Composition helpful for relaxing bowel and application thereof

Abstract

The invention belongs to the technical field of pharmaceutical compositions, and particularly relates to a composition helpful for relaxing bowel and application thereof. The composition comprises fructo-oligosaccharide and curcumin, and can also comprise galactooligosaccharide and oat beta-glucan. The composition disclosed by the invention can achieve the effect of improving constipation, integrates functionality and nutrition, and has a wide application prospect in the aspect of improving the constipation function.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical compositions, and particularly relates to a composition helpful for moistening the intestines and relieving constipation and its application. Background Art

[0002] Constipation is a common disorder of the lower digestive tract function, mainly manifested as difficult defecation, a feeling of incomplete defecation or obstruction, less than 3 spontaneous bowel movements per week without the use of laxatives, and hard stools. The main cause of slow transit constipation is usually related to damage to the colonic smooth muscle or disorder of its enteric nerve function. Constipation not only causes physical discomfort such as abdominal distension, vomiting, intestinal obstruction and perforation, but also increases the risk of anxiety and irritability, thus affecting the quality of life of individuals and causing varying degrees of psychological disorders.

[0003] 5-Hydroxytryptamine (5-HT) is a neurotransmitter, and more than 95% of 5-HT is secreted by enterochromaffin cells. 5-HT can promote intestinal peristalsis by binding to corresponding receptors and plays a key role in intestinal motility and secretion. Serotonin transporter (SERT) is a transmembrane transport protein responsible for reuptaking excessive extracellular 5-HT to terminate its physiological effects and regulating the availability of extracellular 5-HT. Studies have found that the gut microbiota can regulate serotonergic signaling by modulating SERT expression. Gut microecological imbalance may cause abnormal SERT expression, thus slowing down intestinal peristalsis and promoting the occurrence of constipation. Prebiotics refer to dietary components that can selectively promote the growth of beneficial bacteria in the intestine and are fermented by beneficial bacteria in the colon to produce a large amount of short-chain fatty acids (SCFAs). Research shows that SCFAs play an important role in improving intestinal motility, increasing intestinal water absorption and regulating intestinal pH value, and have a significant effect on relieving constipation.

[0004] Conventional clinical methods for treating constipation mainly rely on drug treatment. The main drugs include osmotic laxatives such as lactulose and polyethylene glycol, stimulant laxatives such as bisacodyl and senna, fecal softeners such as mineral oil, and 5-HT4 receptor agonist drugs such as mosapride and prucalopride. Using drug treatment is likely to lead to dependence and is accompanied by adverse reactions such as headache, nausea, abdominal pain, and diarrhea. Research shows that the occurrence of constipation is closely related to gut microbiota imbalance. Therefore, how to find natural, effective and safe compositions to treat constipation from multiple aspects such as gut microbiota regulation while reducing the side effects of traditional drugs has become an urgent problem to be solved. Summary of the Invention

[0005] In view of the deficiencies of the prior art, the present invention aims to provide a composition helpful for moistening the intestines and promoting defecation and its application. By reasonably compounding fructooligosaccharide and curcumin, or reasonably compounding fructooligosaccharide, curcumin, galactooligosaccharide and oat β-glucan, a good effect of moistening the intestines and promoting defecation is exerted. This composition takes into account functionality and nutrition, and shows broad application prospects in moistening the intestines and promoting defecation and improving constipation.

[0006] To achieve the above object, on the one hand, the present invention provides a composition helpful for moistening the intestines and promoting defecation, the composition includes fructooligosaccharide and curcumin, and the mass ratio of the fructooligosaccharide to the curcumin is 40-70:3-10, such as 70:4, 60:3, 50:5, 40:10.

[0007] Further, the mass ratio of the fructooligosaccharide to the curcumin is 10:1.

[0008] On the second hand, the present invention provides a composition helpful for moistening the intestines and promoting defecation, the composition includes fructooligosaccharide, curcumin, galactooligosaccharide and oat β-glucan, and the mass ratio of the fructooligosaccharide, curcumin, galactooligosaccharide and oat β-glucan is 40-70:3-10:10-35:5-18, such as 70:4:10:6, 60:3:15:12, 50:5:25:10, 40:10:35:5, 50:5:17:18, 50:5:20:15.

[0009] Further, the mass ratio of the fructooligosaccharide, curcumin, galactooligosaccharide and oat β-glucan is 10:1:5:2.

[0010] On the third hand, the present invention provides a product, and the composition described in the first aspect or the second aspect is included in the product.

[0011] Further, the product is a health food or a drug, and excipients acceptable for the health food or the drug are further included in the product.

[0012] On the fourth hand, the present invention provides the application of the composition described in the first aspect or the second aspect in the preparation of a product capable of moistening the intestines and promoting defecation, and the product is a health food or a drug.

[0013] Further, the application is the application in the preparation of a product capable of improving constipation, promoting intestinal peristalsis and / or increasing intestinal transit rate.

[0014] Furthermore, the constipation is slow transit constipation.

[0015] Further, the application includes at least one of the following:

[0016] (1) The application in the preparation of a product capable of shortening the defecation time of constipated patients;

[0017] (2) Use in the preparation of a product capable of improving the release of neurotransmitters and gastrointestinal hormones related to constipation;

[0018] (3) Use in the preparation of a product capable of improving the intestinal flora imbalance caused by constipation;

[0019] (4) Use in the preparation of a product capable of improving the intestinal barrier damage caused by constipation

[0020] Furthermore, the use in the product capable of improving the release of neurotransmitters and gastrointestinal hormones related to constipation includes at least one of the following:

[0021] (1) Use in the preparation of a product capable of improving the decrease in the levels of 5-HT and SP caused by constipation;

[0022] (2) Use in the preparation of a product capable of improving the increase in the level of VIP caused by constipation;

[0023] (3) Use in the preparation of a product capable of improving the decrease in the levels of Gas and MTL caused by constipation.

[0024] Compared with the prior art, the present invention has the following advantages

[0025] (1) The present invention proves that the combined intervention of fructooligosaccharide and curcumin can help improve constipation and has a synergistic effect.

[0026] (2) The present invention proves that the combined intervention of fructooligosaccharide, curcumin, galactooligosaccharide and oat β-glucan can help improve constipation and has a synergistic effect.

[0027] (2) The composition of the present invention can effectively improve constipation, shorten the defecation time of constipation patients, increase the intestinal transit rate, and promote intestinal peristalsis; it can improve the decrease in the levels of 5-HT, SP, Gas, and MTL caused by constipation and improve the increase in the level of VIP caused by constipation; it can improve the intestinal flora imbalance caused by constipation and improve the intestinal barrier damage caused by constipation, and has multiple functional curative effects for moistening the intestines and relieving constipation. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 It is a result diagram of the influence of the composition of the embodiment of the present invention on the defecation of mice, where A is the time of the first black stool, B is the fecal weight at 5 h, C is the number of fecal particles at 5 h, D is the fecal water content, and different lowercase letters in the figure indicate significant differences between groups (p < 0.05).

[0029] Figure 2 It is a result diagram of the influence of the composition of the embodiment of the present invention on the ink propulsion rate of mice.

[0030] Figure 3 This is the result graph of the influence of the composition in the embodiment of the present invention on mouse serum neurotransmitters and hormones. Among them, A is the level of 5-hydroxytryptamine (5-HT), B is the level of substance P (SP), C is the level of vasoactive intestinal peptide (VIP), D is the level of motilin (MTL), and E is the level of gastrin (Gas).

[0031] Figure 4 This is the influence of Composition Ⅲ on the α-diversity of the intestinal flora of mice. Among them, A is the Ace index and B is the Simpson index.

[0032] Figure 5 This is the influence of the composition on the β-diversity of the intestinal flora of mice.

[0033] Figure 6 This is the result graph of the influence of the composition on the flora composition of the intestinal flora of mice at the phylum level.

[0034] Figure 7 This is the result graph of the influence of the composition on the flora composition of the intestinal flora of mice at the genus level.

[0035] Figure 8 This is the result graph of the influence of the composition on the short-chain fatty acid level in the feces of mice. Specific Embodiments

[0036] The following further elaborates on the concept and technical effects of the present invention in combination with specific embodiments to fully understand the purpose, features, and effects of the present invention. The methods are conventional methods unless otherwise specified. The materials can be obtained from public commercial channels unless otherwise specified. The illustrative embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute an improper limitation of the present invention. It should be noted that, without conflict, the embodiments in the present invention and the features in the embodiments can be combined with each other.

[0037] In the examples, fructooligosaccharide was purchased from Shandong Bailong Chuangyuan Biotechnology Co., Ltd., galactooligosaccharide was purchased from Shandong Bailong Chuangyuan Biotechnology Co., Ltd., curcumin was purchased from Sigma Company, oat β-glucan was purchased from Xi'an Zhongyan Kangze Biotechnology Co., Ltd., and freeze-dried aloe vera gel was purchased from Shaanxi Baichuan Biotechnology Co., Ltd.

[0038] Example 1 Screening of Composition Raw Materials

[0039] As an important neurotransmitter in the gut-brain axis, 5-HT plays a key role in regulating intestinal motility. SERT is a key protein that regulates the availability of extracellular 5-HT. The present invention uses the SERT mRNA expression level to characterize the potential activity of each substance in improving constipation. The specific steps are as follows:

[0040] Dissolve each test substance in DMEM complete medium (containing 10% fetal bovine serum and 1% penicillin-streptomycin), and filter through a 0.22 μm sterile needle filter to prepare a solution with a concentration of 1 - 200 mg / mL. Seed IEC-6 cells in a 6-well plate at a density of 5×105 / mL, culture overnight at 37°C under 5% CO2, culture for 4 days, and change the cell culture medium on the second day after seeding; before the intervention with the test substance, perform serum starvation for 1 h using serum-free DMEM medium, and then culture with 3 mL of the test substance at different concentrations for 24 h. The blank control group continues to be cultured with DMEM complete medium; after the intervention, discard the medium in the 6-well plate, add 1 mL of Trizol to each well, and then transfer the solution in each well to a 1.5 mL Eppendorf tube. Add 200 μL of chloroform, shake until the solution shows a uniform milky white, then centrifuge at 4°C and 12,000 rpm for 15 min. Transfer the upper aqueous phase to another clean centrifuge tube, add an equal volume of isopropanol, mix well by inverting up and down, let stand at room temperature for 30 min, and then centrifuge at 4°C and 12,000 rpm for 15 min. Discard the supernatant, add 1 mL of pre-cooled 75% ethanol (-20°C, prepared with DEPC water) to wash the precipitate (the precipitate is white); centrifuge at 4°C and 5,000 rpm for 5 min, carefully discard the supernatant, air-dry the precipitate, and dissolve it in 30 μL of DEPC water, then store at -80°C.

[0041] Take 10 μL of RNA in a 200 μL enzyme-free EP tube, add 4 μL of 5×All-In-One RT MasterMix and 6 μL of Nuclease-free H2O. After briefly centrifuging with a hand-held centrifuge, perform the following program: 25°C for 10 min; 42°C for 50 min; 85°C for 5 min to synthesize cDNA. Mix 1 μL of cDNA, 0.2 μL of the upstream primer, 0.2 μL of the downstream primer, 10 μL Premix ExTaq TM and 8.6 μL of sterilized water in an eight-well tube. Set the program as pre-denaturation at 95°C for 180 s, amplification at 95°C for 30 s, 60°C for 30 s, 72°C for 30 s, and the number of cycles is 40. Perform PCR amplification using GAPDH as the internal reference.

[0042] Take the mRNA expression level of the blank control group as 1, and take the ratio of the mRNA expression level of each intervention group to that of the blank control group as the final result.

[0043] As shown in Table 1, fructooligosaccharide, galactooligosaccharide, curcumin, oat β-glucan, and freeze-dried powder of Aloe vera gel all have the activity of downregulating the relative expression level of SERT mRNA.

[0044] Taking into comprehensive consideration factors such as the bioactive functions of raw materials, economic costs, sensory characteristics, safety, sustainability, eco-friendliness and social responsibility, recommended intake, and processing methods, fructooligosaccharide, galactooligosaccharide, oat β-glucan, and curcumin are selected as raw materials in the formula for subsequent research on the composition formula.

[0045] Table 1 Effects of Formulation Raw Materials on the Relative Expression Level of SERT mRNA

[0046]

[0047] Effects of the Composition in Example 2 on the Relative Expression Level of SERT mRNA

[0048] After weighing each raw material according to the following formula, the raw materials are mixed in a double-helix conical mixer for 15 - 30 minutes to obtain the composition.

[0049] Raw materials for Composition Ⅰ: 70 g of fructooligosaccharide, 4 g of curcumin, 10 g of galactooligosaccharide, 6 g of oat β-glucan;

[0050] Raw materials for Composition Ⅱ: 60 g of fructooligosaccharide, 3 g of curcumin, 15 g of galactooligosaccharide, 12 g of oat β-glucan;

[0051] Raw materials for Composition Ⅲ: 50 g of fructooligosaccharide, 5 g of curcumin, 25 g of galactooligosaccharide, 10 g of oat β-glucan;

[0052] Raw materials for Composition Ⅳ: 40 g of fructooligosaccharide, 10 g of curcumin, 35 g of galactooligosaccharide, 5 g of oat β-glucan;

[0053] Raw materials for Composition Ⅴ: 50 g of fructooligosaccharide, 5 g of curcumin, 17 g of galactooligosaccharide, 18 g of oat β-glucan;

[0054] Raw materials for Composition Ⅵ: 50 g of fructooligosaccharide, 5 g of curcumin, 20 g of galactooligosaccharide, 15 g of oat β-glucan;

[0055] Raw materials for Composition Ⅶ: 81.81 g of fructooligosaccharide, 8.18 g of curcumin;

[0056] Raw materials for Comparative Example 1: 90 g of fructooligosaccharide;

[0057] Raw materials for Comparative Example 2: 90 g of curcumin;

[0058] Raw materials for Comparative Example 3: 90 g of galactooligosaccharide;

[0059] Raw materials for Comparative Example 4: 90 g of oat β-glucan.

[0060] The method in Example 1 was used to determine the effects of Compositions I-VII and Comparative Examples 1-4 on the relative expression level of SERT mRNA, so as to characterize the potential effects of each composition in improving constipation.

[0061] Table 2 Effects of Compositions on the Relative Expression Level of SERT mRNA

[0062]

[0063]

[0064] Improvement Effect of the Composition in Example 3 on Constipated Mice

[0065] 1. Experimental Design and Grouping

[0066] Six-week-old SPF-grade male BALB / c mice (body weight 18 - 22 g) were selected, and the breeding environment temperature was 20 ± 2 °C; the relative humidity was 65 ± 5%, and the light-dark cycle was 12 h, with good ventilation maintained. The mice were fed with mouse feed and given free access to water. Before the experiment started, the mice were bred for 1 week to adapt to the environment. The mice were randomly divided into a blank control group, a model group, a low-formula group, a medium-formula group, a high-formula group, a Composition I group, a Composition II group, a Composition IV group, a Composition V group, a Composition VI group, a Composition VII group, a Comparative Example 1 group, a Comparative Example 2 group, a Comparative Example 3 group, a Comparative Example 4 group, and a positive drug group (n = 10), and were fed as follows:

[0067] Blank control group: Intragastric administration of normal saline from day 1 to day 14; from day 15 to day 28, first intragastric administration of normal saline, and 1 h later, intragastric administration of normal saline.

[0068] Model group: Intragastric administration of LOP (10 mg / kg BW) from day 1 to day 14; from day 15 to day 28, first intragastric administration of LOP (10 mg / kg BW), and 1 h later, intragastric administration of normal saline.

[0069] Low-formula group: Intragastric administration of LOP (10 mg / kg BW) from day 1 to day 14; from day 15 to day 28, first intragastric administration of LOP (10 mg / kg BW), and 1 h later, intragastric administration of low-dose Composition III (37 mg / kg BW).

[0070] Medium-formula group: Intragastric administration of LOP (10 mg / kg BW) from day 1 to day 14; from day 15 to day 28, first intragastric administration of LOP (10 mg / kg BW), and 1 h later, intragastric administration of medium-dose Composition III (74 mg / kg BW).

[0071] High-formula group: Intragastric administration of LOP (10 mg / kg BW) from day 1 to day 14; from day 15 to day 28, first intragastric administration of LOP (10 mg / kg BW), and 1 h later, intragastric administration of high-dose Composition III (148 mg / kg BW).

[0072] Group Ⅰ of the composition: LOP (10 mg / kg BW) was intragastrically administered from day 1 to day 14; from day 15 to day 28, LOP (10 mg / kg BW) was first intragastrically administered, and 1 h later, Composition Ⅰ (74 mg / kg BW) was intragastrically administered.

[0073] Group Ⅱ of the composition: LOP (10 mg / kg BW) was intragastrically administered from day 1 to day 14; from day 15 to day 28, LOP (10 mg / kg BW) was first intragastrically administered, and 1 h later, Composition Ⅱ (74 mg / kg BW) was intragastrically administered.

[0074] Group Ⅳ of the composition: LOP (10 mg / kg BW) was intragastrically administered from day 1 to day 14; from day 15 to day 28, LOP (10 mg / kg BW) was first intragastrically administered, and 1 h later, Composition Ⅳ (74 mg / kg BW) was intragastrically administered.

[0075] Group Ⅴ of the composition: LOP (10 mg / kg BW) was intragastrically administered from day 1 to day 14; from day 15 to day 28, LOP (10 mg / kg BW) was first intragastrically administered, and 1 h later, Composition Ⅴ (74 mg / kg BW) was intragastrically administered.

[0076] Group Ⅵ of the composition: LOP (10 mg / kg BW) was intragastrically administered from day 1 to day 14; from day 15 to day 28, LOP (10 mg / kg BW) was first intragastrically administered, and 1 h later, Composition Ⅵ (74 mg / kg BW) was intragastrically administered.

[0077] Group Ⅶ of the composition: LOP (10 mg / kg BW) was intragastrically administered from day 1 to day 14; from day 15 to day 28, LOP (10 mg / kg BW) was first intragastrically administered, and 1 h later, Composition Ⅶ (74 mg / kg BW) was intragastrically administered.

[0078] Group of Comparative Example 1: LOP (10 mg / kg BW) was intragastrically administered from day 1 to day 14; from day 15 to day 28, LOP (10 mg / kg BW) was first intragastrically administered, and 1 h later, fructooligosaccharide (74 mg / kg BW) was intragastrically administered.

[0079] Group of Comparative Example 2: LOP (10 mg / kg BW) was intragastrically administered from day 1 to day 14; from day 15 to day 28, LOP (10 mg / kg BW) was first intragastrically administered, and 1 h later, curcumin (74 mg / kg BW) was intragastrically administered.

[0080] Group of Comparative Example 3: LOP (10 mg / kg BW) was intragastrically administered from day 1 to day 14; from day 15 to day 28, LOP (10 mg / kg BW) was first intragastrically administered, and 1 h later, galactooligosaccharide (74 mg / kg BW) was intragastrically administered.

[0081] Negative control group 4: intragastric administration of LOP (10 mg / kg BW) from day 1 to day 14; from day 15 to day 28, first intragastric administration of LOP (10 mg / kg BW), and 1 h later, intragastric administration of oat β-glucan (74 mg / kg BW).

[0082] Positive drug group: intragastric administration of LOP (10 mg / kg BW) from day 1 to day 14; from day 15 to day 28, first intragastric administration of LOP (10 mg / kg BW), and 1 h later, intragastric administration of polyethylene glycol 4000 (3 g / kg BW).

[0083] 2. Mouse fecal parameter determination experiment

[0084] After the intragastric administration on day 28 was completed, the feces of the mice were collected and stored in a -80 °C refrigerator. The mice were fasted but allowed to drink water for 16 h. On day 29, the fecal parameters of the mice were measured: the blank control group was given normal saline, and the model group, low-formula group, medium-formula group, high-formula group, composition I group, composition II group, composition IV group, composition V group, composition VI group, composition VII group, negative control group 1, negative control group 2, negative control group 3, negative control group 4, and positive drug group were intragastrically administered LOP (10 mg / kg BW). 0.5 h later, the mice in the blank control group and the model group were intragastrically administered ink (0.2 mL), and the low-formula group, medium-formula group, high-formula group, composition I group, composition II group, composition IV group, composition V group, composition VI group, composition VII group, negative control group 1, negative control group 2, negative control group 3, negative control group 4, and positive drug group were given ink (0.2 mL) containing the corresponding test samples. The animals were each housed in a single cage and allowed to drink water and eat normally. Starting from the administration of the ink, the time for the first black feces excretion, the number and weight of black feces excreted within 5 h, and the water content of the feces of each mouse were recorded, and the water content of the feces of the mice was measured.

[0085] The feces were placed in a centrifuge tube of known weight, weighed, and the wet weight of the feces was calculated. The centrifuge tube containing the feces was placed in a vacuum freeze dryer. After drying, the dry weight of the feces was weighed, and the water content of the feces was calculated as follows:

[0086] Fecal water content (%) = (fecal wet weight - fecal dry weight) / fecal wet weight × 100%

[0087] The constipation situation and improvement of the mice were evaluated using the time for the first black feces excretion, the weight and number of black feces within 5 h, and the water content of the feces as indicators. As Figure 1As shown in Figure A, the time for the first black feces in the blank control group of mice was 46.5 ± 1.46 min, but the time for the first black feces in the model group of mice reached 132.33 ± 3.89 min, and the time for the first black feces increased significantly, indicating that the constipation mouse model was successfully constructed in mice after LOP modeling. Compared with the model group, the time for the first black feces in the formula group and the positive drug group of mice decreased significantly. Among them, the time for the first black feces in the medium- and high-formula groups of mice was 69.5 ± 2.10 min and 62 ± 3.00 min respectively, showing no significant difference from the blank control group, indicating that the composition can promote intestinal peristalsis and improve constipation.

[0088] As Figure 1 shown in Figure 1 Figure Figure 1 B,

[0089] 3. Small intestine motility experiment in mice

[0090] After the experiment on the 29th day, the mice were fasted but allowed to drink water for 16 h. On the 30th day, the intestinal propulsion rate of the mice was measured.

[0091] The blank control group was intragastrically administered normal saline, and the model group, low-formula group, medium-formula group, high-formula group, composition I group, composition II group, composition IV group, composition V group, composition VI group, composition VII group, comparative example 1 group, comparative example 2 group, comparative example 3 group, comparative example 4 group, and positive drug group were intragastrically administered LOP (10 mg / kg BW). After 0.5 h, the blank control group and the model group of mice were intragastrically administered ink (0.2 mL), and the low-formula group, medium-formula group, high-formula group, composition I group, composition II group, composition IV group, composition V group, composition VI group, composition VII group, comparative example 1 group, comparative example 2 group, comparative example 3 group, comparative example 4 group, and positive drug group were intragastrically administered ink (0.2 mL) containing the corresponding test samples. The animals were each housed in a single cage and had normal access to water and food. After 25 min, they were sacrificed, the abdominal cavity was opened to separate the mesentery, and the intestinal tube from the upper end at the pylorus to the lower end at the ileocecal junction was cut and placed on graph paper. The small intestine was gently stretched into a straight line, and the length of the intestinal tube was measured as the "total length of the small intestine", and the length from the pylorus to the ink front was measured as the "length of ink propulsion".

[0092] The ink propulsion rate (%) = length of ink propulsion (cm) / total length of small intestine (cm) × 100%

[0093] The ink propulsion rate of each group of mice was measured, and the results are shown in Figure 2As shown in the figure. Compared with the blank control group, the ink propulsion rate of the mice in the model group was significantly decreased; compared with the model group, the ink propulsion rates of the mice in the low-formula group, medium-formula group, high-formula group, and positive drug group were significantly increased, and the medium and high doses of the formula had the best effects. Compared with the low, medium, and high formula groups and each composition group, the effects of increasing the ink propulsion rate of the mice in Comparative Example 1 group, Comparative Example 2 group, Comparative Example 3 group, and Comparative Example 4 group were significantly weakened, indicating that the composition composed of fructooligosaccharide and curcumin and the composition composed of fructooligosaccharide, curcumin, galactooligosaccharide, and oat β-glucan had better intervention effects on constipation than the intervention effects of single-component substances at the same dose. The above results proved that there was a synergistic effect in improving constipation between fructooligosaccharide and curcumin, and there was also a synergistic effect in improving constipation among fructooligosaccharide, curcumin, galactooligosaccharide, and oat β-glucan.

[0094] Effect of the composition in Example 4 on serum neurotransmitters and hormones of constipated mice

[0095] Neurotransmitters such as serotonin (5-HT), substance P (SP), and vasoactive intestinal peptide (VIP) directly affect intestinal peristalsis by regulating intestinal smooth muscle contraction and intestinal secretion function, while gastrointestinal hormones such as motilin (MTL) and gastrin (Gas) play an important role in regulating gastrointestinal motility by regulating gastrointestinal smooth muscle contraction.

[0096] The experimental animals were grouped in the same way as in Example 3. Commercial ELISA kits were used to measure the levels of 5-HT, SP, VIP, MTL, and Gas in the serum of mice. The results were as Figure 3 shown. Compared with the blank control group, after LOP modeling, the levels of 5-HT and SP in the serum of the mice in the model group were significantly decreased, the level of VIP was significantly increased, and the levels of MTL and Gas were significantly decreased. Compared with the model group, the levels of 5-HT and SP in the serum of the mice in the low-formula group, medium-formula group, high-formula group, and positive drug group were significantly increased, the level of VIP was significantly decreased, and the levels of MTL and Gas were significantly increased. After the formula group intervened in constipated mice, the level of the inhibitory neurotransmitter VIP was decreased, and the levels of the excitatory neurotransmitters 5-HT and SP were increased, promoting intestinal peristalsis and effectively improving constipation.

[0097] Effect of the composition in Example 5 on the intestinal flora diversity of constipated mice

[0098] The experimental animals were grouped in the same way as in Example 3. In order to explore the effects of the composition on the diversity and richness of the intestinal flora of constipated mice, the α-diversity of the intestinal flora of the mice was analyzed. The results were as Figure 4As shown, the Ace index reflects the richness of the microbial community. The higher the Ace index, the higher the richness of the microbial community. The Simpson index reflects the diversity of the microbial community. The lower the Simpson index, the higher the diversity of the microbial community. Compared with the blank control group, the Ace index of the model group mice decreased significantly (p < 0.05), and the Simpson index increased significantly (p < 0.05), indicating that constipation led to a significant decrease in the richness and diversity of the intestinal microbiota. Compared with the model group, the Ace index of the formula group (hereinafter, the "formula group" all refers to the "Chinese formula group") increased (p < 0.05), and the Simpson index decreased significantly (p < 0.05), with no significant difference from the positive drug group (p > 0.05), indicating that the formula can effectively restore the α-diversity of the intestinal flora in mice and improve the intestinal flora of constipated mice.

[0099] β-diversity can evaluate the differences between microbial communities in different groups. The results are as Figure 5 shown. The principal component analysis (PCA) based on OUT shows that it can be seen from the figure that there are obvious clusters in the intestinal flora composition of mice in different groups, and each group is clearly distinguished along the PC1 axis, indicating that there are differences in the intestinal flora composition of mice in each group. The blank control group and the model group mice formed independent clusters, and the intestinal flora of the formula group and the positive drug group mice approached the distribution of the blank control group, indicating that this composition can significantly improve the shift of the flora structure caused by constipation.

[0100] Effect of the composition in Example 6 on the species composition of the intestinal flora of constipated mice

[0101] The experimental animals were grouped in the same way as in Example 3. The composition of the intestinal flora of mice in each group at the phylum level is as Figure 6As shown, the intestinal flora of the four groups of mice was mainly composed of Bacteroidota and Fimicutes. Research has shown that a decrease in the abundance of Bacteroidota is an important microbial marker for constipation. Compared with the blank control group, the relative abundance of Bacteroidota in the model group of mice was significantly decreased (p < 0.05). After gavage with the positive drug group, the relative abundance of Bacteroidota was significantly increased (p < 0.05), and there was no significant difference between the formula group and the positive drug group (p > 0.05). Compared with the blank control group, the relative abundance of Fimicutes in the model group of mice was significantly increased. Compared with the model group, the relative abundance of Fimicutes in the formula group and the positive drug group of mice was significantly decreased (p < 0.05). Compared with the model group, the relative abundance of Actinobacteriota in the formula group was significantly increased (p < 0.05). Compared with the blank control group, the abundance of Proteobacteria in the model group of mice was significantly increased (p < 0.05), suggesting that there may be an abnormal increase in the abundance of pathogenic bacteria. After intervention, compared with the model group, the abundance of Proteobacteria in the formula group and the positive drug group of mice was significantly decreased (p < 0.05), and there was no significant difference from the blank control group (p > 0.05), inhibiting the proliferation of pathogenic bacteria such as Proteobacteria.

[0102] The composition of the intestinal flora of mice in each group at the genus level was as Figure 7As shown, compared with the blank control group, the relative abundance of norank_o_Clostridia_UCG-014 (unclassified Clostridia-UCG-014) in the model group (LOP) mice was significantly increased, while the relative abundances of Bacteroides, Ruminococcus, and Alistipes were significantly decreased (p<0.05). Compared with the model group, the relative abundance of norank_o_Clostridia_UCG-014 in the formula group mice was significantly decreased (p<0.05), and there was no significant difference between the formula group and the positive drug group. Compared with the model group, the relative abundance of Ruminococcus in the positive drug group mice was significantly increased (p<0.05), and the relative abundance of Ruminococcus in the formula group mice was also significantly increased, and the recovery effect was significantly better than that of the positive drug group (p<0.05). Some studies have shown that an increase in the relative abundance of the Clostridia UCG-014 flora may lead to inflammation in the body. The increase in pro-inflammatory factors can reduce the synthesis of 5-HT and lower the bioavailability of 5-HT. Bacteroidota is one of the core intestinal flora, which can efficiently decompose dietary fiber and complex polysaccharides to produce short-chain fatty acids (SCFAs). SCFAs can promote the synthesis and release of 5-HT by enterochromaffin cells (EC cells). In addition, it can also enhance the excitability of intestinal neurons, promote intestinal peristalsis reflex, and improve constipation. Bacteroides, Ruminococcus, and Alistipes are all short-chain fatty acid-producing genera, which have an improving effect on constipation.

[0103] Short-chain fatty acids (SCFAs) are metabolites of the intestinal flora, which not only provide necessary nutrients and energy for intestinal epithelial cells, but also are beneficial to the growth and reproduction of beneficial bacteria, while inhibiting the overgrowth of harmful bacteria, and can promote intestinal peristalsis, playing an important role in maintaining intestinal health. The contents of short-chain fatty acids in the feces of mice in each group were measured, and the results are as Figure 8 shown. Compared with the blank control group, the contents of total acid, acetic acid, propionic acid, and butyric acid in the feces of the model group mice were significantly decreased; compared with the model group, the formula group and the positive drug group could significantly increase the contents of total acid, propionic acid, and butyric acid in the feces of mice, and the levels of total acid, propionic acid, and butyric acid in the formula group were significantly higher than those in the positive drug group; the above results indicate that the formula composition can effectively increase the content of short-chain fatty acids in the feces of mice, which is beneficial to intestinal health.

[0104] Through experiments such as measuring the time of the first black stool, the ink propulsion rate, and the fecal water content, the present invention proves that the composition can shorten the defecation time of mice and promote intestinal peristalsis; through the PCR method, it is proved that the composition can down-regulate the expression level of SERT mRNA in IEC-6 cells. At the same time, the contents of serotonin (5-HT), substance P (SP), vasoactive intestinal peptide (VIP), motilin (MTL), and gastrin (Gas) in the serum of mice are detected, proving that the composition can promote the release of neurotransmitters and gastrointestinal hormones related to the improvement of constipation. In addition, by analyzing the intestinal flora of mice and using gas chromatography-mass spectrometry to measure the content of short-chain fatty acids, it is proved that the composition can regulate the intestinal flora, increase its diversity and richness, and can increase the level of short-chain fatty acids, which is beneficial to maintaining intestinal homeostasis.

[0105] The embodiments described above are only a part of the embodiments of the present invention, rather than all embodiments. Based on the embodiments of the present invention, other embodiments obtained by those skilled in the art without creative efforts belong to the scope of protection of the present invention.

Claims

1. A composition that helps moisten the intestines and relieve constipation, characterized in that, The composition comprises fructooligosaccharide and curcumin, and the mass ratio of fructooligosaccharide to curcumin is 40 - 70:3 - 10.

2. The composition according to claim 1, wherein The mass ratio of fructooligosaccharide to curcumin is 10:

1.

3. The composition according to claim 1, wherein The composition further comprises galactooligosaccharide and oat β-glucan, and the mass ratio of fructooligosaccharide, curcumin, galactooligosaccharide and oat β-glucan is 40 - 70:3 - 10:10 - 35:5 - 18.

4. The composition according to claim 3, wherein The mass ratio of fructooligosaccharide, curcumin, galactooligosaccharide and oat β-glucan is 10:1:5:

2.

5. A product, which comprises the composition according to any one of claims 1 - 4.

6. The product according to claim 5, wherein, The product is a health food or a medicine, and the product further comprises excipients acceptable for the health food or the medicine.

7. Use of the composition according to any one of claims 1-4 in the preparation of a product capable of moistening the intestine and relieving constipation, characterized in that, The product is a health food or a medicine.

8. The application according to claim 7, wherein The application is for the preparation of a product capable of improving constipation, promoting intestinal peristalsis and / or increasing intestinal transit rate.

9. The application according to claim 7, wherein The application includes at least one of the following: (1) The application for the preparation of a product capable of shortening the defecation time of constipated patients; (2) The application for the preparation of a product capable of improving the release of neurotransmitters and gastrointestinal hormones related to constipation; (3) The application for the preparation of a product capable of improving the intestinal flora imbalance caused by constipation; (4) The application for the preparation of a product capable of improving the intestinal barrier injury caused by constipation.

10. The application according to claim 9, wherein The application for the preparation of a product capable of improving the release of neurotransmitters and gastrointestinal hormones related to constipation includes at least one of the following: (1) The application for the preparation of a product capable of improving the decrease in the levels of 5-HT and SP caused by constipation; (2) The application for the preparation of a product capable of improving the increase in the level of VIP caused by constipation; (3) The application for the preparation of a product capable of improving the decrease in the levels of Gas and MTL caused by constipation.

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