Use of extract of litsea cubeba in preparation of medicine for preventing and treating functional dyspepsia

Abstract

This invention discloses the application of Amomum villosum extract in the preparation of drugs for the prevention and treatment of functional dyspepsia, belonging to the field of biotechnology. The extract, when applied to the prevention and / or treatment of functional dyspepsia, can increase plasma levels of motilin (MTL) and gastrin (GAS), promote gastrointestinal motility, significantly improve gastric emptying rate in functional dyspepsia model mice, improve pathological damage to gastric tissue, restore mitochondrial function in gastric tissue, and regulate the expression of related factors in gastric tissue. Even at low doses, it can effectively increase plasma MTL and GAS levels in loperamide hydrochloride-induced functional dyspepsia model mice, improve gastric emptying rate, effectively improve pathological damage to gastric tissue in these mice, and regulate the expression of related factors in the gastric tissue. The raw materials of this invention are derived from natural products, are easy to collect, and have high safety.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to the application of Amomum villosum extract in the preparation of drugs for the prevention and treatment of functional dyspepsia. Background Technology

[0002] Functional dyspepsia (FD) is a common functional gastrointestinal disorder affecting approximately 10%–30% of the global population. While the exact causes of FD are unclear, it is likely influenced by psychological, physiological, and dietary factors, involving gastrointestinal motility disorders, visceral hypersensitivity, abnormal brain-gut interactions, dietary habits, psychological factors, infections, and genetic factors. Increased intake of high-fat, high-sugar, and high-protein diets, insufficient dietary fiber, extremely irregular eating habits, and the prevalence of late-night snacks lead to increased digestive burden, disrupted digestive rhythms, reduced digestive efficiency, decreased digestive enzyme function, and weakened protective mechanisms of the digestive system. Symptoms of dyspepsia include various forms of gastrointestinal discomfort, such as upper abdominal pain, upper abdominal heartburn, postprandial fullness, early satiety, nausea, and bloating.

[0003] In recent years, the incidence of this disease has shown an upward trend. Because its pathogenesis is not fully understood and its clinical manifestations are diverse, the treatment outcomes for patients are often unsatisfactory. Currently, medications for functional dyspepsia, such as mosapride, domperidone, and other prokinetic drugs, as well as various acid suppressants, gastric mucosal protectants, digestive enzymes, and antidepressants, have some clinical efficacy. However, most of these medications have certain side effects and a single mode of action. Long-term use can lead to low response rates, easy relapse upon discontinuation, and may also cause adverse side effects such as increased risk of infection, secondary hypergastrinemia, and arrhythmias.

[0004] Amomum villosum is an evergreen herbaceous plant belonging to the genus Amomum in the ginger family (Zingiberaceae). It is the dried, mature fruit of Amomum villosum var. yangchunense, Amomum villosum var. yunnanense, or Amomum villosum var. hainanense. It is mainly produced in Guangdong, Guangxi, Yunnan, and Hainan provinces of my country. As a traditional medicinal and edible herb, its use can be traced back to the Sui and Tang dynasties. On one hand, Amomum villosum is widely used as a seasoning, often steamed, boiled, stewed, or simmered with various poultry, livestock, fish, and meats. It not only effectively removes the fishy and gamey smell of meat and imparts a unique aroma to food, but also has the effect of stimulating appetite and relieving indigestion. On the other hand, Amomum villosum also has high medicinal value. It is pungent and warm in nature, and enters the spleen, stomach, and kidney meridians. Its core functions are to resolve dampness and stimulate appetite, warm the spleen and stop diarrhea, regulate qi and calm the fetus. It is one of the "Four Great Southern Medicines" and one of the "Ten Great Yunnan Medicines."

[0005] Amomum villosum is rich in various active ingredients, including volatile oils (borneol acetate, camphor, etc.), flavonoids, polysaccharides, polyphenols, organic acids, and terpenes. It has been reported that Amomum villosum possesses antibacterial, anti-inflammatory, antioxidant, antitumor, hypoglycemic, uric acid-lowering, and gastric mucosal protective activities. Furthermore, Amomum villosum extract is safe and has no mutagenic effects. However, no domestic or international literature has reported on the effects of Amomum villosum extract on functional dyspepsia. Summary of the Invention

[0006] To address the problems of unsatisfactory efficacy, limited action, easy relapse upon discontinuation, and high production costs of existing drugs for treating functional dyspepsia, this invention provides a novel application of Amomum villosum extract in the prevention and treatment of functional dyspepsia, achieving efficient, safe, and low-cost repair of gastric tissue damage and regulation of gastric motility.

[0007] To achieve the above objectives, the present invention provides the following technical solution: Application of Amomum villosum extract in the preparation of a treatment for the prevention of functional dyspepsia.

[0008] The use of Amomum villosum extract to prevent functional dyspepsia caused by loperamide hydrochloride includes (1)-(4): (1) Significantly increased plasma MTL and GAS levels in mice with functional dyspepsia and significantly improved mitochondrial function in mice with functional dyspepsia; (2) Significantly improves gastric emptying rate in functional dyspepsia model mice and improves gastric tissue pathological damage in functional dyspepsia model mice; (3) Amomum villosum extract significantly improves functional dyspepsia by targeting and regulating the expression of CCKBR and Gnaq proteins; (4) Reduce the risk of developing functional dyspepsia.

[0009] Specifically, the application of Amomum villosum extract in the preparation of drugs for the prevention and treatment of functional dyspepsia, wherein the functional dyspepsia is induced by loperamide hydrochloride.

[0010] Furthermore, the amomum extract can increase the levels of motilin MTL and gastrin GAS in plasma.

[0011] Furthermore, the amomum extract can improve gastric emptying rate and improve pathological damage to gastric tissue.

[0012] Furthermore, the Amomum villosum extract can restore mitochondrial function in gastric tissue, increase ATP and NAD content in gastric tissue. + The NADH ratio and the reduction of ROS levels in gastric tissue.

[0013] Furthermore, the Amomum villosum extract can upregulate the expression of CCKBR and Gnaq proteins in gastric tissue.

[0014] Furthermore, the amomum extract is a 70% ethanol extract of amomum.

[0015] This invention presents a series of studies on the effects of Amomum villosum extract in preventing and treating loperamide-induced functional dyspepsia. The studies confirmed that Amomum villosum extract can significantly increase plasma MTL and GAS levels; significantly improve gastric emptying rate in mice with functional dyspepsia; improve gastric tissue pathological damage in mice with functional dyspepsia; and regulate mitochondrial function and the expression of related factors in mice with functional dyspepsia. Overall, it effectively improves gastric tissue damage in mice with functional dyspepsia. The Amomum villosum or its extract provided by this invention has a significant effect on preventing and treating loperamide-induced functional dyspepsia. Furthermore, Amomum villosum is widely distributed, has high safety, is inexpensive, and the raw materials are readily available. Amomum villosum or its extract has promising development and application prospects in the prevention and treatment of functional dyspepsia.

[0016] Preferably, the amomum extract is an ethanol extract of amomum; the ethanol extract of amomum is an extract obtained by extracting amomum with 70% ethanol by mass concentration.

[0017] Preferably, the extract comprises an ethanol extract of Amomum villosum.

[0018] The Amomum villosum extract was prepared by the following method: (1) Take mature cardamom fruits, dry them in the sun, grind them, and sieve them to obtain cardamom powder; (2) Add 70% ethanol solution to the cardamom powder in step (1) for ultrasonic-assisted extraction, and then centrifuge to obtain cardamom residue and supernatant; (3) Add 70% ethanol to the cardamom residue in step (2), extract with ultrasound 2-3 times, and filter the combined supernatant to obtain the filtrate; (4) The filtrate from step (3) is concentrated by rotary evaporation to recover ethanol until an aqueous phase is obtained; the aqueous phase is further concentrated by rotary evaporation to obtain a concentrated solution; the concentrated solution is pre-frozen in a -20°C freezer and then freeze-dried at -70°C for 72 hours using a vacuum freeze dryer to obtain a 70% ethanol solution extract of Amomum villosum (freeze-dried powder), which is then sealed and stored at 4°C.

[0019] Preferably, the sieving in step (1) is a 60-80 mesh sieve.

[0020] Preferably, the mass-to-volume ratio of the cardamom powder or residue to 70% ethanol in steps (2) and (3) is 1 g: 15 mL.

[0021] Preferably, the centrifugation in steps (2) and (3) is performed by centrifuging at a speed of 4000 r / min for 10 to 20 minutes.

[0022] Preferably, the ultrasonic extraction in steps (2) and (3) is performed by ultrasonication at a power of 60-70W for 25-30 minutes.

[0023] Preferably, the rotary evaporation in step (4) is to concentrate the liquid into a paste at 50°C and 0.07~0.08 MPa.

[0024] Preferably, the dosage of the Amomum villosum extract is 200 mg / kg to 600 mg / kg.

[0025] Preferably, the extract includes Amomum villosum extract; oral preparations containing Amomum villosum extract are ingested into the gastrointestinal tract of mammals.

[0026] This invention provides a treatment for functional dyspepsia using Amomum villosum extract as the active ingredient. One or more formulation-acceptable excipients may be added, or the extract may be combined with other active ingredients to synergistically prevent and treat functional dyspepsia. The extract is an oral preparation, which may be a liquid, granule, pill, tablet, capsule, powder, etc., and its applications include food and pharmaceuticals.

[0027] The beneficial effects of this invention are as follows: 1. The extraction method of the present invention is simple and easy to implement, the raw materials are natural products that are easy to collect, and the safety is high. At the same time, it reduces production costs and is suitable for industrial promotion.

[0028] 2. Compared with the single-direction action of mosapride, a treatment for functional dyspepsia, the Amomum villosum extract of this invention forms a multi-target therapeutic network through the synergistic effect of compound active ingredients, and is more effective in improving mitochondrial function and inhibiting inflammation in functional dyspepsia; it can significantly improve the repair of gastric tissue damage and insufficient gastric motility in functional dyspepsia, greatly reducing the burden of functional dyspepsia on the public health system, and has important pioneering application value.

[0029] 3. The Amomum villosum extract of this invention forms a multi-target regulatory network through the synergistic effect of its compound active ingredients. It can significantly increase plasma motilin (MTL) and gastrin (GAS) levels, promote gastrointestinal motility, significantly improve gastric emptying rate in mice with loperamide hydrochloride-induced functional dyspepsia, and effectively improve gastric tissue pathological damage. At the same time, it can restore mitochondrial function in gastric tissue and increase ATP and NAD content. + It improves the NADH ratio, reduces ROS levels, and upregulates the expression of gastric tissue target proteins CCKBR and Gnaq. Compared with single-target drugs such as mosapride, it is more effective in repairing gastric tissue damage, improving gastric motility, and protecting mitochondrial function. Its mechanism of action is clear and its efficacy is definite.

[0030] 4. The Amomum villosum extract of the present invention provides a new source of drugs for the prevention and treatment of functional dyspepsia. It is characterized by high efficiency, safety and low cost. It can not only fill the gap in clinically effective drugs for functional dyspepsia, but also provide new material options for the research and development of food and medicine. It has extremely high economic value and clinical promotion value. Attached Figure Description

[0031] Figure 1 This is the total ion chromatogram in negative ion mode for principal component analysis of the Amomum villosum ethanol extract of this invention using UPLC-MS / MS. Figure 2 Venn diagram for blood component analysis of Amomum villosum ethanol extract of this invention; Figure 3 The figure shows the changes in MTL and GAS levels in the plasma of model mice after applying the Amomum villosum ethanol extract of the present invention; Figure A shows the plasma MTL level, and Figure B shows the GAS level. Figure 4 This invention illustrates the changes in mitochondrial-related indicators in the gastric tissue of model mice when the ethanol extract of Amomum villosum was applied. Figure A shows the ATP content in the gastric tissue, and Figure B shows the NAD content in the gastric tissue. + / NADH ratio, Figure C shows the expression level of ROS in gastric tissue; Figure 5 The images show the changes in gastric emptying rate and the results of gastric tissue pathological sections of mice treated with the ethanol extract of Amomum villosum in this invention; Figure A shows the changes in gastric emptying rate in mice, and Figure B shows the H&E staining results of gastric tissue in each group of mice. Figure 6 The figures show the expression of target proteins in the gastric tissue of model mice when the Amomum villosum ethanol extract of this invention is applied; Figure A shows the expression of CCKBR and Figure B shows the expression of Gnaq. Detailed Implementation

[0032] The technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of this application, and not all of the embodiments. Based on the embodiments of this application, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of this application.

[0033] Example 1: Preparation and component analysis of Amomum villosum extract

[0034] (1) Preparation of Amomum villosum ethanol extract Mature Amomum villosum was harvested from Xishuangbanna, Yunnan Province. After drying, grinding, and passing through a 60-mesh sieve, Amomum villosum powder was obtained. 200g of Amomum villosum powder was weighed and added to 70% ethanol (mass-volume ratio of Amomum villosum powder to 70% ethanol: 1g:15mL). The mixture was ultrasonically extracted for 30min at 60W, then soaked at 4℃ for 12h, followed by centrifugation for 10min (4000r / min). The supernatant was retained, and the Amomum villosum residue was extracted once more using the same method. After centrifugation, the supernatants were combined and filtered. The combined supernatant was placed in a rotary evaporator and concentrated to a paste-like liquid at 50℃ and 0.07~0.08MPa. The concentrate was pre-frozen at -20℃ and then freeze-dried at -70℃ for 72h using a vacuum freeze dryer. This yielded the freeze-dried Amomum villosum extract powder.

[0035] (2) Qualitative analysis and blood-entry component analysis of the main chemical components of Amomum villosum extract. Qualitative analysis of the main components of Amomum villosum extract was performed using UPLC-MS / MS. An Agilent SB-C18 column (1.8 μm, 2.1 mm × 100 mm) was used; mobile phase A was ultrapure water (containing 0.1% formic acid), and mobile phase B was acetonitrile (containing 0.1% formic acid); the elution program was: 0 min, 5% B; 9 min, 95% B; 10 min, 95% B; 11 min, 5% B. The column temperature was 40℃; the injection volume was 2 μL, and the flow rate was 0.35 mL / min. The analytical results are as follows: Figure 1 As shown in the diagram; the analysis of the blood components of Amomum villosum extract was presented in Venn diagram form, and the results are as follows. Figure 2 As shown. AVE refers to the ethanol extract of Amomum villosum; Control refers to the mouse gavage group administered pure water; AVEH refers to the mouse gavage group administered Amomum villosum extract.

[0036] Depend on Figure 1 It is known that the main compounds contained in the ethanol extract of Amomum villosum include terpenoids such as hydroxycarvone, cis-ocimene, trichoderma A, and curcuminone; flavonoids such as baicalin, apigenin, quercetin, and isoquercitrin; as well as other phenolic acids, amino acids and their derivatives, organic acids, and other compounds.

[0037] Depend on Figure 2 It is known that there are 236 blood-entering components in the ethanol extract of Amomum villosum, and some of these components are shown in Table 1.

[0038]

[0039] Example 2: Evaluation of the activity of Amomum villosum extract in preventing and treating functional dyspepsia 2.1 Establishment and experimental procedure of a mouse model of functional dyspepsia A mouse model of functional dyspepsia was established using male C57BL / 6J mice (8 weeks old, weighing approximately 20g) induced by intraperitoneal injection of loperamide hydrochloride (10mg / kg). After the last gavage, the mice were fasted for 20 hours. Except for the control group, all mice received an intraperitoneal injection of loperamide hydrochloride. Half an hour later, all mice, including the control group, were administered activated charcoal via gavage.

[0040] The efficacy of the 70% ethanol extract of Amomum villosum obtained by gavage in Example 1 against a mouse model of functional dyspepsia induced by loperamide hydrochloride was evaluated. Mice were acclimatized to a standard environment for 7 days after purchase and fed a standard diet. The specific experimental procedure and grouping are as follows: Control group: Administered an equal volume of normal saline by gavage for 7 consecutive days, once a day. After the last gavage, fasted for 20 hours and administered activated charcoal by gavage. Model group: For 7 consecutive days, the same volume of normal saline was administered by gavage once a day. After the last gavage, the patient was fasted for 20 hours and then injected intraperitoneally with loperamide hydrochloride (10 mg / Kg) according to body weight. Activated charcoal was administered by gavage 30 minutes later. The low-dose group of Amomum villosum (AVE(200)): 200 mg / kg of 70% ethanol extract of Amomum villosum was administered by gavage once a day for 7 consecutive days. After the last gavage, the patient was fasted for 20 hours and then injected intraperitoneally with loperamide hydrochloride (10 mg / kg) according to body weight. Activated charcoal was administered by gavage 30 minutes later. High-dose Amomum villosum group (AVE(600)): 600 mg / kg of Amomum villosum 70% ethanol extract was administered by gavage for 7 consecutive days, once a day. After the last gavage, the patient was fasted for 20 hours and injected intraperitoneally with loperamide hydrochloride (10 mg / kg) according to body weight. Activated charcoal was administered by gavage 30 minutes later. Positive control group (MOS): Mosapride 3 mg / kg was administered by gavage once a day for 7 consecutive days. After the last gavage, the patient was fasted for 20 hours and then injected intraperitoneally with loperamide hydrochloride (10 mg / kg) according to body weight. Activated charcoal was administered by gavage 30 minutes later. 2.2 Detection of the effects of active ingredients of Amomum villosum on the levels of MTL and GAS in plasma of model mice and the expression of mitochondrial functional indicators in gastric tissue. 2.2.1 After collecting blood from the eyeballs of mice in each group using anticoagulant tubes, the supernatant was collected by centrifugation at 2000 r / min for 10 min. The MTL and GAS levels in the plasma of each group of mice were detected using MTL and GAS kits. After collecting blood from each group of mice, the stomach was aseptically removed, washed with sterile pre-cooled PBS, and 30 mg of tissue was taken. 100 μL of the corresponding extraction solution from each kit was added, homogenized at 4℃, centrifuged, and the supernatant was collected. The ATP and NAD values ​​in the mitochondria of the stomach tissue of each group of mice were detected using the corresponding kits. + / NADH, ROS expression status.

[0041] 2.2.2 Experimental Results Motilin (MTL) is a 22-amino acid gastrointestinal peptide hormone secreted by Mo cells (endocrine cells) of the duodenal and upper jejunal mucosa. It is a core hormone regulating gastrointestinal motility, gastric emptying, and small intestinal propulsion, and is known as the "gastrointestinal motility initiator." Gastrin (GAS) is a gastrointestinal peptide hormone mainly secreted by G cells in the gastric antrum. It stimulates the apical cells of the stomach to secrete gastric acid, participates in gastric motility, and promotes gastric mucosal growth and repair.

[0042] ATP, as a biological energy currency, is widely present in animals, plants, microorganisms, and cultured cells. It is mainly produced by mitochondria through aerobic respiration and directly provides energy for muscle contraction, substance transport, protein synthesis, and cell repair. Energy charge is a key parameter describing the cellular energy metabolism state; measuring ATP content yields energy charge, indirectly reflecting the energy metabolism state.

[0043] Nicotinamide adenine dinucleotide (NAD) is a coenzyme present in all cells, existing in both oxidized and reduced forms, NAD and NADH. + It is both a coenzyme that transfers electrons in redox reactions and a substrate for many enzymes, participating in intracellular reactions. NAD + It plays an important role in cells and the body; its synthesis and degradation, as well as its products, participate in cell apoptosis, metabolic regulation, and gene expression regulation. Furthermore, NAD+... ＋ The reduction of NAD+ is one of the main factors contributing to cell death. + The importance of NAD+ in regulating cellular redox states and its functions in signaling pathways and transcription make the enzymes synthesized and consumed by NAD+ potential drug targets for various diseases. Therefore, NAD+ can be measured... + The ratio of NADH to gastric tissue reflects the degree of oxidative stress and the level of mitochondrial function and energy metabolism.

[0044] Reactive oxygen species (ROS) play a double-edged sword role in mitochondrial function, exhibiting a concentration-dependent dual function: at physiological concentrations, they are key signaling molecules, while excesses can lead to oxidative damage. Measuring ROS levels is crucial for assessing the effectiveness of improving mitochondrial function.

[0045] Depend on Figure 3 As shown in A and B, compared with the control group, the plasma MTL and GAS levels in the model group mice were significantly decreased. After administration of high-dose and low-dose groups of Amomum villosum extract, the MTL and GAS levels significantly increased. Figure 4 As can be seen from A and B, compared with the control group, the ATP and NAD content in the mitochondria of the gastric tissue of the model group mice was significantly higher. +The NADH / γ ratio was significantly decreased, but recovered markedly after administration of both high-dose and low-dose Amomum villosum extract groups; Figure 4 As can be seen from C, compared with the control group, the mitochondrial ROS level in the gastric tissue of the model group mice was significantly increased, and it decreased significantly after being given high-dose and low-dose groups of Amomum villosum extract. It is statistically significant.

[0046] 2.3 Detection of the effects of active ingredients on gastric emptying rate and gastric histopathology in model mice 2.3.1 Half an hour after gavage with activated charcoal, the intact stomachs of mice in each group were aseptically removed, gently rinsed and dried with sterile pre-cooled PBS, and the total stomach weight was weighed. The stomach was cut open along the greater curvature, and the activated charcoal and other stomach contents were rinsed away with sterile pre-cooled PBS. The net stomach weight was then measured, and the gastric emptying rate of the mice was calculated.

[0047] Gastric emptying rate (%) = [1 - (total gastric weight - net gastric weight) / gavage volume] × 100% 2.3.2 After rinsing the stomachs of mice in each group with sterile pre-cooled PBS, one-third of the rinsed gastric antrum tissue was cut off, fixed, and prepared into paraffin sections. After HE staining, the pathological changes in the gastric tissue structure were observed under a microscope.

[0048] Methods for preparing paraffin sections: Material collection: Fresh gastric tissue was collected and fixed in 10% formalin solution for at least 24 hours. The gastric tissue was removed from the fixative and trimmed in a fume hood using a scalpel. The trimmed tissue and corresponding labels were placed in a dehydration box. Dehydration and wax impregnation: Place the dehydration box into the dehydrator and dehydrate it sequentially with alcohol in a gradient. 75% alcohol for 4 hours, 85% alcohol for 2 hours, 90% alcohol for 2 hours, 95% alcohol for 1 hour, anhydrous ethanol I for 30 minutes, anhydrous ethanol II for 30 minutes, benzene for 5-10 minutes, xylene I for 5-10 minutes, xylene II for 5-10 minutes, molten paraffin I at 65℃ for 1 hour, molten paraffin II at 65℃ for 1 hour, molten paraffin III at 65℃ for 1 hour; Embedding: The tissue impregnated with wax is embedded in an embedding machine. First, the molten wax is placed into the embedding frame. Before the wax solidifies, the tissue is taken out from the dehydration box, placed into the embedding frame according to the requirements of the embedding surface, and labeled accordingly. It is then cooled on a -10℃ freezing stage. After the wax solidifies, the wax block is removed from the embedding frame and trimmed. Sectioning: Place the trimmed wax block on a paraffin microtome and section it to a thickness of 4μm; float the section on 40℃ warm water in a slide spreader to flatten the tissue, pick up the tissue on a glass slide, bake the slide in a 60℃ oven, dry the wax with water and bake it until it melts, then remove it and store it at room temperature for later use.

[0049] The HE staining method is as follows: Dewaxing paraffin sections to water: Immerse the sections in xylene I for 20 min, xylene II for 20 min, anhydrous ethanol I for 5 min, anhydrous ethanol II for 5 min, and 75% ethanol for 5 min, then rinse with tap water; Hematoxylin staining: Immerse the sections in hematoxylin staining solution for 3-5 minutes, wash with tap water, differentiate with differentiation solution, wash with tap water, blue back solution, and rinse with running water. Eosin staining: The sections were dehydrated in 85% and 95% graded alcohol solutions for 5 min each, and then stained in eosin staining solution for 5 min.

[0050] Dehydration and mounting: The sections were sequentially immersed in anhydrous ethanol I for 5 min, anhydrous ethanol II for 5 min, anhydrous ethanol III for 5 min, xylene I for 5 min, and xylene II for 5 min. After clearing, the sections were mounted with neutral resin. Microscopic examination, image acquisition and analysis 2.3.3 Experimental Results Gastric emptying rate is a core indicator for assessing gastric motility and emptying function, reflecting the stomach's contraction, peristalsis, and pyloric regulation capabilities. Mice in each group were weighed 30 minutes after being administered activated charcoal by gavage.

[0051] Gastric emptying rate (%) = [1 - (total gastric weight - net gastric weight) / gavage volume] × 100% HE staining was used to observe the structure of mouse gastric tissue. Figure 5 As shown in Figure A, compared with the control group, the gastric emptying rate in the model group was significantly reduced, and it recovered significantly after administration of both high-dose and low-dose Amomum villosum extract. Figure 5 As shown in Figure B, compared with the control group, the model group exhibited significant pathological damage to the gastric tissue, with loosely arranged gastric glands and marked mucosal damage and inflammatory infiltration. After administration of high-dose and low-dose Amomum villosum extract, the pathological damage was significantly improved, with an improvement effect similar to that of the positive control group. This indicates that Amomum villosum extract can significantly increase the gastric emptying rate and significantly improve gastric tissue pathological damage in mice with functional dyspepsia.

[0052] 2.4 Detection of the effect of active ingredients on the expression of target proteins in the gastric tissue of model mice 2.4.1 The expression of target proteins CCKBR and Gnaq in the gastric tissue of model mice was detected by Western blotting.

[0053] 2.4.2 Experimental Results CCKBR is a G protein-coupled receptor (GPCR) that is highly expressed in the stomach. Belonging to class A GPCRs, it is mainly distributed in gastrointestinal parietal cells, enterochromaffin-like cells, and the central nervous system. It can bind both cholecystokinin (CCK) and gastrin with high affinity, playing a crucial role in regulating gastric acid secretion, gastrointestinal motility, mucosal proliferation, and visceral sensitivity. Gnaq is a core member of the heterotrimeric G protein α subfamily, widely expressed in the gastrointestinal tract and nervous tissue. Upon receptor activation, it undergoes GDP-GTP exchange and dissociation, activating phospholipase (PLCβ), which initiates downstream signal transduction, ultimately regulating cell secretion, contraction, and proliferation, producing a series of biological effects.

[0054] Depend on Figure 6 It can be seen that, compared with the control group, the expression of CCKBR in the model group was significantly reduced, and the expression was significantly restored after administration of high-dose and low-dose groups of Amomum villosum extract; compared with the control group, the expression of Gnaq in the model group was significantly reduced, and the expression was significantly restored after administration of high-dose and low-dose groups of Amomum villosum extract; indicating that the active ingredients of Amomum villosum can significantly regulate the expression of gastric tissue-related factors in functional dyspepsia model mice. It is statistically significant.

[0055] In summary, Amomum villosum extract significantly increased plasma MTL and GAS levels in mice with functional dyspepsia; significantly improved gastric emptying rate and pathological damage in gastric tissue; significantly restored mitochondrial function in gastric tissue of model mice; and regulated the expression of target proteins in gastric tissue of model mice. The Amomum villosum extract provided by this invention has a significant effect on preventing and treating gastric tissue damage caused by functional dyspepsia, and the raw material is derived from natural products, making it highly safe, low-cost, and suitable for industrial production and market application.

[0056] In addition, the preparation method of Amomum villosum ethanol extract, by changing the sieve mesh size of mature Amomum villosum powder to 80 mesh or 100 mesh and ultrasonicating at 70W power for 25 min or at 80W power for 28 min, also yields the above-mentioned Amomum villosum ethanol extract.

[0057] The specific embodiments of the invention have been described in detail above, but these are merely examples, and the invention is not limited to the specific embodiments described above. For those skilled in the art, any equivalent modifications or substitutions to the invention are also within the scope of this invention. Therefore, all equivalent transformations, modifications, and improvements made without departing from the spirit and principles of this invention should be covered within the scope of this invention.

Claims

1. The application of Amomum villosum extract in the preparation of drugs for the prevention and treatment of functional dyspepsia, wherein the functional dyspepsia is induced by loperamide hydrochloride.

2. The application as described in claim 1, characterized in that, The amomum villosum extract can increase the levels of motilin MTL and gastrin GAS in plasma.

3. The application as described in claim 1, characterized in that, The amomum extract can improve gastric emptying rate and improve pathological damage to gastric tissue.

4. The application as described in claim 1, characterized in that, The Amomum villosum extract can restore mitochondrial function in gastric tissue and increase ATP and NAD content in gastric tissue. + The NADH ratio and the reduction of ROS levels in gastric tissue.

5. The application as described in claim 1, characterized in that, The Amomum villosum extract can upregulate the expression of CCKBR and Gnaq proteins in gastric tissue.

6. The application as described in claim 1, characterized in that, The amomum extract is a 70% ethanol extract of amomum.

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