A composition for alleviating oxidative stress damage in the intestinal tract and use thereof

Abstract

The present application belongs to the technical field of biological medicine, and particularly relates to a composition for relieving intestinal oxidative stress injury and application thereof. The licorice extract, kudzu flower extract and dalbergia hupeana flower extract are compounded according to a certain proportion, the antioxidant and anti-inflammatory effects of the licorice extract are complementary to the strong antioxidant and barrier repair functions of the kudzu flower extract and dalbergia hupeana flower extract, and the overall protection effect is enhanced. The synergistic effect can not only more comprehensively reduce the intestinal injury caused by oxidative stress, but also improve the prevention and repair capacity for oxidative damage, so that a more effective relief effect is achieved. On this basis, the gold buckwheat extract is further compounded, so that the overall protection effect is further enhanced, the cell survival rate after H2O2 injury is improved, and the generation of ROS is inhibited. The results of the examples show that the composition provided by the present application can improve the cell survival rate after H2O2 injury, inhibit the generation of ROS, and effectively relieve intestinal oxidative stress injury.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine, and particularly relates to a composition for relieving intestinal oxidative stress injury and application thereof. BACKGROUND

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

[0003] Currently, the treatment methods for intestinal oxidative stress mainly focus on the use of antioxidants and anti-inflammatory drugs. These methods can alleviate the damage caused by oxidative stress to some extent, but there are still some problems, such as low bioavailability of antioxidants, drug side effects, and limited control effect on oxidative stress. Therefore, it is of great clinical significance to develop new and more effective antioxidant and intestinal protection treatment programs. SUMMARY

[0004] The purpose of the present application is to provide a composition for relieving intestinal oxidative stress injury and application thereof, to improve the cell survival rate after intestinal oxidative stress injury, to inhibit the production of ROS, to reduce the intestinal damage caused by oxidative stress, and to improve the prevention and repair ability of oxidative damage, so as to achieve more effective relief effect.

[0005] The present application provides a composition comprising licorice extract, kudzu flower extract, and dalbergia odorifera flower extract; the mass ratio of the licorice extract, kudzu flower extract, and dalbergia odorifera flower extract is (1-10):(1-10):(1-10).

[0006] Preferably, the composition further comprises a goldstring buckwheat extract; the mass ratio of the licorice extract, kudzu flower extract, dalbergia odorifera flower extract, and goldstring buckwheat 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] The licorice powder is soaked with sodium hydroxide solution to obtain a soaking solution;

[0009] The soaking solution is refluxed to adjust the pH of the extraction solution to 1.0-2.0, and the precipitate is collected and dried to obtain the licorice extract.

[0010] The reflux extraction was performed once, at a temperature of 100–120°C, for a time of 1–2 hours.

[0011] Preferably, the mass-to-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 10–24 hours;

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

[0015] Preferably, the preparation method of the kudzu flower extract includes the following steps:

[0016] The kudzu flower powder was mixed with an ethanol solution with a volume concentration of 30-60% and extracted to obtain the extract.

[0017] The extract was concentrated 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 until the ethanol volume concentration is 80% to 90%. After standing for 1 to 4 hours, the mixture is centrifuged and the supernatant is collected.

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

[0020] Preferably, the extraction temperature is 60℃~90℃, the number of extractions is 2~3, and the extraction time for each extraction is 1~2 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 bitter orange flower extract includes the following steps:

[0023] After soaking bitter orange flowers in a 2-8 g / L citric acid aqueous solution for 2-3 hours, microwave extraction was performed 2-3 times, followed by solid-liquid separation and collection of the filtrate. The filtrate was then dried to obtain the bitter orange flower extract.

[0024] The microwave heating extraction method for each extraction is as follows: heating to 60-90℃ with 80-300W microwave power and extracting for 5-20 minutes.

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

[0026] The reflux extraction was performed once, at a temperature of 90–130°C, for a time of 2–5 hours.

[0027] The mass ratio of the 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 above in the preparation of intestinal injury drugs.

[0029] Preferably, the intestinal injury drug includes drugs that alleviate intestinal oxidative stress damage;

[0030] The oxidative stress damage includes H2O2 damage.

[0031] Beneficial effects:

[0032] This invention provides a composition comprising licorice extract, kudzu flower extract, and bitter orange flower extract; the mass ratio of the licorice extract, kudzu flower extract, and bitter orange flower extract is (1-10):(1-10):(1-10). The licorice extract contains glycyrrhizic acid and glycyrrhizin flavonoids, which exert their effects primarily through antioxidant and anti-inflammatory activities. Glycyrrhizic acid can inhibit inflammatory responses induced by oxidative stress, reduce the level of pro-inflammatory factors, and simultaneously increase the activity of antioxidant enzymes in the body, such as superoxide dismutase (SOD), thereby enhancing the antioxidant capacity of cells. Glycyrrhizin flavonoids help repair damaged intestinal barriers and further protect the intestinal mucosa. The main components of kudzu flower extract include puerarin, flavonoids, and isoflavones, which possess strong antioxidant capabilities. Puerarin and flavonoids can scavenge free radicals in the body, reduce oxidative damage, and also have anti-inflammatory effects, inhibiting inflammatory responses caused by oxidative stress. Isoflavones can regulate the intestinal microbiota and improve intestinal health. Bitter orange flower extract is rich in flavonoids and polyphenols, exhibiting significant antioxidant and anti-inflammatory effects. These components reduce oxidative stress-induced intestinal damage by scavenging free radicals and inhibiting inflammatory responses. Furthermore, bitter orange flower extract promotes intestinal mucosal repair and enhances intestinal barrier function. In this invention, the combined use of licorice extract, kudzu flower extract, bitter orange flower extract, and buckwheat extract allows the antioxidant and anti-inflammatory effects of licorice extract to complement the potent antioxidant and barrier repair functions of kudzu flower and bitter orange flower extracts, enhancing the overall protective effect. This synergistic effect not only more comprehensively reduces intestinal damage caused by oxidative stress but also improves the ability to prevent and repair oxidative damage, thereby achieving a more effective alleviating effect.

[0033] This invention further incorporates buckwheat extract into a compound of licorice extract, kudzu flower extract, and bitter orange flower extract. Buckwheat extract is rich in flavonoids, which have antioxidant properties. By neutralizing free radicals and reducing oxidative stress, it protects intestinal cells from oxidative damage, further enhancing the overall protective effect, improving cell survival rate after H2O2 damage, and inhibiting ROS production. Detailed Implementation

[0034] The present invention provides a composition comprising licorice extract, kudzu flower extract and bitter orange flower extract; wherein the mass ratio of the licorice extract, kudzu flower extract and bitter orange flower extract is (1-10):(1-10):(1-10).

[0035] In this invention, the composition comprises licorice extract, kudzu flower extract, and bitter orange flower extract; the preferred mass ratio of the licorice extract, kudzu flower extract, and bitter orange flower extract is 1:1:1.

[0036] In this invention, the preferred method for preparing the licorice extract includes the following steps: soaking licorice powder in sodium hydroxide solution to obtain an soaking solution; reflux extraction of 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, at a temperature of 100-120°C, for a time of 1-2 hours. The preferred mass-to-volume ratio of licorice powder to sodium hydroxide solution in this invention is (1-2) kg:(8-20) L, more preferably 1.5 kg:(10-15) L; the licorice powder is preferably passable through an 80-120 mesh sieve; the concentration of the sodium hydroxide solution is preferably 0.2-0.6 mol / L, more preferably 0.4-0.5 mol / L; the soaking time is preferably 10-24 h, more preferably 15-20 h; the drying temperature is preferably 50-66℃, more preferably 55℃; the drying vacuum degree is preferably ≤-0.08 MPa; the drying time is preferably 3-6 h, more preferably 4-5 h. This invention preferably adjusts the pH of the extract to 1.0-2.0, allows it to stand for 5-12 h, and then collects the precipitate. The preferred standing time in this invention is 8-10 h.

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

[0038] In this invention, the preferred method for preparing the kudzu flower extract includes the following steps: mixing kudzu flower powder with an ethanol solution of 30-60% by volume, extracting to obtain an extract; concentrating the extract under reduced pressure to obtain an extract with a solid content of 15%-40%; mixing the extract with anhydrous ethanol to an ethanol volume concentration of 80-90%, allowing it to stand for 1-4 hours, centrifuging, and collecting 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. In this invention, kudzu flower powder is preferably mixed with an ethanol solution with a volume concentration of 40-50%; the extraction temperature is preferably 60℃-90℃, more preferably 70-80℃; the extraction is preferably performed 2-3 times, and the extraction time for each extraction is preferably 1-2 hours, more preferably 1.5 hours; the mass ratio of kudzu flower powder to an ethanol solution with a volume concentration of 30-60% is preferably 1:(5-15), more preferably 1:(8-12), and even more preferably 1:10. This invention does not have strict requirements for the vacuum concentration step; conventional steps in the art can be used.

[0039] Pueraria lobata extract contains puerarin, flavonoids, polyphenols, and isoflavones. Puerarin has significant antioxidant properties, effectively scavenging free radicals and reducing oxidative damage to cells. It also enhances the activity of antioxidant enzymes such as superoxide dismutase (SOD) and glutathione peroxidase (GPx), helping to improve cellular antioxidant capacity. Flavonoids, such as puerarin and soy isoflavones, possess strong antioxidant properties, effectively neutralizing free radicals and reducing oxidative stress damage to intestinal cells. Furthermore, they inhibit inflammatory responses induced by oxidative stress, thereby protecting the intestinal mucosa. Polyphenols also have antioxidant and anti-inflammatory effects, reducing intestinal inflammation and protecting the intestinal mucosa by inhibiting oxidative stress-induced inflammatory pathways. Isoflavones not only have antioxidant functions but also enhance the intestinal barrier function by regulating the intestinal microbiota, further reducing oxidative stress damage to the intestines. Pueraria lobata extract plays an important role in alleviating intestinal oxidative stress damage.

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

[0041] Bitter orange flower extract is rich in various antioxidants, such as flavonoids. These components can directly eliminate excess free radicals in the intestines, such as superoxide anion radicals and hydroxyl radicals, reducing oxidative damage to intestinal cells. Secondly, it can enhance the intestinal cells' own antioxidant defense system. 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 intestines. Furthermore, bitter orange flower extract also has anti-inflammatory effects. Oxidative stress often triggers intestinal inflammation, and bitter orange 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 regulate the balance of the intestinal microbiota. The growth and reproduction of beneficial bacteria are promoted, while harmful bacteria are inhibited, thereby improving the intestinal microecological environment and indirectly reducing the damage of oxidative stress to the intestines.

[0042] In this invention, the composition preferably further includes buckwheat extract; the mass ratio of licorice extract, kudzu flower extract, bitter orange flower extract and buckwheat extract is (1-10):(1-10):(1-10):(0.5-5), more preferably 1:1:1:0.5.

[0043] In this invention, the preferred method for preparing the buckwheat extract includes the following steps: mixing buckwheat with an ethanol solution of 30-80% (v / v), refluxing to extract, and obtaining an extract; concentrating the extract to obtain the buckwheat extract. The preferred v / v concentration of the ethanol solution is 50-60%; the preferred number of refluxing extractions is once; the preferred refluxing temperature is 90-130°C, more preferably 100-120°C; the preferred refluxing time is 2-5 hours, more preferably 3-4 hours; and the preferred mass ratio of buckwheat to the 30-80% (v / v) ethanol solution is 1:(5-10), more preferably 1:8.

[0044] Buckwheat extract is rich in flavonoids, which have antioxidant properties. By neutralizing free radicals and reducing oxidative stress, these compounds protect intestinal cells from oxidative damage.

[0045] The present invention also provides the application of the composition described above in the preparation of intestinal injury drugs. 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 bitter orange 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 bitter orange flower extract, enhancing the overall protective effect. The addition of buckwheat extract further strengthens this protective effect. This synergistic effect not only more comprehensively reduces intestinal damage caused by oxidative stress but also improves the ability to prevent and repair oxidative damage, thus achieving a more effective relief. The results of the examples show that the combined use of licorice extract, kudzu flower extract, and bitter orange flower extract can improve cell survival rate after H2O2 damage and inhibit ROS production. When combined with buckwheat extract, the cell survival rate after H2O2 damage is further improved, and the effect of inhibiting ROS production is also enhanced.

[0047] To further illustrate the present invention, the following detailed description, in conjunction with embodiments, of a composition for alleviating intestinal oxidative stress damage and its application, 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) Weigh out licorice tablets, crush the licorice tablets and pass them through an 80-120 mesh sieve to obtain licorice powder;

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

[0052] (3) Extraction: The above soaking solution was refluxed at 100℃ for 1.5h and the extract was collected;

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

[0054] (5) Drying: The precipitate was dried and the dried powder was collected to obtain licorice extract; the drying conditions were: temperature 55℃, vacuum degree ≤ -0.08Mpa, and drying time 4h.

[0055] 2. Preparation of Kudzu Flower Extract

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

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

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

[0059] (4) Concentration under reduced pressure: The filtrate is concentrated under reduced pressure to extract, and the solid content of the extract is 30%.

[0060] (5) High concentration ethanol standing: Add anhydrous ethanol to the extract until the ethanol concentration is 85%, and let it stand at room temperature for 2 hours;

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

[0062] (7) Drying: The supernatant was concentrated under reduced pressure to extract. The solid content of the extract was 95%. It was then spray-dried directly to obtain kudzu flower extract.

[0063] 3. Preparation of bitter orange flower extract

[0064] Wash and chop the bitter orange flowers, then add a 5g / L citric acid aqueous solution and soak for 2 hours. Extract twice using microwave heating, filter, and dry the filtrate under reduced pressure to obtain bitter orange flower extract. The microwave heating method for each extraction is as follows: heat to 60℃ at 150W microwave power and extract for 10 minutes.

[0065] 4. Preparation of Buckwheat Extract

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

[0067] Example 2

[0068] The licorice extract, kudzu flower extract and bitter orange 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, bitter orange flower extract and buckwheat 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 viability after H2O2 damage

[0073] The cells (RAW264.7 mouse peritoneal macrophages) were diluted to 1×10⁻⁶. 5 Cells were seeded at a density of 100 μL / well in 96-well plates. After 24 h of cell culture, the cells were randomly divided into four groups: blank group, model group, combination 1 treatment group, combination 2 treatment group, licorice extract treatment group, kudzu flower extract treatment group, bitter orange flower extract treatment group, and buckwheat extract treatment group. The cells were treated as follows:

[0074] Control 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 licorice extract from Example 1;

[0079] Kudzu flower extract treatment group: Cells were treated with DMEM medium containing 25.0 μg / mL kudzu flower extract from Example 1;

[0080] Bitter orange flower extract treatment group: Cells were treated with DMEM medium containing 25.0 μg / mL bitter orange flower extract from Example 1;

[0081] The cells in the buckwheat extract treatment group were treated with DMEM medium containing 25.0 μg / mL of buckwheat extract from Example 1;

[0082] After treating cells for 12 hours, the supernatant was removed from each treatment group. The model group, Composition 1 treatment group, Composition 2 treatment group, licorice extract treatment group, kudzu flower extract treatment group, bitter orange flower extract treatment group, and buckwheat extract treatment group were treated with DMEM medium containing 600 μmol / L H2O2 for 4 hours. The blank group was treated with DMEM medium (without H2O2) for 4 hours. Each group had 6 replicates. 20 μL of MTT assay solution was added to each well. After culturing the 96-well plate in an incubator for another 4 hours, the supernatant was removed, and 150 μL of LDMSO was added to each well. The absorbance of each well was measured using a microplate reader at a detection wavelength of 490 nm. Cell viability was calculated, and the results are shown in Table 1.

[0083] Table 1. Effects of different treatment groups on cell survival rate after H2O2 damage.

[0084] Treatment group Cell survival rate (%) 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* Glycyrrhiza extract treatment group 59.32±6.85* Gou flower extract treatment group 55.45±3.96* Dalbergia extract treatment group 63.13±2.62* Gold buckwheat extract treatment group 53.26±4.13*

[0085] Note: Statistical analysis: Measurements are expressed as mean ± SD. Data analysis was performed using SPSS software. Pairwise comparisons were performed using t-tests. P < 0.05 was considered statistically significant. * P < 0.05 was considered statistically significant compared with the hydrogen peroxide control group.

[0086] As shown in Table 1, the cell viability of the model group treated with 600 μmol / L H2O2 was significantly reduced. Compared with the model group, although the addition of licorice extract, kudzu flower extract, bitter orange flower extract and buckwheat extract could improve the cell survival rate, the improvement was not significant. In contrast, the treatment groups of Composition 1 and Composition 2 could significantly improve the cell survival rate.

[0087] Test Example 2

[0088] Detection of intracellular reactive oxygen species (ROS) levels after H2O2 damage

[0089] The cells (RAW264.7 mouse peritoneal macrophages) were diluted to 1×10⁻⁶. 5Cells were seeded at a density of 2 mL / well in 6-well plates. After 24 h of cell culture, the cells were randomly divided into four groups: blank group, model group, combination 1 treatment group, combination 2 treatment group, licorice extract treatment group, kudzu flower extract treatment group, bitter orange flower extract treatment group, and buckwheat extract treatment group. The cells were treated as follows:

[0090] Control 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 licorice extract from Example 1;

[0095] Kudzu flower extract treatment group: Cells were treated with DMEM medium containing 25.0 μg / mL kudzu flower extract from Example 1;

[0096] Bitter orange flower extract treatment group: Cells were treated with DMEM medium containing 25.0 μg / mL bitter orange flower extract from Example 1;

[0097] The cells in the buckwheat extract treatment group were treated with DMEM medium containing 25.0 μg / mL of buckwheat extract from Example 1;

[0098] After 12 hours of cell treatment, the supernatant was removed from each treatment group. The model group, Composition 1 treatment group, Composition 2 treatment group, licorice extract treatment group, kudzu flower extract treatment group, bitter orange flower extract treatment group, and buckwheat extract treatment group were treated with DMEM medium containing 600 μmol / L H2O2 for 4 hours. The blank group was treated with DMEM medium (without H2O2) for 4 hours. Each group had 6 replicates. After treatment, the supernatant was discarded, and phenol red-free DMEM containing the DCFH-DA fluorescent probe was added. The cells were incubated at 37°C for 30 minutes, and the reactive oxygen species (ROS) content was detected by flow cytometry. The results are shown in Table 2.

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

[0100]

[0101]

[0102] Note: Statistical analysis: Measurements are expressed as mean ± SD. Data analysis was performed using SPSS software. Pairwise comparisons were performed using t-tests. P < 0.05 was considered statistically significant. * P < 0.05 was considered statistically significant compared with the hydrogen peroxide control group.

[0103] As shown in Table 2, the addition of 600 μmol / L H2O2 significantly increased ROS levels in cells. Compared with the model group, the addition of licorice extract, kudzu flower extract, bitter orange flower extract, and buckwheat extract could inhibit ROS production, but the inhibitory effect was not significant. In contrast, the treatment groups of Composition 1 and Composition 2 significantly inhibited ROS production.

[0104] As can be seen from the above, the composition provided by the present invention can improve cell survival rate after H2O2 damage, inhibit ROS production, enhance the ability to prevent and repair oxidative damage, and effectively alleviate intestinal oxidative stress damage.

[0105] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A composition for treating intestinal injury, characterized in that, The composition is made from licorice extract, kudzu flower extract and bitter orange flower extract; the mass ratio of the licorice extract, kudzu flower extract and bitter orange flower extract is 1:1:

1.

2. A composition for treating intestinal injury, characterized in that, The composition is made from licorice extract, kudzu flower extract, bitter orange flower extract and buckwheat extract; the mass ratio of the licorice extract, kudzu flower extract, bitter orange flower extract and buckwheat extract is 1:1:1:0.

5.

3. The composition according to claim 1 or 2, characterized in that, The preparation method of the licorice extract includes the following steps: Licorice powder was soaked in sodium hydroxide solution to obtain an soaking solution; The soaking solution was refluxed for extraction, the pH of the extract was adjusted to 1.0-2.0, the precipitate was collected and dried to obtain the licorice extract; The reflux extraction is performed once, at a temperature of 100-120℃, for a time of 1-2 hours.

4. The composition according to claim 3, characterized in that, The mass-to-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 10-24 hours; The drying temperature is 50~66℃, the vacuum degree is ≤-0.08Mpa, and the time is 3~6h.

5. The composition according to claim 1 or 2, characterized in that, The preparation method of the kudzu flower extract includes the following steps: Mix kudzu flower powder with an ethanol solution with a volume concentration of 30-60%, extract, and obtain the extract; The extract was concentrated 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 until the ethanol volume concentration is 80% to 90%. After standing for 1 to 4 hours, the mixture is centrifuged and the supernatant is collected. The supernatant was concentrated under reduced pressure to obtain an extract with a solid content of 85-95%, which was then spray-dried to obtain the kudzu flower extract.

6. The composition according to claim 5, characterized in that, The extraction temperature is 60℃~90℃, the number of extractions is 2~3, and the extraction time for each extraction is 1~2 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 bitter orange flower extract includes the following steps: After soaking bitter orange flowers in a 2-8 g / L citric acid aqueous solution for 2-3 hours, microwave heating was used to extract the extract 2-3 times. Solid-liquid separation was performed, and the filtrate was collected. The filtrate was then dried to obtain the bitter orange flower extract. The microwave heating extraction method is as follows: use 80~300W microwave power to heat to 60~90℃ and extract for 5~20 minutes.

8. The composition according to claim 2, characterized in that, The preparation method of the buckwheat extract includes the following steps: mixing buckwheat with an ethanol solution with a volume concentration of 30-80%, refluxing and extracting to obtain an extract; concentrating the extract to obtain the buckwheat extract. The reflux extraction is performed once, at a temperature of 90~130℃, for a time of 2~5 hours; The mass ratio of the 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 medicament for intestinal injury.

10. The application according to claim 9, characterized in that, The intestinal injury drugs include drugs that alleviate intestinal oxidative stress damage; the oxidative stress damage includes H2O2 damage.

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