Composition for preventing and treating postoperative constipation in patients and application thereof

Abstract

The present invention relates to the field of biomedicine technology, and in particular to a composition for preventing and treating postoperative constipation in patients and its application, wherein the composition comprises a medicinal rhubarb polypeptide and probiotics; the mass percentages of the medicinal rhubarb polypeptide and the probiotics are 2-10% and 10-30% respectively; the sequence of the medicinal rhubarb polypeptide is shown in SEQ ID NO.1. The present invention identifies the medicinal rhubarb SEVIYWAGDFLK polypeptide for the first time, and compared with the prior art using a combination of multiple traditional Chinese medicines, the components are more streamlined, a new source of active ingredients is developed, and the active ingredients are clear, which is more conducive to large-scale production and preparation using genetic engineering and fermentation engineering, that is, it can reduce costs and is conducive to market promotion; (2) the present invention is used in combination with probiotics, and the effect is better and has a synergistic effect.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedicine, and in particular to a composition for preventing and treating postoperative constipation in patients and an application thereof. Background Art

[0002] Constipation is a common complication after surgery. It not only has a serious adverse effect on postoperative recovery, but some patients may even die suddenly due to excessive cardiopulmonary burden caused by high abdominal pressure, which is life-threatening. There are currently three main causes of postoperative constipation: (1) water loss. After surgery, patients may lose too much water due to excessive sweating, blood loss, or even postoperative fever, which may cause temporary constipation. (2) The influence of drugs. Many patients need to take related drugs for postoperative maintenance treatment. For example, taking calcium carbonate, aluminum hydroxide, morphine, and even other drugs will make patients have difficulty defecating. (3) The food eaten after surgery is too refined, resulting in insufficient dietary nutrients and water in the food, which will make the stool dry and hard, resulting in difficulty defecating.

[0003] In the prior art, compositions for treating constipation generally include traditional Chinese medicine compositions, probiotic compositions, or compositions of probiotics combined with polypeptides.

[0004] For example, the prior art CN 117298184 B discloses a composition for treating constipation and bad breath, which comprises the following ingredients: rhubarb, immature bitter orange, capillaris, ferulic acid, paeonol, bakuchiol, gentianine, kaempferol, oleanolic acid and luteolin, and can effectively treat constipation.

[0005] Prior art CN 114711435 B discloses a dietary fiber probiotic composition and a preparation method thereof. The dietary fiber probiotic composition uses ginseng dietary fiber, oat β-glucan, silkworm pupa protein peptide, and probiotic microcapsules as raw materials. The substances interact with each other and synergize to effectively regulate intestinal flora, promote food digestion, increase intestinal peristalsis, and prevent constipation.

[0006] Prior art CN 114588247 B discloses an active peptide probiotic composition, which comprises bovine liver peptide, soybean peptide and probiotics; the weight ratio of the bovine liver peptide, soybean peptide and probiotics is 1-3:1-3:0.02-0.1; the bovine liver peptide is a bovine liver peptide with a molecular weight less than 1kD; the soybean peptide is a soybean peptide with a molecular weight less than 1kD.

[0007] However, the above-mentioned prior arts all contain multiple components with limited sources, and there are too many components in the composition, which is not convenient for industrial production and cannot meet the growing demand. Therefore, in order to expand new component sources, simplify the components of the composition, and optimize the proportion of the composition, the present application will attempt to obtain a composition with better effect in preventing and treating postoperative constipation in patients. Summary of the invention

[0008] In order to solve the above problems, the present invention first provides a composition for preventing and treating postoperative constipation in patients, comprising medicinal rhubarb polypeptide and probiotics.

[0009] Preferably, the mass percentages of the medicinal rhubarb polypeptide and the probiotics are 2-10% and 10-30% respectively.

[0010] Preferably, the medicinal rhubarb polypeptide is a medicinal rhubarb polypeptide with a molecular weight of less than 4kD.

[0011] Preferably, the medicinal rhubarb polypeptide sequence is shown as SEQ ID NO.1.

[0012] Preferably, the probiotics are selected from one or more of Bifidobacterium, Lactobacillus acidophilus, Lactobacillus casei and Lactobacillus rhamnosus.

[0013] Preferably, the probiotics are bifidobacterium and Lactobacillus rhamnosus.

[0014] Preferably, the mass percentages of the bifidobacterium and the lactobacillus rhamnosus in the probiotics are 10-30% and 10-30% respectively.

[0015] The present invention also provides the use of the above-mentioned composition for preventing and treating postoperative constipation in the preparation of food or medicine having the effects of promoting intestinal peristalsis and / or relieving constipation.

[0016] Finally, the present invention provides a method for preparing the composition for preventing and treating postoperative constipation in patients, wherein the medicinal rhubarb polypeptide is separated by enzymatic hydrolysis, freeze-dried and made into powder; and then evenly mixed with probiotics to obtain the composition for preventing and treating postoperative constipation in patients.

[0017] Preferably, the medicinal rhubarb polypeptide sequence is shown as SEQ ID NO.1.

[0018] Compared with the prior art, the present invention has at least the following beneficial effects:

[0019] (1) The present invention identifies the medicinal rhubarb SEVIYWAGDFLK polypeptide for the first time. Compared with the prior art which uses a combination of multiple traditional Chinese medicines, the components are more streamlined, a new source of active ingredients is developed, and the active ingredients are clear, which is more conducive to large-scale production and preparation using genetic engineering and fermentation engineering, which can reduce costs and facilitate market promotion;

[0020] (2) The present invention is used in combination with probiotics to achieve better effects and a synergistic effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is the peptide mass spectrometry identification diagram. DETAILED DESCRIPTION

[0022] In order to make the technical problems, technical solutions and advantages to be solved by the present invention more clear, a detailed description will be given below with reference to the accompanying drawings and specific embodiments.

[0023] Example 1 Extraction of polypeptides

[0024] (1) Take 100 g of fresh rhizome of medicinal rhubarb (Rheum officinale Baill.), wash it with ddH2O, drain it, cut it into pieces, grind it with an ultrafine grinder to 100 mesh, soak it in 2 volumes of PBS buffer, stir it for 30 min, and filter it with gauze to obtain the residue;

[0025] (2) Transfer the filter residue in step (1) to a large beaker, add 300 mL of ddH2O to the large beaker, and then place the beaker in a water bath, stir at 55°C for 2 h to obtain a colloidal mixed solution; after cooling to 45°C, add cellulase (0.4% w / v) and trypsin (0.2% w / v), stir and hydrolyze for 3 h; centrifuge at 3000g for 15 min to obtain a supernatant;

[0026] (3) The supernatant was filtered and concentrated using a nanofiltration membrane with a membrane molecular cutoff of 4 KDa, and two polypeptide solutions with molecular weights of A (>4 KDa) and B (<4 KDa) were separated and purified. The solutions were desalted using a dialysis bag (100 Da), concentrated by rotary evaporation, and freeze-dried. The components with better effects on preventing and treating constipation were collected and stored in a -10°C refrigerator for later use.

[0027] (4) Component B was separated and purified by Sephadex G-25 to obtain four components, namely, B1, B2, B3, and B4. Each component was concentrated by rotary evaporation and then freeze-dried in a vacuum. The freeze-dried polypeptide powder was collected and prepared into a certain concentration for constipation prevention and treatment experiments;

[0028] Model establishment: Loperamide hydrochloride was diluted with normal saline, mixed, and prepared into loperamide hydrochloride injection. After one week of adaptive feeding, male rats were injected once a day in the morning and evening (single loperamide hydrochloride administration of 5 mg / kg) for one week. During this period, the rats drank water and ate normally. After one week, the feces of the rats were observed. The rats with continuous fecal particle reduction, number reduction, weight reduction, and dry feces were selected as the rats with successful modeling. At the same time, normal mice were injected with the same volume of normal saline.

[0029] Grouping: The mice with successful modeling were treated with drugs (experimental group), and a normal control group (normal mice) and a model group (model mice) were set up at the same time, with 10 mice in each group. The normal control group and the model group were given normal saline by gavage, and the other experimental groups were given the same dose of B1, B2, B3, and B4 components (50 mg of freeze-dried polypeptide powder was added to 1 mL of water and configured as a solution for administration) by gavage, once a day at 1 ml / 100 g body weight, for 14 consecutive days. The spirit, activity, hair, diet, etc. of the rats were observed every day.

[0030] After administration on the 14th day, feces were collected within 4 hours and the feces properties were measured:

[0031] Feces were collected, counted and weighed; feces were collected and freeze-dried to calculate the water content of feces: water content = (feces weight before freeze-drying - feces weight after freeze-drying) / feces weight before freeze-drying × 100%.

[0032] The results of stool properties are shown in Table 1:

[0033] Table 1 Results of preventing and treating constipation

[0034] Grouping Number of feces (pieces) Total weight of feces (g) Fecal water content (%) Normal control group 15.32±0.91 0.51±0.02 60.19±5.31 Model Group 6.29±0.56 0.18±0.02 31.24±6.11 B1 15.41±0.64 0.49±0.01 59.01±4.41 B2 13.41±0.71 0.39±0.03 45.19±5.31 B3 10.29±0.86 0.37±0.02 42.24±5.31 B4 9.99±0.42 0.31±0.01 39.14±5.31

[0035] The results shown in Table 1 indicate that the active peptides in group B1 have the strongest ability to prevent and treat constipation (p<0.01).

[0036] (5): The B1 component was separated and purified by Sephadex G-15, and the obtained B1-1, B1-2, B1-3, and B1-4 components were concentrated by rotary evaporation, and then vacuum freeze-dried. The freeze-dried polypeptide powder was collected and prepared into a certain concentration for the constipation prevention and treatment experiment. The materials and methods are shown in step 4. The results are shown in Table 2;

[0037] Table 2 Results of preventing and treating constipation

[0038]

[0039]

[0040] The results shown in Table 1 indicate that the active peptides in group B1-1 have the strongest ability to prevent and treat constipation (p<0.01).

[0041] (6): The active peptide of group B1-1 was purified by RP-HPLC, and the mass spectrometry results showed that its amino acid sequence was SEVIYWAGDFLK (SEQ ID NO. 1), see Figure 1 .

[0042] Example 3 Preparation of a composition for preventing and treating postoperative constipation in patients

[0043] This embodiment provides a method for preparing a composition for preventing and treating postoperative constipation in patients, which specifically comprises the following steps:

[0044] (1) According to Table 3, the polypeptide freeze-dried powder, animal Bifidobacterium (animal Bifidobacterium Bb-12 purchased from Denmark Co., Ltd.) and Lactobacillus rhamnosus (Lactobacillus rhamnosus LGG purchased from China General Microbiological Culture Collection Administration Center) were weighed and the solution was prepared at room temperature.

[0045] Table 3 Composition of each experimental group (mass percentage)

[0046] Group B1-1 lyophilized powder % Bifidobacterium animalis % Lactobacillus rhamnosus % margin Composition 1 5 10 10 water Composition 2 5 20 water Composition 3 5 20 water Composition 4 10 10 water Composition 5 5 water Composition 6 20 water Composition 7 20 water

[0047] Refer to step 4 of Example 2 to conduct a constipation treatment experiment. See Table 4 for the results.

[0048] Table 4 Experimental results of different compositions for treating constipation

[0049] Grouping Number of feces (pieces) Total weight of feces (g) Fecal water content (%) Normal control group 15.92±0.91 0.53±0.02 61.01±4.61 Model Group 6.31±0.56 0.19±0.02 31.74±5.21 Composition 1 18.21±0.64 0.55±0.01 63.04±6.12 Composition 2 17.13±0.47 0.54±0.01 61.42±4.49 Composition 3 16.93±0.49 0.53±0.01 61.12±4.72 Composition 4 14.51±0.61 0.48±0.03 55.19±5.21 Composition 5 16.13±0.52 0.52±0.01 60.02±4.52 Composition 6 12.53±0.56 0.42±0.02 46.69±5.31 Composition 7 12.31±0.62 0.39±0.02 45.49±5.42

[0050] Table 4 shows that Composition 1 of the present invention has the best effect in preventing and treating constipation (p<0.01), which means that the components of Composition 1 have achieved a synergistic effect and can be better used in preventing and treating constipation and clinical promotion.

[0051] The above is a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A composition for preventing and treating postoperative constipation in patients, characterized in that: It comprises medicinal rhubarb polypeptide and probiotics; the mass percentages of the medicinal rhubarb polypeptide and probiotics are 2-10% and 10-30% respectively; the sequence of the medicinal rhubarb polypeptide is shown in SEQ ID NO.1; and the probiotics are bifidobacterium and lactobacillus rhamnosus.

2. Use of the composition for preventing and treating postoperative constipation in patients according to claim 1 in the preparation of food or medicine having the effect of promoting intestinal peristalsis and / or relieving constipation.

3. The method for preparing the composition for preventing and treating postoperative constipation in patients according to claim 1, characterized in that: The medicinal rhubarb polypeptide is separated by enzymatic hydrolysis, freeze-dried and made into powder; and then mixed evenly with probiotics to obtain the composition for preventing and treating postoperative constipation in patients. The sequence of the medicinal rhubarb polypeptide is shown in SEQ ID NO.1.

Images