A Chinese medicine and probiotic composition for improving constipation and preparation method thereof

Abstract

A Chinese medicine and probiotic composition for improving constipation, prepared from the following raw materials in parts by weight: 18-22g of whole Trichosanthes kirilowii, 10-14g of safflower, 10-14g of rhubarb, 10-14g of stir-fried Citrus aurantium, 7-11g of Magnolia officinalis, 7-11g of Licorice and a concentration of 1.2×10 9 CFU / mL of rhamnosus lactobacillus 50 μL. The invention has abundant raw materials, a simple preparation method, and is easy to produce. It effectively improves colon pathology on the basis of improving constipation symptoms, has good safety, and has huge social and economic benefits.

Description

Technical Field

[0001] The invention relates to the field of traditional Chinese medicines, in particular to a traditional Chinese medicine and probiotic composition for improving constipation and a preparation method thereof. Background Art

[0003] Existing treatment methods include drug therapy, dietary adjustment and probiotic therapy. However, existing treatment methods have problems such as large side effects and unstable efficacy. Western medicine treatment can increase intestinal smooth muscle contraction, thereby promoting gastrointestinal motility and defecation. However, adverse reactions such as drug dependence, electrolyte disorders, and irreversible intestinal nerve damage and liver and kidney damage caused by long-term use limit their application in the treatment of constipation. Traditional Chinese medicine improves constipation while enhancing the patient's physique through syndrome differentiation and treatment. It is suitable for many types of constipation, but it depends on the doctor's dialectic and the effect varies from person to person. Probiotics can regulate intestinal microecology, inhibit the reproduction of pathogenic bacteria, and have a wider range of applications, but they are slow to take effect. Therefore, in order to reduce side effects during treatment and improve constipation, it is of great significance to develop a safe and effective method for treating constipation. Summary of the invention

[0004] In view of the above situation, in order to overcome the defects of the prior art, the purpose of the present invention is to provide a Chinese medicine and probiotic composition for improving constipation and a preparation method thereof, which can effectively solve the problems of existing drugs for treating constipation, such as large side effects, poor effects and slow onset of action.

[0005] To achieve the above object, the technical solution provided by the present invention is a Chinese medicine and probiotic composition for improving constipation, which is prepared from the following raw materials in parts by weight: whole Trichosanthes kirilowii 18-22g, safflower 10-14g, rhubarb 10-14g, stir-fried Citrus aurantium 10-14g, Magnolia officinalis 7-11g, liquorice 7-11g and a concentration of 1.2 × 10 9 CFU / mL of Lactobacillus rhamnosus 50μL.

[0006] The preparation method of the Chinese medicine and probiotic composition for improving constipation comprises the following steps: (1) Mix whole Trichosanthes kirilowii, safflower, rhubarb, stir-fried Citrus aurantium, Magnolia officinalis and licorice, soak in water and boil twice. For the first time, add 8 times the volume of water of the total weight of the raw materials and boil for 1 hour to obtain the first decoction. For the second time, add 6 times the volume of water of the total weight of the raw materials and boil for 0.5 hour to obtain the second decoction. Combine the two decoctions and filter to obtain the Chinese medicine solution. The weight and volume are measured in grams for solids and in milliliters for liquids; (2) Take 50 μL of 1.2 × 10 9CFU / mL of Lactobacillus rhamnosus was evenly spread on an MRS agar plate and inverted cultured at 37°C for 24 hours to obtain a traditional Chinese medicine and probiotic composition.

[0007] The invention has abundant raw materials, a simple preparation method, and is easy to produce. It effectively improves colon pathological conditions on the basis of improving constipation symptoms, has good safety, and has huge social and economic benefits. BRIEF DESCRIPTION OF THE DRAWINGS

[0008] Figure 1 This is an observation diagram of the co-cultivation of the Chinese medicine composition of the present invention and the bacterial strain.

[0009] Figure 2 This is a line graph showing the weight and food intake of mice in an embodiment of the present invention (wherein A. mouse weight; B. mouse food intake (n=3)).

[0010] Figure 3 The figure is a graph showing the change of fecal moisture content and the time of the first red stool in mice in the present invention (wherein, A. Fecal moisture content of mice; B. Time of the first red stool in mice (n=3); Note: Compared with the NC group, *P <0.05; compared with the HFD group, # P <0.05).

[0011] Figure 4 This is a diagram of the changes in the mouse colon structure according to an embodiment of the present invention (H&E staining of mouse colon (n=3)).

[0012] Figure 5 1 is a diagram of the changes in the colon structure of mice according to an embodiment of the present invention (wherein A. mouse serum SP content; B. mouse serum VIP content (n=3)). DETAILED DESCRIPTION

[0013] The specific implementation of the present invention is described in detail below with reference to the accompanying drawings and examples.

[0014] The specific implementation of the present invention is given by the following examples. Example 1

[0015] A Chinese medicine and probiotic composition for improving constipation is prepared from the following raw materials in parts by weight: 18g of whole Trichosanthes kirilowii, 10g of safflower, 10g of rhubarb, 10g of stir-fried immature bitter orange, 7g of Magnolia officinalis, 7g of liquorice and a concentration of 1.2 × 10 9CFU / mL of Lactobacillus rhamnosus 50μL; whole Trichosanthes kirilowii, safflower, rhubarb, stir-fried immature bitter orange, Magnolia officinalis and liquorice were mixed, soaked in water and decocted twice. The first time, 8 times the volume of water of the total weight of the raw materials was added and decocted for 1 h to obtain the first decoction. The second time, 6 times the volume of water of the total weight of the raw materials was added and decocted for 0.5 h to obtain the second decoction. The two decoctions were combined and filtered to obtain the traditional Chinese medicine solution; 50μL of 1.2 × 10 9 CFU / mL of Lactobacillus rhamnosus was evenly spread on an MRS agar plate and inverted cultured at 37°C for 24 hours to obtain a traditional Chinese medicine and probiotic composition. Example 2

[0016] A Chinese medicine and probiotic composition for improving constipation is prepared from the following raw materials in parts by weight: 22g of whole Trichosanthes kirilowii, 14g of safflower, 14g of rhubarb, 14g of stir-fried Citrus aurantium, 11g of Magnolia officinalis, 11g of Licorice and a concentration of 1.2 × 10 9 CFU / mL of Lactobacillus rhamnosus 50 μL. The preparation method is the same as Example 1. Example 3

[0017] A Chinese medicine and probiotic composition for improving constipation is prepared from the following raw materials in parts by weight: 20g of whole Trichosanthes kirilowii, 12g of safflower, 12g of rhubarb, 12g of stir-fried Citrus aurantium, 9g of Magnolia officinalis, 9g of Licorice and a concentration of 1.2 × 10 9 CFU / mL of Lactobacillus rhamnosus 50 μL. The preparation method is the same as Example 1. Example 4

[0018] A Chinese medicine and probiotic composition for improving constipation is prepared from the following raw materials in parts by weight: 18g of whole Trichosanthes kirilowii, 14g of safflower, 10g of rhubarb, 14g of stir-fried Citrus aurantium, 7g of Magnolia officinalis, 11g of liquorice and a concentration of 1.2 × 10 9 CFU / mL of Lactobacillus rhamnosus 50 μL. The preparation method is the same as Example 1. Example 5

[0019] A Chinese medicine and probiotic composition for improving constipation is prepared from the following raw materials in parts by weight: 19g of whole Trichosanthes kirilowii, 13g of safflower, 11g of rhubarb, 12g of stir-fried Citrus aurantium, 8g of Magnolia officinalis, 10g of liquorice and a concentration of 1.2 × 10 9 CFU / mL of Lactobacillus rhamnosus 50 μL. The preparation method is the same as Example 1.

[0020] In the above-mentioned drug composition, among them: Whole Trichosanthes: cold in nature, sweet in taste, slightly bitter. It clears away heat and eliminates phlegm, has the effects and functions of clearing heat and resolving phlegm, moistening the lungs and relieving cough, widening the chest and dispersing stagnation, moistening the intestines and relieving constipation, reducing swelling and detoxifying.

[0021] Safflower: Spicy, warm. Enters the heart and liver meridians. Good at promoting blood circulation and removing blood stasis, relieving swelling and pain. Strong medicinal power. Can be used to treat all symptoms of blood stasis, especially those with cold. It has the effects of promoting blood circulation and relieving pain.

[0022] Rhubarb: bitter, cold. Enters the spleen, stomach, and large intestine meridians. Bitter and cold, it settles, clears and relieves constipation. When taken internally, it is good at clearing away gastrointestinal stagnation and heat, relieving heat and relieving constipation, guiding damp-heat evil out of the stool, promoting bile and relieving jaundice, "removing the firewood from the bottom of the pot" and removing heat and toxins from the blood, relieving heat and toxins from the blood, relieving heat and toxins from the blood, activating blood circulation and removing blood stasis, cooling blood and stopping bleeding, and reducing swelling.

[0023] Stir-fried Citrus aurantium: Citrus aurantium is bitter, pungent, sour, and slightly cold in nature. It enters the spleen and stomach meridians. It has the effects of breaking up qi and eliminating stagnation, resolving phlegm and dispersing lumps. After being stir-fried, its effect is more significant.

[0024] Magnolia officinalis: warm in nature, bitter and pungent in taste. Enters the spleen meridian, stomach meridian, lung meridian, and large intestine meridian. It dries dampness and eliminates phlegm, relieves qi and relieves fullness. It is a dampness-removing drug. It is used to treat dampness stagnation, abdominal distension, vomiting and diarrhea, food accumulation and qi stagnation, abdominal distension and constipation, phlegm and cough.

[0025] Licorice: sweet and neutral in nature, it enters the heart, lung, spleen, and stomach meridians. Licorice is sweet, nourishing, moistening, and slowing down. It is cool when raw, and warm when roasted. It mainly enters the spleen and lung meridians, and also enters the stomach and heart meridians. It can replenish qi and tonify the middle, relieve pain, moderate the properties of the medicine, and also eliminate phlegm, relieve cough, and detoxify. Roasted with honey, it has strong effects of replenishing qi and relieving urgency; raw use can purge fire and detoxify.

[0026] The above-mentioned effective combination of drugs supports each other, with whole Trichosanthes as the main drug to clear away heat, relieve chest tightness, and moisten the intestines. Rhubarb, Magnolia officinalis, and fried Citrus aurantium are used as ministers to relieve fullness, eliminate accumulation and relieve constipation, and safflower is used as an adjuvant to promote blood circulation, promote menstruation, and relieve blood stasis. Licorice is used as a guiding drug to nourish and moisten the body and harmonize the drugs. The main, minister, adjuvant, and guiding drugs are scientifically and rationally matched, and they support and cooperate with each other, exerting the efficacy of the drugs while reducing side effects on the body.

[0027] As can be seen from the above, the Chinese medicine and probiotic composition for improving constipation of the present invention has mutually supportive drugs, reasonable compatibility, and scientific preparation. It is used in combination with Lactobacillus rhamnosus to achieve a synergistic effect, is effectively used for constipation, and has achieved very good beneficial technical effects through experiments. Taking Example 3 as an example, the relevant experimental data are as follows: 1. Experimental Materials 1.1 Experimental animals Thirty 6-week-old SPF male C57BL / 6J mice were purchased from Liaoning Changsheng Biotechnology Co., Ltd., with the experimental animal production license number SCXK (Liao) 2020-0001. The mice were acclimatized at a constant temperature (22 ± 1 °C) with a light / dark cycle of 12 h and free access to food and water.

[0028] 1.2 Experimental strains Lactobacillus rhamnosus (Beina Biotech - Henan Industrial Microbial Strain Engineering Technology Research Center, strain number BNCC186192).

[0029] 1.3 Drugs and reagents MRS broth and MRS agar ready-to-use plates were purchased from Beina Bio-Henan Industrial Microbiology Engineering Technology Research Center (No. BNCC370433, BNCC352238); loperamide hydrochloride capsules were purchased from Xi'an Janssen Pharmaceutical Co., Ltd. (Batch No. NAJ070H); mosapride citrate tablets were purchased from Chengdu Kanghong Pharmaceutical Group Co., Ltd. (Batch No. 4C011); magenta (Beijing Solebold Technology Co., Ltd., Product No. C8540); normal saline (Henan Kelun Pharmaceutical Co., Ltd., Batch No. A2100602A).

[0030] 1.4 Instruments Electric constant temperature incubator (Shanghai Boxun Medical Biological Instrument Co., Ltd.), vertical intelligent precision shaker (Shanghai Boxun Medical Biological Instrument Co., Ltd.), electronic balance (Shandong Olaibo Instrument Co., Ltd.), and air drying oven (Shandong Olaibo Instrument Co., Ltd.).

[0031] 2. Experimental Methods 2.1 Preparation of Chinese medicine solution 20g of whole Trichosanthes kirilowii, 12g of safflower, 12g of rhubarb, 12g of stir-fried Citrus aurantium, 9g of Magnolia officinalis and 9g of Licorice; wherein, the above components are mixed, soaked in water and decocted twice, the first time, 8 times the volume of water of the total weight of the raw materials is added and decocted for 1 hour to obtain the first decoction, the second time, 6 times the volume of water of the total weight of the raw materials is added and decocted for 0.5h to obtain the second decoction, the two decoctions are combined and filtered to obtain the traditional Chinese medicine liquid.

[0032] 2.2 Preparation of bacterial suspension According to the instructions, the lyophilized powder of the strain was activated. After the colony grew, a single colony was picked and inoculated into a sterilized liquid culture medium. The culture was shaken at 37°C overnight, 180rpm, and 30% glycerol was used to preserve the culture at a ratio of 1:1. Take 50uL of the bacterial solution for coating and inverted culture at 37°C for 24h. Pick a single colony and dilute it into normal saline, vortex and mix it, and measure OD600. After multiple dilution, select the last two concentrations and apply 50uL to the plate respectively, and count them after inverted culture at 37°C for 24h. Take the plate with a colony count between 20-200 for calculation to determine the concentration of the bacterial solution. Take an appropriate amount of second-generation glycerol bacteria and inoculate it into a liquid culture medium, and incubate it at 37°C for 24h. After centrifugation, wash it three times with normal saline, then resuspend it with normal saline, and adjust it to the required dosage concentration with normal saline or Chinese medicine after measuring OD600.

[0033] 2.3 Co-cultivation of microorganisms and traditional Chinese medicine Take 50 μL of the solution with a concentration of 1.2 × 10 9 CFU / mL of Lactobacillus rhamnosus was evenly spread on the MRS agar plate, and 50 μL of 1.2 × 10 9 CFU / mL of Lactobacillus rhamnosus was evenly spread on MRS agar plates and incubated at 37°C for 24 h before observation.

[0034] 2.4 Animal grouping, modeling and drug administration After one week of adaptive feeding, C57BL / 6J mice were randomly divided into two groups: 5 mice in the normal control (NC) group and 25 mice in the loperamide (Lop) modeling group. Loperamide 10 mg / kg bw / d was administered by intragastric gavage to establish the model.

[0035] Three weeks later, the Lop modeling group was randomly divided into five groups, with five rats in each group, namely, the Lop group, the Mosapride (Mos) group, the Chinese medicine group, the Lactobacillus rhamnosus group, and the Chinese medicine + Lactobacillus rhamnosus group. The bacterial suspension was adjusted to 1.2 × 10 9 CFU / mL, and the NC group and the Lop group were given the same volume of normal saline for 2 consecutive weeks.

[0036] 3. Observation indicators 3.1 Observation of general condition of mice and measurement of body weight and food intake The mental state, hair gloss, activity, defecation, weight changes, and food intake of each group of mice were observed at the same time every day, and the results were recorded and counted for subsequent analysis.

[0037] 3.2 Determination of feces moisture content Collect mouse feces once a week, weigh and record the wet weight of the feces, put it in a forced air drying oven until the weight no longer changes, weigh it again and record it as the dry weight of the feces. The water content of feces can be calculated according to the following formula: Fecal moisture content (%) = [(wet weight - dry weight) / wet weight] × 100%.

[0038] 3.3 Determination of the time of first red stool Each mouse was gavaged with 0.2 mL of magenta solution and placed individually in a clean cage. This time was recorded as the start time, and the time when the mouse excreted the first red stool was the end time. The interval between the two times was the time of the first red stool.

[0039] 3.4 HE staining The fixed colon tissue was dehydrated and embedded, and 5 μm thin slices were cut using a manual rotary slicer. The paraffin sections were dewaxed with xylene solution and hydrated with gradient ethanol. After hematoxylin staining, they were soaked in hydrochloric acid and counterstained with eosin. After dehydration, they were soaked in ethanol, xylene carbolic acid solution, and xylene. After sealing, the slides were placed under a microscope to collect images.

[0040] IV. Results 4.1 Observation on co-culture of traditional Chinese medicine and Lactobacillus rhamnosus The results are as follows Figure 1 As shown, compared with the single strain culture group, more colonies grew on the plate of the Chinese medicine + Lactobacillus rhamnosus group, indicating that Chinese medicine can promote colony growth to a certain extent.

[0041] 4.2 Observation of general condition of mice During the modeling period of the Lop group, compared with the NC group, the mice in the Lop group showed poor fur gloss and reduced activity. The weight of mice in each group increased steadily, but there was no significant change in food intake, which ruled out the influence of weight and food intake on mice. After intervention in the Chinese medicine group, Lactobacillus rhamnosus group, and Chinese medicine + Lactobacillus rhamnosus group, the activity of constipated mice improved. Figure 2 .

[0042] 5.3 Changes in fecal moisture content and time to first red stool in mice During the modeling period, compared with the NC group, the fecal water content of mice in the Lop group that were gavaged showed a significant decrease (P < 0.05). After administration, each group showed a different degree of recovery trend, among which the fecal water content of mice in the Chinese medicine + Lactobacillus rhamnosus group increased significantly (P < 0.05). The time to the first red stool also showed the same trend, indicating that the combination of Chinese medicine and Lactobacillus rhamnosus can effectively improve the constipation of mice. The results are as follows Figure 3 shown.

[0043] 5.4 Changes in the structure of mouse colon like Figure 4 As shown in the figure, the colon epithelial cells and smooth muscle layer structures of mice in the NC group were intact, the boundaries between the crypt structure and goblet cells were clear, and the tissue layer was thicker. Compared with the NC group, the colon muscle layer of mice in the Lop group was significantly thinner, accompanied by a decrease in goblet cells and a decrease in colon mucus secretion. After treatment, the colon muscle layer of mice in each drug group thickened to varying degrees, and the number of goblet cells also increased. Among them, the colon muscle layer of mice in the Lactobacillus rhamnosus group was significantly thickened, and the number of goblet cells increased significantly.

[0044] 5.5 Changes of active peptides in mice The present invention focuses on the changes in the content of excitatory gastrointestinal active peptides - substance P (SP) and inhibitory gastrointestinal active peptides - vasoactive intestinal peptide (VIP), and studies the effects of Lactobacillus rhamnosus and its combination with traditional Chinese medicine on the content of peptide hormones in constipated mice. Figure 5 As shown in the figure, the SP content in the serum of the Lop group mice was significantly lower than that of the NC group, and the Lactobacillus rhamnosus and its combination with Chinese medicine group were able to effectively increase the SP content in the serum of mice. The VIP content in the serum of the Lop group mice increased significantly, and the Chinese medicine group and the Lactobacillus rhamnosus + Chinese medicine group significantly reversed the upward trend. This result shows that the combination of Lactobacillus rhamnosus and Chinese medicine can effectively increase the SP content in the serum of constipated mice and reduce the VIP content.

[0045] VI. Conclusion In summary, the present invention will screen out Lactobacillus rhamnosus with good constipation prevention and treatment effect, and use it with Chinese medicine, provide Chinese medicine and probiotic composition with synergistic prevention and treatment of functional constipation. Experimental findings: Chinese medicine preparations, Lactobacillus rhamnosus intervention alone can improve constipation symptoms to varying degrees, and when Lactobacillus rhamnosus is used in conjunction with Chinese medicine, the feces moisture content of mice significantly increases, and the first red stool is discharged as early as possible, indicating that Chinese medicine and Lactobacillus rhamnosus can effectively improve the constipation situation of mice, and effectively improve the colon pathology on the basis of symptom relief. Provide reference indicators for the industrial production of probiotics and Chinese medicine compositions, and also provide theoretical basis and data support for the promotion and application of probiotics and Chinese medicine compositions in the field of preventing and treating functional constipation, with significant social and economic benefits.

[0046] It should be pointed out that the above is only a preferred embodiment of the present invention and does not limit the present invention in any form. Any technician familiar with the profession can make changes or modify the technical contents disclosed above into equivalent embodiments with equivalent changes without departing from the scope of the technical solution of the present invention, which all fall within the protection scope of the present invention.

Claims

1. A Chinese medicine and probiotic composition for improving constipation, characterized in that: The invention is prepared from the following raw materials in parts by weight: whole Trichosanthes root 18-22g, safflower 10-14g, rhubarb 10-14g, stir-fried Citrus aurantium 10-14g, Magnolia officinalis 7-11g, liquorice 7-11g and a concentration of 1.2 × 10 9 CFU / mL of Lactobacillus rhamnosus 50μL.

2. The Chinese medicine and probiotic composition for improving constipation according to claim, characterized in that: The following raw materials were included by weight: whole Trichosanthes root 18g, safflower 10g, rhubarb 10g, stir-fried Citrus aurantium 10g, Magnolia officinalis 7g, liquorice 7g and a concentration of 1.2 × 10 9 CFU / mL of Lactobacillus rhamnosus 50μL.

3. The Chinese medicine and probiotic composition for improving constipation according to claim, characterized in that: The following raw materials are included by weight: whole Trichosanthes root 22g, safflower 14g, rhubarb 14g, stir-fried Citrus aurantium 14g, Magnolia officinalis 11g, liquorice 11g and a concentration of 1.2 × 10 9 CFU / mL of Lactobacillus rhamnosus 50μL.

4. The Chinese medicine and probiotic composition for improving constipation according to claim, characterized in that: The following raw materials are included by weight: whole Trichosanthes root 20g, safflower 12g, rhubarb 12g, stir-fried Citrus aurantium 12g, Magnolia officinalis 9g, Licorice root 9g and a concentration of 1.2 × 10 9 CFU / mL of Lactobacillus rhamnosus 50μL.

5. The Chinese medicine and probiotic composition for improving constipation according to claim, characterized in that: The raw materials include the following weight: whole Trichosanthes root 18g, safflower 14g, rhubarb 10g, stir-fried Citrus aurantium 14g, Magnolia officinalis 7g, licorice 11g and a concentration of 1.2 × 10 9 CFU / mL of Lactobacillus rhamnosus 50μL.

6. The Chinese medicine and probiotic composition for improving constipation according to claim, characterized in that: The raw materials include the following weight: whole Trichosanthes root 19g, safflower 13g, rhubarb 11g, stir-fried Citrus aurantium 12g, Magnolia officinalis 8g, licorice 10g and a concentration of 1.2 × 10 9 CFU / mL of Lactobacillus rhamnosus 50μL.

7. The method for preparing the composition of traditional Chinese medicine and probiotics for improving constipation according to any one of claims 1 to 6, characterized in that: The following steps are involved: (1) Mix whole Trichosanthes kirilowii, safflower, rhubarb, stir-fried Citrus aurantium, Magnolia officinalis and licorice, soak in water and boil twice. For the first time, add 8 times the volume of water of the total weight of the raw materials and boil for 1 hour to obtain the first decoction. For the second time, add 6 times the volume of water of the total weight of the raw materials and boil for 0.5 hour to obtain the second decoction. Combine the two decoctions and filter to obtain the Chinese medicine solution. The weight and volume are measured in grams for solids and in milliliters for liquids; (2) Take 50 μL of 1.2 × 10 9 CFU / mL of Lactobacillus rhamnosus was evenly spread on an MRS agar plate and inverted cultured at 37°C for 24 hours to obtain a traditional Chinese medicine and probiotic composition.

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