Preparation method and application of oral inulin gel

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a preparation method and application of oral inulin gel. The oral gel is prepared on the basis of prebiotic inulin capable of adjusting intestinal flora, the effect of repairing intestinal mucosa can be achieved, and the purpose of effectively treating the alcoholic liver disease is achieved. The gel provided by the invention is stable in structure and good in rheological property; in-vivo experiments show that the traditional Chinese medicine can effectively relieve chronic alcoholic liver diseases. The preparation process is simple, the stability is good, the biocompatibility is good, and the preparation method has a good application prospect.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a preparation method and application of oral inulin gel. Background Art

[0002] Chronic alcohol consumption is one of the most common causes of death worldwide. Alcohol abuse can lead to damage to multiple end organs, primarily the liver, intestines, and brain, resulting in multisystem impairment. Alcoholic liver disease (ALD) is a liver injury disease caused by chronic or acute alcohol abuse and includes alcoholic fatty liver disease, steatosis, alcoholic steatohepatitis, fibrosis, cirrhosis, and even liver cancer. ALD is one of the leading causes of cirrhosis and liver disease-related mortality worldwide, posing a serious threat to human health and social development. Currently, treatment strategies for ALD primarily include alcohol abstinence, nutritional support, medication, and liver transplantation. Abstinence is a cornerstone of treatment for ALD, but many patients struggle to adhere to this regimen. Regarding pharmacological treatment, glucocorticoids can improve short-term survival in patients with severe ALD, but their long-term effects are limited and are associated with side effects. Furthermore, metadoxine can accelerate alcohol metabolism and improve symptoms of alcoholism. Medications such as silymarin and glycyrrhizic acid preparations have antioxidant, anti-inflammatory, and hepatocellular protective properties. However, existing drug treatments have limited efficacy, and some have side effects such as hepatotoxicity. For advanced patients, liver transplantation is the last resort, but it faces problems such as donor shortage and high cost.

[0003] Recent studies have found that long-term alcohol consumption can significantly alter the composition and function of the intestinal flora, manifesting as a decrease in the number of beneficial bacteria and an increase in the number of harmful bacteria, leading to intestinal dysbiosis. This dysbiosis can disrupt the integrity of the intestinal mucosa, enhance intestinal permeability, and lead to the translocation of endotoxins and other substances into the liver. These toxins can activate macrophages in the liver, stimulate the release of proinflammatory cytokines, generate reactive oxygen species and oxidative stress, further aggravate the liver's inflammatory response, and perpetuate inflammation and fibrosis within the liver. Therefore, improving intestinal barrier function and alleviating inflammatory responses by regulating the intestinal flora has gradually become a new direction for the treatment of ALD.

[0004] As a natural prebiotic, inulin can modulate the composition and function of the intestinal flora, promoting the proliferation of beneficial bacteria and inhibiting the growth of harmful bacteria. In the colon, inulin is hydrolyzed and fermented by beneficial intestinal microorganisms to produce short-chain fatty acids (such as acetate, propionate, and butyrate), thereby regulating the composition and activity of the intestinal flora. However, direct oral administration of inulin has a short retention time in the intestine, typically requiring frequent dosing to maintain its effects on regulating the intestinal flora. Therefore, preparing inulin into inulin gel can prolong its retention time in the intestine, thereby enhancing its regulatory effects on the intestinal flora. Summary of the Invention

[0005] Therefore, the present invention aims to provide a prebiotic inulin oral gel. The inulin gel of the present invention can regulate intestinal flora and effectively treat alcoholic liver disease.

[0006] To achieve the above objectives, the first aspect of the present invention provides a method for preparing inulin gel.

[0007] Furthermore, a certain amount of inulin is dissolved in a solvent, heated, and then left at room temperature to obtain inulin gel.

[0008] Furthermore, the inulin concentration is 10-1000 mg / mL.

[0009] Furthermore, the solvent is water, phosphate buffer, citrate buffer, Hank's balanced salt solution, HEPES buffer, Tris buffer, etc.

[0010] Furthermore, the heating temperature is 30°C~90°C.

[0011] Furthermore, the heating time is 1 min to 1 h.

[0012] Furthermore, the room temperature storage time is 12 to 48 hours.

[0013] The second aspect of the present invention provides use of the inulin gel described in the first aspect for treating alcohol-related diseases.

[0014] Furthermore, the alcohol-related diseases include one or more of alcoholic pancreatitis, alcoholic ketoacidosis, alcoholic liver disease, peptic ulcer, esophageal varices, alcoholic peripheral neuropathy, alcoholic dementia, alcoholic cardiomyopathy, alcoholic gout, alcoholic depression, and alcoholic osteoporosis.

[0015] Furthermore, the alcoholic liver disease includes one or more of chronic alcoholic liver disease, acute alcoholic liver disease, and acute alcohol poisoning.

[0016] The present invention has the beneficial effects of preparing an oral inulin gel that can treat alcoholic liver disease by regulating intestinal flora. The inulin gel provided by the present invention has a stable structure, excellent centrifugal stability, injectability, and rheological properties, and can prolong drug retention in the intestine, making it suitable for the treatment of acute and chronic alcoholic liver disease. The present invention also features a simple preparation method, good biocompatibility, and promising industrial application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings, in which: Figure 1 This is a scanning electron microscope (SEM) image of the inulin gel obtained in one embodiment of the present invention, with a scale of 20 μm; Figure 2 This is a diagram showing the injectability of inulin gel obtained in one embodiment of the present invention; Figure 3 Graph showing the rheological properties of inulin gel obtained according to one embodiment of the present invention: A. Storage modulus (G') and loss modulus (G'') of inulin gel measured; B. Variation of gel viscosity with shear rate; Figure 4 This figure shows the results of an investigation of plasma liver function indicators using the inulin gel obtained in one embodiment of the present invention for the treatment of chronic alcoholic liver disease: A. Alanine aminotransferase (ALT) level measurement; B. Aspartate aminotransferase (AST) level measurement; Figure 5 This figure shows the results of an investigation into hepatic lipid accumulation in the treatment of chronic alcoholic liver disease using inulin gel obtained in one embodiment of the present invention: A. Triglyceride (TG) level measurement; B. Total cholesterol (TC) level measurement. DETAILED DESCRIPTION

[0018] The present invention will be described in further detail below with reference to the examples, but the embodiments of the present invention are not limited thereto. The present invention will be further described in detail below with reference to the examples, but it should be understood by those skilled in the art that the present invention is not limited to these examples and the preparation methods used. Moreover, modifications and substitutions made to the method, steps or conditions of the present invention are within the scope of the present invention without departing from the spirit and substance of the present invention.

[0019] Example 1. Preparation of oral inulin gel This example is an illustrative description of the method for preparing the oral inulin gel of the present invention.

[0020] Take inulin, dissolve it in ultrapure water to prepare a 100 mg / mL inulin aqueous solution, then place it in a constant temperature water bath at 60°C and heat it for 5 minutes. After heating, cool it to room temperature for 12-24 hours to obtain inulin gel.

[0021] Example 2: Preparation of oral inulin gel This example is another exemplary description of the method for preparing the oral inulin gel of the present invention.

[0022] The preparation methods of Examples 2 to 8 are the same as those of Example 1, except for Table 1.

[0023] Table 1 Preparation method of inulin gel in Examples 2 to 8

[0024] Table 1 Example 3: Appearance of oral inulin gel This example shows the appearance of the oral inulin gel of the present invention.

[0025] The gelling time of the inulin gels prepared in Examples 1 to 7 was observed and recorded, and the appearance was recorded by taking photos. Table 2 Appearance of inulin gel prepared in Examples 1 to 7

[0026] Table 2 Example 4: Evaluation of centrifugal stability of inulin gel A sample of oral inulin gel was centrifuged at 3000 rpm for 30 minutes and the gel state was observed. No changes in appearance, delamination, or water separation were observed, indicating good centrifugal stability.

[0027] Example 5: Microstructure Observation of Inulin Gel The morphology of the dried gel was observed using a scanning electron microscope (SEM). The gel sample was quickly frozen in liquid nitrogen for 30 minutes, then transferred to a vacuum freeze dryer and dried overnight at -80°C. After drying, a small amount of sample was taken with tweezers and fixed to the SEM stage using conductive adhesive. Nitrogen was used to remove any excess sample, and the sample was then gold-sputtered using an ion sputtering instrument. Finally, the sample was scanned and imaged at an accelerating voltage of 5 kV.

[0028] Figure 1 The results showed that the gel had a reticular porous structure.

[0029] Example 6: Investigation of the injectability of inulin gel Take the gel sample into a 1 mL syringe and squeeze the gel into a dish and a vial filled with water. Simultaneously observe the continuity of the sample to see if there is any interruption or spraying, and take photos to record the morphology of the extruded gel.

[0030] Figure 2 The results showed that the prepared inulin gel could be continuously extruded from a 1 mL syringe without discontinuous ejection, and the word "SWU" could be accurately written through pattern forming experiments. These results demonstrated that the gel had good injectability.

[0031] Example 7: Investigation of rheological properties of inulin gel The gel sample was transferred to the rheometer plate and equilibrated at 25 °C for 3 min. The plate diameter was 40 mm. In the flow scanning mode, the shear rate was measured from 0.1 to 100 s. -1 In the oscillation frequency mode, the frequency was set to 1.0 Hz and the strain rate was 5.0%, and the storage modulus (G') and loss modulus (G'') were measured in the frequency range of 0.1-100 rad / s.

[0032] The results in Figure 3 show that the storage modulus (G') of the prepared inulin gel is consistently higher than the loss modulus (G''), and the loss tangent is less than 1. This indicates that the gel system is primarily elastic and exhibits solid-like behavior, which is typical of physically cross-linked hydrogels. The relationship between viscosity and shear rate shows that the gel viscosity decreases with increasing shear rate, exhibiting typical shear-thinning behavior. This indicates that under the action of shear force, the internal network structure of the gel is gradually destroyed, the intermolecular interactions are weakened, and the fluidity is enhanced.

[0033] Example 8: Investigation of the therapeutic effect of inulin gel on chronic alcoholic liver disease A mouse model of chronic alcoholic liver disease was established and the efficacy of inulin gel was investigated. After 3 days of free adjustment of the Lieber-DeCarli diet, 20 C57BL / 6J mice were randomly divided into 4 groups (n = 5): (1) Control group: Control diet was given throughout the whole period; (2) Model group: Lieber-DeCarli diet containing ethanol was used without restriction except for adaptation to the control diet; (3) Free inulin group: Compared with the positive control group, 4 g / kg of inulin solution was gavaged daily in the third period; (4) Inulin gel group: Compared with the positive control group, 4 g / kg of inulin gel was gavaged daily in the third period. Phase 1, days 1-5: All mice were adapted to the Lieber-DeCarli diet (i.e., feed containing maltodextrin) for 5 days. Phase 2, days 6-10: Lieber-DeCarli diet containing ethanol was used, in which the alcohol content was gradually increased from 1% to 5%, and the mice were fed for 5 days; healthy mice continued to eat the control diet. During the third phase (days 11–20), the control group received a paired isocaloric control diet and a daily control liquid gavage, supplemented with 4 g / kg of phosphate-buffered saline. All other model groups received unrestricted access to an ethanolic Lieber-DeCarli diet containing 5% ethanol for 10 days. Simultaneously, the free inulin-treated group and the inulin gel-treated group received daily oral gavages of 4 g / kg of inulin solution and inulin gel, respectively. During the fourth phase (days 21), mice were fasted and gavaged with 6 g / kg of 50% ethanol, except for the negative control group, which received an isocaloric dose of maltodextrin. Six hours later, orbital blood was drawn, and the mice were sacrificed for measurement of relevant biomarkers. Figure 4 and Figure 5 It shows that after oral administration of inulin gel to mice with chronic alcoholic liver disease model, the levels of ALT and AST in plasma can be significantly reduced, the levels of TG and TC in the liver can be significantly reduced, the accumulation of lipids in the liver can be reduced, and chronic alcoholic liver disease can be effectively alleviated.

Claims

1. A method for preparing oral inulin gel, characterized in that: The inulin gel preparation method comprises the following steps: dissolving a certain amount of inulin in a solvent, heating the solvent, and then leaving the solvent at room temperature to obtain the inulin gel.

2. The preparation method according to claim 1, characterized in that The inulin concentration is 10 mg / mL-1000 mg / mL.

3. The preparation method according to claim 2, characterized in that The heating temperature is 30°C to 90°C.

4. The preparation method according to claim 2, characterized in that The heating time is 1 min to 1 h.

5. The preparation method according to claim 2, characterized in that The room temperature placement time is 12 to 48 hours.

6. The use of the oral inulin gel according to any one of claims 1 to 5, characterized in that The oral gel is used to treat one or more of alcoholic pancreatitis, alcoholic ketoacidosis, alcoholic liver disease, peptic ulcer, esophageal varices, alcoholic peripheral neuropathy, alcoholic dementia, alcoholic cardiomyopathy, alcoholic gout, alcoholic depression, and alcoholic osteoporosis.

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