wheat flour and Jimai 38 whole wheat flour as raw materials (hereinafter referred to as low ferulic acid whole wheat flour culture media).

[0086] IV. In vitro antioxidant activity assay of Drosophila culture medium containing materials with different ferulic acid contents

[0087] The in vitro antioxidant activity of the Drosophila culture medium prepared in step 2 using Zhengdan 958 corn flour, Yunong 907 whole wheat flour, Shangmai 189 whole wheat flour, Zhoumai 18 whole wheat flour, Jimai 38 whole wheat flour, and Jimai 22 refined flour was determined using the ABTS, DPPH, and FRAP methods, respectively. The specific steps for determining in vitro antioxidant activity using the ABTS, DPPH, and FRAP methods refer to the method in the reference "Zhang GL, Zhou PC, Gong YL, et al. Boosting the antioxidant potential of pasta by a premature stop mutation in wheat keto-acythiolase-2[J]. Food Chemistry, 2022, 385:132634."

[0088] The results are as follows Figure 1 As shown in Figure CE, the results indicate that the in vitro antioxidant activity of Drosophila culture medium prepared with corn flour was slightly higher than that of culture medium containing high-ferulic acid whole wheat flour, but there was no significant difference between the two. The in vitro antioxidant activity of culture medium containing high-ferulic acid whole wheat flour was significantly higher than that of culture medium containing low-ferulic acid whole wheat flour. Furthermore, no data were available for culture medium prepared with refined flour, as the in vitro antioxidant capacity was below the detection limit.

[0089] 5. Experiment on feeding preference of Drosophila culture medium containing materials with different ferulic acid content

[0090] A feeding experiment was conducted on the fruit fly culture media prepared in step 2 using Zhengdan 958 corn flour, Yunong 907 whole wheat flour, Shangmai 189 whole wheat flour, Zhoumai 18 whole wheat flour, Jimai 38 whole wheat flour, and Jimai 22 refined flour as raw materials. The specific steps are as follows:

[0091] 1. Passaging of Drosophila

[0092] 1) Under an optical microscope, select w 118 For the strain of fruit flies, anesthetize the fruit flies with a CO2 anesthesia needle. Transfer 30 ml of fruit fly culture medium to each propagation bottle, place a layer of sterile gauze on the tube mouth, let it stand for 12 hours, and then plug the tube mouth with a high-density sponge plug disinfected with ethanol. Place 15 female flies and 10 male flies in each propagation bottle containing fruit fly culture medium, and lie the propagation bottle horizontally. After the fruit flies wake up, place the propagation bottle containing fruit fly culture medium upright, record the number and date, and place it in an incubator. Cultivate at room temperature (25°C), set the light cycle to 12 hours of light and 12 hours of darkness, and wipe the incubator with 75% ethanol for disinfection every 3 days.

[0093] 2) After 10 days, anesthetize the parent fruit fly with a CO2 anesthesia needle and remove it. Place a layer of toilet paper on the bottom of the propagation bottle to provide a dry place for the second-generation fruit fly larvae to pupate.

[0094] 3) Once the first generation fruit flies become adults, the generation is complete and feeding experiments can be performed.

[0095] 2. Drosophila feeding experiment

[0096] 1) Propagate Drosophila according to the "Passage of Drosophila" method. After the first generation develops into adults, collect flies every 8 hours and transfer them to culture tubes containing Drosophila culture medium. Place 20 female flies and 5 male flies in each culture tube. Collect flies continuously for up to 3 days, for a total of 10 tubes. Record the tube numbers and dates, and place them in an incubator. Incubate at room temperature (25°C) with a photoperiod of 12 hours light and 12 hours dark. Disinfect the incubator with 75% ethanol every 3 days.

[0097] 2) After the first generation fruit flies were cultured for 7 days, female fruit flies were picked with a brush under an optical microscope and transferred to a culture tube containing 3 mL of 1% agarose solution and starved for 24 hours.

[0098] 3) After starvation treatment, transfer the flies to 1% blue assay medium. After 15 minutes, quickly collect the flies into 1.5 ml centrifuge tubes and freeze them at -20°C to kill. 1% blue assay medium is a fruit fly culture medium containing 1% Brilliant Blue dye (Catalog #229730050, Thermo Fisher Scientific, China).

[0099] 4) Place dead fruit flies under a light microscope. Remove any fruit flies with no blue pigment in their guts (this is to exclude fruit flies that died of starvation before ingesting the bright blue food). Use forceps to remove the heads of the remaining fruit flies (which contain pigment) under a light microscope. Use a brush to transfer groups of 20 fruit flies to 2 mL thick-walled tubes. Add three 1 mm steel balls and 500 μL of 1x phosphate buffer. Grind using a high-throughput tissue grinder at 60 Hz for 5 seconds. Repeat once.

[0100] 5) Centrifuge the sample at 13,500 rpm for 20 minutes, aspirate 400 μL of the supernatant into a 1.5 mL centrifuge tube. Centrifuge the sample at 13,500 rpm for 20 minutes, aspirate 100 μL of the supernatant into a 96-well plate, shake and mix in a microplate reader, and read the OD at 320 nm.

[0101] The results are as follows Figure 1 As shown in F, the results showed that the above 6 culture media were effective in female wild-type fruit flies (w 118 ) There was no significant difference in food preference, which ruled out the influence of calorie restriction on subsequent experiments.

[0102] Example 2: Effects of Ferulic Acid-Different Material Culture Medium on Drosophila Lifespan Under 5% Hydrogen Peroxide Induction

[0103] This example uses the six culture media prepared in step 2 of Example 1 to explore their effects on the wild-type female Drosophila melanogaster (W 118 ) lifespan. The specific steps are as follows:

[0104] 1. Propagate Drosophila according to the "Drosophila Passage" method in Example 1. After the first generation develops into adults, collect flies every 8 hours and transfer them to culture tubes containing Drosophila culture medium. Place 20 female flies and 2 male flies in each culture tube. Collect 150 flies per experimental group for up to 3 consecutive days. Record the numbers and dates. Incubate at 25°C with a photoperiod of 12 hours of light and 12 hours of darkness. Disinfect the incubator with 75% ethanol every 3 days.

[0105] 2. After the first generation of fruit flies were cultured for 7 days, female fruit flies were picked with a brush under an optical microscope and transferred to a culture bottle containing 3 mL of 1% agarose solution and starved for 24 h.

[0106] 3. After starvation treatment, transfer the fruit flies to culture tubes containing 5% hydrogen peroxide Drosophila medium for continued culture. Place 10 female flies in each culture tube, for a total of 10 tubes. The 5% hydrogen peroxide Drosophila medium is prepared as follows: 1) Prepare 5% hydrogen peroxide Drosophila medium according to the following recipe: 0.8 g agarose, 10 g sucrose, 10 ml 10x phosphate buffer (pH 7.0), and dilute to 100 ml with water. 2) Heat the above components in a microwave oven until completely dissolved. After cooling to 40°C, add 16.6 ml of 30% hydrogen peroxide and stir thoroughly. 3) Use a syringe to dispense the medium into culture tubes, 2 ml per tube. Place a layer of sterile gauze at the tube opening. Once the medium becomes clear and transparent, plug the tube opening with an ethanol-sterilized sponge plug. The tube is ready for use.

[0107] 4. Count the number of dead fruit flies every 12 hours and add 3 ml of 5% hydrogen peroxide fruit fly culture medium into the bottle with a syringe to prevent the decomposition of hydrogen peroxide until all fruit flies are dead.

[0108] The results are as follows Figure 2 As shown in Table 1, the results showed that fruit flies fed with cornmeal had the longest maximum survival time (156 hours), while those fed with refined flour had the shortest maximum survival time (96 hours). Compared with the median survival time (72 hours) of fruit flies fed with low-ferulic acid whole wheat flour and refined flour, the median survival time of fruit flies fed with high-ferulic acid Yunong 907 whole wheat flour (96 hours) and Shangmai 189 whole wheat flour (84 hours) increased by 33% and 17%, respectively.

[0109] Under the conditions of 5% hydrogen peroxide-induced oxidative damage, there was no significant difference in the median survival time among the three groups of fruit flies after being fed with corn flour, Yunong 907 whole wheat flour and Shangmai 189 whole wheat flour medium. However, compared with fruit flies fed with Zhoumai 18 whole wheat flour, Jimai 38 whole wheat flour and refined flour medium, they had significantly higher resistance to 5% hydrogen peroxide-induced oxidative damage. The results showed that compared with fruit flies fed with low ferulic acid whole wheat flour medium and refined flour medium, fruit flies fed with high ferulic acid whole wheat flour medium enhanced the survival rate of wild-type female fruit flies (w 118 ) to oxidative damage and increased the resistance of wild-type female Drosophila (w 118 ) median survival time.

[0110] Table 1. Survival analysis of Drosophila culture medium fed with materials containing different levels of ferulic acid under the 5% hydrogen peroxide injury model

[0111]

[0112]

[0113] Note: The numbers in brackets are the total number of female fruit flies. The p value and chi-square were calculated by log rank test.

[0114] Example 3: Effects of Ferulic Acid-Different Material Culture Medium on Drosophila Lifespan under High Temperature Stress

[0115] As the global climate gradually warms, temperature changes will have a negative impact on human health. High temperature environments can induce the accumulation of ROS, causing oxidative damage to the body. This example uses the six culture media prepared in Example 1 to explore their effects on wild-type female Drosophila melanogaster (W 118 ) lifespan. The specific steps are as follows:

[0116] 1. Propagate Drosophila according to the "Drosophila Passage" method in Example 1. After the first generation develops into adults, collect flies every 8 hours and transfer them to culture tubes containing Drosophila culture medium. Place 20 female flies and 5 male flies in each culture tube. Collect flies continuously for up to 3 days, for a total of 10 tubes. Record the tube numbers and dates. Incubate at 29°C with a photoperiod of 12 hours of light and 12 hours of darkness. Disinfect the incubator with 75% ethanol every 3 days.

[0117] 2. Replace the culture medium every 2-3 days, count the number of dead female flies, and replenish the number of male flies to keep the number of male flies at 5 until all female flies die.

[0118] The results are as follows Figure 3 As shown in Table 2, the results showed that fruit flies fed with cornmeal had the longest maximum survival time (40 days) and median survival time (35 days), while those fed with refined flour had the shortest maximum survival time (36 days). Compared with the median survival times of fruit flies fed with low-ferulic acid whole-wheat flour (30 days) and refined flour (30 days), the median survival times of fruit flies fed with Yunong 907 whole-wheat flour (34 days) and Shangmai 189 whole-wheat flour (33 days) increased by 33% and 11%, respectively. Under high-temperature stress conditions, fruit flies fed with cornmeal had the best ability to cope with heat stress. The median survival time of fruit flies fed with low-ferulic acid whole-wheat flour was significantly lower than that of fruit flies fed with high-ferulic acid whole-wheat flour. The above results showed that compared with feeding low-ferulic acid whole wheat flour medium and refined flour medium, feeding high-ferulic acid whole wheat flour medium enhanced the ability of wild-type female fruit flies to resist high temperature (29°C) stress and increased the median survival time of fruit flies.

[0119] Table 2. Survival analysis of Drosophila fed with culture media containing different ferulic acid contents under high temperature (29°C) stress conditions

[0120]

[0121]

[0122] Note: The numbers in brackets are the total number of female fruit flies. The p value and chi-square were calculated by log rank test.

[0123] Example 4: Effects of culture media containing different ferulic acid contents on the lifespan of wild-type female Drosophila in a natural aging model

[0124] With advances in medical care, life expectancy is increasing year by year, inevitably leading to an aging population. Delaying aging has become a hot topic in research on this aging society. The concept of "medicine and food share the same origin" is a key tenet of Traditional Chinese Medicine (TCM). While dosage differences often create a distinction between traditional Chinese medicine and food, the two are closely linked. Proper dietary adjustments can also help slow aging.

[0125] This example uses the six culture media prepared in Example 1 to explore their effects on the wild-type female Drosophila melanogaster (w 118 ) lifespan. The specific steps are as follows:

[0126] 1. Propagate Drosophila according to the "Drosophila Passage" method in Example 1. After the first generation develops into adults, collect flies every 8 hours and transfer them to culture tubes containing Drosophila culture medium. Place 20 female flies and 5 male flies in each culture tube, collecting 10 tubes in total. Collect flies continuously for up to 3 days, recording the number and date. Incubate at 25°C with a photoperiod of 12 hours of light and 12 hours of darkness. Disinfect the incubator with 75% ethanol every 3 days.

[0127] 2. Replace the culture medium every 2-3 days. Each time you change the food, count the number of dead female flies and replenish the number of male flies to keep the number of male flies at 5 until all the female flies die.

[0128] The results are as follows Figure 4As shown in Table 3, the results showed that fruit flies fed on cornmeal had the longest maximum survival time (72 days), while those fed on refined flour had the shortest maximum survival time (68 days). Fruit flies fed on high-ferulic acid whole wheat flour had a maximum survival time (71 days), which was 2% and 1% higher than those fed on low-ferulic acid Jimai 38 whole wheat flour (69 days) and Zhoumai 18 whole wheat flour (70 days), respectively. Fruit flies fed on cornmeal had the longest median survival time (58 days), while those fed on refined flour had the shortest median survival time (49 days). The median survival time of fruit flies fed with Shangmai 189 whole wheat flour medium (56 days) and Yunong 907 whole wheat flour medium (55 days) was extended by 5% and 4% respectively compared with that fed with low ferulic acid Jima 38 whole wheat flour medium (52 ​​days), and by 8% and 6% respectively compared with that fed with low ferulic acid Zhoumai 18 whole wheat flour medium (53 days). At the same time, under the natural aging model (25℃), there was no significant difference in the average lifespan among the three groups of fruit flies fed with corn flour, Shangmai 189 whole wheat flour and Yunong 907 whole wheat flour medium, but compared with the median survival time of fruit flies fed with low ferulic acid Jima 38 whole wheat flour and Zhoumai 18 whole wheat flour medium, they were all significantly improved. The above results show that compared with feeding with low ferulic acid whole wheat flour medium and refined flour medium, feeding with high ferulic acid whole wheat flour medium increased the lifespan of wild-type female fruit flies (w 118 ) Median survival time in a natural aging (25°C) model.

[0129] Table 3. Survival analysis of Drosophila fed with culture media containing different ferulic acid contents under the natural aging model (25°C)

[0130]

[0131]

[0132] Note: The numbers in brackets are the total number of fruit flies. The p value and chi-square were calculated by log rank test.

[0133] The present invention has been described in detail above. It will be apparent to those skilled in the art that the present invention may be practiced over a wide range of parameters, concentrations, and conditions without departing from the spirit and scope of the present invention and without unnecessary experimentation. Although specific embodiments have been given herein, it should be understood that further modifications may be made to the present invention. In summary, this application is intended to encompass any variations, uses, or improvements to the present invention, including those made by conventional techniques known in the art that depart from the scope of the present invention. Applications of the essential features may be made within the scope of the following claims.

Claims

1. Use of cereal flour or cereal flour products in any of the following A1)-A4): A1) preparing products for delaying animal aging; A2) preparing products that extend animal lifespan; A3) preparing products for enhancing the resistance of animals to high temperature stress; A4) preparing products for enhancing the resistance of animals to oxidative damage; The ferulic acid content in the cereal flour is at least 0.96 mg / g; The ferulic acid content in the cereal flour product is at least 0.37 mg / g.

2. The use according to claim 1, characterized in that: The cereal flour is made from cereal grains with a ferulic acid content of at least 0.96 mg / g.

3. The use according to claim 1 or 2, characterized in that: The cereal flour is corn flour or wheat flour.

4. The use according to claim 3, characterized in that: The corn flour is made from corn kernels having a ferulic acid content of at least 0.96 mg / g; Alternatively, the wheat flour is made from wheat grains having a ferulic acid content of at least 0.96 mg / g.

5. The use according to any one of claims 1 to 4, characterized in that: The cereal flour product is food or feed.

6. The use according to any one of claims 1 to 5, characterized in that: The high temperature stress is 29°C stress.

7. The use according to any one of claims 1 to 6, characterized in that: The oxidative damage is damage induced by hydrogen peroxide.

8. The use according to any one of claims 1 to 7, characterized in that: The animals include mammals and insects.

9. A cereal flour product, which is food or feed; the cereal flour product comprises the cereal flour according to claims 1-8; the ferulic acid content in the cereal flour product is at least 0.37 mg / g; the function of the cereal flour product is any one of the following B1) to B4): B1) Delaying animal aging; B2) Extend animal lifespan; B3) Enhance the animal's resistance to high temperature stress; B4) Enhance the animal body's resistance to oxidative damage.

10. Any of the following methods D1)-D4): D1) A method for delaying aging of an animal, comprising the step of causing the animal to eat or feed the cereal flour product according to any one of claims 1 to 9 as food or feed; D2) A method for prolonging the lifespan of an animal, comprising the step of causing the animal to eat or feed the cereal flour product according to any one of claims 1 to 9 as food or feed; D3) A method for enhancing the resistance of an animal to high temperature stress, comprising the step of causing the animal to consume or feed the cereal flour product according to any one of claims 1 to 9 as food or feed; D4) A method for enhancing the resistance of an animal body to oxidative damage, comprising the step of allowing the animal to eat or feed the cereal flour product according to any one of claims 1 to 9 as food or feed.

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