Application of colorant and composition in preparation of products for relieving gastrointestinal mucosa stimulation and preventing and relieving gastrointestinal diseases

Abstract

The invention discloses application of a colorant and a composition in preparation of a product for relieving gastrointestinal mucosa stimulation and preventing and relieving gastrointestinal diseases, and relates to the technical field of application of colorants. Cell experiments prove that the chromosome has a relieving and protecting effect on gastrointestinal mucosa stimulation injury caused by chili products, cytotoxicity experiments prove that the chromosome has no influence on cell activity, in addition, the chromosome can relieve inflammatory response caused by capsaicin, and compared with carotenoid, the chromosome has the advantage that the effect is better. And the chromosome can better relieve oxidative stress caused by capsaicin. Therefore, the colored body has a good application prospect in preparation of products for relieving gastrointestinal mucosa stimulation.

Description

Technical Field

[0001] The present invention relates to the technical field of chromoplast application, and in particular to the application of a chromoplast and a composition in the preparation of a product for relieving gastrointestinal mucosal irritation and preventing and relieving gastrointestinal diseases. Background Art

[0002] Chili peppers are widely consumed vegetables, condiments and spices in daily life. Capsaicin is the main source of chili peppers' pungent properties, accounting for about 69% of capsaicinoids and is considered to be the main pungent ingredient. Currently, chili pepper extracts and their main ingredients have been reported to have a variety of pharmacological effects, such as antibacterial, anti-obesity, and analgesic effects. However, the role of its main ingredient, capsaicin, remains controversial. [1] Studies have shown that capsaicin is a double-edged sword, with potential for biological activity at low concentrations, but often causing adverse effects at high concentrations. Life experience shows that after consuming a large amount of capsaicin, the gastrointestinal tract can produce a strong burning sensation. [2] , and even gastrointestinal pain and diarrhea [3] . Carotenoids are the most abundant natural pigments in nature besides chlorophyll. They are tetraterpenoids that are yellow, orange or red in color. There are many types of carotenoids, with a total of more than 750 species, which are widely found in animals, plants, microorganisms and algae. As one of the pigment components, carotenoids can not only play a key role in evaluating the appearance and nutritional quality of food, but also promote human health and reduce the risk of disease. They have active functions such as being a provitamin A, anti-oxidant, immune regulation, and prevention of cardiovascular disease. They are a class of substances with important nutritional value and functional activity for the visual and nervous systems. [4-5] Therefore, natural carotenoids are expected to become promising functional additives in the food industry due to their coloring properties and high nutritional value for human health.

[0003] Chromoplasts are organelles with phospholipid bilayer membranes that biosynthesize and accumulate carotenoids in plants. They have certain advantages in improving the water solubility and stability of carotenoids. A recent study found that under different storage conditions, tomato chromoplasts are more stable than free carotenoid extracts in tomatoes in terms of color properties and carotenoid preservation. [6] Therefore, chromoplasts show great potential as natural water-soluble colorants replacing carotenoids and functional food additives due to the advantages of their natural structure.

[0004] Patent CN201310050957.2 discloses a tomato chili sauce for regulating immunity and its preparation method, which comprises grinding the whole tomato colloid, boiling and fermenting it, grinding the chili colloid into glutinous chili and frying it, and then adding ingredients such as mushrooms, licorice and ginger powder, mixing and stir-frying the components, sterilizing, etc. The patent directly adds the whole tomato, which contains pectin, fiber and other food matrices that hinder the release and efficacy of carotenoids (i.e. lycopene) in the tomato; in addition, the patent does not verify the functional activity in vivo and in vitro.

[0005] Currently, there is a lack of gastrointestinal protective chili products with added chromoplasts as natural water-soluble carotenoids on the market.

[0006] In view of this, the present invention is proposed. Summary of the invention

[0007] The purpose of the present invention is to provide a chromoplast and a composition for use in preparing products for relieving gastrointestinal mucosal irritation and preventing and relieving gastrointestinal diseases to solve the above technical problems.

[0008] The present invention is achieved in that:

[0009] In a first aspect, the present invention provides the use of chromoplasts in the preparation of products for relieving gastrointestinal mucosal irritation, wherein the products are food additives, foods, pharmaceutical excipients, health products, and feed additives.

[0010] In a second aspect, the present invention also provides the use of chromoplasts in the preparation of drugs for preventing and alleviating gastrointestinal diseases, wherein the gastrointestinal diseases are selected from: heartburn, inflammatory bowel disease, Crohn's disease, ulcerative colitis, peptic ulcer, stress ulcer, colitis, or any combination of the above diseases.

[0011] In a third aspect, the present invention further provides a composition comprising: chromoplasts and capsaicin.

[0012] The present invention has the following beneficial effects:

[0013] Chromoplasts are organelles in plants that biosynthesize and accumulate carotenoids and have a phospholipid bilayer membrane structure. Using chromoplasts for product preparation overcomes the problems of poor stability and lipid solubility of individual carotenoids. Chromoplasts can be widely used in the preparation of water-soluble products.

[0014] The present invention confirms through cell experiments that chromoplasts have a protective effect on the irritation and damage of the gastrointestinal mucosa caused by pepper products. Cytotoxicity experiments prove that chromoplasts have no effect on cell viability. In addition, chromoplasts can alleviate the inflammatory response caused by capsaicin. Compared with carotenoids, chromoplasts can better alleviate the oxidative stress caused by capsaicin.

[0015] Therefore, chromoplasts have good application prospects in the preparation of products that relieve gastrointestinal mucosal irritation or the preparation of drugs for preventing and relieving gastrointestinal diseases. Experiments have shown that adding chromoplasts to chili products (such as chili sauce, spicy strips, etc.) not only does not affect the sensory color, but also has functional activity. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0017] Figure 1 Microscopic observations of chromosomes (A: polarized light micrograph of chromosomes under bright field, scale bar = 25 μm; B: transmission electron micrograph of chromosomes, scale bar = 500 nm, arrows mark tubular substructures in chromosomes).

[0018] Figure 2 This is the statistical result of detecting cell activity by CCK-8 method (A: capsaicin; B: chromoplasts; C: carotenoids);

[0019] Figure 3 It is a statistical diagram of the experimental results of anti-oxidative stress (A is the relative level of ROS, B is the level of SOD enzyme activity, C is the level of CAT enzyme activity, and D is the level of GSH-1 enzyme activity);

[0020] Figure 4 It is a statistical chart of the experimental results of the efficacy of inhibiting cellular inflammation (A is the level of IL-1β, B is the level of IL-6, and C is TNFα-1);

[0021] Figure 5 This is a product compounded with chromoplasts and chili sauce. Figure ①: added chromoplasts; ②: blank control. DETAILED DESCRIPTION

[0022] In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0023] Glossary:

[0024] Chromoplast: It is an organelle with a phospholipid bilayer membrane that biosynthesizes and accumulates carotenoids in plants. It is mainly found in the flowers and fruits of plants, and also in the nutritional organs of plants, such as carrot roots. Chromoplasts can be transformed from leucoplasts or chloroplasts.

[0025] Methods for extracting chromoplasts include but are not limited to: sucrose density centrifugation, etc.

[0026] In a first aspect, the present invention provides the use of chromoplasts in the preparation of products for relieving gastrointestinal mucosal irritation, wherein the products are food additives, foods, pharmaceutical excipients, health products, and feed additives.

[0027] The present invention finds that chromoplasts can effectively relieve gastrointestinal mucosal irritation and gastrointestinal mucosal irritation damage caused by pepper products. It can be used to prepare food additives, food, health products, feed additives. Pharmaceutical excipients include but are not limited to veterinary (use) drug excipients and human drug excipients.

[0028] Food additives include but are not limited to any one of flavoring agents, sweeteners, and acidulants.

[0029] In a preferred embodiment of the present invention, when the above-mentioned product is a pharmaceutical excipient, the pharmaceutical excipient also includes other pharmaceutically acceptable excipients.

[0030] In a preferred embodiment of the present invention, the pharmaceutically acceptable excipient is selected from at least one of a filler, a disintegrant, a lubricant, a flavoring agent, a binder, a suspending agent and a flavor.

[0031] Pharmaceutically acceptable excipients include but are not limited to: pharmaceutically acceptable carriers, auxiliary substances or solvents. Pharmaceutically acceptable excipients include various organic or inorganic carriers and / or auxiliary materials because they are commonly used for pharmaceutical purposes, especially for solid pharmaceutical preparations. Examples include: excipients, such as sucrose, starch, mannitol, sorbitol, lactose, glucose, cellulose, talc, calcium phosphate, calcium carbonate; binders, such as cellulose, methylcellulose, hydroxypropyl cellulose, polypropyl pyrrolidone, gelatin, gum arabic, polyethylene glycol, sucrose, starch; disintegrants, such as starch, hydrolyzed starch, carboxymethyl cellulose, carboxymethyl cellulose calcium salt, hydroxypropyl starch, ethylene glycol starch sodium, sodium bicarbonate, calcium phosphate, calcium citrate; lubricants, such as magnesium stearate, talc, sodium lauryl sulfate; flavors, such as citric acid, menthol, glycine , orange powder; preservatives, such as sodium benzoate, sodium bisulfite, parabens (such as methyl paraben, ethyl paraben, propyl paraben, butyl paraben); stabilizers, such as citric acid, sodium citrate, acetic acid and polycarboxylic acids from the titriplex series, such as diethylenetriaminepentaacetic acid (DTPA); suspending agents, such as methylcellulose, polyvinylpyrrolidone, aluminum stearate; dispersants; diluents, such as water, organic solvents; waxes, fats and oils, such as beeswax, cocoa butter; polyethylene glycol; white petrolatum, etc.

[0032] The health care product is selected from hard capsules, tablets, granules, pills or oral liquids.

[0033] In a preferred embodiment of the present invention, the feed further comprises one or more of complex vitamins, enzyme preparations, probiotic preparations, and basic feed.

[0034] In a preferred embodiment of the present invention, the chromoplasts are from at least one of the following parts of the plant: flowers, fruits, roots, leaves, juice sacs and peels, such as roots of carrots, juice sacs and leaves of citrus fruits, peels of peppers, melons and tomatoes, petals of daffodils and sunflowers, and sepals of nasturtiums.

[0035] In a preferred embodiment of the present invention, the chromoplasts are derived from wolfberry, physalis, marigold, sweet potato, daffodil, papaya, watermelon, corn, pepper, citrus, pumpkin, tomato, carrot, mango or palm fruit.

[0036] In a preferred embodiment of the present invention, the food is beverages, meat, sausages, bread, candy, snacks, noodles, ice cream, dairy products, soups, electrolyte drinks, drinking water, alcoholic beverages, chewing gum, tea, vitamin complexes, jams and chili sauces.

[0037] In one embodiment, the food is selected from the group consisting of spreadable food products, candy products, savory products, dairy products, dairy substitutes, and beverages. In one embodiment, the food is selected from the group consisting of jams, jellies, nut butters, hard candies, gummy candies including soft gel candies, chocolate syrups, flavored syrups, fruit snacks, fruit gel bars, gelatin substitutes, aspics, creamers, yogurts, cheeses, cream cheeses, sour creams, low-fat dairy products, non-dairy creamers, non-dairy yogurts, non-dairy cream cheeses, non-dairy sour creams, low-fat non-dairy products, protein shakes, meal replacement shakes, and any food described herein.

[0038] In a preferred embodiment of the present invention, the application includes any of the following uses:

[0039] (1) Reduce cellular oxidative stress levels;

[0040] (2) Inhibit cellular inflammation.

[0041] In a preferred embodiment of the present invention, reducing the cellular oxidative stress level includes but is not limited to: increasing the activity of at least one of the following enzymes in the cell: SOD, CAT and GSH-1 enzyme.

[0042] In a preferred embodiment of the present invention, inhibiting cellular inflammation includes but is not limited to: inhibiting the level of cellular pro-inflammatory factors.

[0043] In a preferred embodiment of the present invention, the cellular proinflammatory factor is selected from IL-1β, IL-6 and TNFα-1.

[0044] In a second aspect, the present invention also provides the use of chromoplasts in the preparation of drugs for preventing and alleviating gastrointestinal diseases, wherein the gastrointestinal diseases are selected from: heartburn, inflammatory bowel disease, Crohn's disease, ulcerative colitis, peptic ulcer, stress ulcer, colitis, or any combination of the above diseases.

[0045] In a third aspect, the present invention further provides a composition comprising: chromoplasts and capsaicin.

[0046] Capsaicin, also known as capsaicin, is an organic compound with a molecular formula of C 18 H 27 NO3, CAS number: 404-86-4.

[0047] The form of the composition includes but is not limited to: liquid, solid or semi-solid.

[0048] In a preferred embodiment of the present invention, the mixing ratio of capsaicin and chromoplasts is 0-50 μg: 0.07-0.35 nmoL; such as 1-10 μg: 0.07-0.3 nmoL, or 2-30 μg: 0.07-0.3 nmoL, or 10-50 μg: 0.07-0.35 nmoL; or 20-50 μg: 0.1-0.35 nmoL.

[0049] In a preferred embodiment of the present invention, the mixing ratio of capsaicin and chromoplasts is 50 μg:0.35 nmol / L.

[0050] In a preferred embodiment of the present invention, the composition is a chili product;

[0051] The composition is a condiment.

[0052] Condiments are tomato-based, white sauce-based, fat-based, oil-based, cheese-based, mayonnaise-based, meat-based, fish-based, vegetable-based, hot sauce-based or mixtures thereof.

[0053] In a preferred embodiment of the present invention, the chili products are chili sauce, chili oil, chili powder and spicy strips.

[0054] In a preferred embodiment of the present invention, the chromoplasts are from at least one of the following parts of the plant: flowers, fruits, roots, leaves, juice sacs and peels;

[0055] In a preferred embodiment of the present invention, the chromoplasts are derived from wolfberry, physalis, marigold, sweet potato, daffodil, papaya, watermelon, corn, pepper, citrus, pumpkin, tomato, carrot, mango or palm fruit.

[0056] The features and performance of the present invention are further described in detail below in conjunction with the embodiments.

[0057] Example 1

[0058] This embodiment provides a method for preparing a chromoplast, which is as follows:

[0059] Sucrose density gradient centrifugation was used to obtain chromoplasts (chr) from carotenoid-rich plant tissues (wolfberry). Briefly, the washed carotenoid-rich plant tissues were weighed, pre-cooled PBS was added at a ratio of 1:10, and the homogenization was stirred using a spiral juicer (power 300W), with intermittent pulsation 4 times, each time for 1s. After simple filtration through two layers of absorbent cotton cloth, the filtrate was collected, centrifuged at 4°C, 3000g for 20min, the supernatant was discarded, and after removing the supernatant, the crude chromoplast extract was resuspended in PBS buffer and the resuspended solution was collected. Prepare 20% and 50% sucrose solutions and divide them into 50mL centrifuge tubes. The specific operation is as follows: first add 5mL of 50% sucrose solution, then add 5mL of 20% sucrose solution, and finally take 5mL of resuspension and add it to the sucrose solution, centrifuge horizontally at 5000g, 30min, 4℃, carefully pipette the chromoplast enriched layer (i.e. the middle layer, the chromoplast density is higher than 20% sucrose but lower than 50% sucrose) into a beaker, and dilute it with PBS buffer at a ratio of 1:1, centrifuge at 3000g, 15-20min, 4℃, discard the supernatant, and the bottom precipitate is the chromoplast (chr). Then re-dissolve it with DMEM containing 0.1% DMSO for subsequent cell experiments.

[0060] Optical Microscopy Reference for Chromoplasts Figure 1 As shown, the results showed that the chromoplasts of wolfberry fruit appeared orange under bright field, were spherical or elliptical in shape, and had a diameter of 4-6 μm.

[0061] Comparative Example 1

[0062] This comparative example provides a method for preparing plant puree, which is as follows:

[0063] Using carotenoid-rich plant tissue as raw material, DMEM containing 0.1% DMSO was added at a ratio of 1:10 (the reason for selecting this solvent: to be used for functional verification in subsequent cell experiments) and homogenized (300W), and then simply filtered with absorbent cotton cloth to prepare plant puree.

[0064] Comparative Example 2

[0065] This comparative example provides a method for preparing carotenoids (CAR), which is as follows:

[0066] About 10 mL of extract (methanol / ethyl acetate / petroleum ether, 1:1:1, v / v / v, both containing 0.1 g / L BHA (butylated hydroxyanisole) and BHT (butylated hydroxytoluene)) was added to 0.1 g of carotenoid-rich plant tissue, and extracted for 30 s using an ultrasonic disruptor. The sample was placed in an ice bath, and after ultrasonic extraction, centrifuged (3000 g, 5 min), and the supernatant was pipetted into a brown bottle. Repeat the extraction until the sample is colorless, combine the supernatants, blow nitrogen to dryness, and prepare carotenoids (car). Subsequently, it was re-dissolved in DMEM containing 0.1% DMSO for subsequent cell experiments.

[0067] BHA (butylated hydroxyanisole) and BHT (butylated hydroxytoluene) are two commonly used food additives, which are mainly used to prevent or delay the oxidation process of food, thereby improving the stability of food and extending the storage period.

[0068] Comparative Example 3

[0069] Weigh a certain amount of capsaicin standard (source leaf, item number A10027) and dissolve it in 0.1% DMSO in DMEM to prepare a certain concentration of capsaicin solution (cap) for subsequent cell experiments.

[0070] Experimental Example 1

[0071] This experimental example provides a quantitative analysis method for carotenoids, as follows:

[0072] About 10 mL of extract (methanol / ethyl acetate / petroleum ether, 1:1:1, v / v / v, each containing 0.1 g / L BHA and BHT) was added to the color body prepared in Example 1 or the plant puree prepared in Comparative Example 1, respectively, and extracted for 30 seconds using an ultrasonic crusher. The sample was ice-bathed, and after ultrasonic extraction, centrifuged (3000 g, 5 min), and the supernatant was pipetted into a brown bottle. The extraction was repeated until the sample was colorless, the supernatant was combined, and nitrogen was blown to dryness. The dried extract was dissolved in a mixture of methanol and methyl tert-butyl ether (1:1, v / v), filtered through a 0.45 μm PTFE membrane into a brown vial, and analyzed using HPLC.

[0073] A YMC C30 reverse phase column (250 mm × 4.6 mm, 3 μm) plus a YMC C30 guard column of the same material was used for liquid chromatography separation. The analysis conditions were: temperature 40°C, mobile phase rate 1 mL / min, mobile phase A was methanol / tBME / water (80:18:2, v / v / v); mobile phase B was methanol / tBME / water (8:90:2, v / v / v).

[0074] The elution gradient was as follows: the concentration of mobile phase B increased from 0% to 30% in 5 min, from 30% to 86% in 30 min, from 86% to 100% in 2 min, and decreased from 100% to 0% in 4 min, and stayed at 0% for 4 min, with a total time of 45 min. The total amount of carotenoids was recorded as the sum of the contents of zeaxanthin (ZEA), zeaxanthin monopalmitate (ZMP) and zeaxanthin dipalmitate (ZDP). Among them, ZEA and ZDP were qualitatively determined by comparing the retention time with the standard and UV-visible absorption spectra, and quantitatively determined by the standard curve of their standards, while ZMP was quantitatively determined by UV-visible absorption spectra and reference to Hempel et al. [7] The quantitative analysis was performed using the standard curve of ZEA standard.

[0075] To ensure that the carotenoid content in the chromoplasts used in the cell experiment and their control ratio (plant puree, free carotenoids) is consistent.

[0076] Experimental Example 2

[0077] In order to determine the safe concentration range of the samples when the survival rate of GES-1 cells is greater than 80%, the cytotoxicity of capsaicin, chromoplasts and carotenoids was evaluated using CCK-8 assay in this experiment.

[0078] Human gastric mucosal cells (GES-1) grown to 80-90% confluence in a cell culture flask were plated at 1×10 5 cell / mL, 100 μL per well was inoculated in a 96-well plate, 5 replicate wells were set for each treatment, the culture plate was placed in an incubator for pre-culture for 12 to 18 hours, and the cells in each well grew by attachment to the wall and the density reached 70 to 80%; the liquid in the well was aspirated, and the PBS buffer was washed twice, and different concentrations of the micellar phase to be tested (prepared or diluted with basal culture medium) were added to the culture plate, and incubated in an incubator for 24 hours; 10 μL of CCK-8 solution was added to each well, and care was taken not to generate bubbles; incubated in an incubator for 1 to 2 hours; the absorbance of each well at 450 nm was measured by an enzyme marker.

[0079] Calculation formula: Cell survival rate = [(As-Ab) / (Ac-Ab)] × 100%

[0080] As: absorbance of experimental wells (containing cells, culture medium, CCK-8 solution, and drug solution);

[0081] Ac: absorbance of control well (containing cells, culture medium, CCK-8 solution, but no drug);

[0082] Ab: absorbance of blank well (containing culture medium and CCK-8 solution, but not cells or drugs).

[0083] Cell activity results reference Figure 2 As shown, the original concentration of capsaicin is 100 μg / mL. Figure 2 As shown in Figure A, the cell survival rate was greater than 90% after the capsaicin was diluted 2 times, so 50 μg / mL was selected for subsequent experimental exploration. The original concentrations of chromoplasts and carotenoid samples were both 1.42 nmol / mL. Figure 2 As can be seen from Figures B and C, the carotenoid sample had no effect on cell viability after being diluted 2 times, while the chromoplast had no effect on cell viability after being diluted 4 times. In order to compare the efficacy of carotenoids and chromoplasts at the same concentration, samples diluted 4 times, i.e., a concentration of 0.35 nmol / mL, were selected for subsequent experiments.

[0084] Experimental Example 3

[0085] This experimental example verifies the anti-oxidative stress efficacy. The chromoplasts prepared in Example 1, the plant puree prepared in Comparative Example 1, the carotenoids prepared in Comparative Example 2, and the capsaicin prepared in Comparative Example 3 are used to verify the anti-oxidative stress efficacy.

[0086] Dilute the cell suspension (GES-1 cells) to 1×10 6 cell / mL, 1mL was inoculated into 12-well plates per well, 3 replicates were set for each treatment, and the culture plates were placed in an incubator for 24h until the cells adhered to the wall and grew and the density reached 70-80%; the liquid in the wells was aspirated, washed twice with PBS buffer, and the cells were treated with 1mL 0.1% DMSO in DMEM, 50μg / mL capsaicin solution (cap), 50μg / mL capsaicin solution + 0.35nmol / mL chromoplasts (cap-chr), and 50μg / mL capsaicin solution + 0.35nmol / mL carotenoids (cap-car) for 24 hours, respectively, and the solution was removed and washed twice with PBS. Subsequently, the subsequent operations were carried out according to the ROS, SOD, CAT, and GSH-Px kits to verify the preventive and protective effects of carotenoids in different food matrices on the oxidative stress of gastric mucosal cells induced by capsaicin. The ROS, SOD, CAT, and GSH-Px test kits were all from Nanjing Jiancheng Bioengineering Research Institute, and the catalog numbers are as follows: ROS (E004-1-1), SOD (A001-3-2), CAT (A007-1-1), and GSH-PX (A006-2-1).

[0087] Depend on Figure 3It can be seen that compared with the blank group, after capsaicin treatment, the SOD, CAT and GSH-1 enzyme activities in the cells were significantly reduced, while the enzyme activity was significantly increased after adding chromoplasts and carotenoids; although the ROS content did not change significantly before and after addition, the antioxidant-related enzyme activity was significantly increased, indicating that both chromoplasts and carotenoids can reduce the cell oxidative stress caused by capsaicin, and the CAT in the cells treated with chromoplasts was significantly higher than that in the carotenoid treatment group, indicating that chromoplasts are significantly better than carotenoids in preventing and alleviating the oxidative stress caused by capsaicin.

[0088] Experimental Example 4

[0089] This experimental example verifies the efficacy of preventing inflammation. The chromoplasts prepared in Example 1, the plant puree prepared in Comparative Example 1, the carotenoids prepared in Comparative Example 2, and the capsaicin prepared in Comparative Example 3 are used to verify the efficacy of preventing inflammation.

[0090] Dilute the cell suspension (GES-1 cells) to 1×10 6 cell / mL, 1mL was inoculated into 12-well plates per well, 3 replicates were set for each treatment, and the culture plates were placed in an incubator for 24h, until the cells grew and adhered to the wall and the density reached 70-80%; the liquid in the wells was aspirated, washed twice with PBS buffer, and the cells were treated with 1mL 0.1% DMSO in DMEM, 50μg / mL capsaicin solution (cap), 50μg / mL capsaicin solution + 0.35nmol / mL chromoplasts (cap-chr), and 50μg / mL capsaicin solution + 0.35nmol / mL carotenoids (cap-car) for 24 hours, respectively, and the solution was removed and washed twice with PBS. Subsequently, the subsequent operations were carried out according to the ELISA kits of IL-6, IL-1β, and TNF-α to verify the preventive and protective effects of carotenoids in different food matrices on gastric mucosal cell inflammation caused by capsaicin.

[0091] The ELISA kits for IL-6, IL-1β, and TNF-α were all from Nanjing Jiancheng Bioengineering Institute, and the catalog numbers are as follows: IL-6 (H007-1-1), IL-1b (H002-1-2), and TNF-a (H052-1-2).

[0092] Results reference Figure 4 As shown by Figure 4 It can be seen that compared with the blank group, after capsaicin treatment, the levels of IL-1β, IL-6 and TNFα-1 inflammatory factors secreted by cells were significantly increased, while the levels of inflammatory factors were significantly reduced after the addition of chromoplasts and carotenoids, indicating that both chromoplasts and carotenoids can alleviate the inflammatory response caused by capsaicin, and there is no significant difference in the effects of chromoplasts and carotenoids.

[0093] In summary, both chromoplasts and carotenoids can prevent and alleviate the cell oxidative stress and inflammatory response caused by capsaicin, and there is no significant difference in the effects between the two. However, considering the limitations of the application of carotenoids due to fat solubility, the subsequent selection of 50 μg of capsaicin products and 0.35 nmol of chromoplasts. It should be clear that within the range of non-toxicity to cells, the addition of carotenoids and chromoplasts can have an effect. This experimental example only shows the effect after the maximum amount of chromoplasts and carotenoids are added. In other embodiments, it is also possible to choose to mix capsaicin products with chromoplasts according to other compounding ratios.

[0094] Experimental Example 5

[0095] In this experimental example, a chili product including chromoplasts and capsaicin was prepared, including red chopped pepper sauce, yellow lantern pepper sauce, spicy strips, chili powder, and chili oil chili products. Taking the extraction of chromoplasts from wolfberry and adding them to red chopped pepper sauce as an example, evaluation and identification were carried out. A compound product of chromoplasts and chopped pepper sauce was prepared by mixing 50μg of capsaicin product with 0.35nmoL of chromoplasts. Take a commercially available chopped pepper product, determine its capsaicin content, and add 50μg of capsaicin product and 0.35nmoL of chromoplasts to a commercially available chopped pepper product (Xiangjunfu red chopped pepper sauce) according to the optimized optimal ratio of 50μg of capsaicin product and 0.35nmoL of chromoplasts. The national standard "GBT21266-2007" is used for quantitative analysis of the capsaicin content in chili products.

[0096] The inspection team for sensory evaluation of the differences between chopped pepper samples by the triangle test method consists of 16 evaluators. The evaluators are required to observe and taste three chopped pepper samples in turn, one of which is chopped pepper with wolfberry chromoplasts added, and the other two are pure chopped pepper. The evaluators can use bread and rinse their mouths with water when evaluating the samples.

[0097] After the assessment, please give the corresponding answers on the triangle test question and answer form of the Doujiao difference test (see Table 1).

[0098] When analyzing, consider:

[0099] H0: Null hypothesis, there is no significant difference between samples.

[0100] H1: Alternative hypothesis: There are significant differences between samples.

[0101] α is 0.05

[0102] Table 1: Question and answer table for the triangle test method for the difference test of chopped pepper with added chromoplasts

[0103]

[0104] 877, 899 and 865 are sample numbers.

[0105] In the tasting experiment, the number of valid questionnaires is 16, of which 5 are correct answers and 11 are wrong answers. For this result, X 2 =0.03<<3.84,

[0106] Use chi-square test to calculate X 2 , if the error rate is too high, that is, X 2 It is much smaller than 3.84, indicating that there is no significant difference between the samples.

[0107] This proves that the evaluators cannot detect whether chromoplasts are added to the chopped pepper sauce samples.

[0108] Results reference Figure 5 As shown, the addition of chromoplasts has no significant change on the appearance of the chopped pepper sauce, and the addition of chromoplasts will not have any effect on the appearance and taste of the chopped pepper sauce.

[0109] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

[0110] References

[0111] [1]Srinivasan K. Biological activities of red pepper(Capsicum annuum) and its pungent principle capsaicin:A Review[J].Critical Reviews inFoodScience and Nutrition, 2016, 56(9):1488-1500.

[0112] [2]Van Avesaat M, Troost FJ, Westerterp-Plantenga MS, et al. Capsaicin-induced satiety is associated with gastrointestinal distress but not with the release of satiety hormones[J]. American Journal of Clinical Nutrition, 2016, 103(2):305-313.

[0113] [3]Hammer J,Vogelsang H.Characterization of sensations inducedbycapsaicin in the upper gastrointestinal tract[J].NeurogastroenterologyandMotility,2007,19(4):279-287.

[0114] [4]Izumi-Nagai,K.,Nagai,N.&Ohgami,K.(2007).Macular pigmentlutein isanti-inflammatory in preventing.Arteriosclerosis,Thrombosis,andVascularBiology,27,2555-2562.DOI:10.1161 / ATVBAHA.107.151431.

[0115] [5]Lopresti,A.L.,Smith,S.J.&Drummond,P.D.(2022).The effects ofluteinand zeaxanthin supplementation on cognitive function in adults withself-reported mild cognitive complaints:A randomized,double-blind,placebo-controlled study.Frontiers in Nutrition,9,843512.DOI:10.3389 / fnut.2022.843512.

[0116] [6]Wang Y Q,Yang Y,Fei Z J,et al.Proteomic analysis ofchromoplastsfrom six crop species reveals insights into chromoplast functionanddevelopment[J].Journal of Experimental Botany,2013,64(4):949-61.DOI:10.1093 / jxb / ers375.

[0117] [7]Hempel J, N C,Sprenger J,et al.Ultrastructuraldepositionforms and bioaccessibility of carotenoids and carotenoidesters from gojiberries(Lycium barbarum L.)[J].Food Chemistry,2017,218:525-533.DOI:10.1016 / j.foodchem.2016.09.065。

Claims

1. Application of chromoplasts in the preparation of products for relieving gastrointestinal mucosal irritation, It is characterized in that The products are food additives, foods, pharmaceutical excipients, health products, and feed additives.

2. The use according to claim 1, It is characterized in that The chromoplasts are from at least one of the following parts of the plant: flowers, fruits, roots, leaves, juice sacs and peels; Preferably, the chromoplasts are derived from wolfberry, physalis, marigold, sweet potato, daffodil, papaya, watermelon, corn, pepper, citrus, pumpkin, tomato, carrot, mango or palm fruit.

3. The use according to claim 1, It is characterized in that The food products are drinks, meat, sausages, bread, candy, snacks, noodles, ice cream, dairy products, soups, electrolyte drinks, drinking water, alcoholic beverages, chewing gum, tea, vitamin complexes, jams and chili sauce.

4. The use according to claim 1, It is characterized in that The application includes any of the following uses: (1) Reduce cellular oxidative stress levels; (2) Inhibit cellular inflammation.

5. The use according to claim 4, It is characterized in that The reducing the cellular oxidative stress level comprises: increasing the activity of at least one of the following enzymes in the cell: SOD, CAT and GSH-1 enzyme; Preferably, the inhibiting of cellular inflammation comprises: inhibiting the level of cellular pro-inflammatory factors; Preferably, the cellular proinflammatory factor is selected from IL-1β, IL-6 and TNFα-1.

6. Application of chromoplasts in the preparation of drugs for preventing and alleviating gastrointestinal diseases. It is characterized in that The gastrointestinal disease is selected from the group consisting of heartburn, inflammatory bowel disease, Crohn's disease, ulcerative colitis, peptic ulcer, stress ulcer, colitis, or any combination thereof.

7. A composition, It is characterized in that It includes: Chromoplasts and capsaicin.

8. The composition according to claim 7, It is characterized in that The mixing ratio of capsaicin and chromoplasts is 0-50 μg: 0.07-0.35 nmoL; Preferably, the mixing ratio of capsaicin and chromoplasts is 50 μg:0.35 nmoL.

9. The composition according to claim 8, It is characterized in that The composition is a condiment; Preferably, the condiment is selected from chili products; Preferably, the chili product is chili sauce, chili oil, chili powder and spicy strips.

10. The composition according to claim 8, It is characterized in that The chromoplasts are from at least one of the following parts of the plant: flowers, fruits, roots, leaves, juice sacs and peels; Preferably, the chromoplasts are derived from wolfberry, physalis, marigold, sweet potato, daffodil, papaya, watermelon, corn, pepper, citrus, pumpkin, tomato, carrot, mango or palm fruit.

Images