Dha-gynostemma pentaphyllum-panax notoginseng compound for repairing intestinal mucosal damage of celiac disease

Abstract

The present application relates to the technical field of biological medicine, and provide DHA-gardenia oil-gynostemma pentaphyllum compound for repairing celiac disease intestinal mucosa damage, the DHA-gardenia oil-gynostemma pentaphyllum compound is composed of docosahexaenoic acid, gardenia oil and gynostemma pentaphyllum extract, and the mass ratio of docosahexaenoic acid, gardenia oil and gynostemma pentaphyllum extract is 1-10:10-25:20-30.DHA-gardenia oil-gynostemma pentaphyllum compound is prepared by mixing docosahexaenoic acid and gardenia oil under nitrogen protection, and then mixing with gynostemma pentaphyllum extract, which can repair the small intestinal villus atrophy and crypt hyperplasia caused by gluten protein, enhance the mucus barrier and epithelial tight junction, promote the production of short-chain fatty acids and regulate the local immune response of intestinal tract, and has auxiliary protection effect on celiac disease intestinal mucosa damage.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, and in particular relates to a DHA-gardenia oil-Gynostemma pentaphyllum compound for repairing celiac disease intestinal mucosal damage. Background Technology

[0002] Celiac disease is a chronic inflammatory autoimmune bowel disease caused by gluten intake in genetically susceptible individuals. Typical pathological changes include atrophy of small intestinal villi, crypt hyperplasia, disruption of tight junctions between intestinal epithelial cells, and loss of intestinal mucosal barrier function. Currently, a gluten-free diet is the main intervention for celiac disease, but adherence is poor and complete repair of the intestinal mucosa is difficult to achieve; many patients still experience persistent intestinal mucosal damage.

[0003] Studies have shown that sprouted barley enzymes can efficiently degrade secalin and gliadin peptides in rye, and enzyme pretreatment can improve secalin-induced increased epithelial cell permeability and damage to tight junction proteins. However, enzymatic hydrolysis is costly, complex, and cannot directly repair damaged intestinal mucosal structures. Other studies have found that glycosaminoglycans can reduce gliadin-induced interleukin-1β production, but lack direct repair capabilities for existing intestinal epithelial cell membrane damage and tight junction structure disruption. Summary of the Invention

[0004] To address the shortcomings of existing technologies, this invention provides a DHA-gardenia oil-Gynostemma pentaphyllum compound for repairing celiac disease intestinal mucosal damage, with the aim of solving the problems mentioned in the background art.

[0005] In a first aspect, the present invention provides a DHA-gardenia oil-Gynostemma pentaphyllum compound for repairing celiac disease intestinal mucosal damage, wherein the DHA-gardenia oil-Gynostemma pentaphyllum compound is composed of docosahexaenoic acid, gardenia oil and Gynostemma pentaphyllum extract, and the mass ratio of docosahexaenoic acid:gardenia oil:Gynostemma pentaphyllum extract is 1-10:10-25:20-30.

[0006] Furthermore, by mass ratio, docosahexaenoic acid : gardenia oil : gynostemma pentaphyllum extract = 5 : 20 : 30.

[0007] Furthermore, the active ingredient of the Gynostemma pentaphyllum extract is Gynostemma pentaphyllum saponin.

[0008] Furthermore, the preparation method of the Gynostemma pentaphyllum extract specifically includes the following steps: after pulverizing the whole herb of Gynostemma pentaphyllum, it is extracted twice by heating and reflux with 70% ethanol as solvent, the extracts are combined and concentrated; after purification by macroporous adsorption resin, the fraction rich in Gynostemma pentaphyllum saponins is collected, and after concentration and drying, the Gynostemma pentaphyllum extract is obtained.

[0009] Furthermore, the reflux extraction temperature is 90°C, and the reflux extraction time is 1.5 hours.

[0010] Furthermore, the gardenia oil is an oil obtained by supercritical CO2 extraction of gardenia fruit, wherein the crocin content is ≥2.0% and the geniposide content is ≤0.5%.

[0011] Furthermore, the conditions for supercritical CO2 extraction are: extraction pressure 25-35 MPa, extraction temperature 40-55℃, and extraction time 2-4 hours.

[0012] Secondly, the present invention provides a method for preparing a DHA-gardenia oil-Gynostemma pentaphyllum compound for repairing celiac disease intestinal mucosal damage, comprising the following steps: first, mixing the prescribed amount of docosahexaenoic acid and gardenia oil evenly under nitrogen protection to obtain a mixed oil phase; then, adding Gynostemma pentaphyllum extract to the mixed oil phase, grinding or stirring thoroughly to ensure that the Gynostemma pentaphyllum extract is evenly dispersed and adsorbed into the mixed oil phase, and sieving to obtain the DHA-gardenia oil-Gynostemma pentaphyllum compound.

[0013] Thirdly, the present invention provides the application of a DHA-gardenia oil-Gynostemma pentaphyllum compound for repairing celiac disease in the preparation of a drug that helps protect against celiac disease intestinal mucosal damage.

[0014] Furthermore, the drug is formulated into clinically acceptable tablets, pills, capsules, suspensions, gels, solutions, emulsions, ointments, or lotions.

[0015] Furthermore, the drug also includes pharmaceutically acceptable excipients.

[0016] Furthermore, the excipients include microcrystalline cellulose, sodium carboxymethyl starch, magnesium stearate, or silicon dioxide.

[0017] The present invention has the following beneficial effects: (1) DHA-gardenia oil-Gynostemma pentaphyllum compound can repair intestinal villus atrophy and crypt hyperplasia caused by gluten protein, improve the orderly arrangement of intestinal epithelial cells, increase goblet cell density, and enhance intestinal mucosal barrier function.

[0018] (2) The DHA-gardenia oil-Gynostemma pentaphyllum compound contains three core active ingredients, each with specific physicochemical properties and biological activities to synergistically protect against celiac disease-induced intestinal mucosal damage. Docosahexaenoic acid (DHA) is incorporated into the phospholipid bilayer of the cell membrane, improving membrane fluidity and antioxidant capacity, and inhibiting lipid peroxidation. Gardenia oil is rich in carotenoids such as crocin, which can not only directly scavenge reactive oxygen species in the intestine and reduce oxidative stress damage to the intestinal epithelium, but also upregulate the expression of tight junction proteins Occludin and ZO-1, reduce intestinal mucosal permeability, and block the penetration of gluten protein fragments into the submucosa; more importantly, crocin, as a natural antioxidant, can effectively protect DHA from oxidative damage during preparation and storage, ensuring the stability of the compound from a chemical perspective. Gynostemma pentaphyllum extract (with a ginsenoside content ≥80%) possesses an amphiphilic structure, enabling it to integrate into the intestinal epithelial cell membrane, enhance membrane stability, promote the proliferation of damaged epithelial cells, and accelerate villous repair. Simultaneously, its amphiphilic nature allows it to act as a natural emulsifier in formulations, promoting the uniform dispersion of docosahexaenoic acid (DHA) and gardenia oil. These three components work synergistically to provide auxiliary protection against celiac disease-related intestinal mucosal damage from four levels: membrane stability, antioxidant activity, barrier repair, and formulation stability.

[0019] (3) The preparation method of DHA-gardenia oil-Gynostemma pentaphyllum compound is to first mix the two oils under nitrogen protection and then mix them with Gynostemma pentaphyllum extract powder. This process minimizes the oxidation of docosahexaenoic acid during the preparation process. At the same time, the amphiphilicity of Gynostemma pentaphyllum saponins promotes the uniform dispersion of the oil phase and improves the physical stability and bioavailability of the compound. Attached Figure Description

[0020] Exemplary embodiments of the present invention can be more fully understood by referring to the following figures: Figure 1 This is an H&E staining (hematoxylin-eosin) staining image of an intestinal section from Example 4 of the present invention. In the image, the red line segment represents the villus height measurement line, and the blue line segment represents the crypt depth measurement line.

[0021] Figure 2 This is a statistical chart of the intestinal villus-crypt ratio from Example 4 of the present invention. The NDP.view2 software was used to measure the villus height and crypt depth of all morphologically intact and well-oriented villi and their adjacent crypts in H&E-stained sections. The villus height to crypt depth ratio was calculated by dividing the villus height by the corresponding crypt depth. Five mice were used in each group for measurement. Different letters represent statistically significant differences.

[0022] Figure 3This is an AB-PAS (Alcian Blue-Periodic Acid Schiff) staining image of an intestinal section from Example 4 of the present invention. After AB-PAS staining, the acidic mucus in the goblet cells turns dark blue. The dark blue circular or oval structures distributed along the intestinal villi epithelium can be used as markers for identifying goblet cells and are indicated by arrows in the image.

[0023] Figure 4 This is a statistical graph showing the relative expression levels of intestinal Muc2 (mucin 2) mRNA in Example 4 of the present invention. Different letters represent statistically significant differences.

[0024] Figure 5 This is a statistical chart of the relative mRNA expression levels of intestinal Tjp1 (tight junction protein 1) in Example 4 of the present invention. Different letters represent statistical differences.

[0025] Figure 6 This is a statistical graph showing the relative mRNA expression levels of Ocln (closure protein) in the intestine in Example 4 of this invention. Different letters represent statistically significant differences.

[0026] Figure 7 This is a statistical chart of acetic acid content in the cecal contents of Example 4 of the present invention. The test results are expressed as the mass of short-chain fatty acids contained in each milligram of cecal contents, in ng / mg. Different letters represent statistical differences.

[0027] Figure 8 This is a statistical chart of butyric acid content in the cecal contents of Example 4 of the present invention. The test results are expressed as the mass of short-chain fatty acids contained in each milligram of cecal contents, in ng / mg. Different letters represent statistical differences.

[0028] Figure 9 It is the mesenteric lymph node (MLN) in Embodiment 4 of the present invention. The cell proportion chart shows that different letters represent statistically significant differences.

[0029] Figure 10 It is the mesenteric lymph node (MLN) in Embodiment 4 of the present invention. The cell proportion chart shows that different letters represent statistically significant differences. Detailed Implementation

[0030] To make the technical problems to be solved, the technical solutions, and the beneficial effects of the present invention clearer, the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the present invention and are not intended to limit the present invention.

[0031] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains; the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the invention.

[0032] Example 1: Preparation of Gynostemma pentaphyllum extract One kilogram of whole Gynostemma pentaphyllum herb was pulverized and passed through a 40-mesh standard sieve. Using 70% ethanol as the extraction solvent, the extract was added at a material-to-liquid ratio of 1 g:15 mL and refluxed at 90℃ for 1.5 h. This extraction was repeated twice. All extracts were combined and filtered. The filtrate was concentrated to a thick extract state after ethanol recovery under reduced pressure. The resulting extract was dissolved in water to prepare a loading solution with a concentration of 1 g / mL (based on crude drug). This solution was loaded onto a pretreated AB-8 macroporous adsorption resin column (2 L column volume). The column was eluted sequentially with 3 column volumes of deionized water, 5 column volumes of 50% ethanol, and 4 column volumes of 70% ethanol. The eluent corresponding to the 70% ethanol concentration was collected, concentrated under reduced pressure, and spray-dried to obtain 125 g of Gynostemma pentaphyllum extract. HPLC analysis showed that the mass fraction of Gynostemma pentaphyllum saponins in the extract was 83.6%.

[0033] Example 2: Preparation of Gardenia Oil One kilogram of dried gardenia fruit was crushed and passed through a 40-mesh sieve. The powder was then placed in a supercritical CO2 extraction vessel and extracted for 3 hours at an extraction pressure of 30 MPa and an extraction temperature of 45°C. The extract was collected and separated under two stages of reduced pressure to obtain 168 g of gardenia oil. HPLC analysis showed that the crocin content was 2.5% and the geniposide content was 0.25%.

[0034] Example 3: Preparation of DHA-Gardenia Oil-Gynostemma pentaphyllum compound (1) Weigh out docosahexaenoic acid, gardenia oil, and gynostemma pentaphyllum extract according to formulas A, B, and C respectively: Formula A: docosahexaenoic acid: gardenia oil: gynostemma pentaphyllum extract = 1:10:20; Formula B, docosahexaenoic acid: gardenia oil: gynostemma pentaphyllum extract = 5:20:30; Formula C, docosahexaenoic acid: gardenia oil: gynostemma pentaphyllum extract = 10:25:30; (2) First, put the prescribed amount of docosahexaenoic acid and gardenia oil into a mixer, stir at 25 r / min for 15 minutes under nitrogen protection, mix evenly, and obtain a mixed oil phase; (3) Under continuous nitrogen protection, slowly add the Gynostemma pentaphyllum extract powder to the mixed oil phase, stir or grind for 30 minutes to make the Gynostemma pentaphyllum extract evenly dispersed and fully adsorbed into the mixed oil phase, and pass through an 80-mesh sieve to obtain the DHA-gardenia oil-Gynostemma pentaphyllum compound. (4) Mix the DHA-gardenia oil-Gynostemma pentaphyllum compound with excipients (microcrystalline cellulose, sodium carboxymethyl starch, magnesium stearate and silicon dioxide) evenly and compress it into tablets.

[0035] The specific weight of each tablet for each formula is as follows: Formula A: Docosahexaenoic acid 8.9mg, Gardenia oil 88.7mg, Gynostemma pentaphyllum extract 177.4mg, Microcrystalline cellulose 140mg, Sodium carboxymethyl starch 30mg, Magnesium stearate 15mg, Silicon dioxide 40mg; Formula B: Docosahexaenoic acid 25.0mg, Gardenia oil 100.0mg, Gynostemma pentaphyllum extract 150.0mg, Microcrystalline cellulose 150mg, Sodium carboxymethyl starch 30mg, Magnesium stearate 15mg, Silica 30mg; Formula C: Docosahexaenoic acid 42.3mg, Gardenia oil 105.8mg, Gynostemma pentaphyllum extract 126.9mg, Microcrystalline cellulose 155mg, Sodium carboxymethyl starch 30mg, Magnesium stearate 15mg, Silicon dioxide 25mg; In Formula A, the proportion of Gynostemma pentaphyllum extract powder is relatively high (powder / oil ratio is about 2.0). The powder itself can adsorb most of the oil. Appropriately increasing the amount of silica (40mg) can further improve the powder flowability and compressibility. In Formula C, the total amount of oil is the highest (about 148.1mg / tablet), and the amount of Gynostemma pentaphyllum extract powder is relatively low (powder / oil ratio is about 0.86). The powder itself has insufficient oil adsorption capacity. Therefore, the amount of microcrystalline cellulose (155mg) is appropriately increased to enhance the adsorption of excess oil and balance the tablet weight. At the same time, the amount of silica is adjusted to 25mg to play a flow-aiding role. Example 4: Protective effect of DHA-gardenia oil-Gynostemma pentaphyllum compound on intestinal mucosal damage in celiac disease mice (1) Animal model establishment: 80 healthy C57 female mice were randomly divided into 8 groups: normal control group (blank group), model control group, docosahexaenoic acid control group, gardenia oil control group, gynostemma pentaphyllum extract control group and three experimental groups (formulas A, B and C in Example 3).

[0036] Except for the normal control group, mice in the other groups were administered 500 μg of gliadin digestible peptides and 25 μg of cholera toxin by gavage once a week for three consecutive weeks. For the next two weeks, they were administered 200 μL of 0.5% CMC-Na suspension containing 10 mg of gliadin by gavage three times a week, while the normal control group was administered an equal volume of 0.5% CMC-Na solution (200 μL) by gavage.

[0037] (2) Administration method: During the modeling period, each group was administered the corresponding drug by gavage once a day. The gavage volume for each group was calculated at 10 mL / kg body weight. For example, for a mouse weighing 20 g, the single gavage volume was 200 μL. The normal control group and the model control group were administered 0.5% CMC-Na solution by gavage; the dose of docosahexaenoic acid control group was 25 mg / kg, calculated based on the actual DHA content; the dose of gardenia oil control group was 100 mg / kg, calculated based on the actual gardenia oil content; the dose of Gynostemma pentaphyllum extract control group was 150 mg / kg, calculated based on the actual Gynostemma pentaphyllum extract content; the experimental groups of formulations A, B, and C were all administered the drug at a total tablet weight of 500 mg / kg as described in Example 3. The tablets were thoroughly crushed before administration. The three single-component control groups and the experimental groups of formulations A, B, and C were all prepared into suspensions using 0.5% CMC-Na solution as the solvent. The dosage unit mg / kg refers to milligrams per kilogram of mouse body weight, which is the amount of drug administered per kilogram of mouse body weight. The dosages for the three single-component control groups were determined based on Formulation B tablets: each Formulation B tablet has a total mass of 500 mg, containing 25.0 mg of docosahexaenoic acid, 100.0 mg of gardenia oil, and 150.0 mg of Gynostemma pentaphyllum extract. Therefore, when Formulation B tablet powder is administered at 500 mg / kg, the actual dosages of DHA, gardenia oil, and Gynostemma pentaphyllum extract are 25, 100, and 150 mg / kg, respectively, which are the same as the dosages of the corresponding single-component control groups. Formulation B was used to compare with the three single-component control groups to evaluate the protective effect of the compound under the condition of the same dosage of each component; Formulation A and Formulation C were used to examine the protective effects of compound formulations with different component ratios.

[0038] (3) Detection indicators: 1) Morphological examination of ileum tissue: Pathological sections were prepared from mouse ileum tissue, and the morphology of ileal villi and crypts was observed using hematoxylin-eosin (H&E) staining. Villi with intact tissue structure and good cut direction and their corresponding crypts were selected, and villi height and crypt depth were measured using NDP.view2 software. Among them, villi height is the distance from the villi tip to the villi-crypt junction, and crypt depth is the distance from the crypt opening to the crypt base. The ratio of villi height to crypt depth was calculated according to the following formula: villi-crypt ratio = villi height ÷ corresponding crypt depth. 2) Pathological sections were prepared from mouse ileum tissue, and the morphology and distribution of goblet cells were observed using AB-PAS staining. After staining, the acidic mucus in the goblet cells turned dark blue, and the dark blue circular or oval structures distributed along the intestinal villi epithelium can be used as a marker for identifying goblet cells. Representative fields of view were selected to acquire images, and the location of goblet cells was marked with arrows in the images. 3) Detection of intestinal barrier-related gene expression: Mouse ileum tissue was collected, and the relative mRNA expression levels of Muc2, Tjp1, and Ocln were detected using real-time quantitative polymerase chain reaction (RT-qPCR). 4) Detection of short-chain fatty acids: Mouse cecal contents were collected, and the content of short-chain fatty acids acetic acid and butyric acid was detected. The results are expressed as the mass of short-chain fatty acids per milligram of cecal contents, in ng / mg. 5) Detection of mesenteric lymph node immune cells: Mouse MLNs were collected, single-cell suspensions were prepared, and flow cytometry was used for detection. Cells and The proportion of cells.

[0039] Effects of different formulations on intestinal mucosal structure and barrier function in mice, such as Figures 1-10 As shown, the results indicated that the model control group exhibited significant pathological damage, primarily manifested as atrophy or breakage of small intestinal villi, crypt hyperplasia, and structural disorder. Compared to the model control group, all three experimental groups (formulas A, B, and C) were able to repair gluten-induced atrophy of small intestinal villi and crypt hyperplasia, and improve mucosal tissue structure. Figure 1 By calculating the villus-crypt ratio, it was further verified that all three formulations could significantly increase the ratio of villus height to crypt depth, with better effects than the docosahexaenoic acid control group, gardenia oil control group, and gynostemma pentaphyllum extract control group, and no significant difference compared with the normal control group. Figure 2 AB-PAS staining results showed that, compared with the model control group, the number of goblet cells in the small intestinal tissue of mice in all three experimental groups (formulas A, B, and C) was increased. Figure 3 Furthermore, the relative expression level of Muc2 mRNA in mouse intestinal tissue was significantly increased. Figure 4Muc2 is an important mucin constituting the intestinal mucus layer. Its increased expression level was consistent with the increase in goblet cells shown in AB-PAS staining, indicating that the three compound formulations can promote the formation and recovery of the intestinal mucus barrier. Furthermore, compared with the model control group, the relative mRNA expression levels of Tjp1 and Ocln in the intestinal tissue of mice in the three experimental groups (formulas A, B, and C) were significantly increased. Figure 5 and Figure 6 The increased expression levels of Tjp1 and Ocln indicate that the three compound formulations can enhance the tight junctions between intestinal epithelial cells and improve the integrity of the intestinal epithelial barrier. Furthermore, the acetic acid and butyric acid contents in the cecal contents of mice in all three experimental groups (formulas A, B, and C) were significantly increased, with the most significant increase in acetic acid content observed in formulation B. Figure 7 and Figure 8 The increased levels of acetic acid and butyric acid helped maintain an acidic intestinal environment, providing energy for intestinal epithelial cells and promoting the maintenance of intestinal mucosal barrier function, suggesting that the three compound formulations may exert their protective effect on the intestinal mucosa by promoting the production of short-chain fatty acids in the intestine. Finally, the levels of MLN in the three experimental groups (formulas A, B, and C) of mice were... Cells and The proportion of cells was significantly reduced in all groups, with the most significant reduction observed in formulation B. Figure 9 and Figure 10 ). Cells and The reduced cell ratio indicates that the three compound formulations can inhibit Th1-like and Th17-like inflammatory responses induced by gluten, and alleviate local intestinal immune inflammation. In summary, the three compound formulations can play an auxiliary protective role against gluten-induced intestinal mucosal damage by improving intestinal mucosal tissue structure, strengthening the mucus barrier and epithelial tight junctions, promoting the production of short-chain fatty acids, and regulating local intestinal immune responses. Furthermore, the effects of the three compound formulations are superior to those of docosahexaenoic acid (DHA), gardenia oil, and Gynostemma pentaphyllum extract used alone. The experimental group (Formula B, DHA: Gardenia oil: Gynostemma pentaphyllum extract = 5:20:30) showed the most significant effect; that is, when the ratio of DHA: Gardenia oil: Gynostemma pentaphyllum extract is 5:20:30, this compound exhibits the best auxiliary protective effect against celiac disease-induced intestinal mucosal damage.

[0040] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A DHA-gardenia oil-Gynostemma pentaphyllum compound for repairing intestinal mucosal damage in celiac disease, characterized in that, The DHA-gardenia oil-Gynostemma pentaphyllum compound is composed of docosahexaenoic acid, gardenia oil, and Gynostemma pentaphyllum extract, with a mass ratio of docosahexaenoic acid: gardenia oil: Gynostemma pentaphyllum extract = 1-10: 10-25: 20-30.

2. The DHA-gardenia oil-Gynostemma pentaphyllum compound for repairing celiac disease intestinal mucosal damage as described in claim 1, characterized in that, By mass ratio, docosahexaenoic acid: gardenia oil: gynostemma pentaphyllum extract = 5:20:

30.

3. The DHA-gardenia oil-Gynostemma pentaphyllum compound for repairing celiac disease intestinal mucosal damage as described in claim 1, characterized in that, The active ingredient in the Gynostemma pentaphyllum extract is Gynostemma pentaphyllum saponin.

4. The DHA-gardenia oil-Gynostemma pentaphyllum compound for repairing celiac disease intestinal mucosal damage as described in claim 3, characterized in that, The preparation method of the Gynostemma pentaphyllum extract is as follows: after the whole herb of Gynostemma pentaphyllum is pulverized, it is extracted twice by heating and reflux with 70% ethanol as solvent, the extracts are combined and concentrated; after purification by macroporous adsorption resin, the fraction rich in Gynostemma pentaphyllum saponins is collected, and after concentration and drying, the Gynostemma pentaphyllum extract is obtained.

5. The DHA-gardenia oil-Gynostemma pentaphyllum compound for repairing celiac disease intestinal mucosal damage as described in claim 4, characterized in that, The reflux extraction temperature is 90°C, and the reflux extraction time is 1.5 hours.

6. The DHA-gardenia oil-Gynostemma pentaphyllum compound for repairing celiac disease intestinal mucosal damage as described in claim 1, characterized in that, The gardenia oil is an oil obtained by supercritical CO2 extraction of gardenia fruit, wherein the content of crocin is ≥2.0% and the content of geniposide is ≤0.5%.

7. The DHA-gardenia oil-Gynostemma pentaphyllum compound for repairing celiac disease intestinal mucosal damage as described in claim 6, characterized in that, The conditions for supercritical CO2 extraction are: extraction pressure 25-35 MPa, extraction temperature 40-55℃, and extraction time 2-4 hours.

8. The preparation method of the DHA-gardenia oil-Gynostemma pentaphyllum compound for repairing celiac disease intestinal mucosal damage as described in any one of claims 1-7, characterized in that, The process includes the following steps: First, the prescribed amount of docosahexaenoic acid and gardenia oil are mixed evenly under nitrogen protection to obtain a mixed oil phase; then, Gynostemma pentaphyllum extract is added to the mixed oil phase, and the mixture is thoroughly ground or stirred to ensure that the Gynostemma pentaphyllum extract is evenly dispersed and adsorbed into the mixed oil phase. The mixture is then sieved to obtain the DHA-gardenia oil-Gynostemma pentaphyllum compound.

9. The use of the DHA-gardenia oil-Gynostemma pentaphyllum compound as described in any one of claims 1-7 in the preparation of a medicament for the auxiliary protection of celiac disease intestinal mucosal damage.

10. The application as described in claim 9, characterized in that, The drug is formulated as a clinically acceptable tablet, pill, capsule, suspension, gel, solution, emulsion, ointment, or lotion.

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