Traditional Chinese medicine composition for preventing constipation due to spleen deficiency and preparation method and application thereof, and traditional Chinese medicine preparation

Abstract

The invention relates to a traditional Chinese medicine composition for preventing spleen deficiency constipation, a preparation method and application thereof and a traditional Chinese medicine preparation, and belongs to the technical field of traditional Chinese medicine. The traditional Chinese medicine composition provided by the invention comprises rhizoma polygonati and poria cocos. From the perspective of'prevention before illness', through the concept of'prevention before illness' of traditional Chinese medicine, spleen deficiency constipation is prevented by utilizing compatibility of rhizoma polygonati and poria cocos, the possibility of illness is reduced, and constipation can be prevented from the source.

Description

Technical Field

[0001] The invention relates to a traditional Chinese medicine composition for preventing spleen deficiency constipation, a preparation method and application thereof, and a traditional Chinese medicine preparation, belonging to the technical field of traditional Chinese medicines. Background Art

[0002] Constipation is a common digestive disorder. Constipated patients typically experience symptoms such as decreased bowel movement frequency, lumpy, dry stool, and difficulty or straining during defecation. The prevalence of constipation in the Chinese adult population ranges from approximately 3% to 11%, with variations depending on region, occupation, and culture. The prevalence of constipation increases with age, reaching over 22% among the elderly over 60 years old. This imposes a significant economic burden and diverts public healthcare resources. Constipation has become a prominent topic in public life, and products to prevent and treat constipation hold broad market potential.

[0003] Spleen deficiency is one of the underlying conditions for constipation, affecting food digestion and absorption, as well as the formation and excretion of stool. Traditional Chinese Medicine considers the spleen and stomach the foundation of acquired constitution, the source of Qi and blood metabolism, and the hub for the rise and fall of Qi in the human body. They are closely linked to the body's digestion, absorption, and immune defenses. Spleen and stomach Qi deficiency impairs digestion and absorption, accompanied by Qi blockage. This in turn impairs the large intestine's ability to propel food, weakening and slowing intestinal transit, ultimately leading to spleen deficiency constipation.

[0004] Modern treatments for spleen deficiency constipation are numerous, including single-use and modified Chinese herbal formulas, combined treatments, and combined treatments with other physical therapies. However, research on its prevention is currently scarce. However, there are numerous studies on the prevention of other diseases in the prior art, with promising results. Therefore, research on the prevention of spleen deficiency constipation with Chinese herbal medicine is of great significance. Summary of the Invention

[0005] In view of the problems existing in the above-mentioned prior art, one of the purposes of the present invention is to provide a traditional Chinese medicine composition for preventing spleen deficiency constipation, which can be used to prevent spleen deficiency constipation.

[0006] A second object of the present invention is to provide a method for preparing a Chinese medicine for preventing spleen deficiency constipation.

[0007] The third object of the present invention is to provide an application of a Chinese medicine composition in the preparation of a Chinese medicine preparation for preventing spleen deficiency constipation.

[0008] A fourth object of the present invention is to provide a Chinese medicine preparation for preventing spleen deficiency constipation.

[0009] In order to achieve the above object, the first aspect of the present invention is to provide a traditional Chinese medicine composition for preventing spleen deficiency constipation, wherein the traditional Chinese medicine composition comprises Polygonatum sibiricum and Poria cocos.

[0010] The Yellow Emperor's Classic of Internal Medicine states, "Drink enters the stomach, where it overflows with essence and qi, which then ascends to the spleen... The five meridians run parallel." The spleen governs transportation and transformation, serving as the source of qi and blood production. Normal spleen and stomach function ensures the internal organs are well-filled with qi and blood. If the spleen fails to function properly, qi and blood production are depleted, qi deficiency weakens the large intestine, and blood deficiency depletes the intestines, leading to constipation. The spleen also transports and transforms water and moisture. When water metabolism is normal, it is distributed throughout the body, nourishing the five internal organs and six bowels. Conversely, when the spleen fails to function properly, water is not properly distributed, the intestines lose moisture, and constipation occurs. Furthermore, the spleen and stomach, located in the middle jiao (center burner), are responsible for ascending the lucid and descending the turbid, forming the hubs of the body's qi movement. A healthy middle jiao (center burner) function, with normal ascending and descending functions, promotes orderly qi movement, harmony within the five internal organs, unobstructed bowel movements, and normal excretion of waste products. If the spleen and stomach are weak, clear qi fails to ascend, turbid yin fails to descend, and the large intestine loses its function, impairing waste excretion. Therefore, the core pathogenesis of spleen deficiency constipation lies in the spleen's dysfunctional function, malfunctioning ascending and descending functions, and malfunctioning large intestine conduction. The effects of Polygonatum sibiricum and Poria cocos align with the pathogenesis of spleen deficiency constipation, and their combination adheres to the principle of "both tonifying and purging, ascending and descending in tandem." Polygonatum sibiricum primarily tonifies the spleen and nourishes spleen yin, aligning with the principle of "tonifying all deficiencies and replenishing the essence" as stated in the Compendium of Materia Medica. It can improve intestinal fluid deficiency caused by spleen deficiency. Poria cocos, on the other hand, is known for its ability to "drain water and dampness, strengthen the spleen and harmonize the stomach." Together, the two herbs achieve the effect of "tonifying without stagnation, benefiting without harming the body." Furthermore, Polygonatum sibiricum promotes the ascending of spleen qi to distribute body fluids, while Poria cocos clears the waterways to reduce turbidity. Together, they restore the ascending and descending Qi of the middle jiao and promote intestinal conduction.

[0011] The present invention starts from the perspective of "prevention before illness occurs" and adopts the concept of "prevention before illness occurs" in traditional Chinese medicine. It uses the combination of Polygonatum sibiricum and Poria cocos to prevent spleen deficiency constipation, reduce the possibility of illness, and prevent the occurrence of constipation from the source.

[0012] As a preferred solution, the mass ratio of the polygonatum to poria is 1:0.2~3.

[0013] As a more preferred embodiment, the mass ratio of Polygonatum sibiricum to Poria cocos is 1:1-3. The inventors have found that this preferred embodiment is more effective in improving stool characteristics, intestinal flora imbalance, and enzyme activity disorders in spleen deficiency constipation, and has a better preventive effect on spleen deficiency constipation. A further preferred ratio is 1:1. The inventors have found that a 1:1 ratio of Polygonatum sibiricum to Poria cocos is the most effective.

[0014] As a preferred solution, the traditional Chinese medicine composition is composed of Polygonatum sibiricum and Poria cocos in a mass ratio of 1:0.2-3.

[0015] The second aspect of the present invention is to provide a method for preparing a traditional Chinese medicine for preventing spleen deficiency constipation, which is obtained by mixing and crushing the composition described in the first aspect.

[0016] As a preferred solution, the pulverization is performed so that the particle size of the mixture is ≤0.25 mm.

[0017] As a preferred solution, the total dosage of Polygonatum sibiricum and Poria cocos is 15-30 g / day for an adult, more preferably 20 g / day.

[0018] As a preferred solution, the Chinese medicine composition is taken orally with water.

[0019] The third aspect of the present invention is to provide the use of the Chinese medicine composition described in the first aspect in the preparation of a Chinese medicine preparation for preventing spleen deficiency constipation.

[0020] The fourth aspect of the present invention is to provide a Chinese medicine preparation for preventing spleen deficiency constipation, comprising the Chinese medicine composition described in the first aspect and pharmaceutically acceptable excipients.

[0021] As a preferred embodiment, the Chinese medicine preparation is in the form of granules, powders, decoctions, pills, tablets, capsules, mixtures or syrups.

[0022] Compared with the prior art, the present invention has at least the following advantages:

[0023] The present invention takes the perspective of "prevention before illness occurs" and provides an experimental basis for using Chinese medicine combinations to prevent spleen deficiency constipation, providing a new idea and theoretical basis for the prevention and early clinical application of such functional diseases.

[0024] The traditional Chinese medicine composition provided by the invention has the effects of improving stool properties, intestinal flora imbalance and enzyme activity disorder of spleen deficiency type constipation. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 The effect of the combination of Polygonatum sibiricum and Poria cocos on the average food intake and average water intake of mice. Figure 1 A is the average food intake, Figure 1 B in the figure is the average water intake;

[0026] Note: CN: normal control group; MD: model group; HFA: group with a ratio of Polygonatum sibiricum to Poria cocos A; HFB: group with a ratio of Polygonatum sibiricum to Poria cocos B; HFC: group with a ratio of Polygonatum sibiricum to Poria cocos C. The following figure and table are the same.

[0027] Figure 2 The effect of the combination of Polygonatum sibiricum and Poria cocos on the body weight of mice;

[0028] Note: Compared with CN group: represents p<0.05, = (#) indicates p < 0.01; compared with the MD group: # indicates p < 0.05, ## indicates p < 0.01; compared with the HFA group: △ indicates p < 0.05, △△ indicates p < 0.01; compared with the HFB group: ▲ indicates p < 0.05, ▲▲ indicates p < 0.01. The following figures and tables are the same.

[0029] Figure 3 The effect of the combination of Polygonatum sibiricum and Poria cocos on the organ indexes of mice;

[0030] Figure 4 The effect of the combination of Polygonatum sibiricum and Poria cocos on the fecal characteristics of mice;

[0031] Figure 5 The effect of the combination of Polygonatum sibiricum and Poria cocos on the water content of mouse feces; Figure 5 A in the figure is the change of water content in mouse feces. Figure 5 B in the equation is 21 days. Figure 5 C in the equation is 23 days. Figure 5 D in the equation is 25 days. Figure 5 The E in the equation is 27 days. Figure 5 F in is 29 days;

[0032] Figure 6 The effect of the combination of Polygonatum sibiricum and Poria cocos on the D-xylose content in mouse serum;

[0033] Figure 7 The effect of the combination of Polygonatum sibiricum and Poria cocos on the microbial activity in the intestine of mice;

[0034] Figure 8 The effect of the combination of Polygonatum sibiricum and Poria cocos on the behavior of mice, Figure 8 A in is the activity time, Figure 8 B in the equation is the stationary time. DETAILED DESCRIPTION

[0035] The endpoints of the ranges and any values ​​disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoints of each range, the endpoints of each range and 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 considered to be specifically disclosed herein.

[0036] The present invention is further described below with reference to specific embodiments, but the scope of protection of the present invention is not limited to the following specific embodiments. Obviously, the embodiments described below are only a part of the embodiments, and all other embodiments obtained by professionals in this field without making creative efforts are still within the scope of protection of the present invention.

[0037] Unless otherwise specified, various raw materials, reagents, instruments and equipment used in the present invention can be purchased from the market or prepared by existing methods.

[0038] The present invention establishes a mouse animal model of spleen deficiency constipation, observes the general signs of the mice, evaluates their intestinal microbial activity and enzyme activity, determines the number of culturable microbial colonies in the intestine, and detects the serum D-xylose level of the mice, to explore the effect of the combination of Polygonatum sibiricum and Poria cocos in preventing spleen deficiency constipation.

[0039] There are many methods for modeling spleen deficiency constipation in mice, such as swimming plus hunger and satiety disorder modeling, loperamide hydrochloride suspension gavage modeling, senna leaf plus controlled diet and water intake, and other composite factor modeling methods. Based on the analysis of multiple indicators such as general behavioral characteristics of mice, time to first black stool, number and quality of stools in 6 hours, serum D-xylose content, intestinal propulsion rate, and colon mucosal tissue pathological characteristics, the senna leaf plus controlled diet and water intake modeling method is more in line with the characteristics of the animal model of spleen deficiency constipation syndrome in traditional Chinese medicine. Therefore, this method is selected for modeling in the present invention.

[0040] Example 1: Animal efficacy experiment

[0041] In clinical practice of traditional Chinese medicine, Poria cocos is generally not used to treat constipation. Inspired by the fact that Poria cocos strengthens the spleen and promotes diuresis, enters the spleen, kidney, and heart meridians, and replenishes spleen qi; and Polygonatum sibiricum nourishes yin, enters the spleen, kidney, and lung essence, and nourishes kidney essence, the present invention conducted a mouse experiment on the effects of Poria cocos and Polygonatum sibiricum on spleen deficiency constipation. While establishing a mouse model of spleen deficiency constipation, Poria cocos, Polygonatum sibiricum, and Poria cocos + Polygonatum sibiricum were used for intervention, respectively.

[0042] 1.1 Materials

[0043] 1.1.1 Experimental animals and breeding environment

[0044] Experimental Animals: Fifty male SPF Kunming mice (weight 20 ± 2 g) were purchased from Hunan Slake Jingda Laboratory Animal Co., Ltd. (License No. SCXK (Xiang) 2019-0004). They were housed in a barrier environment at the Animal Experimental Center of Hunan University of Chinese Medicine (License No. SYXK (Xiang) 2019-0009) at a room temperature of 23–25°C and a relative humidity of 50%–70%. Animal Ethics Welfare Approval Number: SLBH-202505150004, issued by the Animal Ethics Committee of Hunan University of Chinese Medicine.

[0045] 1.1.2 Experimental drugs and preparation

[0046] Polygonatum sibiricum and Poria cocos solution: Grind Xinhua Polygonatum sibiricum and Jingzhou Poria cocos separately, pass through a No. 4 sieve (65 mesh), and store at 4°C. Weigh a certain amount of Polygonatum sibiricum and Poria cocos powder, add appropriate amounts of water, and boil for 30 minutes to prepare Polygonatum sibiricum, Polygonatum sibiricum + Poria cocos, and Poria cocos aqueous solutions. Store without filtration at 4°C. Dilute the solution before use, administering 0.3 ml to each mouse twice daily by oral gavage.

[0047] The above dosages of Polygonatum and Poria are approximately equivalent to 20 grams of Polygonatum per day for adults, 10 grams of Polygonatum per day + 10 grams of Poria per day, and 20 grams of Poria per day.

[0048] Senna Leaf Decoction: Weigh 180 g of senna leaves (Anhui Jinren Chinese Medicine Pieces Co., Ltd., batch number 2407032) and soak in water for 30 minutes. Then, add five times the amount of water and boil for 30 minutes. Filter through gauze. Add an appropriate amount of water to the remaining residue and continue boiling for 15 minutes. Filter through gauze and combine the two filtrates. Concentrate the filtrate to 1 g / mL using a rotary evaporator at 60°C and 15 rpm / min. Store the senna leaf decoction at 4°C.

[0049] 1.2 Methods

[0050] 1.2.1 Modeling method

[0051] After three days of adaptive feeding, 50 KM mice were randomly divided into a normal group, a model group, a Polygonatum sibiricum group, a Polygonatum sibiricum + Poria cocos group, and a Poria cocos group, with 10 mice in each group. A spleen deficiency-induced constipation model was established in mice using a combination of senna leaf gavage, altered satiety, water restriction, and a low-fiber diet. The model lasted for 15 days. For the spleen deficiency model, a decoction of senna leaves (20 g / kg / day, once daily) was administered orally for seven days, followed by a normal diet and water intake to induce spleen deficiency. For the constipation model, senna leaf administration was discontinued on day eight, and altered satiety and water restriction (a low-fiber diet consisting of 4-8 g raw rice per mouse with free access to water for 0.5 hours per day) were used to maintain spleen deficiency and constipation for eight days.

[0052] 1.2.2 Dosage regimen

[0053] For prophylactic administration, the three treatment groups were given 1.95 g / (kg·d) of Polygonatum sibiricum per mouse, 1.95 g / (kg·d) of Polygonatum sibiricum + Poria cocos per mouse, and 1.95 g / (kg·d) of Poria cocos per mouse, respectively. The drug solution was diluted before use and administered orally twice daily. The normal and model groups were given an equal volume of sterile water for 14 days. Subsequently, the three treatment groups and the model group were established. During the establishment of the model, the treatment groups were continuously given the corresponding Polygonatum sibiricum and Poria cocos solution for prophylactic administration, while the normal and model groups were given an equal volume of sterile water for 15 days. The oral dose of the Polygonatum sibiricum and Poria cocos solution was calculated based on the body surface area of ​​humans and mice.

[0054]

[0055] 1.2.3 Determination of fecal moisture content

[0056] Fresh feces of mice were collected on the 1st, 4th, 7th, 8th, 12th and 15th day after the modeling experiment. The wet weight of the feces was recorded, dried at 110°C to a constant weight and weighed to calculate the moisture content of the feces.

[0057] Fecal moisture content (%) = (fecal wet weight - fecal dry weight) / fecal wet weight × 100%.

[0058] 1.3 Results and Analysis

[0059] 1.3.1 Effects of Polygonatum sibiricum and Poria cocos on general signs of mice

[0060] During the spleen deficiency modeling phase, mice in the model group huddled together, while those in the drug-treated groups showed less clumping. Mice gavaged with senna leaf experienced severe diarrhea and lethargy, with yellow feces clinging to the perianal area. Their coats were ragged, erect, and dull, and their bedding was damp and smelly. Their ears and paws were pale and bloodless, but the drug-treated groups fared better than the model group. This suggests that Polygonatum sibiricum and Poria cocos are effective in alleviating spleen deficiency. During the constipation modeling phase, mice in the model group were more irritable, while the three drug-treated groups were closer to the normal group. Mice in the model, Poria cocos, and Polygonatum groups had fewer bowel movements, while the Polygonatum sibiricum + Poria cocos group had more bowel movements, better spirits, smoother and more lustrous coats, and significantly increased food and water intake, approaching that of the normal group.

[0061] 1.3.2 Effects of Polygonatum sibiricum and Poria cocos on mouse feces

[0062]

[0063] As can be seen from the above table, the results showed that the use of Poria cocos and Polygonatum sibiricum alone had no effect on spleen deficiency constipation, while the combined intervention of Polygonatum sibiricum and Poria cocos could relieve spleen deficiency constipation.

[0064] Based on the above experiments, the inventors considered the possibility that Poria cocos can clear water and Polygonatum sibiricum can nourish yin and provide water sources from the perspective of the body's water source, and proposed the possibility of using Poria cocos and Polygonatum sibiricum to treat spleen deficiency constipation by harmonizing the prenatal and postnatal constitutions (the spleen is the prenatal foundation, and the kidney is the acquired foundation).

[0065] Example 2: Animal efficacy experiment

[0066] 2.1 Materials

[0067] 2.1.1 Experimental animals and breeding environment

[0068] Experimental Animals: Fifty male SPF Kunming mice (weight 20 ± 2 g) were purchased from Hunan Slake Jingda Laboratory Animal Co., Ltd. (License No. SCXK (Xiang) 2019-0004). They were housed in a barrier environment at the Animal Experimental Center of Hunan University of Chinese Medicine (License No. SYXK (Xiang) 2019-0009) at a room temperature of 23–25°C and a relative humidity of 50%–70%. Animal Ethics Welfare Approval Number: SLBH-202505150004, issued by the Animal Ethics Committee of Hunan University of Chinese Medicine.

[0069] 2.1.2 Experimental drugs and preparation

[0070] Polygonatum sibiricum and Poria cocos solution: Xinhua Polygonatum sibiricum and Jingzhou Poria cocos were ground separately, passed through a No. 4 sieve (65 mesh), and stored at 4°C. A certain amount of Polygonatum sibiricum and Poria cocos powder was weighed and prepared into Polygonatum sibiricum and Poria cocos powders at ratios A (Poria cocos sibiricum: Poria cocos = 3:1), B (Poria cocos sibiricum: Poria cocos = 1:1), and C (Poria cocos sibiricum: Poria cocos = 1:3). Appropriate amounts of water were added to the Polygonatum sibiricum and Poria cocos powders at each ratio, and the solution was prepared by boiling for 30 minutes. The solution was stored at 4°C and diluted to a total of 1.95 g Polygonatum sibiricum and Poria cocos per kg / day per mouse, with a 0.3 ml oral gavage volume for twice daily administration.

[0071] The dosage ratio A of the above-mentioned Polygonatum and Poria is (3:1, approximately equivalent to 15g Polygonatum and 5g Poria per day for adults), the ratio B is (1:1, approximately equivalent to 10g Polygonatum and 10g Poria per day for adults), and the ratio C is (1:3, approximately equivalent to 5g Polygonatum and 15g Poria per day for adults).

[0072] Senna Leaf Decoction: Weigh 180 g of senna leaves (Anhui Jinren Chinese Medicine Pieces Co., Ltd., batch number 2407032) and soak in water for 30 minutes. Then, add five times the amount of water and boil for 30 minutes. Filter through gauze. Add an appropriate amount of water to the remaining residue and continue boiling for 15 minutes. Filter through gauze and combine the two filtrates. Concentrate the filtrate to 1 g / mL using a rotary evaporator at 60°C and 15 rpm / min. Store the senna leaf decoction at 4°C.

[0073] 2.1.3 Microbial culture media and kits

[0074] Bacterial culture media: Beef Extract Peptone Agar (Haibo Biotechnology Co., Ltd., Qingdao High-Tech Industrial Park, HB0109-11); Escherichia coli culture: Eosin-Methylene Blue Agar (EMB) (Shanghai Shengsi Biochemical Technology Co., Ltd., MSS065); Bifidobacterium culture: Bifidobacterium Medium (Haibo Biotechnology Co., Ltd., Qingdao High-Tech Industrial Park, HB0396-1); Lactobacillus culture: MRS Agar Medium (Haibo Biotechnology Co., Ltd., Qingdao High-Tech Industrial Park, HB0384-51). D-xylose Elisa kit (Jiangsu Jingmei Biotechnology Co., Ltd., JM-12962M2) was used.

[0075] 2.1.4 Experimental instruments

[0076] LC-RE-301 rotary evaporator (Shanghai Lichen Bangsi Instrument Technology Co., Ltd.), ultrapure water meter (ELGA), 5810R desktop large-capacity refrigerated centrifuge (Eppendorf, Germany), SHZ-82 gas bath constant temperature oscillator (Jintan Ronghua Instrument Manufacturing Co., Ltd.), SPAPK multifunctional microplate reader (Tecan, Sweden), HH·BII·500-S electric constant temperature incubator (Shanghai Botai Laboratory Equipment Co., Ltd.), 722 visible spectrophotometer (Shanghai Sunny Hengping Scientific Instrument Co., Ltd.), JA2003 electronic balance (Shanghai Sunny Hengping Scientific Instrument Co., Ltd.), SQ510C high pressure sterilizer (YAMATO).

[0077] 2.2 Methods

[0078] 2.2.1 Modeling method

[0079] After three days of adaptive feeding, 50 KM mice were randomly divided into a normal group, a model group, a Polygonatum-Poria cocos A ratio group, a Polygonatum-Poria cocos B ratio group, and a Polygonatum-Poria cocos C ratio group, with 10 mice in each group. A spleen deficiency-induced constipation mouse model was established using a combination of factors, including oral gavage with senna leaves, altered satiety, water restriction, and a low-fiber diet. The model lasted for 15 days. For the spleen deficiency model, a decoction of senna leaves (20 g / kg / day, once daily) was administered orally for seven days, followed by a normal diet and water intake to induce spleen deficiency. For the constipation model, on the eighth day, senna leaves were discontinued, and altered satiety and water restriction (a low-fiber diet consisting of 4-8 g raw rice per mouse with free access to water for 0.5 hours per day) were used to maintain spleen deficiency and constipation for eight days.

[0080] 2.2.2 Dosage regimen

[0081] For prophylactic administration, the three treatment groups were given a corresponding ratio of Polygonatum sibiricum and Poria cocos solution by gavage at 1.95 g / (kg·d), 0.3 ml / time, twice daily. The normal and model groups were given an equal volume of sterile water by gavage for 14 days. Subsequently, the three treatment groups and the model group were subjected to model establishment. During the model establishment period, the treatment groups were given continuous prophylactic administration of the Polygonatum sibiricum and Poria cocos solution by gavage, while the normal and model groups were given an equal volume of sterile water by gavage for 15 days. The gavage dose of the Polygonatum sibiricum and Poria cocos solution was calculated based on the body surface area of ​​humans and mice.

[0082]

[0083] 2.2.3 Evaluation criteria for spleen deficiency constipation

[0084] The modeling method was evaluated by the following indicators: the mice had a shriveled, thin appearance, erect hair, arched back, reduced activity, weight loss, dry and hard stools, reduced stool quantity, and small particles; the dissection showed that the stool was accumulated in the colon, in a spherical or beaded shape, and there was no obvious fecal residue in the jejunum and ileum; the intestinal propulsion rate was reduced and the serum D-xylose level was reduced.

[0085] 2.2.4 Measurement of food and water intake, body weight, and feces moisture content

[0086] The mice were weighed on days 1, 5, 9, 13, 17, 21, 25, and 29 of the experiment. Fresh feces were collected on days 21, 23, 25, 27, and 29. The wet weight of the feces was recorded, dried at 110°C to a constant weight, and weighed to calculate the fecal moisture content. Food and water intake were recorded on days 4, 8, 12, 16, and 20 of the experiment. Because diet and water restriction were required on day 8 of modeling, food and water intake was not recorded thereafter.

[0087] Fecal moisture content (%) = (fecal wet weight - fecal dry weight) / fecal wet weight × 100%.

[0088] 2.2.5 Organ index measurement

[0089] The complete liver, spleen, kidney and thymus of mice were collected, and after removing excess fat and blood from the surface of the organs, they were weighed, recorded and the organ index was calculated to evaluate the effects of Polygonatum sibiricum and Poria cocos on the liver, spleen, kidney and thymus of mice with spleen deficiency constipation.

[0090] Organ index (%) = organ weight / body weight × 100%.

[0091] 2.2.6 Serum D-xylose determination

[0092] After modeling, mice were fasted for 24 hours but not water. A 3% D-xylose solution (10 ml / kg / day) was administered orally. One hour later, blood was collected from the eyeballs. After the blood rested for 3 hours, it was centrifuged at 3000 rpm for 10 minutes at 4°C. The supernatant was aspirated to obtain serum samples. Serum D-xylose levels were measured using an enzyme-linked immunosorbent assay (ELISA, Jiangsu Jingmei Biotechnology Co., Ltd.) in strict accordance with the D-xylose kit instructions.

[0093] 2.2.7 Intestinal microbial culture

[0094] Spleen deficiency constipation is a specific syndrome type of chronic constipation in Traditional Chinese Medicine theory. Its course, symptoms, and pathological mechanisms all conform to the modern medical definition of chronic constipation. Clinical and basic research combining traditional Chinese and Western medicine has further confirmed the correlation between the two. Studies have shown that the main mechanism of chronic constipation is colonic sensory and motor dysfunction, and colonic transit contraction defects may be one of the core mechanisms of slow-transit constipation, indicating that the colon and constipation are highly correlated. Therefore, the present invention mainly uses the colon for a series of studies. The colon contents are removed by aseptic operation, and sterile water is added in proportion (sample: sterile water = 3:50). The sample is shaken in an air bath constant temperature oscillator for 30 minutes to completely release the microorganisms in the contents, and then diluted into samples of different concentrations. Samples of different concentrations were inoculated onto beef extract peptone agar, eosin-methylene blue agar, bifidobacterium agar, and MRS agar using the spreading method. Bacteria and Escherichia coli were cultured in a 37°C constant temperature incubator for 24 h and then counted. Bifidobacteria and Lactobacilli were cultured anaerobically in a 37°C constant temperature incubator for 48 h and then counted.

[0095] 2.2.8 Determination of microbial activity

[0096] Mix the contents obtained according to 2.2.7 with sterile water and centrifuge at 3000 rpm / min and 4°C for 15 minutes. Collect the supernatant as the crude enzyme solution. To prepare the fluorescein diacetate (FDA) stock solution, add 2 mg of FDA to 1 mL of acetone to prepare a 2 mg / mL FDA stock solution. Store at -20°C in the dark. To prepare the FDA reaction solution, pipette the FDA stock solution into PBS to a concentration of 10 μg / mL. Under aseptic conditions, add 2 mL of FDA reaction solution and 10 μL of crude enzyme solution to a sample tube. Shake at 24°C for 90 minutes, then terminate the reaction by adding 2 mL of acetone. To a blank control tube, add 2 mL of FDA reaction solution, 2 mL of acetone, and 10 μL of crude enzyme solution, and shake at 24°C for 90 minutes. After the reaction, measure the microbial activity of the sample tube using a UV-Vis spectrophotometer (wavelength 490 nm).

[0097] 2.2.9 Intestinal enzyme activity assay

[0098] The supernatant collected in 2.2.8 was used as the crude enzyme solution. The activities of the various enzymes in the crude enzyme solution were measured using a UV-Vis spectrophotometer. One unit of enzyme activity (U) is defined as the amount of reducing sugar produced by 1 g of intestinal contents at a specific temperature and for a specific time. Amylase activity was measured using the DNS method at 520 nm, cellulase activity was measured using the DNS method at 540 nm, xylanase activity was measured using the DNS method at 550 nm, and protease activity was measured using the Folin-phenol method at 660 nm.

[0099] 2.2.10 Behavioral assessment

[0100] On the 15th day after modeling, after gavage, the mice were placed in an open-field testing room. After acclimation for 30 minutes, they were placed in an open-field box (50 cm × 50 cm × 50 cm) and tested using the Smart 3.0 system. The mice's immobility, activity, and movement trajectory were observed within 1 minute. After each experiment, the box was cleaned of feces and urine, and sprayed with 75% ethanol to remove mouse odor. The experiment was continued after the ethanol evaporated.

[0101] 2.2.11 Serum 5-HT, CCK, MTL, VIP, MDA, and SOD determination

[0102] After modeling, mice were fasted for 24 hours, but not water, and blood was collected from their eyeballs. After the blood rested for 3 hours, it was centrifuged at 3000 rpm / min for 10 minutes at 4°C. The supernatant was aspirated to obtain serum samples. The serum levels of 5-HT, CCK, MTL, VIP, MDA, and SOD in each mouse were measured using enzyme-linked immunosorbent assays (ELISA kits for 5-HT, CCK, MTL, VIP, MDA, and SOD, Jiangsu Jingmei Biotechnology Co., Ltd.) in strict accordance with the instructions of the 5-HT, CCK, MTL, VIP, MDA, and SOD kits.

[0103] 2.2.12 Statistical analysis and graphing

[0104] Statistical analysis was performed using IBM SPSS Statistics 25.0 software, and data are presented as mean ± standard deviation. If data across multiple groups conformed to a normal distribution and homogeneity of variance, one-way analysis of variance was performed; otherwise, the Kruskal-Wallis rank-sum test (Kruskal-Wallis H test) was used. P < 0.05 indicated a significant difference, and P < 0.01 indicated a strongly significant difference. Graphs were generated using Origin 2024 software.

[0105] 2.3 Results and Analysis

[0106] 2.3.1 Effects of Polygonatum sibiricum and Poria cocos combination on general signs, average food intake, and average water intake of mice

[0107] After oral administration of senna leaves, mice began to huddle together, with the drug-treated group showing less clumping compared to the model group. The mice also developed severe diarrhea, lethargy, yellow feces clinging to the perianal area, ragged, erect, and dull fur, and their litter was damp and strongly smelly. The ears and paws of the model group mice were pale and bloodless. During the constipation phase, the model group mice were more irritable, while the three drug-treated groups were closer to the normal group. The model group mice were thin, had infrequent bowel movements, arched backs, and dull, dry fur. Compared to the model group, the three drug-treated groups had more bowel movements, were in better spirits, and had smoother, more lustrous fur.

[0108] Figure 1 The effect of the combination of Polygonatum sibiricum and Poria cocos on the average food intake and average water intake of mice. Figure 1 A is the average food intake, Figure 1 B in the figure is the average water intake. Figure 1It can be seen that during the early and middle stages of the preventive phase, the average food intake and water intake of the three treatment groups were lower than those of the normal and model groups. These intakes gradually caught up with and even surpassed those of the control and model groups in the late stages of the preventive phase. Following oral administration of senna leaves, the average food intake of the model and three treatment groups first decreased and then increased, while the average water intake significantly increased compared to the normal group.

[0109] 2.3.2 Effect of Polygonatum sibiricum and Poria cocos combination on body weight of mice

[0110] Figure 2 The effect of the combination of Polygonatum sibiricum and Poria cocos on the body weight of mice. Figure 2 As can be seen, during the preventive administration phase, the weight of mice in the three treatment groups was lower than that of the normal group and the model group. At the end of the first week, the weight of mice in the Polygonatum sibiricum and Poria cocos B and C ratio groups was extremely significantly lower than that of the normal group (p < 0.01) and significantly lower than that of the model group (p < 0.05). After oral administration of senna leaves, the weight of mice in the model group began to decrease compared with the normal group. The weight of mice in the Polygonatum sibiricum and Poria cocos A ratio group continued to increase, while the weight of mice in the Polygonatum sibiricum and Poria cocos B and C ratio groups remained lower than that of the normal group, showing significant differences (p < 0.05). During the constipation phase, the weight of mice in the model group began to decrease, while the weight of mice in the three treatment groups remained unchanged.

[0111] 2.3.3 Effects of Polygonatum sibiricum and Poria cocos on organ indices in mice

[0112] Figure 3 The effect of the combination of Polygonatum sibiricum and Poria cocos on mouse organ indices was observed. The spleen indices of mice in all three treatment groups were lower than those in the normal and model groups. The spleen indices in the Polygonatum sibiricum and Poria cocos B ratio group showed significant differences from those in the normal and model groups (p<0.05). The kidney, thymus, and liver indices of mice in the groups were not statistically significant, with no significant differences. The liver index in the model group was significantly elevated, approaching that of the normal group in all three treatment groups. The kidney index of mice in the Polygonatum sibiricum and Poria cocos B ratio group was higher than that in the other two treatment groups.

[0113] 2.3.4 Effects of Polygonatum sibiricum and Poria cocos on fecal characteristics of mice

[0114] Figure 4The study examined the effects of the combination of Polygonatum and Poria on the fecal characteristics of mice. During the preventive phase, there were no significant differences in feces between the groups. Following oral administration of senna leaves, the model group and the three treatment groups experienced severe diarrhea, with completely unformed, watery feces. The next morning, before oral administration, the feces of the mice were barely formed but difficult to pick up. The feces of the model group mice adhered to gloves and did not slide off easily, while the feces of the treatment groups mice slid off smoothly. During the constipation phase, the feces of the model group mice decreased in volume and were yellow, with a hard texture and smaller particles. The feces of the three treatment groups were yellow-black, soft, and easy to break apart. The feces of the mice in the Polygonatum / Poria B ratio group had larger, better-formed particles, closer to those of the normal group. This suggests that the combination of Polygonatum / Poria B has the most effective preventive effect on spleen deficiency constipation in mice.

[0115] 2.3.5 Effect of Polygonatum sibiricum and Poria cocos on fecal moisture content in mice

[0116] Figure 5 This is the effect of the combination of Polygonatum and Poria on the water content of mouse feces. As can be seen from the figure, on the 21st day of the experiment, due to the oral administration of senna leaves, the fecal water content of the mice in the model group and the three drug-treated groups was higher than that of the normal group. In the early stage of constipation, except for the normal group, the fecal water content of the mice in the other groups dropped sharply. However, as time went on, the fecal water content of the drug-treated groups gradually increased. Among them, the fecal water content of the mice in the Polygonatum and Poria B ratio group was closest to that of the normal group, and the effect was the best ( Figure 5 A). In the late stage of constipation, the fecal water content of the model group mice was significantly different from that of the normal group (p<0.01), and the fecal water content of the Huangjing Fuling B ratio group mice was significantly different from that of the model group (p<0.05), and was close to that of the normal group. The Huangjing Fuling A ratio group and the Huangjing Fuling C ratio group had certain effects, but the Huangjing Fuling B ratio combination had a better effect on the prevention of spleen deficiency constipation in mice ( Figure 5 E. Figure 5 F).

[0117] 2.3.6 Effect of Polygonatum sibiricum and Poria cocos on Serum D-xylose Content in Mice

[0118] Figure 6 The study examined the effect of a combination of Polygonatum sibiricum and Poria cocos on serum D-xylose levels in mice. D-xylose levels in mouse serum are a classic indicator of spleen deficiency and are used to assess intestinal absorption in mice. The results showed that serum D-xylose levels in the model group were significantly lower than those in the normal group (p<0.01). The Polygonatum sibiricum and Poria cocos B and C ratio groups showed significant differences from the model group (p<0.05), approaching those in the normal group. The Polygonatum sibiricum and Poria cocos A ratio group showed some improvement, but the difference was not significant. This suggests that the combination of Polygonatum sibiricum and Poria cocos B and C ratios is more effective in preventing spleen deficiency in mice.

[0119] 2.3.7 Effect of Polygonatum sibiricum and Poria cocos combination on the number of culturable bacterial colonies in the intestinal tract of mice

[0120]

[0121] Table 4 shows that the intestinal flora of mice with spleen deficiency constipation underwent significant changes, with highly significant increases in Escherichia coli, bacteria, Lactobacilli, and Bifidobacteria (p < 0.01), suggesting intestinal dysbiosis in the model group. The intestinal E. coli counts of mice in the Polygonatum sibiricum and Poria cocos B ratio group were closer to those of the normal group, while the other two treatment groups were closer to the model group. The intestinal bacterial counts of mice in the Polygonatum sibiricum and Poria cocos B and C ratio groups were significantly lower than those in the model group (p < 0.05), approaching those in the normal group. The intestinal Lactobacilli counts of mice in the three treatment groups were highly significant different from those in the model group (p < 0.01). The Polygonatum sibiricum and Poria cocos B ratio group had the highest Lactobacilli counts in the normal group, while the other two treatment groups had significantly lower counts (p < 0.05). The number of bifidobacteria in the intestines of mice treated with the A, B, and C ratios of Polygonatum and Poria showed significant differences compared to the model group (p<0.05). The number of bifidobacteria in the B and C ratios was almost identical to that in the normal control group, while the number in the C ratio group was lower than that in the normal control group. In terms of the number of culturable colonies in the intestine, the three treatment groups were closer to the normal group. Among them, the combination of Polygonatum and Poria in the B ratio had the best effect in preventing spleen deficiency constipation in mice.

[0122] 2.3.8 Effect of Polygonatum sibiricum and Poria cocos combination on microbial activity in the intestinal contents of mice

[0123] Fluorescein diacetate (FDA) can be hydrolyzed by nonspecific enzymes in bacteria and fungi, releasing a colored fluorescein product. The absorbance of the product can be measured using a UV-spectrophotometer, reflecting the fluorescein content. The activity of this hydrolase can, to a certain extent, reflect the metabolic capacity of intestinal microorganisms. Figure 7 The effect of Polygonatum sibiricum and Poria cocos on the microbial activity in the intestine of mice was investigated. Figure 7 As can be seen, the intestinal microbial activity of the model group mice was significantly increased compared to the normal group (p < 0.01). This is consistent with the culturable colony counts reported in Section 3.7, suggesting that spleen deficiency constipation may promote the growth of certain microorganisms. The three treatment groups were closer to the normal group. The intestinal microbial activity of the mice in the Polygonatum sibiricum / Poria cocos B ratio group was significantly different from that of the model group (p < 0.01), being closest to the normal group. This suggests that the Polygonatum sibiricum / Poria cocos B ratio combination may be more effective in preventing spleen deficiency constipation in mice.

[0124] 2.3.9 Effect of Polygonatum sibiricum and Poria cocos on intestinal enzyme activity in mice

[0125]

[0126] As shown in Table 5, intestinal protease activity in the model group was significantly higher than that in the normal group. The protease activity in the Polygonatum-Poriacocos A and Poriacocos C ratio groups was closer to that in the normal group, but the protease activity in the Polygonatum-Poriacocos B ratio group was significantly higher than that in the normal group (p < 0.05). Amylase activity in the model and Polygonatum-Poriacocos A ratio groups was significantly higher than that in the normal group (p < 0.05). The amylase activity in the Polygonatum-Poriacocos B ratio group was higher than that in the normal group, but lower than that in the model group. The amylase activity in the Polygonatum-Poriacocos C ratio group was significantly lower than that in the normal group and showed a highly significant difference from the model and Polygonatum-Poriacocos A ratio groups (p < 0.01). Intestinal cellulase activity in the model and Polygonatum-Poriacocos C ratio groups was significantly lower than that in the normal group (p < 0.01), while it was significantly higher in the Polygonatum-Poriacocos A ratio group (p < 0.01). The Polygonatum-Poriacocos B ratio group was close to that in the normal group. The intestinal xylanase activity of mice in the model group was significantly higher than that in the normal group (p<0.01). The activity of the Polygonatum sibiricum and Poria cocos B and C ratio groups was close to that of the normal group, and was significantly different from the model group and the Polygonatum sibiricum and Poria cocos A ratio group (p<0.01). The Polygonatum sibiricum and Poria cocos A ratio group was lower than that of the normal group, and the difference was also significant (p<0.01). This shows that the intestinal enzyme activity of mice with spleen deficiency constipation has undergone significant changes. All three drug groups have a certain preventive effect on spleen deficiency constipation in mice, and the Polygonatum sibiricum and Poria cocos B ratio combination has the best preventive effect on spleen deficiency constipation in mice.

[0127] 2.3.10 Effects of Polygonatum sibiricum and Poria cocos on the Behavior of Mice

[0128] Generally speaking, healthy mice prefer to stay near the "open field," that is, the peripheral area. When mice have cognitive impairment, they are more willing to explore the central area in the new closed environment. Figure 8 The effects of a combination of Polygonatum sibiricum and Poria cocos on mouse behavior are shown in the figure. Mice with spleen deficiency-induced constipation exhibited more agitation in their new, enclosed environment, while mice in the normal and drug-treated groups were more docile. Mice in the normal group moved more along the periphery, occasionally crossing the center, with a regular pattern. Mice in the model group exhibited disorganized movements, primarily confined to the center. Mice in the three drug-treated groups tended to move more toward the periphery, with improved movement patterns compared to the model group. In terms of both active and inactive time, the drug-treated groups showed improved activity compared to the model group, with mice in the Polygonatum sibiricum and Poria cocos B ratio group more similar to the normal group.

[0129] 2.3.11 Effects of Polygonatum sibiricum and Poria cocos on the brain-gut axis, gastrointestinal motility, and oxidative stress in mice

[0130] Serotonin (5-HT) is a neurotransmitter that plays an important role in the intestine, primarily secreted by enterochromaffin cells. 5-HT regulates gastrointestinal motility and secretory function. Decreased 5-HT levels often indicate decreased intestinal motility and delayed defecation, a key mechanism of functional constipation.

[0131] Furthermore, cholecystokinin (CCK) is widely present in the digestive and nervous systems of animals. It is a brain-gut peptide that acts directly on the intestinal mucosa, stimulating intestinal fluid secretion and enhancing intestinal motility. D-xylose, a pentose sugar, is absorbed in the small intestine after oral administration, and its absorption rate reflects the small intestine's absorptive function. In cases of spleen deficiency, small intestinal absorptive function may be impaired, resulting in a reduced D-xylose absorption rate, making it considered a classic indicator of spleen deficiency.

[0132] Motilin (MTL) primarily promotes gastric emptying and intestinal motility and is a key hormone in gastrointestinal motility. Reduced MTL levels slow intestinal motility, prolonging food retention in the gastrointestinal tract, leading to excessive water absorption and constipation. MTL can be used as a marker for assessing gastrointestinal motility dysfunction, particularly in the context of spleen deficiency-induced metabolic disorders.

[0133] Vasoactive intestinal peptide (VIP) is an inhibitory neurotransmitter that slows gastrointestinal motility and inhibits the secretion of gastric acid and pancreatic enzymes. When VIP levels are elevated, it inhibits intestinal motility and aggravates constipation.

[0134] Malondialdehyde (MDA) is a product of lipid peroxidation that reflects oxidative stress and tissue damage. Spleen deficiency constipation is often accompanied by chronic inflammation and metabolic disorders, often accompanied by liver oxidative damage. Elevated MDA indicates increased oxidative stress.

[0135] Superoxide dismutase (SOD) primarily scavenges free radicals, provides antioxidant protection, and maintains cellular homeostasis. In constipation caused by spleen deficiency, SOD activity may decrease, suggesting a weakened antioxidant capacity.

[0136] As shown in Table 6, the model group mice showed decreased 5-HT, CCK, MTL, and SOD levels, while increased VIP and MDA levels, with significant differences compared to the normal group (p < 0.01). This suggests that mice with spleen deficiency constipation exhibit weakened gastrointestinal motility, increased oxidative stress, and decreased antioxidant capacity, consistent with the symptoms of spleen deficiency constipation. Compared to the model group, the three drug-treated groups showed increased 5-HT, CCK, MTL, and SOD levels, while decreased VIP and MDA levels (p < 0.05), approaching the levels of the normal group mice. This suggests that the combination of Polygonatum sibiricum and Poria cocos has a good preventive effect on spleen deficiency constipation.

[0137]

[0138] Note: CN: normal group; MD: model group; HFA: Polygonatum sibiricum and Poria cocos A ratio group; HFB: Polygonatum sibiricum and Poria cocos B ratio group; HFC: Polygonatum sibiricum and Poria cocos C ratio group.

[0139] Compared with the CN group: represents p<0.05, Represents p < 0.01; compared with the MD group: # represents p < 0.05, ## represents p < 0.01; compared with the HFA group: △ represents p < 0.05, △△ represents p < 0.01; compared with the HFB group: ▲ represents p < 0.05, ▲▲ represents p < 0.01.

[0140] Spleen deficiency constipation is often caused by insufficient spleen transport and transformation, which prevents the distribution of food and beverage essences, weakens intestinal conduction, and inadequate fluid production. This results in dry, hard stools, accompanied by difficulty defecation, lack of energy, and abdominal distension. Based on the above characteristics, the present invention tested the food intake, water intake, body weight, behavior, organ indexes, fecal characteristics, and water content of mice.

[0141] The results showed that during the spleen deficiency phase, after oral administration of senna leaves, the mice's water intake increased significantly, but their food intake initially decreased and then increased. The change in water intake may be related to compensatory drinking behavior triggered by the purgative effect of senna leaves, while the change in food intake may be due to the body's adaptation to the stimulation of senna leaves through self-regulatory mechanisms. After oral administration of senna leaves, the model group mice showed a trend of weight gain, which may be caused by obesity caused by spleen deficiency and dampness. Since spleen-tonifying and dampness-removing drugs are significantly effective in patients with spleen deficiency and dampness-removing obesity, the weight of the treated group remained essentially unchanged and even decreased slightly due to the spleen-tonifying and dampness-removing effects, which also reveals the spleen-tonifying effect of Polygonatum sibiricum and Poria cocos.

[0142] During the constipation phase, due to dietary restriction and water restriction, all mice in all groups, except the normal group, experienced weight loss, with the model group experiencing significant weight loss and minimal weight fluctuation in the drug-treated group. Following modeling, the feces of the model group were yellow, firm, with small, and reduced particles. In contrast, the feces of the drug-treated groups were softer, with a yellowish-black tint, and some even exhibited black feces. Feces in all three drug-treated groups were yellow-black, soft, and easy to break apart. The feces of the group containing the Polygonatum sibiricum and Poria cocos B ratio were most similar to those of the normal group. Fecal moisture content also showed that the Polygonatum sibiricum and Poria cocos B ratio group was the most effective, closer to the normal group and significantly different from the model group (p < 0.01). Organ index is an important marker in toxicology research, primarily used to assess organ toxicity. Organ damage is often accompanied by pathological changes such as edema and congestion. This index, by quantifying changes in the ratio of organ weight to body mass, can indirectly reflect changes in immune function.

[0143] From the behavioral perspective, there were differences in the activity time and immobility time of mice in the drug-treated group and the model group, but there was no statistical difference. Among them, the ratio of Polygonatum sibiricum to Poria cocos B had a better effect on improving the behavior of mice with spleen deficiency constipation.

[0144] The human intestinal microecological system is a complex symbiotic network composed of a large number of microbial flora, and its stability is of great significance for maintaining host homeostasis. Chinese herbal medicine has a significant impact on the structure of the intestinal flora. This influence can further affect the metabolites of intestinal microorganisms, and these products can in turn affect the transformation and absorption of Chinese herbal medicine. This two-way interaction mechanism is of great significance for explaining the mechanism of action of Chinese medicine. Escherichia coli (non-pathogenic strains) normally exist in the intestines of humans and animals, and are harmless when the intestinal microecological system is balanced. Once the microecological system is unbalanced due to diseases or other reasons, enteropathogenic Escherichia coli (harmful strains) will multiply in large numbers. In the present invention, the intestinal colony count of mice with spleen deficiency constipation increased significantly (p<0.01), showing obvious dysbacteriosis, and the combination of Polygonatum sibiricum and Poria cocos effectively alleviated this situation. The number of Escherichia coli, bacteria and Lactobacillus in the intestines of the mice in the drug-treated group approached the normal physiological range, and the number of Bifidobacterium in the intestine was no different from normal. This shows that the combination of Polygonatum and Poria has a certain protective effect on spleen deficiency constipation, among which the combination of Polygonatum and Poria in the ratio of B has the best comprehensive effect.

[0145] Gastrointestinal microbes produce and secrete enzymes that facilitate nutrient digestion and absorption by breaking down fats, proteins, and carbohydrates. Regulating enzyme levels can be used to treat a variety of intestinal diseases, and monitoring the enzyme activity of intestinal microbes can also be used to assess drug efficacy. However, higher enzyme activity is not always necessarily better; excessive activity of certain enzymes may indicate potential health issues. Studies have shown that while intestinal proteases mediate digestion and immune signaling, increased activity can lead to intestinal barrier damage and visceral hypersensitivity, manifested as increased intestinal permeability and more intense responses to stimuli. Humans do not produce cellulase; instead, intestinal microbes secrete it to break down dietary fiber into short-chain fatty acids (SCFAs), such as acetate, propionate, and butyrate. These SCFAs not only provide energy for intestinal epithelial cells but also stimulate intestinal motility and promote defecation. In the present invention, the protease activity of the model group increased compared with the normal group, the amylase activity increased significantly (p<0.05), the cellulase activity decreased significantly (p<0.01), and the xylanase activity increased significantly (p<0.01), indicating that the intestinal enzyme activity of the model group was disordered. The enzyme activity of the drug-treated group approached the normal range, among which the combination of Polygonatum sibiricum and Poria cocos B and C performed better, indicating that it is more effective in preventing spleen deficiency constipation. The results of intestinal microbial activity in mice showed that the activity of the model group was significantly increased compared with the normal group (p<0.01), and the Polygonatum sibiricum and Poria cocos B ratio group had a very significant difference compared with the model group (p<0.01), approaching the normal group. This reveals that the combination of Polygonatum sibiricum and Poria cocos B has the best preventive effect on spleen deficiency constipation.

[0146] In summary, the combination of Polygonatum and Poria has a certain effect on the prevention of spleen deficiency constipation in mice, among which the combination of Polygonatum and Poria B has the best effect in improving the general characteristics, fecal characteristics, intestinal enzyme activity and intestinal microbial environment of mice. The present invention successfully verified the effectiveness of the combination of Polygonatum and Poria in preventing spleen deficiency constipation in an animal model through preventive administration, based on the traditional Chinese medicine concept of "preventing disease before it occurs", providing new ideas and potential solutions for the early intervention and health management of such functional diseases, and reflecting the unique value of traditional Chinese medicine's "preventive treatment" in maintaining intestinal health.

[0147] The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited thereto. Within the technical concept of the present invention, various simple variations of the technical solution of the present invention may be made, including combining the various technical features in any other appropriate manner. These simple variations and combinations should also be regarded as disclosed in the present invention and fall within the scope of protection of the present invention.

[0148] Example 3: Animal efficacy experiment

[0149] 3.1 Materials

[0150] 3.1.1 Experimental animals and breeding environment

[0151] Experimental Animals: Thirty SPF-grade Kunming mice (male, weighing 20 ± 2 g) were purchased from Hunan Slake Jingda Laboratory Animal Co., Ltd. (License No. SCXK (Xiang) 2019-0004). They were housed in a barrier environment at the Animal Experimental Center of Hunan University of Chinese Medicine (License No. SYXK (Xiang) 2019-0009) at a room temperature of 23–25°C and a relative humidity of 50%–70%. Animal Ethics Welfare Approval Number: SLBH-202505150004, issued by the Animal Ethics Committee of Hunan University of Chinese Medicine.

[0152] 3.1.2 Experimental drugs and preparation

[0153] Polygonatum sibiricum and Poria cocos solution: Grind Xinhua Polygonatum sibiricum and Jingzhou Poria cocos separately, pass through a No. 4 sieve (65 mesh), and store at 4°C. Weigh a certain amount of Polygonatum sibiricum and Poria cocos powder to prepare a Polygonatum sibiricum and Poria cocos powder with a ratio of 1:1 (Poria cocos:Poria cocos). Add an appropriate amount of water to the Polygonatum sibiricum and Poria cocos powder and boil for 30 minutes to prepare the solution. Store at 4°C. Dilute the solution to a total of 1.95 g Polygonatum sibiricum and Poria cocos per kg / day per mouse, using a 0.3 ml oral gavage volume, for a twice-daily dose.

[0154] The dosage ratio B of the above-mentioned Polygonatum sibiricum and Poria cocos is (1:1, approximately equivalent to 10g Polygonatum sibiricum and 10g Poria cocos per day for adults).

[0155] 3.2 Methods

[0156] 3.2.1 Modeling method

[0157] After 3 days of adaptive feeding, 30 KM mice were randomly divided into a normal group, a yang deficiency constipation model group, and a yang deficiency constipation medication group, with 10 mice in each group. The yang deficiency constipation model was replicated according to "Yue Qi, Liu Cong, Wang Yu'e, et al. Establishment and evaluation of a yang deficiency constipation rat model. World Latest Medical Information Digest, 2017, 17(45):143-145+153". The modeling time was 18 days.

[0158] 3.2.2 Dosage regimen

[0159] For preventive administration, the drug-treated group was given 1.95 g / (kg·d) of Polygonatum sibiricum and Poria cocos solution (Poria cocos:Poria cocos = 1:1) by oral gavage, 0.3 ml / time, twice / day. See the table below for details.

[0160]

[0161] 3.2.3 Determination of fecal moisture content

[0162] Fresh feces of mice were collected on the 14th, 18th, 25th and 32nd days of the experiment. The wet weight of the feces was recorded, dried at 110°C to a constant weight and weighed to calculate the moisture content of the feces.

[0163] Fecal moisture content (%) = (fecal wet weight - fecal dry weight) / fecal wet weight × 100%.

[0164] 3.3 Results and Analysis

[0165]

[0166] As can be seen from the table, the medicinal solution with a ratio of Polygonatum and Poria B has no preventive effect on yang deficiency constipation.

Claims

1. A Chinese medicine composition for preventing spleen deficiency constipation, characterized in that: The traditional Chinese medicine composition comprises polygonatum and tuckahoe.

2. A Chinese medicine composition for preventing spleen deficiency constipation according to claim 1, characterized in that: The mass ratio of the polygonatum to poria is 1:0.2-3.

3. A Chinese medicine composition for preventing spleen deficiency constipation according to claim 2, characterized in that: The mass ratio of the polygonatum to poria is 1:1-3.

4. A Chinese medicine composition for preventing spleen deficiency constipation according to any one of claims 1 to 3, characterized in that: The traditional Chinese medicine composition consists of polygonatum and poria in a mass ratio of 1:0.2-3.

5. A method for preparing a Chinese medicine for preventing spleen deficiency constipation, characterized in that: The composition according to any one of claims 1 to 4 is mixed and crushed to obtain the product.

6. A method for preparing a Chinese medicine for preventing spleen deficiency constipation according to claim 5, characterized in that: The pulverization is performed so that the particle size of the mixture is ≤0.25 mm.

7. Use of the traditional Chinese medicine composition according to any one of claims 1 to 4 in the preparation of a traditional Chinese medicine preparation for preventing spleen deficiency constipation.

8. A Chinese medicine preparation for preventing spleen deficiency constipation, characterized by: The invention comprises the traditional Chinese medicine composition according to any one of claims 1 to 4 and pharmaceutically acceptable excipients.

9. The Chinese medicine preparation according to claim 8, characterized in that: The Chinese medicine preparation is in the form of granules, powders, decoctions, pills, tablets, capsules, mixtures or syrups.

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