Animal bifidobacterium and radish seed composition with synergistic effect of preventing and treating functional constipation

Abstract

The present invention provides a composition of animal bifidobacterium and radish seeds with the synergistic effect of preventing and treating functional constipation. The animal bifidobacterium is named YRK-12, which is deposited in the General Microbiological Center of the China Microbiological Culture Management Committee and is classified as animal bifidobacterium. Bifidobacterium animalum , the deposit number is CGMCC NO.28597, and the deposit date is October 9, 2023. The animal Bifidobacterium and Radish Seed Composition described in this application have potential interactions and can cooperate with each other to exert synergistic effects. By increasing the number of fecal particles and fecal water content, reducing defecation time, and improving small intestinal propulsion rate, it provides reference indicators for the industrial production of probiotics and Chinese medicine extract compound compositions, and also provides theoretical basis and data support for the promotion and application of probiotics and Chinese medicine extract compound compositions in the field of preventing and treating functional constipation.

Description

Technical Field

[0001] The invention belongs to the technical field of new application of microorganisms, and particularly relates to an animal bifidobacterium and radish seed composition capable of synergistically preventing and treating functional constipation. Background Art

[0002] Constipation is a common clinical gastrointestinal disease characterized by difficult, low-frequency or incomplete defecation. It has a great impact on the patient's quality of life. About 5.5% of people worldwide suffer from functional constipation. At present, the main treatment methods are: ① Diet and lifestyle changes: eat more high-fiber foods and exercise appropriately. ② Drug treatment: such as laxatives containing laxative ingredients, drugs that promote intestinal peristalsis, etc., but these drugs are prone to long-term dependence. ③ Biofeedback therapy: This method uses instrument detection to guide training and exercise to improve muscle contraction and intestinal defecation function. ④ Psychotherapy: Psychotherapy is used to relieve constipation caused by emotional problems, stress and other factors.

[0003] With the increasing incidence of constipation and the limitations and side effects of various treatments, it is particularly important to find new and effective treatment methods. In terms of improving constipation, probiotics and medicinal and edible substances each have unique advantages, and the combination of the two can complement each other to achieve a more comprehensive therapeutic effect. Probiotics can regulate the balance of intestinal flora, increase the number of intestinal peristalsis, and promote the frequency of natural defecation and the normal formation of feces. They produce short-chain fatty acids by decomposing dietary fiber, which can stimulate intestinal peristalsis and help soften feces, thereby relieving constipation symptoms. In addition, probiotics can also help repair damaged intestinal mucosa, enhance intestinal barrier function, and reduce the invasion of harmful bacteria. Medicinal and edible substances refer to those substances that can be used as food and have certain medicinal value. For example, the ingredients such as mulberry, honey, and hemp seeds in Shuangren Runchang Oral Liquid are all medicinal and edible materials. They have the effect of moisturizing the intestines and promoting bowel movements, which can help improve constipation symptoms. These substances are usually mild in nature, have a wide range of applications, and are suitable for long-term use by people of different constitutions. However, existing research lacks relevant research on the compounding of bifidobacteria with traditional Chinese medicine ingredients. Summary of the invention

[0004] In view of the problems existing in the prior art, the purpose of the present invention is to screen and obtain probiotics with good efficacy in preventing and treating constipation, and to explore the synergistic therapeutic effect between probiotics and Chinese medicine ingredients, and to provide a composition of animal bifidobacterium and radish seeds with synergistic prevention and treatment of functional constipation. In order to provide reference indicators for the industrial production of probiotics and Chinese medicine extract composite compositions, and also to provide theoretical basis and data support for the promotion and application of probiotics and Chinese medicine extract composite compositions in the field of preventing and treating functional constipation.

[0005] The objective of the present invention is achieved through the following technical solutions:

[0006] Due to the complex composition of Chinese herbal extracts, the reasons for their effectiveness in helping to improve constipation symptoms are not clear, resulting in different probiotics compounded with different Chinese herbal extracts, and the combined effect may show three different results: antagonism, synergy or addition. Therefore, the inventors have conducted research on this, hoping to find a Chinese herbal extract that can be used in combination with the animal Bifidobacterium YRK-12 described in this application to achieve synergistic enhancement. The medicinal and edible materials screened in the early stage of the study include: mulberry, honey, hemp seed and radish seed. However, it was found through experiments that the above four substances can improve constipation symptoms when intervened alone, but after compounding with animal Bifidobacterium YRK-12, the composition formed by YRK-12+mulberry extract has an inhibitory effect on the intervention of constipation mice; the composition of YRK-12+honey basically shows an additive combined effect on the intervention of constipation mice. The extracts of hemp seed and radish seed are used in combination with animal Bifidobacterium YRK-12, respectively, and then show obvious synergistic effects.

[0007] The first aspect of the present invention provides an animal Bifidobacterium and radish seed composition having the synergistic effect of preventing and treating functional constipation, the composition comprising animal Bifidobacterium and radish seed extract;

[0008] The animal bifidobacterium is named YRK-12, which is deposited in the General Microbiological Center of the China Microbiological Culture Management Committee and is classified as animal bifidobacterium. Bifidobacterium animalis , the deposit number is CGMCC NO.28597, and the deposit date is October 9, 2023;

[0009] In the composition, the final therapeutic dose of animal Bifidobacterium YRK-12 is a viable bacterial count of not less than 1.0×10 10 CFU / d / kg body weight.

[0010] Furthermore, the radish seed extract is a water extract, and the extraction method of the radish seed water extract is:

[0011] The radish seeds are crushed into powder by a grinder, and 8 to 15 times the weight of water is added for decocting for 2 to 5 times, each time for 0.5 to 1.5 hours. The decocted seeds are filtered, and the filtrates are combined and filtered, and then rotary evaporated at 45 to 55° C. until the medicinal material: water = 1:1 to 2, and freeze-dried into powder to obtain the radish seed water extract.

[0012] Furthermore, in the composition, the final therapeutic dose of animal Bifidobacterium YRK-12 is 1.0-3.0×10 10 CFU / d / kg.

[0013] Furthermore, in the composition, the final therapeutic dose of the radish seed extract is 0.11-0.44 g / d / kg body weight.

[0014] Furthermore, the dosage form of the composition includes lyophilized powder, capsule, tablet or granule.

[0015] Another aspect of the present invention provides the use of the above-mentioned animal Bifidobacterium and radish seed composition in the preparation of a product for preventing and treating functional constipation.

[0016] Furthermore, the product is any one of food, health care product, medicine or feed.

[0017] Furthermore, the application is one or more of the following:

[0018] (1) The application can reduce the time of defecation;

[0019] (2) The application can increase the number of fecal particles;

[0020] (3) The application can increase the water content of feces;

[0021] (4) The application can improve the small intestinal propulsion rate.

[0022] Another aspect of the present invention provides a probiotic composition, which comprises the above-mentioned animal Bifidobacterium and radish seed composition having the synergistic effect of preventing and treating functional constipation.

[0023] Furthermore, the probiotic composition also includes a medically acceptable carrier.

[0025] The animal Bifidobacterium was named YRK-12, and the strain was deposited in the General Microbiology Center of China Microbiological Culture Management Committee (CGMCC), address: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing; it was classified and named Animal Bifidobacterium Bifidobacterium animalis , the deposit number is CGMCC NO.28597, and the deposit date is October 9, 2023.

[0027] 1. The present application compounds animal Bifidobacterium YRK-12 with radish seed water extract to achieve synergistic synergy, and achieves the treatment of functional constipation by increasing the number of fecal particles and fecal water content, reducing defecation time, and improving small intestinal propulsion rate. The intervention effect on improving mouse constipation is better than the intervention effect of animal Bifidobacterium YRK-12 and radish seed water extract as a single substance, reaching the level of drug intervention, which has important practical significance for the development of efficient functional constipation drugs;

[0028] 2. Compared with the prior art, the present invention combines probiotics with edible and medicinal substances, which can work together on intestinal health from two aspects. The composition of the animal bifidobacterium and radish seeds composition is clear and safe, and has a significant therapeutic effect on functional constipation due to the positive control bacteria BB12. Among them, animal bifidobacteria are responsible for regulating intestinal flora and improving the intestinal environment, while the edible and medicinal substance-radish seeds extract directly relieves constipation symptoms through its laxative properties. This combination can not only improve the therapeutic effect, but also reduce dependence on a single treatment method and reduce the risk of side effects.

[0030] The present invention will be further described below in conjunction with the accompanying drawings and embodiments:

[0031] Figure 1 The results of the successful modeling of functional constipation mice described in Test Example 1 are shown; wherein A is the time of first red pellet discharge, B is the number of feces pellets in 6 hours, and C is the water content of feces;

[0032] Figure 2 The comparison results of different groups of mice with functional constipation treated in Test Example 1 are shown; wherein A is the time of first red pellet discharge, B is the number of feces pellets in 6 hours, and C is the water content of feces;

[0033] Figure 3 The results showed that the animal Bifidobacterium YRK-12 and radish seed water extract were 1~3×10 10 Results of the treatment of functional constipation mice after the combination of CFU / d / kg and 0.22 g / d / kg; A is the time of first red pellet excretion, B is the number of fecal pellets in 6 hours, C is the fecal water content, and D is the intestinal propulsion rate;

[0034] Figure 4 The results showed that the animal bifidobacterium and radish seed water extract were 1~3×10 10 Synergistic enhancement results after the combination of CFU / d / kg and 0.22 g / d / kg (combined normalized equivalent hologram); where A is animal Bifidobacterium YRK-12 and B is animal Bifidobacterium BB-12.

[0036] The implementation of the present invention is not limited to the following embodiments, and any form of modification and / or change made to the present invention will fall within the scope of protection of the present invention. In the present invention, unless otherwise specified, all equipment and raw materials can be purchased from the market or are commonly used in the industry. Unless otherwise specified, the methods used in the embodiments are common techniques in the art.

[0037] For numerical ranges, the endpoints of each range, the endpoints of each range and the individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be deemed to be specifically disclosed herein.

[0038] In the following examples, animal Bifidobacterium YRK-12 was provided by Shaanxi Yiruikang Biotechnology Co., Ltd., and the strain was deposited in the General Microbiological Center of China Microbiological Culture Collection Administration. The strain originated from the human intestine. The BB12 strain originated from Chr. Hansen of Denmark. Other chemical reagents used were purchased from regular chemical reagent suppliers and were analytically pure.

[0039] In the following examples, the culture medium formulations involved include:

[0040] The modified MRS liquid culture medium has a formula of: 10 g of peptone, 5 g of yeast extract powder, 20 g of glucose, 10 g of beef extract, 2 g of K2HPO4, 2 g of diammonium citrate, 0.58 g of MgSO4·7H2O, 0.25 g of MnSO4·4H2O, 5 g of anhydrous sodium acetate, 1 mL of Tween 80, 0.5 g of cysteine ​​hydrochloride, distilled water to 1 L, pH adjusted to 6.5, and sterilized at 121°C for 15 min.

[0042] 1) Preparation of Bifidobacterium animalis YRK-12 bacteria:

[0043] A probiotic strain was obtained by isolating human intestinal flora and was identified as Bifidobacterium animalis Bifidobacterium animalis , deposited in the General Microbiology Center of China Microbiological Culture Management Committee, with the deposit number CGMCC NO.28597 and the deposit date of October 9, 2023.

[0044] The isolated animal Bifidobacterium YRK-12 was subcultured twice in a modified MRS liquid medium, then inoculated into a modified MRS liquid medium at a 2% inoculum and cultured for 12 h at 37 ± 1 °C and 0% oxygen concentration. The supernatant was removed by centrifugation (4000 rpm, 10 min), and the bacteria were washed twice with PBS. When gavage was performed, the bacteria were resuspended in 0.9% saline and the concentration of the bacterial suspension was adjusted to 1-3 × 10 10 CFU / g.

[0045] 2) Preparation of animal Bifidobacterium BB-12 bacteria:

[0046] After animal Bifidobacterium BB-12 was subcultured twice in modified MRS liquid medium, it was inoculated in modified MRS liquid medium at a 2% inoculum and cultured for 12 h at 37 ± 1 °C and 0% oxygen concentration. The supernatant was removed by centrifugation (4000 rpm, 10 min), and the bacteria were washed twice with PBS. When gavage was performed, the bacteria were resuspended in 0.9% saline and the concentration of the bacterial suspension was adjusted to 1~3×10 10 CFU / g.

[0048] This embodiment provides a method for preparing a water extract of radish seeds, the specific steps comprising:

[0049] Radish seeds were crushed into powder by a grinder, decocted 3 times with 10 times the weight of water, each time for 1 hour, the filtrates were combined, filtered through 8 layers of gauze, and rotary evaporated at 50°C until the ratio of medicinal material to water was 1:1 (mass concentration was 1 g / mL), and then freeze-dried into powder and weighed.

[0050] The concentration multiple between the radish seed extract and radish seed was calculated, and the extract was resuspended with 0.9% saline and adjusted to a concentration of 0.04 g / mL, 0.08 g / mL, and 0.16 g / mL during oral administration.

[0052] SPF BALB / c (6-week-old, female) mice were used as experimental animals and purchased from Beijing Huafukang Biotechnology Co., Ltd. The animal experiments were conducted in two batches.

[0053] The first batch of animal experiments were conducted to screen the dose of radish seed extract, and were divided into 8 groups, namely, blank group, model group, drug group, radish seed extract high-dose group, radish seed extract medium-dose group, radish seed extract low-dose group, animal Bifidobacterium YRK-12 group, and animal Bifidobacterium BB-12 group. There were 8 mice in each group, for a total of 64 mice.

[0054] The second batch of animal experiments were conducted to evaluate the synergistic combination of animal Bifidobacterium YRK-12 and radish seed extract, which were divided into 7 groups, namely blank group, model group, drug group, animal Bifidobacterium YRK-12 group, animal Bifidobacterium BB-12 group, animal Bifidobacterium YRK-12 and radish seed extract medium dose mixed group and animal Bifidobacterium BB-12 and radish seed extract medium dose mixed group. Each group had 8 mice, a total of 56 mice.

[0055] The mice were kept under the following conditions: temperature 24°C, 12h light, 12h dark, and bedding was changed every three days. After one week of adaptive feeding, each mouse was gavaged with 15 mg / kg bw of loperamide hydrochloride every day, and weighed once a week.

[0056] The blank group was gavaged with 0.9% saline until the end of the experiment;

[0057] The model group and each intervention group were first gavaged with 0.1 mL of 15 mg / kg bw loperamide hydrochloride until the model was successful (7-14 days). After the model was successfully established, the model group and the intervention group were gavaged with 0.1 mL of 15 mg / kg bw loperamide hydrochloride every other day until the end of the experiment; at the same time, the drug group was gavaged with polyethylene glycol 4000 powder, and each intervention group was gavaged with low, medium, and high doses of radish seed extract (low dose 0.11 g / d / kg, medium dose 0.22 g / d / kg, high dose 0.44 g / d / kg), 1-3×10 10 CFU / d / kg of Bifidobacterium animalis YRK-12, 1~3×10 10 CFU / d / kg of animal Bifidobacterium BB-12, 1~3×10 10 CFU / d / kg of animal Bifidobacterium YRK-12 and radish seeds (0.22 g / d / kg) and 1~3×10 10 CFU / d / kg of animal Bifidobacterium BB-12 and radish seeds (0.22g / d / kg) were mixed in the group, with an intragastric volume of 0.1 mL and the administration lasted for 2-3 weeks.

[0059] This test example measures the feces water content, the number of stool pellets within 6 hours and the time of first red stool pellet in each group of experimental animals in Example 3. The specific method and results are as follows:

[0060] Determination of fecal water content: On the 29th day, after gavage, mice were placed individually in a cage lined with absorbent paper to collect feces. The wet weight was obtained by weighing, and the dry weight was obtained after freeze-drying. The fecal water content was calculated according to formula (1): ;

[0061] Number of stool particles within 6 hours: On the 30th day, the blank group was given 0.1 mL of 0.9% saline, and the model group and intervention group were given 0.1 mL of loperamide hydrochloride solution (15 mg / kg bw). After 1 hour, each mouse was placed in a cage lined with absorbent paper, and feces were collected every 30 minutes for a total of 6 hours.

[0062] Time of first red stool: On the 30th day, the blank group was given 0.1 mL 0.9% saline, and the model group and intervention group were given 0.1 mL loperamide hydrochloride solution (15 mg / kg bw). One hour later, each group was gavaged with 0.1 mL 0.6% (m:v) carmine. Starting from the gavage of carmine, the time of the first red stool for each mouse was recorded.

[0063] like Figure 1The results showed that 7 days after modeling, the time to first red pellet excretion, number of feces pellets and feces water content of mice in the model group were 2.5 times, 2.75 times and 1.4 times that of the blank group, respectively, and there were significant differences, indicating that the constipation model was successfully established.

[0064] The performance of the intervention group was compared with that of the model group, e.g. Figure 2 The results showed that after 14 days of intervention with different doses of radish seeds, low-dose radish seed extract (0.11g / d / kg), medium-dose radish seed extract (0.22g / d / kg), high-dose radish seed extract (0.44g / d / kg), animal Bifidobacterium YRK-12 and animal Bifidobacterium BB-12 all improved the time to first red pellet, number of fecal pellets and fecal water content. Among them, the intervention effect of medium-dose radish seed extract (0.22g / d / kg) was the best. Compared with the model group, the time to first red pellet was significantly reduced by 100 minutes ( Figure 2 Middle A); the number of fecal particles increased significantly by 10 ( Figure 2 B) and significantly increased fecal water content by 21% ( Figure 2 C). The time of first red discharge of Bifidobacterium animalis YRK-12 was 72 minutes shorter than that of Bifidobacterium animalis radish seed BB-12 ( Figure 2 Middle A); the number of fecal particles increased significantly by 3 ( Figure 2 B) and significantly increased fecal water content by 7% ( Figure 2 C). This indicates that animal Bifidobacterium YRK-12 is better than animal Bifidobacterium radish seed BB-12 in improving constipation in mice, and the medium dose of radish seed extract (0.22 g / d / kg) has the best effect on improving constipation in mice.

[0065] like Figure 3 The results showed that after 14 days of co-intervention of constipation mice with animal Bifidobacterium YRK-12 and radish seed extract (0.22g / d / kg), the constipation symptoms of mice were significantly improved. Compared with the model group, the time to first red pill was significantly reduced by 132 minutes ( Figure 3 Middle A); the number of fecal particles increased significantly by 26 ( Figure 3 B) and significantly increased fecal water content by 28% ( Figure 3 C), and the effect was not significantly different from the drug intervention group. Animal Bifidobacterium radish seed BB-12 and radish seed extract (0.22g / d / kg) co-intervention of constipated mice for 14 days also significantly improved the constipation symptoms of mice. Compared with the model group, the time to first red pill was significantly reduced by 64 minutes ( Figure 3 Middle A); the number of fecal particles increased significantly by 18 ( Figure 3 B) and significantly increased fecal water content by 15% ( Figure 3In the middle C), the intervention effect did not reach the intervention effect of the drug group. This shows that the combination of animal Bifidobacterium YRK-12 and radish seed water extract (0.22g / d / kg) is better than the combination of animal Bifidobacterium radish seed BB-12 and radish seed water extract (0.22g / d / kg) in improving constipation in mice.

[0067] This test example further measured the small intestinal propulsion rate of each group of experimental animals in Example 3. The specific method and results are as follows:

[0068] On the 30th day, mice in each group were fasted overnight but not water-free. At 8:00 a.m. on the 31st day, the blank group was given 0.1 mL of 0.9% saline solution, and the model group and the bacterial injection group were given 0.1 mL of loperamide hydrochloride solution (15 mg / kg bw). 30 minutes later, the blank group and the model group were gavaged with 0.6% (m:v) carmine, and the bacterial injection group was gavaged with carmine containing the respective gavage contents. After 30 minutes, the mice were killed, the abdominal cavity was opened, and the upper end from the pylorus to the lower end to the cecum was cut off. The total length of the small intestine was measured as the "total length of the small intestine", and the length from the pylorus to the front of the carmine was the "carmine advancement length". The small intestine advancement rate was calculated according to formula (2) ;

[0069] like Figure 3 The intestinal propulsion rate results shown in D show that after 14 days of intervention of constipated mice with animal Bifidobacterium YRK-12, the small intestinal propulsion rate of mice was significantly improved (increased by 1.7 times) compared with the model group, and was significantly better than 1.4 times of animal Bifidobacterium radish seed BB12. Compared with the model group, the combination of animal Bifidobacterium YRK-12 and radish seed extract (0.22g / d / kg) significantly improved the small intestinal propulsion rate of mice (increased by 2 times), and there was no difference with the blank group. The propulsion rate of the combination of animal Bifidobacterium YRK-12 and radish seed extract (0.22g / d / kg) was significantly higher than the propulsion rate of a single substance of animal Bifidobacterium YRK-12 and radish seed water extract. Meanwhile, the advancement rate of the combination of Bifidobacterium animalis YRK-12 and radish seed extract (0.22 g / d / kg) was significantly higher than that of the combination of Bifidobacterium animalis radish seed BB-12 and radish seed extract (0.22 g / d / kg).

[0070] Based on the Median-effect Principle (or Chou-Talalay combination index method), CompuSyn statistical software was used to draw the combination index curve under different effects, and the relationship between the combined effect of the two substances and the combination index was used to quantitatively evaluate whether there was a synergistic effect between the two substances.

[0071] Combination index CI = D1 / D X1 + D2 / D X2; D1 and D2 are the concentrations of the two substances when they produce the X effect when used alone, and D X1 , D X2 They are the concentrations of the two substances when the two substances produce the X effect when used together. When the CI is greater than 1, it indicates inhibition (i.e., the effect of the two substances when used together is antagonistic); CI=1 indicates that the effect of the two substances when used together is additive; when the CI is less than 1, it indicates promotion (i.e., the effect of the two substances when used together is synergistic), and the smaller the value, the stronger the synergistic effect.

[0072] like Figure 4 The results showed that CompuSyn software was used to analyze the red discharge time of constipated mice treated with Bifidobacterium animalis YRK-12, Bifidobacterium animalis BB12, and Radish Seed Extract alone or together. 10 CFU / d / kg) and radish seed extract (0.22g / d / kg) had a CI of 0.919 ( Figure 4 A), while Bifidobacterium animalis subsp. lactis BB-12 (1~3×10 10 CFU / d / kg) and radish seed extract (0.22g / d / kg) had a CI of 1.340 ( Figure 4 The results showed that the combination of animal Bifidobacterium YRK-12 and radish seed extract (0.22 g / d / kg) had a promoting effect on the intervention of constipation in mice, while the combination of animal Bifidobacterium lactis subsp. BB-12 and radish seed extract (0.22 g / d / kg) had an inhibitory effect on the intervention of constipation in mice.

[0073] In summary, animal Bifidobacterium YRK-12, radish seed water extract, and the combination of animal Bifidobacterium YRK-12 and radish seed water extract can experimentally treat functional constipation by increasing the number of fecal particles and fecal water content, reducing defecation time, and improving small intestinal propulsion rate. The intervention effect of animal Bifidobacterium YRK-12 combined with radish seed water extract in improving mouse constipation is better than the intervention effect of animal Bifidobacterium YRK-12 and radish seed water extract alone, and has a synergistic effect.

[0074] Finally, it should be noted that the above is only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred arrangement scheme, a person skilled in the art should understand that the technical solution of the present invention may be modified or replaced by equivalents without departing from the spirit and scope of the technical solution of the present invention.

Claims

1. An animal bifidobacterium and radish seed composition having the synergistic effect of preventing and treating functional constipation, characterized in that: The active ingredients of the composition are animal bifidobacterium and radish seed extract; The animal bifidobacterium is named YRK-12, which is deposited in the General Microbiological Center of the China Microbiological Culture Management Committee and is classified as animal bifidobacterium. Bifidobacterium animalis , the deposit number is CGMCC NO.28597, and the deposit date is October 9, 2023; In the composition, the final therapeutic dose of animal Bifidobacterium YRK-12 is a viable bacterial count of not less than 1.0×10 10 CFU / d / kg body weight.

2. The composition according to claim 1, characterized in that The radish seed extract is a water extract, and the extraction method of the radish seed water extract is: The radish seeds are crushed into powder by a grinder, and 8 to 15 times the weight of water is added for decocting for 2 to 5 times, each time for 0.5 to 1.5 hours. The decocted seeds are filtered, and the filtrates are combined and filtered, and then rotary evaporated at 45 to 55° C. until the medicinal material: water = 1:1 to 2, and freeze-dried into powder to obtain the radish seed water extract.

3. The composition according to claim 1, characterized in that In the composition, the final therapeutic dose of animal Bifidobacterium YRK-12 is 1.0-3.0×10 10 CFU / d / kg body weight.

4. The composition according to claim 1, characterized in that In the composition, the final therapeutic dose of the radish seed extract is 0.11-0.44 g / d / kg body weight.

5. The composition according to any one of claims 1 to 4, characterized in that The dosage form of the composition includes lyophilized powder, capsule, tablet or granule.

6. Use of the composition according to any one of claims 1 to 5 in preparing a product for preventing and treating functional constipation.

7. The use according to claim 6, characterized in that The product is any one of food, health care product, medicine or feed.

8. The use according to claim 6, characterized in that The application is one or more of the following: (1) The application can reduce the time of defecation; (2) The application can increase the number of fecal particles; (3) The application can increase the water content of feces; (4) The application can improve the small intestinal propulsion rate.

9. A probiotic composition, characterized in that: The invention comprises the animal Bifidobacterium and radish seed composition having the synergistic effect of preventing and treating functional constipation as described in any one of claims 1 to 5.

10. The probiotic composition according to claim 9, characterized in that Medically acceptable carriers are also included.

Images