Composition for relieving oxidative stress injury of intestinal tract and application thereof

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to a composition for relieving oxidative stress injury of intestinal tracts and application of the composition. The liquorice extract, the pueraria flower extract and the citrus aurantium flower extract are compounded according to a certain proportion, the antioxidant and anti-inflammatory effects of the liquorice extract are complementary with the powerful antioxidant and barrier repair functions of the pueraria flower extract and the citrus aurantium flower aqueous extract, and the overall protection effect is enhanced. The synergistic effect not only can more comprehensively relieve intestinal injury caused by oxidative stress, but also can improve the capability of preventing and repairing oxidative injury, so that a more effective relieving effect is achieved. On the basis, the wild buckwheat rhizome extract is further compounded, so that the overall protection effect can be further enhanced, the cell survival rate after H2O2 damage is improved, and ROS generation is inhibited. Results of the embodiment show that the composition provided by the invention can improve the cell survival rate after H2O2 injury, inhibit the generation of ROS (reactive oxygen species) and effectively relieve the oxidative stress injury of the intestinal tract.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to a composition for alleviating intestinal oxidative stress damage and an application thereof. Background Art

[0002] Intestinal oxidative stress refers to the oxidative damage to intestinal cells and tissues caused by excessive production of free radicals and other oxidative substances, which exceeds the clearance capacity of the intestinal antioxidant system. This damage not only affects the structure and function of the intestine, but may also lead to a series of serious health problems, including inflammation, infection, immune dysfunction, and systemic diseases. In recent years, intestinal oxidative stress has been considered to be closely related to the occurrence and progression of many gastrointestinal diseases, such as enteritis, inflammatory bowel disease (IBD), irritable bowel syndrome (IBS), and intestinal cancer.

[0003] Currently, the treatment of intestinal oxidative stress mainly focuses on the use of antioxidants and anti-inflammatory drugs. These methods can alleviate the damage caused by oxidative stress to a certain extent, but there are still some problems, such as low bioavailability of antioxidants, drug side effects, and limited control of oxidative stress. Therefore, the development of new and more effective anti-oxidation and intestinal protection treatment options is of great clinical significance. Summary of the invention

[0004] The purpose of the present invention is to provide a composition for alleviating intestinal oxidative stress damage and its application, to improve the cell survival rate after intestinal oxidative stress damage, to inhibit the production of ROS, to reduce intestinal damage caused by oxidative stress, to improve the prevention and repair capabilities of oxidative damage, thereby achieving a more effective relief effect.

[0005] The present invention provides a composition, which comprises liquorice extract, kudzu flower extract and dahliae flower extract; the mass ratio of the liquorice extract, kudzu flower extract and dahliae flower extract is (1-10): (1-10): (1-10).

[0006] Preferably, the composition further comprises a fagopyrum serrata extract; the mass ratio of the licorice extract, the kudzu flower extract, the agave clover flower extract and the fagopyrum serrata extract is (1-10): (1-10): (1-10): (0.5-5).

[0007] Preferably, the preparation method of the licorice extract comprises the following steps:

[0008] Soaking liquorice powder in sodium hydroxide solution to obtain a soaking solution;

[0009] Reflux extraction is performed on the soaking solution, the pH value of the extract is adjusted to 1.0-2.0, and the precipitate is collected and dried to obtain the licorice extract;

[0010] The reflux extraction is performed once, the temperature is 100-120° C., and the time is 1-2 hours.

[0011] Preferably, the mass volume ratio of the licorice powder to the sodium hydroxide solution is (1-2) kg: (8-20) L;

[0012] The concentration of the sodium hydroxide solution is 0.2-0.6 mol / L;

[0013] The soaking time is 10 to 24 hours;

[0014] The drying temperature is 50-66° C., the vacuum degree is ≤-0.08Mpa, and the drying time is 3-6 hours.

[0015] Preferably, the preparation method of the Pueraria lobata flower extract comprises the following steps:

[0016] The kudzu flower powder is mixed with an ethanol solution having a volume concentration of 30 to 60%, and extracted to obtain an extract;

[0017] Concentrating the extract under reduced pressure to obtain an extract with a solid content of 15% to 40%;

[0018] The extract with a solid content of 15% to 40% is mixed with anhydrous ethanol to a volume concentration of 80 to 90% ethanol, and the mixture is allowed to stand for 1 to 4 hours and then centrifuged to collect the supernatant;

[0019] The supernatant is concentrated under reduced pressure to obtain an extract with a solid content of 85-95%, and the extract is spray-dried to obtain the kudzu flower extract.

[0020] Preferably, the extraction temperature is 60°C to 90°C, the number of times is 2 to 3 times, and the time of each extraction is 1 to 2 hours;

[0021] The mass ratio of the kudzu flower powder to the ethanol solution with a volume concentration of 30-60% is 1:(5-15).

[0022] Preferably, the preparation method of the daidai flower extract comprises the following steps:

[0023] After soaking the daidai flower in a 2-8 g / L citric acid aqueous solution for 2-3 hours, extracting by microwave heating for 2-3 times, separating the solid and the liquid, and collecting the filtrate; drying the filtrate to obtain the daidai flower extract;

[0024] The microwave heating extraction method each time is: heating to 60-90° C. with a microwave power of 80-300 W and extracting for 5-20 minutes.

[0025] Preferably, the preparation method of the golden buckwheat extract comprises the following steps: mixing golden buckwheat with an ethanol solution having a volume concentration of 30 to 80%, and refluxing to obtain an extract; concentrating the extract to obtain the golden buckwheat extract;

[0026] The reflux extraction is performed once at a temperature of 90 to 130° C. for 2 to 5 hours.

[0027] The mass ratio of the golden buckwheat to the ethanol solution with a volume concentration of 30-80% is 1:(5-10).

[0028] The present invention also provides the use of the composition described in the above technical solution in the preparation of intestinal injury drugs.

[0029] Preferably, the intestinal injury drug includes a drug that alleviates intestinal oxidative stress damage;

[0030] The oxidative stress damage includes H2O2 damage.

[0031] Beneficial effects:

[0032] The present invention provides a composition, which comprises a licorice extract, a kudzu flower extract and a dahliae flower extract; the mass ratio of the licorice extract, the kudzu flower extract and the dahliae flower extract is (1-10): (1-10): (1-10). The licorice extract contains ingredients such as glycyrrhizic acid and glycyrrhizin flavonoids, and exerts its effects mainly through anti-oxidation and anti-inflammatory effects. Glycyrrhizic acid can inhibit the inflammatory response caused by oxidative stress, reduce the level of pro-inflammatory factors, and at the same time increase the activity of antioxidant enzymes in the body, such as superoxide dismutase (SOD), and enhance the antioxidant capacity of cells. Glycyrrhizin flavonoids help repair damaged intestinal barriers and further protect intestinal mucosa. The main components of the kudzu flower extract include puerarin, flavonoids and isoflavones, which have strong antioxidant capacity. Puerarin and flavonoids can scavenge free radicals in the body and reduce oxidative damage. At the same time, they also have anti-inflammatory effects and inhibit the inflammatory response caused by oxidative stress. Isoflavones can regulate intestinal microbiota and improve intestinal health. The extract of the daisy flower is rich in flavonoids and polyphenols, and has significant antioxidant and anti-inflammatory effects. These ingredients reduce the damage of oxidative stress to the intestine by scavenging free radicals and inhibiting inflammatory reactions. In addition, the daisy flower extract can also promote the repair of intestinal mucosa and enhance the intestinal barrier function. When the present invention uses licorice extract, kudzu flower extract, daisy flower extract and golden buckwheat extract in combination, the antioxidant and anti-inflammatory effects of the licorice extract complement the potent antioxidant and barrier repair functions of the kudzu flower extract and the daisy flower extract, thereby enhancing the overall protective effect. This synergistic effect can not only more comprehensively reduce the intestinal damage caused by oxidative stress, but also improve the ability to prevent and repair oxidative damage, thereby achieving a more effective relief effect.

[0033] The present invention further compounds golden fagopyrum extract on the basis of compounding licorice extract, kudzu flower extract and safflower flower extract. The golden fagopyrum extract is rich in flavonoids. These compounds have antioxidant properties, can neutralize free radicals, reduce oxidative stress, thereby protecting intestinal cells from oxidative damage, can further enhance the overall protective effect, improve the cell survival rate after H2O2 damage, and inhibit the generation of ROS. DETAILED DESCRIPTION

[0034] The present invention provides a composition, which comprises liquorice extract, kudzu flower extract and dahliae flower extract; the mass ratio of the liquorice extract, kudzu flower extract and dahliae flower extract is (1-10): (1-10): (1-10).

[0035] In the present invention, the composition comprises licorice extract, kudzu flower extract and elaeagnus flower extract; the mass ratio of the licorice extract, kudzu flower extract and elaeagnus flower extract is preferably 1:1:1.

[0036] In the present invention, the preparation method of the licorice extract preferably includes the following steps: soaking licorice powder in sodium hydroxide solution to obtain a soaking solution; refluxing the soaking solution, adjusting the pH of the extract to 1.0-2.0, collecting the precipitate and drying it to obtain the licorice extract; the reflux extraction is performed once, the temperature is 100-120°C, and the time is 1-2h. The mass volume ratio of the licorice powder to the sodium hydroxide solution of the present invention is preferably (1-2) kg: (8-20) L, and more preferably 1.5 kg: (10-15) L; the licorice powder can preferably pass through an 80-120 mesh sieve; the concentration of the sodium hydroxide solution is preferably 0.2-0.6 mol / L, and more preferably 0.4-0.5 mol / L; the soaking time is preferably 10-24 h, and more preferably 15-20 h; the drying temperature is preferably 50-66 ° C, and more preferably 55 ° C; the vacuum degree of the drying is preferably ≤-0.08 Mpa; the drying time is preferably 3-6 h, and more preferably 4-5 h. The present invention preferably adjusts the pH of the extract to 1.0-2.0, and then stands for 5-12 h to collect the precipitate. The standing time of the present invention is preferably 8-10 h.

[0037] Licorice extract contains rich bioactive ingredients, such as glycyrrhizic acid and glycyrrhizin, which have significant antioxidant effects and reduce the damage caused by oxidative stress through multiple mechanisms. First, licorice extract can directly remove free radicals in the body and reduce the oxidative damage of these free radicals to intestinal cells. Secondly, licorice extract can increase the activity of antioxidant enzymes in the body, such as superoxide dismutase (SOD) and glutathione peroxidase (GPx), and enhance the body's antioxidant capacity. Licorice extract can also regulate inflammatory responses in the intestine and reduce the level of pro-inflammatory factors, thereby further protecting the intestinal mucosa from oxidative stress. In addition, licorice extract further reduces intestinal damage by promoting the repair of intestinal barrier function, enhancing the integrity of the mucosal barrier, and reducing the penetration of harmful substances.

[0038] In the present invention, the preparation method of the kudzu flower extract preferably includes the following steps: mixing kudzu flower powder and an ethanol solution with a volume concentration of 30-60%, extracting to obtain an extract; concentrating the extract under reduced pressure to obtain an extract with a solid content of 15% to 40%; mixing the extract with a solid content of 15% to 40% with anhydrous ethanol to an ethanol volume concentration of 80-90%, standing for 1-4 hours and then centrifuging to collect the supernatant; concentrating the supernatant under reduced pressure to obtain an extract with a solid content of 85-95%, and spray drying to obtain the kudzu flower extract. The present invention preferably mixes kudzu flower powder and an ethanol solution with a volume concentration of 40-50%; the extraction temperature is preferably 60°C-90°C, more preferably 70-80°C; the number of extractions is preferably 2-3 times, and the time for each extraction is preferably 1-2 hours, more preferably 1.5 hours; the mass ratio of the kudzu flower powder to the ethanol solution with a volume concentration of 30-60% is preferably 1:(5-15), more preferably 1:(8-12), and more preferably 1:10. The present invention has no strict requirements on the reduced pressure concentration step, and conventional steps in the art can be used.

[0039] Pueraria extract contains puerarin, flavonoids, polyphenols and isoflavones; puerarin has significant antioxidant effects. It can remove free radicals in the body and reduce the oxidative damage to cells caused by free radicals. Puerarin can also enhance the activity of antioxidant enzymes in the body, such as superoxide dismutase (SOD) and glutathione peroxidase (GPx), helping to improve the antioxidant capacity of cells; flavonoids, such as pueraria flavonoids and soy isoflavones, have strong antioxidant properties and can effectively neutralize free radicals and reduce the damage to intestinal cells caused by oxidative stress. In addition, they can also inhibit the inflammatory response caused by oxidative stress, thereby protecting the intestinal mucosa; polyphenols also have antioxidant and anti-inflammatory effects, which can reduce the inflammatory response of the intestine and protect the intestinal mucosa by inhibiting the inflammatory pathway induced by oxidative stress; isoflavones not only have antioxidant functions, but also can regulate the intestinal microbial community, enhance the function of the intestinal barrier, and further reduce the damage to the intestine caused by oxidative stress. Pueraria extract plays an important role in alleviating intestinal oxidative stress damage.

[0040] In the present invention, the preparation method of the daidai flower extract preferably includes the following steps: soaking the daidai flower in a 2-8g / L citric acid aqueous solution for 2-3h, extracting by microwave heating 2-3 times, separating the solid and liquid, and collecting the filtrate; drying the filtrate to obtain the daidai flower extract; each time the microwave heating extraction is performed by heating to 60-90°C with a microwave power of 80-300W and extracting for 5-20min. The concentration of the citric acid aqueous solution of the present invention is preferably 4-5g / L; the microwave power is preferably 100-250W, more preferably 150-200W; the extraction time is preferably 10-15min.

[0041] The extract of dai dai flower is rich in a variety of antioxidant ingredients, such as flavonoids. These ingredients can directly remove excessive free radicals in the intestine, such as superoxide anion free radicals, hydroxyl free radicals, etc., and reduce the oxidative damage of free radicals to intestinal cells. Secondly, it can enhance the antioxidant defense system of intestinal cells themselves. By upregulating the activity of antioxidant enzymes, such as superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), it promotes the body's ability to scavenge free radicals and maintains the redox balance in the intestine. Furthermore, dai dai flower extract also has anti-inflammatory effects. Oxidative stress often triggers intestinal inflammatory reactions, while dai dai flower extract can inhibit the release of inflammatory mediators, reduce the damage of inflammation to intestinal tissues, and protect the integrity of the intestinal mucosal barrier. In addition, it can also regulate the balance of intestinal microbial communities. The growth and reproduction of beneficial bacteria are promoted, and harmful bacteria are inhibited, thereby improving the intestinal microecological environment and indirectly reducing the damage of oxidative stress to the intestine.

[0042] In the present invention, the composition preferably also includes a golden fagopyrum extract; the mass ratio of the licorice extract, the kudzu flower extract, the agave clover flower extract and the golden fagopyrum extract is (1-10): (1-10): (1-10): (0.5-5), and is further preferably 1:1:1:0.5.

[0043] In the present invention, the preparation method of the golden buckwheat extract preferably comprises the following steps: mixing golden buckwheat with an ethanol solution with a volume concentration of 30-80%, refluxing extraction, and obtaining an extract; concentrating the extract to obtain the golden buckwheat extract. The volume concentration of the ethanol solution of the present invention is preferably 50-60%; the number of reflux extractions is preferably 1; the temperature of the reflux extraction is preferably 90-130°C, more preferably 100-120°C; the time of the reflux extraction is preferably 2-5h, more preferably 3-4h; the mass ratio of the golden buckwheat to the ethanol solution with a volume concentration of 30-80% is preferably 1:(5-10), more preferably 1:8.

[0044] Fagopyrum serrata extract is rich in flavonoids, which have antioxidant properties and protect intestinal cells from oxidative damage by neutralizing free radicals and reducing oxidative stress.

[0045] The present invention also provides the use of the composition described in the above technical solution in the preparation of an intestinal injury drug. In the present invention, the intestinal injury drug preferably includes a drug for alleviating intestinal oxidative stress damage; the oxidative stress damage preferably includes H2O2 damage.

[0046] When licorice extract, kudzu flower extract and dai dai flower extract are used in combination, the antioxidant and anti-inflammatory effects of licorice extract complement the potent antioxidant and barrier repair functions of kudzu flower extract and dai dai flower extract, enhancing the overall protective effect. Compounding with golden buckwheat extract can further enhance the protective effect. This synergistic effect can not only more comprehensively reduce the intestinal damage caused by oxidative stress, but also improve the ability to prevent and repair oxidative damage, thereby achieving a more effective mitigation effect. The results of the embodiment show that the combined use of licorice extract, kudzu flower extract and dai dai flower extract can improve the cell survival rate after H2O2 damage and inhibit the generation of ROS. When compounding with golden buckwheat extract, the cell survival rate after H2O2 damage is further improved, and the effect of inhibiting the generation of ROS is also improved.

[0047] In order to further illustrate the present invention, a composition for alleviating intestinal oxidative stress damage and its application provided by the present invention are described in detail below in conjunction with examples, but they should not be construed as limiting the scope of protection of the present invention.

[0048] Example 1

[0049] 1. Preparation of Licorice Extract

[0050] (1) Weighing licorice tablets, crushing the licorice tablets and passing them through a 80-120 mesh sieve to obtain licorice powder;

[0051] (2) Pretreatment: adding licorice powder to a sodium hydroxide solution for soaking to obtain a soaking solution; the concentration of the sodium hydroxide solution is 0.5 mol / L, the solid-liquid ratio of licorice powder to the sodium hydroxide solution is 1.5 kg:15 L; the soaking time is 20 h;

[0052] (3) Extraction: reflux the above soaking solution at 100°C for 1.5 h, and collect the extract;

[0053] (4) Precipitation: adjust the pH value of the extract to 1.0 with 2 mol / L hydrochloric acid, cool to room temperature, let stand for 8 h, discard the supernatant; then wash with 2 mol / L hydrochloric acid, and filter to obtain the precipitate;

[0054] (5) Drying: Drying the obtained precipitate, collecting the dried powder, and obtaining the licorice extract; wherein the drying conditions are: temperature 55° C., vacuum degree ≤-0.08 MPa, and drying time 4 h.

[0055] 2. Preparation of Pueraria lobata Flower Extract

[0056] (1) Raw material preparation: Take dried kudzu flower and make it into coarse powder;

[0057] (2) Extraction: Take the above crude powder raw material, add 50% (v / v) ethanol solution with a weight 10 times the total weight of the raw material, stir and extract at 75°C for 2 hours, extract 3 times, and combine the extracts;

[0058] (3) Filtration: The extract is filtered to separate the residue;

[0059] (4) Concentrating under reduced pressure: Concentrating the filtrate under reduced pressure to obtain an extract having a solid content of 30%;

[0060] (5) High concentration ethanol standing: add anhydrous ethanol to the extract until the ethanol concentration reaches 85%, and stand at room temperature for 2 h;

[0061] (6) Centrifugation: centrifuge the ethanol solution in step (5) at 1500-6000 r / min to obtain a supernatant;

[0062] (7) Drying: The supernatant is concentrated under reduced pressure to an extract having a solid content of 95%, and directly spray-dried to obtain the kudzu flower extract.

[0063] 3. Preparation of the extract of the daisy flower

[0064] Wash and chop the jasmine flower, then add a 5 g / L citric acid aqueous solution, soak for 2 hours, extract twice using microwave heating, filter, and dry the filtrate under reduced pressure to obtain the jasmine flower extract; wherein, the method of each microwave heating extraction is: heating to 60°C at a microwave power of 150 W and extracting for 10 minutes.

[0065] 4. Preparation of Fagopyrum truncatum Extract

[0066] Add 8 times the weight of 60% (v / v) ethanol solution to the golden buckwheat, reflux extract at 120°C for 3 hours and then concentrate to obtain the golden buckwheat extract.

[0067] Example 2

[0068] The licorice extract, kudzu flower extract and agave clover flower extract obtained in Example 1 were mixed in a mass ratio of 1:1:1 to obtain composition 1.

[0069] Example 3

[0070] The licorice extract, kudzu flower extract, agave clover flower extract and golden fagopyrum extract obtained in Example 1 were mixed in a mass ratio of 1:1:1:0.5 to obtain composition 2.

[0071] Test Example 1

[0072] Detection of cell survival rate damaged by H2O2

[0073] The cells (RAW264.7 mouse peritoneal macrophages) were diluted to 1×10 5 / mL, and were seeded in a 96-well plate at a density of 100 μL / well. After 24 hours of cell culture, the cells were randomly divided into a blank group, a model group, a composition 1 treatment group, a composition 2 treatment group, a liquorice extract treatment group, a kudzu flower extract treatment group, a dai dai flower extract treatment group, and a golden buckwheat extract treatment group, and treated as follows:

[0074] Blank group: cells were treated with DMEM medium;

[0075] Model group: cells were treated with DMEM medium;

[0076] Composition 1 treatment group: cells were treated with DMEM medium containing 25.0 μg / mL of Composition 1 prepared in Example 2;

[0077] Composition 2 treatment group: cells were treated with DMEM medium containing 25.0 μg / mL of Composition 2 prepared in Example 3;

[0078] Licorice extract treatment group: cells were treated with DMEM medium containing 25.0 μg / mL of the licorice extract in Example 1;

[0079] Pueraria flower extract treatment group: cells were treated with DMEM medium containing 25.0 μg / mL of the Pueraria flower extract in Example 1;

[0080] The group treated with the extract of the daidai flower: the cells were treated with DMEM medium containing 25.0 μg / mL of the daidai flower extract in Example 1;

[0081] The Fagopyrum truncatum extract-treated group treated cells with DMEM medium containing 25.0 μg / mL of the Fagopyrum truncatum extract of Example 1;

[0082] After treating the cells in each treatment group for 12 hours, the supernatant was removed respectively. The model group, the composition 1 treatment group, the composition 2 treatment group, the licorice extract treatment group, the kudzu flower extract treatment group, the dai dai flower extract treatment group and the golden buckwheat extract treatment group were respectively added with DMEM medium containing 600 μmol / L H2O2 to treat the cells for 4 hours; the blank group was added with DMEM medium (without H2O2) to treat the cells for 4 hours, and 6 replicates were set for each group. 20 μL of MTT detection solution was added to each well. After the 96-well plate was placed in the incubator and continued to be cultured for 4 hours, the supernatant was removed, 150 μL DMSO was added to each well, and the absorbance of each well was detected at a detection wavelength of 490 nm using an enzyme reader, and the cell survival rate was counted. The results are shown in Table 1.

[0083] Table 1 Effects of different treatment groups on the survival rate of cells damaged by H2O2

[0084] Treatment Group Cell viability (%) Blank group 103.86±7.23 Model Group 50.26±5.21 Composition 1 treatment group 78.96±5.68* Composition 2 treatment group 79.06±5.72* Licorice extract treatment group 59.32±6.85* Pueraria flower extract treatment group 55.45±3.96* Group treated with daisy flower extract 63.13±2.62* Fagopyrum truncatum extract treatment group 53.26±4.13*

[0085] Note: Statistical analysis: The measurement was M±SD, and the data were analyzed using SPSS software. The t-test was used for pairwise comparison, and p<0.05 was considered statistically significant; compared with the hydrogen peroxide control group, * indicates p<0.05.

[0086] According to Table 1, it can be seen that in the model group treated with 600 μmol / L H2O2, the cell viability was significantly reduced. Compared with the model group, although the addition of licorice extract, kudzu flower extract, dahlia flower extract and golden buckwheat extract could improve the cell survival rate, the degree of improvement was not obvious. The composition 1 treatment group and the composition 2 treatment group could significantly improve the cell survival rate.

[0087] Test Example 2

[0088] Detection of intracellular reactive oxygen species (ROS) levels in H2O2-damaged cells

[0089] The cells (RAW264.7 mouse peritoneal macrophages) were diluted to 1×10 5 / mL, and were inoculated in a 6-well plate at a density of 2mL / well. After 24h of cell culture, the cells were randomly divided into a blank group, a model group, a composition 1 treatment group, a composition 2 treatment group, a liquorice extract treatment group, a kudzu flower extract treatment group, a dai dai flower extract treatment group, and a golden buckwheat extract treatment group, and treated as follows:

[0090] Blank group: cells were treated with DMEM medium;

[0091] Model group: cells were treated with DMEM medium;

[0092] Composition 1 treatment group: cells were treated with DMEM medium containing 25.0 μg / mL of Composition 1 prepared in Example 2;

[0093] Composition 2 treatment group: cells were treated with DMEM medium containing 25.0 μg / mL of Composition 2 prepared in Example 3;

[0094] Licorice extract treatment group: cells were treated with DMEM medium containing 25.0 μg / mL of the licorice extract in Example 1;

[0095] Pueraria flower extract treatment group: cells were treated with DMEM medium containing 25.0 μg / mL of the Pueraria flower extract in Example 1;

[0096] The group treated with the extract of the daidai flower: the cells were treated with DMEM medium containing 25.0 μg / mL of the daidai flower extract in Example 1;

[0097] The Fagopyrum truncatum extract-treated group treated cells with DMEM medium containing 25.0 μg / mL of the Fagopyrum truncatum extract of Example 1;

[0098] After treating the cells in each treatment group for 12 hours, the supernatant was removed respectively, and the model group, the composition 1 treatment group, the composition 2 treatment group, the liquorice extract treatment group, the kudzu flower extract treatment group, the dai dai flower extract treatment group and the golden buckwheat extract treatment group were respectively added with DMEM medium containing 600 μmol / L H2O2 to treat the cells for 4 hours; the blank group was added with DMEM medium (without H2O2) to treat the cells for 4 hours, and 6 replicates were set for each group. After the treatment of each treatment group, the supernatant of each group was discarded and the phenol red-free DMEM containing the DCFH-DA fluorescent probe was added and cultured in a 37°C incubator for 30 minutes, and then the reactive oxygen species (ROS) content was detected by flow cytometry, and the results are shown in Table 2.

[0099] Table 2 Effects of different treatment groups on ROS in cells damaged by H2O2

[0100]

[0101]

[0102] Note: Statistical analysis: The measurement was M±SD, and the data were analyzed using SPSS software. The t-test was used for pairwise comparison, and p<0.05 was considered statistically significant; compared with the hydrogen peroxide control group, * indicates p<0.05.

[0103] It can be seen from Table 2 that the addition of 600 μmol / L H2O2 caused a significant increase in ROS in the cells. Compared with the model group, the addition of licorice extract, kudzu flower extract, dahlia flower extract and golden buckwheat extract could inhibit the generation of ROS, but the inhibitory effect was not obvious, while the composition 1 treatment group and the composition 2 treatment group could significantly inhibit the generation of ROS.

[0104] Based on the above content, it can be seen that the composition provided by the present invention can improve the survival rate of cells after H2O2 damage, inhibit the generation of ROS, improve the ability to prevent and repair oxidative damage, and effectively alleviate intestinal oxidative stress damage.

[0105] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A composition, characterized in that The composition comprises liquorice extract, kudzu flower extract and dahliae flower extract; the mass ratio of the liquorice extract, kudzu flower extract and dahliae flower extract is (1-10): (1-10): (1-10).

2. The composition according to claim 1, characterized in that The composition also includes a golden fagopyrum extract; the mass ratio of the liquorice extract, the kudzu flower extract, the sedge flower extract and the golden fagopyrum extract is (1-10): (1-10): (1-10): (0.5-5).

3. The composition according to claim 1 or 2, characterized in that The preparation method of the licorice extract comprises the following steps: Soaking liquorice powder in sodium hydroxide solution to obtain a soaking solution; Reflux extraction is performed on the soaking solution, the pH value of the extract is adjusted to 1.0-2.0, and the precipitate is collected and dried to obtain the licorice extract; The reflux extraction is performed once, the temperature is 100-120° C., and the time is 1-2 hours.

4. The composition according to claim 3, characterized in that The mass volume ratio of the licorice powder to the sodium hydroxide solution is (1-2) kg: (8-20) L; The concentration of the sodium hydroxide solution is 0.2-0.6 mol / L; The soaking time is 10 to 24 hours; The drying temperature is 50-66° C., the vacuum degree is ≤-0.08Mpa, and the drying time is 3-6 hours.

5. The composition according to claim 1 or 2, characterized in that The preparation method of the kudzu flower extract comprises the following steps: The kudzu flower powder is mixed with an ethanol solution having a volume concentration of 30 to 60%, and extracted to obtain an extract; Concentrating the extract under reduced pressure to obtain an extract with a solid content of 15% to 40%; The extract with a solid content of 15% to 40% is mixed with anhydrous ethanol to a volume concentration of 80 to 90% ethanol, and the mixture is allowed to stand for 1 to 4 hours and then centrifuged to collect the supernatant; The supernatant is concentrated under reduced pressure to obtain an extract with a solid content of 85-95%, and the extract is spray-dried to obtain the kudzu flower extract.

6. The composition according to claim 5, characterized in that The extraction temperature is 60°C to 90°C, the number of times is 2 to 3 times, and the time of each extraction is 1 to 2 hours; The mass ratio of the kudzu flower powder to the ethanol solution with a volume concentration of 30-60% is 1:(5-15).

7. The composition according to claim 1 or 2, characterized in that The preparation method of the daidai flower extract comprises the following steps: After soaking the daidai flower in a 2-8 g / L citric acid aqueous solution for 2-3 hours, extracting by microwave heating for 2-3 times, separating the solid and the liquid, and collecting the filtrate; drying the filtrate to obtain the daidai flower extract; The microwave heating extraction method each time is: heating to 60-90° C. with a microwave power of 80-300 W and extracting for 5-20 minutes.

8. The composition according to claim 2, characterized in that The preparation method of the golden buckwheat extract comprises the following steps: mixing golden buckwheat with an ethanol solution with a volume concentration of 30 to 80%, refluxing and extracting to obtain an extract; concentrating the extract to obtain the golden buckwheat extract; The reflux extraction is performed once at a temperature of 90 to 130° C. for 2 to 5 hours. The mass ratio of the golden buckwheat to the ethanol solution with a volume concentration of 30-80% is 1:(5-10).

9. Use of the composition according to any one of claims 1 to 8 in the preparation of a drug for intestinal injury.

10. The use according to claim 9, characterized in that: The intestinal injury drugs include drugs for alleviating intestinal oxidative stress damage; the oxidative stress damage includes H2O2 damage.

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