Application of compound radix astragali spleen-tonifying oral liquid in improvement of functional dyspepsia of young rats

Abstract

The invention discloses application of compound radix astragali spleen-tonifying oral liquid to improvement of functional dyspepsia of young rats, and belongs to the technical field of medicines. Comprising the following steps: model construction and grouping: establishing a young rat functional dyspepsia model, randomly dividing successfully molded animals into a model control group, a positive drug control group and at least one compound radix astragali spleen-tonifying oral liquid administration group, and setting a blank control group; dosing in groups; performing pharmacodynamic evaluation; and analyzing data. The invention systematically discloses a multi-target action mechanism of the compound radix astragali spleen-tonifying oral liquid for treating functional dyspepsia for the first time, and provides an experimental basis for clinical popularization and application of the compound radix astragali spleen-tonifying oral liquid. The compound radix astragali spleen-tonifying oral liquid has the advantages of definite curative effect, comprehensive action mechanism, small adverse reaction and the like, and is particularly suitable for treating young functional dyspepsia patients.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to the application of Compound Astragalus Spleen-Strengthening Oral Liquid in improving functional dyspepsia in juvenile rats. Background Technology

[0002] Functional dyspepsia (FD) is a common gastrointestinal disorder characterized by symptoms such as upper abdominal pain, early satiety, and bloating, significantly impacting patients' quality of life. While commonly used medications like domperidone offer some efficacy, they also present with significant adverse reactions and a high relapse rate. Traditional Chinese medicine (TCM) possesses unique advantages in regulating gastrointestinal function. Compound Astragalus Spleen-Strengthening Oral Liquid, a TCM compound preparation, has the effects of strengthening the spleen and replenishing qi, harmonizing the stomach and intestines. However, its mechanism of action in treating FD, especially juvenile FD, remains unclear, lacking systematic experimental research and application evidence. Summary of the Invention

[0003] The purpose of this invention is to provide the application of Compound Astragalus Spleen-Strengthening Oral Liquid (hereinafter referred to as FFHQ) in improving functional dyspepsia in juvenile rats. Through experiments using a juvenile rat FD model, it was confirmed that FFHQ can significantly improve FD-related symptoms.

[0004] The technical solution adopted in this invention is as follows: The application of Compound Astragalus Spleen-Strengthening Oral Liquid in improving functional dyspepsia in juvenile rats includes the following steps: S1. Model Construction and Grouping: A functional dyspepsia model was established in juvenile rats, and the successfully modeled animals were randomly divided into a model control group, a positive drug control group, and at least one compound astragalus spleen-strengthening oral liquid administration group. A blank control group was also set up. S2. Grouped administration: Domperidone was administered by gavage to the positive drug control group, different doses of Compound Astragalus Spleen-Strengthening Oral Liquid were administered by gavage to the Compound Astragalus Spleen-Strengthening Oral Liquid administration group, and the same volume of solvent was administered by gavage to the model control group and the blank control group. The administration was carried out once a day for a predetermined period of time. S3. Pharmacodynamic evaluation: After administration, the general condition of rats in each group was scored, and their gastrointestinal motility index, visceral sensitivity index, serum gastrointestinal hormone level and hypothalamic-pituitary-adrenal axis-related hormone level were measured. S4. Data Analysis: Statistical analysis was performed on the data obtained in step S3 to evaluate the effect of Compound Astragalus Spleen-Strengthening Oral Liquid on improving functional dyspepsia in juvenile rats.

[0005] In step S1, the functional dyspepsia model in juvenile rats was constructed using the following method: Six-day-old SD rat pups were randomly divided into two groups after 7 days of adaptive feeding: a blank control group (n=16) and a model group (n=48). The model group was orally administered 1 mg / mL iodoacetamide solution containing 2% sucrose at a dose of 10 mg / kg. The blank control group was administered an equal volume of 2% sucrose solution daily by gavage for one week. At four weeks of age, the model group SD rat pups were subjected to fatigue-induced sleep deprivation using a modified multi-platform sleep deprivation method at regular intervals for two consecutive weeks. The blank control group was fed normally.

[0006] In step S1, after the modeling was completed, 8 SD rats were randomly selected from the model group and the blank control group for model evaluation. The SD rats in the model group were randomly divided into 5 groups according to their body weight, with 8 rats in each group. These groups were the model control group, the low-dose compound astragalus and spleen-strengthening oral liquid group, the medium-dose compound astragalus and spleen-strengthening oral liquid group, the high-dose compound astragalus and spleen-strengthening oral liquid group, and the positive drug control group.

[0007] In step S2, the drug doses in the low-dose, medium-dose, and high-dose groups of the compound astragalus spleen-strengthening oral liquid administration group were 2.52 mL / kg, 5.04 mL / kg, and 10.08 mL / kg, respectively; the dosage of the positive drug control group was 2.7 mg / kg of domperidone per day.

[0008] In step S3, the gastrointestinal motility index is evaluated by measuring gastric emptying rate and small intestinal propulsion rate; the visceral sensitivity index is evaluated by abdominal wall withdrawal reflex score.

[0009] The method for determining the gastric emptying rate and small intestinal propulsion rate includes: administering a semi-solid paste containing carbon powder to each group of rats by gavage, anesthetizing them after a specified time, and collecting samples. The results are calculated by measuring the amount of residual gastric contents and the propulsion distance of the carbon powder in the small intestine.

[0010] In step S3, the determination of serum gastrointestinal hormone levels includes detecting at least one of ghrelin, motilin, gastrin, cholecystokinin, and vasoactive intestinal peptide; the determination of hypothalamic-pituitary-adrenal axis-related hormone levels includes detecting at least one of corticotropin-releasing hormone, adrenocorticotropic hormone, and corticosterone.

[0011] In step S3, the determination of serum inflammatory factor levels includes detecting at least one of interleukin-1β, interleukin-6, and tumor necrosis factor-α.

[0012] In step S2, the predetermined dosing cycle is 21 days.

[0013] In summary, due to the adoption of the above technical solution, the beneficial effects of the present invention are: This invention systematically reveals for the first time the multi-target mechanism of action of Compound Astragalus Spleen-Strengthening Oral Liquid in treating functional dyspepsia, providing experimental evidence for its clinical application. Compound Astragalus Spleen-Strengthening Oral Liquid has advantages such as clear efficacy, comprehensive mechanism of action, and few side effects, making it particularly suitable for the treatment of young patients with functional dyspepsia. Attached Figure Description

[0014] Figure 1 This is a flowchart of the experimental process of the present invention; Figure 2 This is a graph showing the effect of FFHQ on rat body weight and food intake in this invention; Figure 3 This is a graph showing the effects of FFHQ on gastrointestinal motility, visceral sensitivity, and serum hormone levels in FD juvenile rats in this invention. Figure 4 This is a graph showing the effects of FFHQ on gastrointestinal motility, visceral sensitivity, and serum hormone levels in FD juvenile rats in this invention. Figure 5 In the middle, A and A' are the TIC overlay maps of the mass spectra of the quality control samples in positive and negative ion modes; B and B' are the PCA loading maps of the metabolic spectra in positive and negative ion modes, respectively; CD and C'-D' are the OPLS-DA loading maps of the metabolic spectra in positive and negative ion modes, respectively; EF and E'-F' are the OPLS-DA verification maps of the metabolic spectra in positive and negative ion modes, respectively. Figure 6 This is a graph showing the effect of FFHQ on serum metabolites in FD juvenile rats in this invention; Figure 6 In the middle section, A and A' are Volcano plots of Control vs FD metabolic profiles under positive and negative ion modes, respectively; B and B' are Volcano plots of FD vs FD_FFHQ metabolic profiles under positive and negative ion modes, respectively; C and C' are heatmaps of the changes in the content of differential metabolites under positive and negative ion modes (blue to red represents an increase in content); D is the metabolic pathway related to the differential metabolites; n=3. Detailed Implementation

[0015] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. The components of the embodiments of the present invention described and shown in the accompanying drawings can generally be arranged and designed in various different configurations.

[0016] Therefore, the following detailed description of the embodiments of the invention provided in the accompanying drawings is not intended to limit the scope of the claimed invention, but merely to illustrate selected embodiments of the invention. All other embodiments obtained by those skilled in the art based on the embodiments of the invention without inventive effort are within the scope of protection of the invention. Example

[0017] In this embodiment, the reagents and materials were as follows: Compound Astragalus Spleen-Strengthening Oral Liquid (batch number: 22035) was provided by Lanzhou Foci Pharmaceutical Co., Ltd. Domperidone (NCJ3300) and iodoacetamide (I105563) were purchased from Xian Janssen Pharmaceutical Co., Ltd. and Aladdin Reagent (Shanghai) Co., Ltd., respectively. Sixty-four healthy SD suckling rats (6 days old, half male and half female, 15-20g) were purchased from the Experimental Animal Center of Lanzhou University, production license number: SCXK(Gan)-2023-0003. All procedures were approved by the Animal Ethics Committee of the School of Pharmacy of Lanzhou University.

[0018] This embodiment provides the application of Compound Astragalus Spleen-Strengthening Oral Liquid in improving functional dyspepsia in juvenile rats, including the following steps: 1. The functional dyspepsia model was established, and the animal grouping and drug administration regimen was as follows: During the experiment, suckling mice drank breast milk before 3 weeks of age, and were given free access to water and feed after weaning at 3 weeks of age. The room temperature was controlled at 22℃, the humidity was controlled at 40%-70%, and the circadian rhythm was maintained at 24h.

[0019] Six-day-old suckling mice were randomly divided into two groups after seven days of acclimatization: a blank control group (n=16) and a model group (n=48). The model group was administered 1 mg / mL iodoacetamide solution (containing 2% sucrose) orally via gavage at a dose of 10 mg / kg, while the blank control group was administered an equal volume of 2% sucrose solution daily via gavage for one week. At four weeks of age, the model group rats underwent fatigue induction using a modified multiple platform method (MMPM) at regular intervals for two weeks, while the blank control group was fed normally. After model establishment, eight rats from each of the model group (FD, IA+MMPM) and the blank control group were randomly selected for model evaluation.

[0020] After modeling, the rats in the model group were randomly divided into 5 groups of 8 rats each, based on their body weight: model control group (FD), low-dose FFHQ administration group (FD_FFHQ-L), medium-dose FFHQ administration group (FD_FFHQ-M), high-dose FFHQ administration group (FD_FFHQ-H), and positive control group (domperidone, (FD_DOM)). Drug dosages were converted to rat dosages based on the ratio of body surface area between children over 3 years old and rats. The drug dosages in the low-dose, medium-dose, and high-dose groups of the compound astragalus spleen-strengthening oral liquid administration group were 2.52 mL / kg, 5.04 mL / kg, and 10.08 mL / kg, respectively. The positive control group received domperidone 2.7 mg / kg daily by gavage, while the blank control group and FD group received an equal volume of distilled water by gavage once daily for 21 days. The experimental procedure is as follows: Figure 1 .

[0021] 2. General condition observation Referring to the "Principles of Clinical Research Indicators for Traditional Chinese Medicine Treatment of Spleen Deficiency Syndrome", the general condition of rats was observed at 13:00 on the last three days. According to the table below, each of the four indicators was given a weight of 25% and a comprehensive score was calculated.

[0022] Table 1 Rating criteria for general condition of rats

[0023] 3. Gastrointestinal motility measurement Gastric emptying rate: Rats were administered a nutritional semi-solid paste mixed with carbon powder (2 mL / rat) by gavage. 40 minutes later, the rats were anesthetized, and the upper edge of the cardia and the lower edge of the pylorus were ligated, and the stomach was removed. The stomach surface was cleaned and dried, and the total stomach weight was measured. The stomach contents were cleaned and dried, and the empty stomach weight was measured. Gastric emptying rate % = 1 - (total stomach weight - empty stomach weight) / total stomach weight × 100%.

[0024] Small intestinal propulsion rate: The mesentery was separated, the small intestine was removed and laid flat on a piece of white paper, and the total length from the pylorus to the cecum and the distance from the pylorus to the leading edge of the carbon semi-solid paste were measured. Small intestinal propulsion rate = (distance of carbon semi-solid paste propulsion in the small intestine / total length of the small intestine) × 100%.

[0025] Isolated muscle strip perfusion experiment: Rats were fasted for 12 hours but given free access to water. They were euthanized by cervical dislocation, and the gastric body and antrum tissues were harvested, rinsed, and placed in Krebs solution, through which a mixture of 95% oxygen and 5% carbon dioxide was introduced. Longitudinal muscle of the gastric body and circular muscle of the gastric antrum were prepared. The muscle strips were placed in a 37°C perfusion tank continuously filled with the mixed gas, with an external circulation heating device maintaining a constant temperature. A tension transducer and a biosignal acquisition system were connected to record the contraction and motion curves of the muscle strips.

[0026] 4. Visceral sensitivity testing Visceral sensitivity was determined using the abdominal withdrawal reflex (AWR) scoring test. The procedure was as follows: Rats were fasted for 24 hours, and their intestines were emptied by pressing on the anus. Subsequently, the rats were anesthetized by an intraperitoneal injection of 30 mg / kg of 1% sodium pentobarbital. A glycerin-coated catheter was inserted 2 cm into the rat's rectum and secured with medical tape. After the rats regained consciousness and acclimatized to the experimental environment for 30 minutes, the intestines were then distended using the catheter inflation method. The rats' responses were recorded at inflation volumes of 1.0 mL and 2.0 mL, and scored using the method shown in the table below. This procedure was repeated three times, with a 10-minute interval between each repetition.

[0027] Table 2. Scoring criteria for abdominal wall withdrawal reflex

[0028] 5. Measurement of gastrointestinal hormone levels The serum levels of amylase (AMS) activity, D-lactic acid, D-xylose, ghrelin, motilin (MTL), gastrin (GAS), cholecystokinin (CCK), vasoactive intestinal peptide (VIP), corticotropin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH), and corticosterone (CORT) were measured according to the ELISA kit instructions.

[0029] 6. Rat serum metabolomics analysis Serum samples from rats in the Control, FD, and FD_FFHQ groups were analyzed by UPLC-Q-TOF-MS. After transformation, peak extraction, alignment, and correction, the raw data were used to identify metabolites by integrating multiple databases, with a quality control sample (QC) included for every 10 samples tested. PCA, OPLS-DA, permutation test, and OPLS-DA / S-plots loading analysis were performed using R software. Potential biomarkers were identified based on precise molecular weights from the Human Metabolomics Database (HMDB) and the METLIN database. Differential metabolite pathway analysis was performed using the MetaboAnalyst database. Multi-omics analysis was assisted by Wuhan Metawell Biotechnology Co., Ltd. (Wuhan, China).

[0030] 7. Statistical Analysis Data were processed using Graph Pad Prism 8.0 software. One-way ANOVA was used for comparisons between groups. Data are expressed as mean ± standard deviation (x±SD), and p < 0.05 indicates a significant difference.

[0031] 8. Test Results 8.1 General Condition Observation The effects of FFHQ on the general condition of rats are shown in the table below. The model control group rats exhibited symptoms such as lethargy, lack of movement, disheveled fur, and loose stools, with symptom scores significantly higher than the blank control group (P<0.01). The positive control group and all FFHQ dosage groups showed significant improvement in symptoms, with shiny fur, essentially normal stools, and better mental state. Except for the FFHQ-L group, the improvement effects were significant in all other groups (P<0.05).

[0032] Table 3 Comparison of general condition scores of rats in each group

[0033] 8.2 Weight and food intake The effects of FFHQ on rat body weight and food intake are shown in the figure. Figure 2 The body weight of rats in the medium and high dose FFHQ groups was significantly higher than that in the FD group (P<0.05), while there was no significant difference in body weight between the low dose FFHQ group and the DPLT group and the FD group (P>0.05). The food intake of rats in the FD group was significantly lower than that in the Control group (P<0.01), and except for the low dose FFHQ group, the food intake of rats in all other treatment groups was significantly higher than that in the FD group (P<0.05).

[0034] Figure 2 Table A shows the change in body weight and the comparison of the average body weight at the last feeding (n=8); Table B shows the food intake of the rats (n=8). Compared with the control group: * P<0.05, ** P<0.01; Compared with the FD group: # P<0.05, ## P<0.01; Compared with the DOM group: & P<0.05, && P<0.01; Mean±SD.

[0035] 8.3 Effects of FFHQ on gastrointestinal motility and visceral sensitivity in juvenile FD rats Gastric emptying rate and intestinal propulsion rate were improved in all treatment groups. The DOM and FFHQ-H groups significantly increased gastric emptying rate and intestinal propulsion rate in FD juvenile rats. Figure 3 In the A and B groups, both P < 0.05; the low-dose FFHQ group only significantly improved gastric emptying rate (P < 0.01). Normal isolated muscle strips exhibit rhythmic contractile activity, such as... Figure 3As shown in C and D, compared with the Control group, the contractile activity of the longitudinal muscle of the gastric body and the circular muscle of the gastric antrum in the FD group was significantly weakened, and the amplitude of the rhythmic contraction wave generated per gram of muscle strip tissue was significantly reduced (P<0.01). In contrast, the high-dose DOM and FFHQ groups significantly enhanced the contractile activity of isolated muscle strips in the model rats (P<0.01).

[0036] AWR score results are as follows Figure 3 As shown in Figure E, compared with the Control group, the AWR score of the FD group was significantly increased at different inflation volumes (P<0.01), indicating increased visceral sensitivity in juvenile FD rats. The AWR scores of all treatment groups decreased significantly, especially in the FFHQ groups at 2.0 mL, and the AWR score of the FFHQ-H group basically recovered to the control group level (all P<0.05).

[0037] Figure 3 In the middle section, A represents gastric emptying rate (n=5); B represents small intestinal transit rate (n=5); C represents representative rhythmic contractions of the longitudinal muscles of the gastric body, with the contraction amplitude quantified by (C') (n=3); D represents representative rhythmic contractions of the circular muscles of the gastric antrum, with D' representing the quantified contraction amplitude (n=3); and E represents the abdominal withdrawal reflex score (n=8). Compared with the Control group: * P<0.05, ** P<0.01; Compared with the FD group: # P<0.05, ## P<0.01; Compared with the DOM group: &P<0.05, &&P<0.01; mean±SD.

[0038] 8.4 Effects of FFHQ on serum hormone levels in juvenile FD rats like Figure 4 As shown in the AE, serum Ghrelin, MTL, and GAS levels in the FD group were significantly lower than those in the Control group (P<0.01), while CCK and VIP levels were significantly higher in the FD group (P<0.01). Compared with the FD group, all treatment groups showed some improvement in the above indicators. The positive control group significantly increased serum Ghrelin, MTL, and GAS levels in juvenile FD rats (P<0.01), and the FFHQ-H group showed a similar effect to the positive control group (P<0.01); the FFHQ-M group had a similar effect on MTL and GAS as the positive control group (P<0.01). The positive control group only significantly reduced serum CCK levels (P<0.01), while all three FFHQ dose groups significantly reduced serum CCK and VIP levels (all P<0.01). These results suggest that FFHQ can regulate gastrointestinal hormone secretion and maintain intestinal homeostasis.

[0039] Compared to the Control group, the FD group showed significantly increased serum levels of hypothalamic-pituitary-adrenal (HPA) cascade hormones CRH, ACTH, and CORT. Figure 4 In the FH group, P<0.05 or P<0.01. All treatment groups reduced the levels of the three hormones and improved HPA axis hyperactivity, but only the FFHQ-H group significantly reversed the increase in the levels of the three hormones (P<0.05 or P<0.01).

[0040] The above experimental results indicate that FFHQ has the effect of regulating gastrointestinal hormone secretion and improving HPA axis hyperactivity.

[0041] Figure 4 The figure shows the effects of FFHQ on gastrointestinal motility, visceral sensitivity, and serum hormone levels in FD juvenile rats. Figure 4 The AEs in the data represent serum Ghrelin, MTL, GAS, CCK, and VIP levels (n=8). Figure 4 FH in the table represents serum CRH, ACTH, and CORT levels (n=8). Compared with the Control group: * P<0.05, ** P<0.01; Compared with the FD group: # P<0.05, ## P<0.01; Compared with the DOM group: & P<0.05, && P<0.01; Mean±SD.

[0042] 7.6. FFHQ regulates serum metabolites in juvenile FD rats The repeatability results of the analytical method are as follows: Figure 5 As shown in Figure A, the curves for detecting the total ion current of metabolites showed high overlap in both positive and negative ion modes, indicating good instrument stability and reasonable experimental data during sample detection. Meanwhile, an internal standard of known concentration was added to the QC sample, and the detection results are shown in Table 4. The CV values ​​and coefficients of variation of the eight internal standards under both positive and negative ion modes were all less than 3%, indicating strong data stability and suitability for subsequent analysis.

[0043] Table 4. Stability of internal standard in quality control samples

[0044] To investigate the effects of FFHQ on endogenous metabolites in FD juvenile rats, unsupervised PCA was used to examine serum metabolic profiles in the Control, FD, and FFHQ groups under both positive and negative ion modes, and the degree of dispersion among the samples was analyzed. Figure 5(B in the text). The results showed that the three groups were clearly separated under both positive and negative ion modes, indicating that the serum metabolites of FD juvenile rats were significantly altered, and that FFHQ had a certain regulatory effect.

[0045] To better reflect the differences between groups and further explore the changes in serum metabolites under FD disease conditions, supervised OPLS-DA method was used to perform pattern recognition analysis on the three groups of serum samples. Figure 5 (C, D). OPLS-DA score plots showed significant differences in serum metabolic profiles between the Control and FD groups, and between the FD and FFHQ groups under positive and negative ion modes, with Tscore1 values ​​of 48.7%, 52%, 47.8%, and 46.6%, respectively. This suggests a clear separation between FD juvenile rats and healthy rats, indicating impaired metabolism. FFHQ administration also altered metabolism, showing a clear separation from the FD group. OPLS-DA model validation results ( Figure 5 The EF values ​​are as follows: Control group and FD group: positive ion mode (R2X=0.593, R2Y=1, Q2=0.932), negative ion mode (R2X=0.652, R2Y=1, Q2=0.945); FD group and FFHQ administration group: positive ion mode (R2X=0.637, R2Y=1, Q2=0.905), negative ion mode (R2X=0.652, R2Y=1, Q2=0.896). This indicates that the established OPLS-DA model has good predictive ability and reliability and can be used for the analysis of differential metabolism.

[0046] Further screening of differentially expressed metabolites was conducted using volcano plots, with the following screening criteria: a) metabolites with VIP > 1; b) metabolites with P-value < 0.05; and c) metabolites with FC > 1.5. S-plot loading analysis revealed the upregulated and downregulated metabolites between the Control and FD groups, and between the FD and FFHQ dosing groups, under positive and negative ion modes. Figure 6 (AB).

[0047] To investigate the changes in differential metabolites after drug administration, the intersection of differential metabolites between two control groups (Control group and FD group, FD group and FFHQ group) was collected under both positive and negative ion modes. This resulted in 36 and 42 differential metabolites identified under positive and negative ion modes, respectively. Preliminary annotation of the intersecting differential metabolites was performed using the KEGG and HMDB databases, providing detailed information on the differential metabolites among the Control group, FD group, and FFHQ group under both positive and negative ion modes, as shown in Table 5.

[0048] Further, based on the relative content of differentially metabolites, a cluster analysis heatmap was constructed. Figure 6(C in the text). In the positive ion mode, the levels of 36 metabolites, including corticosterone, were significantly reversed in the FFHQ group; in the negative ion mode, the levels of 42 metabolites, including flavin mononucleotide, were significantly reversed in the FFHQ group.

[0049] Metabolic pathway enrichment analysis was performed based on the screened differential metabolites to explore the mechanism by which FFHQ improves FD. The results are as follows: Figure 6 As shown in D and Table 6. A total of 23 pathways were enriched in the screened differential metabolites, of which 13 had Impact > 0. These pathways are: steroid hormone biosynthesis, riboflavin metabolism, cysteine ​​and methionine metabolism, tyrosine metabolism, phenylalanine metabolism, ascorbate and aldarate metabolism, histidine metabolism, pentose and glucuronate interconversions, fructose and mannose metabolism, the citric acid cycle (TCA cycle), beta-Alanine metabolism, pyruvate metabolism, and glycolysis or gluconeogenesis.

[0050] Table 5. Identification results of potential differentially expressed metabolites in serum No. RT (min) Differential metabolites Formula Adduct HMDB KEGG Mode FD / Control FD_FFHQ / FD 1 0.65 342.12 4-O-beta-D-Mannopyranosyl-D-mannopyranose <![CDATA[C 12 H 22 O 11 ]]> <![CDATA[[M+Cl] - ]]> HMDB0029933 - neg up down 2 0.78 182.08 Sorbitol <![CDATA[C6H 14 O6]]> <![CDATA[[M-2H+3Na] + ]]> HMDB0000247 C00794 pos down up 3 0.81 318.13 Tyr-His <![CDATA[C 15 H 18 N4O4]]> <![CDATA[[M+] + ]]> HMDB0029107 - pos down up 4 0.82 309.17 Lys-Tyr <![CDATA[C 15 H 23 N3O4]]> <![CDATA[[M+Cl] - ]]> HMDB0028963 - neg down up 5 0.82 226.11 Carnosine <![CDATA[C9H 14 N4O3]]> <![CDATA[[M+Na+HCOOH] + ]]> HMDB0000033 C00386 pos down up 6 0.96 194.04 Glucuronic Acid <![CDATA[C6H 10 O7]]> <![CDATA[[M-H2O-H] - ]]> HMDB0000127 C00191 neg up down 7 0.96 176.03 Ascorbicacid <![CDATA[C6H8O6]]> <![CDATA[[M-H2O-H] - ]]> HMDB0000044 C00072 neg down up 8 1.27 219.13 Ritalinicacid <![CDATA[C 13 H 17 NO2]]> <![CDATA[[M+H-H2O] + ]]> HMDB0042008 - pos down up 9 1.28 559.07 ADP-ribose <![CDATA[C 15 H 23 N5O 14 P2]]> <![CDATA[[M+H] + ]]> HMDB0001178 C00301 pos down up 10 1.39 88.02 PyruvicAcid <![CDATA[C3H4O3]]> <![CDATA[[2M-H] - ]]> HMDB0000243 C00022 neg up down 11 1.42 285.11 Vidarabinemonohydrate <![CDATA[C 10 H 15 N5O5]]> <![CDATA[[M+H] + ]]> HMDB0014340 C07195 pos down up 12 1.64 153.04 Aminosalicylic Acid <![CDATA[C7H7NO3]]> <![CDATA[[M+H] + ]]> HMDB0014378 C02518 pos down up 13 1.66 135.04 Methylcysteine <![CDATA[C4H9NO2S]]> <![CDATA[[M+H] + ]]> HMDB0002108 C22040 pos down up 14 1.73 325.08 Dihyroxy-1H-indoleglucuronide I <![CDATA[C 14 H 15 NO8]]> <![CDATA[[M+H] + ]]> HMDB0059997 - pos down up 15 2.21 157.07 1-Acetylproline <![CDATA[C7H 11 NO3]]> <![CDATA[[M-H] - ]]> HMDB0242126 - neg down up 16 2.22 333.13 Trp-glu <![CDATA[C 16 H 19 N3O5]]> <![CDATA[[M+H] + ]]> HMDB0029082 - pos up down 17 2.29 108.06 p-Cresol <![CDATA[C7H8O]]> <![CDATA[[M+NH4] + ]]> HMDB0001858 C01468 pos down up 18 2.65 297.09 5'-Deoxy-5'-(Methylthio)Adenosine <![CDATA[C 11 H 15 N5O3S]]> <![CDATA[[M+H] + ]]> HMDB0001173 C00170 pos down up 19 2.65 223.08 N-Acetyl-L-tyrosine <![CDATA[C 11 H 13 NO4]]> <![CDATA[[M-H] - ]]> HMDB0000866 C01657 neg down up 21 2.68 205.04 Xanthurenic acid <![CDATA[C 10 H7NO4]]> <![CDATA[[M+H] + ]]> HMDB0000881 C02470 pos down up 22 2.95 244.03 Apraclonidine <![CDATA[C9H 10 Cl2N4]]> <![CDATA[[M+] + ]]> HMDB0015099 C07668 pos up down 23 3.00 189.04 Kynurenicacid <![CDATA[C 10 H7NO3]]> <![CDATA[[M+] + ]]> HMDB0000715 C01717 pos down up 24 3.02 207.04 Pterin-6-carboxylic acid <![CDATA[C7H5N5O3]]> <![CDATA[[M+Cl] - ]]> HMDB0033136 - neg up down 25 3.07 464.10 Isoquercitrin <![CDATA[C 21 H 20 O 12 ]]> <![CDATA[[M+H] + ]]> HMDB0037362 C05623 pos down up 26 3.07 288.06 (+ / -)-Eriodictyol <![CDATA[C 15 H 12 O6]]> <![CDATA[[M+H] + ]]> HMDB0005810 C05631 pos up down 27 3.11 342.13 Coniferin <![CDATA[C 16 H 22 O8]]> <![CDATA[[M-H] - ]]> HMDB0013682 C00761 neg up down 28 3.22 448.10 Isoorientin <![CDATA[C 21 H 20 O 11 ]]> <![CDATA[[M+HCOO] - ]]> HMDB0301981 C01821 neg up down 29 3.37 283.12 Anaxagoreine <![CDATA[C 17 H 17 NO3]]> <![CDATA[[M+] + ]]> HMDB0034669 - pos down up 30 3.47 225.10 6-Benzylaminopurine <![CDATA[C 12 H 11 N5]]> <![CDATA[[M+] + ]]> HMDB0039238 C11263 pos down up 31 3.53 193.07 Methylhippurate <![CDATA[C 10 H 11 NO3]]> <![CDATA[[M+H-H2O] + ]]> HMDB0000859 - pos down up 32 3.68 556.18 Cassitoroside <![CDATA[C 25 H 32 O 14 ]]> <![CDATA[[M-H] - ]]> HMDB0041289 - neg down up 33 3.70 590.16 5,7-dihydroxy-2-(4-hydroxy-3-methoxyphenyl)-6-[(2R,3S,4S,6S)-4-hydroxy-6-methyl-5-oxo-3-[(2S,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxyoxan-2-yl]chromen-4-one <![CDATA[C 28 H 30 O 14 ]]> <![CDATA[[M-H] - ]]> HMDB0037420 - neg down up 34 3.81 137.08 1-Hydroxylamino-2-phenylethane <![CDATA[C8H 11 NO]]> <![CDATA[[M+H-H2O] + ]]> HMDB0001065 C02735 pos down up 35 3.82 133.05 Indoxyl <![CDATA[C8H7NO]]> <![CDATA[[M+H] + ]]> HMDB0004094 C05658 pos down up 36 3.89 232.08 Phenobarbital <![CDATA[C 12 H 12 N2O3]]> <![CDATA[[M-H] - ]]> HMDB0015305 C07434 neg down up 37 3.90 207.09 N-Acetyl-L-phenylalanine <![CDATA[C 11 H 13 NO3]]> <![CDATA[[M-H] - ]]> HMDB0000512 C03519 neg down up 38 4.00 478.11 3,4,5-trihydroxy-6-{[7-hydroxy-2-(3-hydroxy-4-methoxyphenyl)-4-oxo-3,4-dihydro-2H-1-benzopyran-5-yl]oxy}oxane-2-carboxylic acid <![CDATA[C 22 H 22 O 12 ]]> <![CDATA[[M-H] - ]]> HMDB0128063 - neg up down 39 4.03 340.16 Citalopram N-oxide <![CDATA[C 20 H 21 FN2O2]]> <![CDATA[[M+Cl] - ]]> HMDB0060654 C16607 neg up down 40 4.06 475.18 Codeine-6-glucuronide <![CDATA[C 24 H 29 NO9]]> <![CDATA[[M+NH4-2H] - ]]> HMDB0060464 C16577 neg up down 41 4.09 164.05 Phenylpyruvic acid <![CDATA[C9H8O3]]> <![CDATA[[2M-H] - ]]> HMDB0000205 C00166 neg up down 42 4.33 244.00 (E)-3-(4-sulfooxyphenyl)prop-2-enoicacid <![CDATA[C9H8O6S]]> <![CDATA[[M-H] - ]]> HMDB0125166 - neg up down 43 4.34 416.11 Daidzin <![CDATA[C 21 H 20 O9]]> <![CDATA[[M+HCOO] - ]]> HMDB0033991 C10216 neg down up 44 4.35 298.10 Amlexanox <![CDATA[C 16 H 14 N2O4]]> <![CDATA[[M-H] - ]]> HMDB0015160 - neg up down 45 4.37 448.10 Quercitrin <![CDATA[C 21 H 20 O 11 ]]> <![CDATA[[M-H] - ]]> HMDB0033751 C01750 neg down up 46 4.42 470.09 (Z)-Resveratrol 3-glucoside5-sulfate <![CDATA[C 20 H 22 O 11 S]]> <![CDATA[[M-H] - ]]> HMDB0036043 - neg up down 47 4.78 352.23 PGF3α <![CDATA[C 20 H 32 O5]]> <![CDATA[[M+NH4] + ]]> HMDB0002122 C06476 pos up down 48 4.78 434.12 Naringenin-7-O-glucoside <![CDATA[C 21 H 22 O 10 ]]> <![CDATA[[M-H] - ]]> HMDB0140590 - neg up down 49 4.78 456.10 Flavinmononucleotide <![CDATA[C 17 H 21 N4O9P]]> <![CDATA[[M-H] - ]]> HMDB0001520 C00061 neg down up 50 4.78 519.10 Dabrafenib <![CDATA[C 23 H 20 F3N5O2S2]]> <![CDATA[[M-H] - ]]> HMDB0250818 - neg up down 51 4.84 216.08 O-Desmethylnaproxen <![CDATA[C 13 H 12 O3]]> <![CDATA[[M+HCOO] - ]]> HMDB0013989 - neg down up 52 4.85 308.10 Warfarin <![CDATA[C 19 H 16 O4]]> <![CDATA[[M+Cl] - ]]> HMDB0001935 C01541 neg down up 53 4.96 147.07 Indole-3-carbinol <![CDATA[C9H9NO]]> <![CDATA[[M+H-H2O] + ]]> HMDB0005785 - pos up down 54 5.15 222.13 2-Hydroxyibuprofen <![CDATA[C 13 H 18 O3]]> <![CDATA[[M+H] + ]]> HMDB0060920 - pos down up 55 5.30 160.11 3-Hydroxyvalproicacid <![CDATA[C8H 16 O3]]> <![CDATA[[M+Na] + ]]> HMDB0013899 C16651 pos up down 56 5.35 182.09 Dihydroconiferylalcohol <![CDATA[C 10 H 14 O3]]> <![CDATA[[M-H] - ]]> HMDB0303757 C10448 neg up down 57 5.39 466.26 Androsteroneglucuronide <![CDATA[C 25 H 38 O8]]> <![CDATA[[M-H2O-H] - ]]> HMDB0002829 C11135 neg down up 58 5.45 346.21 Corticosterone <![CDATA[C 21 H 30 O4]]> <![CDATA[[M+H] + ]]> HMDB0001547 C02140 pos up down 59 5.51 424.25 Pravastatin <![CDATA[C 23 H 36 O7]]> <![CDATA[[M-H2O-H] - ]]> HMDB0005022 C01844 neg down up 60 5.51 449.31 Chenodeoxyglycocholic acid <![CDATA[C 26 H 43 NO5]]> <![CDATA[[M-] - ]]> HMDB0006898 C05462 neg up down 61 5.53 402.30 Sorbitan,monohexadecanoate <![CDATA[C 22 H 42 O6]]> <![CDATA[[M+HCOO] - ]]> HMDB0029887 - neg up down 62 5.74 240.17 L-Menthylacetoacetate <![CDATA[C 14 H 24 O3]]> <![CDATA[[M-H] - ]]> HMDB0032369 - neg down up 63 5.81 436.33 2-Phytyl-1,4-naphthoquinone <![CDATA[C 30 H 44 O2]]> <![CDATA[[M+Cl] - ]]> HMDB0004649 C13309 neg down up 64 5.84 290.22 Androsterone <![CDATA[C 19 H 30 O2]]> <![CDATA[[M+H-H2O] + ]]> HMDB0000031 C00523 pos down up 65 5.84 350.21 Prostaglandin D3 <![CDATA[C 20 H 30 O5]]> <![CDATA[[M-H2O-H] - ]]> HMDB0003034 C13802 neg down up 66 5.84 330.24 9,10,13-Trihydroxy-11-octadecenoic acid <![CDATA[C 18 H 34 O5]]> <![CDATA[[M-H] - ]]> HMDB0004710 C14835 neg up down 67 5.84 314.22 Progesterone <![CDATA[C 21 H 30 O2]]> <![CDATA[[M+H] + ]]> HMDB0001830 C00410 pos up down 68 5.85 394.20 T863-(DGAT-3) <![CDATA[C 22 H 26 N4O3]]> <![CDATA[[M+H] + ]]> HMDB0247300 - pos down up 69 5.88 432.25 (1R,2R,5R,10R,14R,16R,17S,18R,21S)-17,18-dihydroxy-2,5-dimethyl-8-propan-2-yl-15,20,22-trioxapentacyclo[12.8.0.02,10.05,9.016,21]docosa-8,12-diene-13-carbaldehyde <![CDATA[C 25 H 36 O6]]> <![CDATA[[M-H] - ]]> HMDB0040390 - neg down up 70 5.89 534.26 Pyropheophorbide a <![CDATA[C 33 H 34 N4O3]]> <![CDATA[[M+K-2H] - ]]> HMDB0031150 C18064 neg down up 71 5.89 678.25 3-(2,4-dihydroxyphenyl)-1-{3-[5-(2,4-dihydroxyphenyl)-6-(5-hydroxy-2,2-dimethyl-2H-chromene-6-carbonyl)-3-methylcyclohex-2-en-1-yl]-2,4-dihydroxyphenyl}propan-1-one <![CDATA[C 40 H 38 O 10 ]]> <![CDATA[[M-H] - ]]> HMDB0126375 - neg down up 72 5.89 348.23 5beta-Dihydrocorticosterone <![CDATA[C 21 H 32 O4]]> <![CDATA[[M+H] + ]]> HMDB0006757 C05475 pos up down 73 6.00 371.22 Tamoxifen <![CDATA[C 26 H 29 NO]]> <![CDATA[[M+NH4] + ]]> HMDB0014813 C07108 pos down up 74 6.21 332.24 16alpha-Hydroxypregnenolone <![CDATA[C 21 H 32 O3]]> <![CDATA[[M+H-H2O] + ]]> HMDB0000315 C06390 pos down up 75 6.28 449.31 Glycodeoxycholicacid <![CDATA[C 26 H 43 NO5]]> <![CDATA[[M-H+2Na] + ]]> HMDB0000631 C05464 pos up down 76 6.29 709.44 Fasciculic acid C <![CDATA[C 38 H 63 NO 11 ]]> <![CDATA[[M+H] + ]]> HMDB0036440 NA pos up down 77 6.70 202.03 Xanthotoxol <![CDATA[C 11 H6O4]]> <![CDATA[[M-H] - ]]> HMDB0029457 C00841 neg up down 78 7.02 994.69 Dodecaprenyldiphosphate <![CDATA[C 60 H 100 O7P2]]> <![CDATA[[M+] + ]]> HMDB0012217 NA pos up down Table 6. Metabolic pathway analysis of differentially metabolites Pathway Total Hits Raw -lg() Holm adjust FDR Impact Steroid hormone biosynthesis 87 5 0.0016 2.7970 0.1277 0.1277 0.0967 Riboflavin metabolism 4 1 0.0421 1.3760 1.0000 0.7468 0.5000 Cysteine and methionine metabolism 33 2 0.0467 1.3309 1.0000 0.7468 0.0209 Tyrosine metabolism 42 2 0.0720 1.1429 1.0000 0.9232 0.0004 Phenylalanine metabolism 8 1 0.0825 1.0837 1.0000 0.9232 0.2619 Ascorbate and aldarate metabolism 9 1 0.0923 1.0347 1.0000 0.9232 0.5238 Histidine metabolism 16 1 0.1585 0.7999 1.0000 1.0000 0.0902 Pentose and glucuronate interconversions 19 1 0.1855 0.7317 1.0000 1.0000 0.0964 Fructose and mannose metabolism 20 1 0.1943 0.7116 1.0000 1.0000 0.0331 Citrate cycle (TCA cycle) 20 1 0.1943 0.7116 1.0000 1.0000 0.0463 beta-Alanine metabolism 21 1 0.2030 0.6925 1.0000 1.0000 0.0560 Pyruvate metabolism 23 1 0.2202 0.6573 1.0000 1.0000 0.1914 Glycolysis or Gluconeogenesis 26 1 0.2452 0.6104 1.0000 1.0000 0.0979

Claims

1. The application of Compound Astragalus Spleen-Strengthening Oral Liquid in improving functional dyspepsia in juvenile rats, characterized in that, Includes the following steps: S1. Model Construction and Grouping: A functional dyspepsia model was established in juvenile rats, and the successfully modeled animals were randomly divided into a model control group, a positive drug control group, and at least one compound astragalus spleen-strengthening oral liquid administration group. A blank control group was also set up. S2. Grouped administration: Domperidone was administered by gavage to the positive drug control group, different doses of Compound Astragalus Spleen-Strengthening Oral Liquid were administered by gavage to the Compound Astragalus Spleen-Strengthening Oral Liquid administration group, and the same volume of solvent was administered by gavage to the model control group and the blank control group. The administration was carried out once a day for a predetermined period of time. S3. Pharmacodynamic evaluation: After administration, the general condition of rats in each group was scored, and their gastrointestinal motility index, visceral sensitivity index, serum gastrointestinal hormone level and hypothalamic-pituitary-adrenal axis-related hormone level were measured. S4. Data Analysis: Statistical analysis was performed on the data obtained in step S3 to evaluate the effect of Compound Astragalus Spleen-Strengthening Oral Liquid on improving functional dyspepsia in juvenile rats.

2. The application of the compound astragalus spleen-strengthening oral liquid according to claim 1 in improving functional dyspepsia in juvenile rats, characterized in that, In step S1, the functional dyspepsia model in juvenile rats was constructed using the following method: Six-day-old SD rats were randomly divided into two groups after 7 days of adaptive feeding: a blank control group (n=16) and a model group (n=48). The model group was orally administered 1 mg / mL iodoacetamide solution containing 2% sucrose at a dose of 10 mg / kg. The blank control group was administered an equal volume of 2% sucrose solution by gavage daily for one week. When the rats were 4 weeks old, the model group SD rats were subjected to fatigue induction using a modified multi-platform sleep deprivation method at regular intervals for two consecutive weeks. The blank control group was fed normally.

3. The application of the compound astragalus spleen-strengthening oral liquid according to claim 2 in improving functional dyspepsia in juvenile rats, characterized in that, In step S1, after the modeling was completed, 8 SD rats were randomly selected from the model group and the blank control group for model evaluation. The SD rats in the model group were randomly divided into 5 groups according to their body weight, with 8 rats in each group. These groups were the model control group, the low-dose compound astragalus and spleen-strengthening oral liquid group, the medium-dose compound astragalus and spleen-strengthening oral liquid group, the high-dose compound astragalus and spleen-strengthening oral liquid group, and the positive drug control group.

4. The application of the compound astragalus spleen-strengthening oral liquid according to claim 3 in improving functional dyspepsia in juvenile rats, characterized in that, In step S2, the drug doses in the low-dose, medium-dose, and high-dose groups of the compound astragalus spleen-strengthening oral liquid administration group were 2.52 mL / kg, 5.04 mL / kg, and 10.08 mL / kg, respectively; the dosage of the positive drug control group was 2.7 mg / kg of domperidone per day.

5. The application of the compound astragalus spleen-strengthening oral liquid according to claim 1 in improving functional dyspepsia in juvenile rats, characterized in that, In step S3, the gastrointestinal motility index is evaluated by measuring gastric emptying rate and small intestinal propulsion rate; the visceral sensitivity index is evaluated by abdominal wall withdrawal reflex score.

6. The application of the compound astragalus spleen-strengthening oral liquid according to claim 5 in improving functional dyspepsia in juvenile rats, characterized in that, The method for determining the gastric emptying rate and small intestinal propulsion rate includes: administering a semi-solid paste containing carbon powder to each group of rats by gavage, anesthetizing them after a specified time, and collecting samples. The results are calculated by measuring the amount of residual gastric contents and the propulsion distance of the carbon powder in the small intestine.

7. The application of the compound astragalus spleen-strengthening oral liquid according to claim 1 in improving functional dyspepsia in juvenile rats, characterized in that, In step S3, the determination of serum gastrointestinal hormone levels includes detecting at least one of ghrelin, motilin, gastrin, cholecystokinin, and vasoactive intestinal peptide; the determination of hypothalamic-pituitary-adrenal axis-related hormone levels includes detecting at least one of corticotropin-releasing hormone, adrenocorticotropic hormone, and corticosterone.

8. The application of the compound astragalus spleen-strengthening oral liquid according to claim 1 in improving functional dyspepsia in juvenile rats, characterized in that, In step S2, the predetermined dosing cycle is 21 days.

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