Morinda officinalis polysaccharide-inulin composite gel and its preparation method and application

Abstract

The present invention belongs to the field of biomedicine technology, and specifically relates to a Morinda officinalis polysaccharide-inulin composite gel, a preparation method thereof, and an application thereof. The Morinda officinalis polysaccharide-inulin composite gel provided by the present invention is composed of Morinda officinalis polysaccharide, inulin, and water. The Morinda officinalis polysaccharide-inulin composite gel has good biocompatibility and multifunctionality, and can intervene in the intestinal microenvironment and renal fibrosis at the same time, filling the gap in existing treatment methods. Its potential therapeutic effects in intestinal barrier repair, inflammation regulation, microbial regulation, and renal fibrosis inhibition provide new treatment options for clinical practice. The raw material of the composite gel is based on natural polysaccharides, has good safety and degradability, and meets the high requirements of the medical field for the safety of biomaterials. Compared with existing single-component hydrogels, its composite design can achieve multi-target effects, significantly improve the therapeutic effect, and has a greater market competitive advantage.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and in particular relates to a Morinda officinalis polysaccharide-inulin composite gel and a preparation method and application thereof. Background Art

[0002] Renal interstitial fibrosis (RIF) is a hallmark and ultimate outcome of various progressive chronic kidney diseases (CKD). With changes in lifestyle and environmental factors, the incidence of CKD has increased annually, becoming a major health concern and cause of death. It is also considered a key cause of renal failure. Renal fibrosis is caused by the accumulation of connective tissue in renal tissue. This process gradually disrupts the structure and function of the renal tubules, glomeruli, and interstitium, thereby affecting the normal functioning of the kidneys.

[0003] In-depth research into the pathophysiological mechanisms of renal infarction (RIF) in recent years has revealed that the immune response plays a crucial role in the development and progression of renal infarction (RIF). Modulating the immune response may be a key pathway for altering the course of RIF. Immunoregulatory imbalance is closely associated with the development of RIF. In CKD, persistent immune activation leads to massive inflammatory cell infiltration, renal parenchymal cell damage, or fibroblast activation, promoting persistent renal fibrosis and ultimately, progression to chronic renal failure. During RIF, the immune response is often abnormally activated, resulting in significant changes in the renal immune environment. This persistent immune activation is caused by pathological inflammation and is closely associated with the involvement of multiple immune cells in the kidney. For example, the infiltration and persistent activation of immune cells such as T cells, B cells, macrophages, and dendritic cells promotes the exacerbation of the local inflammatory response. Immune cell infiltration and the cytokines and chemokines they secrete further exacerbate renal parenchymal cell damage and dysfunction. Therefore, modulating the renal immune response has become a potential therapeutic target for altering the course of RIF.

[0004] On the other hand, the development of CRF can affect intestinal nutrient absorption and metabolic function, disrupting intestinal ecological stability. Simultaneously, intestinal dysbiosis can increase the accumulation of CRF enterogenic toxins, leading to an imbalance in the intestinal mucosal immune system and subsequently affecting the systemic inflammatory system, causing intestinal endocrine regulation dysfunction. Because enterogenic metabolites are difficult to remove through hemodialysis, accumulated toxins can directly damage the intestinal mucosal barrier. Bacterial translocation and the entry of metabolites into the bloodstream can cause endotoxemia and sepsis. These can also directly contact and activate inflammatory signaling pathways such as TLRs, leading to increased intestinal mucosal permeability and functional impairment.

[0005] Microbiome manipulation promotes mucosal immune homeostasis and intestinal epithelial barrier repair by altering microbiome composition and metabolites, and interacts with the mucosal immune system. Compared with fecal transplantation and probiotics, regulating the intestinal microbiota with diet and prebiotics is safe and low-cost, and may affect a broad spectrum of microorganisms.

[0006] In recent years, research both domestically and internationally has focused on the immunomodulatory and renal protective effects of Morinda officinalis and its polysaccharide components. Morinda officinalis (MO) is a traditional Chinese medicine commonly used to nourish kidney yang and enhance immune function. Its active ingredients include polysaccharides, steroids, and flavonoids. Morinda officinalis polysaccharides (MOPs), one of its main active substances, have demonstrated significant immunomodulatory and renal protective potential. Studies have shown that MO polysaccharides can improve immune status in mice by increasing peripheral white blood cell counts and immune organ weights. Furthermore, MO polysaccharides have been found to be effective in the treatment of chronic kidney injury, promoting kidney repair and regeneration by reducing oxidative stress in the kidneys, inhibiting the release of inflammatory factors, and improving renal tubular function.

[0007] However, it is worth noting that it is difficult to achieve comprehensive reconstruction of intestinal homeostasis and sustained maintenance of long-term remission based on a single-drug treatment strategy. From a pharmacological and physiological perspective, drugs face extremely complex and harsh physical and chemical environmental challenges during their transit through the gastrointestinal tract. The gastrointestinal tract is rich in a variety of digestive enzymes, such as trypsin and pancreatic amylase, which can enzymatically degrade the structure of drugs. The diversity and complexity of the intestinal flora should not be underestimated, and their metabolites and the bacteria themselves may interact with drugs, affecting drug activity. In addition, different sections of the gastrointestinal tract show significant pH gradient changes, from the highly acidic environment in the stomach to the weakly alkaline environment in the small intestine, which poses a severe test to the stability and solubility characteristics of the drug. In addition, intestinal peristalsis is rhythmic and propels at a fast speed, which shortens the effective residence time of the drug in the intestine, thereby adversely affecting the absorption and efficacy of the drug.

[0008] Therefore, providing a drug that can regulate the immune response and intestinal microbiome and deeply analyzing the relevant mechanism of action of the key active ingredients of the drug in the process of renal fibrosis can provide a material basis and translational value for the clinical treatment of renal fibrosis. Summary of the Invention

[0009] In view of the above-mentioned deficiencies, the present invention provides a Morinda officinalis polysaccharide-inulin composite gel and its preparation method and application. The Morinda officinalis polysaccharide-inulin composite gel provided by the present invention is composed of Morinda officinalis polysaccharide, inulin and water. The Morinda officinalis polysaccharide-inulin composite gel has good biocompatibility and multifunctionality, and can intervene in the intestinal microenvironment and renal fibrosis at the same time, filling the gap in existing treatment methods. Its potential therapeutic effects in intestinal barrier repair, inflammation regulation, microbial regulation and renal fibrosis inhibition provide new treatment options for clinical practice. The raw material of the composite gel is based on natural polysaccharides, has good safety and degradability, and meets the high requirements of the medical field for the safety of biomaterials. Compared with existing single-component hydrogels, its composite design can achieve multi-target effects, significantly improve the therapeutic effect, and has a greater market competitive advantage.

[0010] The technical solution of the present invention is:

[0011] In one aspect, the present invention provides a Morinda officinalis polysaccharide-inulin composite gel, which is composed of Morinda officinalis polysaccharide, inulin and water.

[0012] Specifically, the Morinda officinalis polysaccharide-inulin composite gel comprises, by weight, 0.5-1.5 parts of Morinda officinalis polysaccharide, 5-15 parts of inulin and 5-15 parts of water.

[0013] Preferably, the Morinda officinalis polysaccharide-inulin composite gel comprises, by weight, 0.5-0.6 parts, 0.6-0.7 parts, 0.7-0.8 parts, 0.8-0.9 parts, 0.9-1.0 parts, 1.0-1.1 parts, 1.1-1.2 parts, 1.2-1.3 parts, 1.3-1.4 parts or 1.4-1.5 parts of Morinda officinalis polysaccharide.

[0014] Preferably, the Morinda officinalis polysaccharide-inulin composite gel comprises, by weight, 5-6 parts, 6-7 parts, 7-8 parts, 8-9 parts, 9-10 parts, 10-11 parts, 11-12 parts, 12-13 parts, 13-14 parts or 14-15 parts of inulin.

[0015] Preferably, the Morinda officinalis polysaccharide-inulin composite gel comprises, by weight, 5-6 parts, 6-7 parts, 7-8 parts, 8-9 parts, 9-10 parts, 10-11 parts, 11-12 parts, 12-13 parts, 13-14 parts or 14-15 parts of water.

[0016] Further preferably, the Morinda officinalis polysaccharide-inulin composite gel comprises, by weight, 0.8 parts of Morinda officinalis polysaccharide, 10 parts of inulin and 10 parts of water.

[0017] On the other hand, the present invention provides a method for preparing the Morinda officinalis polysaccharide-inulin composite gel described in any one of the above items, the preparation method comprising the following steps:

[0018] S1. Mixing Morinda officinalis polysaccharide and inulin according to parts by weight to obtain a mixture;

[0019] S2. Heat water and add the mixture obtained in step S1 to the water according to parts by weight;

[0020] S3. Stir until the mixture becomes homogeneous and viscous, cool down, and let it stand to obtain a Morinda officinalis polysaccharide-inulin composite gel.

[0021] Specifically, the preparation method of Morinda officinalis polysaccharide described in step S1 includes:

[0022] (1) Slice Morinda officinalis and mix it with water in a certain proportion to obtain a mixed solution;

[0023] (2) After the mixed solution is refluxed for extraction, the extract is collected, centrifuged, the supernatant is collected, and filtered to obtain filtrate 1;

[0024] (3) Repeat step (2) to obtain filtrate 2;

[0025] (4) Combine filtrate 1 and filtrate 2, add ethanol for precipitation, and collect the precipitate;

[0026] (5) The precipitate is washed with ethanol, dialyzed, decolorized, and protein removed to obtain polysaccharide, which is then freeze-dried to obtain Morinda officinalis polysaccharide.

[0027] Preferably, the mass ratio of Morinda officinalis to water in step (1) is 1:1-5.

[0028] Further preferably, the mass ratio of Morinda officinalis to water in step (1) is 1:1.

[0029] Preferably, the reflux extraction conditions in step (2) are reflux extraction at 90-100° C. for 1-3 h.

[0030] Further preferably, the reflux extraction condition in step (2) is reflux extraction at 90° C. for 2 h.

[0031] Preferably, the ethanol described in step (4) is ethanol with a volume fraction of 70-90%.

[0032] Further preferably, the ethanol described in step (4) is ethanol with a volume fraction of 80%.

[0033] Preferably, the precipitation time in step (4) is 24-48 hours.

[0034] Further preferably, the precipitation time in step (4) is 24 hours.

[0035] Preferably, the ethanol described in step (5) is ethanol with a volume fraction of 90-95%.

[0036] Further preferably, the ethanol in step (5) is ethanol with a volume fraction of 95%.

[0037] Preferably, the washing in step (5) is performed 1-5 times;

[0038] More preferably, the washing in step (5) is performed twice.

[0039] Preferably, the dialysis in step (5) is performed using distilled water.

[0040] Preferably, the dialysis in step (5) is performed using a dialysis bag with a molecular weight cutoff value of 3.0-5.0 kDa.

[0041] Further preferably, the dialysis in step (5) is performed using a dialysis bag with a molecular weight cutoff value of 3.5 kDa.

[0042] Preferably, the decolorization in step (5) is carried out by using activated carbon decolorization.

[0043] Preferably, the protein removal method in step (5) is the Savage method.

[0044] Specifically, the heating temperature in step S2 is 65-85°C.

[0045] Preferably, the heating temperature in step S2 is 65-66°C, 66-67°C, 67-68°C, 68-69°C, 69-70°C, 70-71°C, 71-72°C, 72-73°C, 73-74°C, 74-75°C, 75-76°C, 76-77°C, 77-78°C, 78-79°C, 79-80°C, 80-81°C, 81-82°C, 82-83°C, 83-84°C or 84-85°C.

[0046] More preferably, the heating temperature in step S2 is 72°C.

[0047] Specifically, the cooling temperature in step S3 is 20-30°C.

[0048] Preferably, the cooling temperature in step S3 is 20-21°C, 21-22°C, 22-23°C, 23-24°C, 24-25°C, 25-26°C, 26-27°C, 27-28°C, 28-29°C or 29-30°C.

[0049] More preferably, the cooling temperature in step S3 is 26°C.

[0050] Specifically, the standing temperature in step S3 is 0-4° C., and the standing time is ≥24 h.

[0051] Preferably, the standing temperature in step S3 is 0-1°C, 1-2°C, 2-3°C or 3-4°C.

[0052] More preferably, the standing temperature in step S3 is 4°C.

[0053] Preferably, the standing time in step S3 is 24-48 hours.

[0054] Still more preferably, the standing time in step S3 is 24 hours.

[0055] In another aspect, the present invention provides the use of the Morinda officinalis polysaccharide-inulin composite gel described in any one of the above items or the Morinda officinalis polysaccharide-inulin composite gel obtained by any one of the above preparation methods in the preparation of drugs for preventing or treating renal fibrosis.

[0056] In another aspect, the present invention provides a drug for preventing or treating renal fibrosis, wherein the drug comprises the Morinda officinalis polysaccharide-inulin composite gel described in any one of the above items or the Morinda officinalis polysaccharide-inulin composite gel obtained by any one of the above preparation methods.

[0057] Specifically, the dosage form of the drug includes any one or more of: decoction, pills, powders, powders, ointments, pills, wine preparations, tablets, capsules, suppositories, aerosols, films or injections.

[0058] Specifically, the medicine also includes pharmaceutically acceptable excipients.

[0059] Preferably, the pharmaceutically acceptable excipients include: any one or more combinations of gas generating agents, fillers, disintegrants, binders, plasticizers, anti-adherents, wetting agents, lubricants, solvents, suspending agents, solubilizers, diluents, buffers, emulsifiers, preservatives, antioxidants, thickeners, stabilizers, sweeteners and flavorings.

[0060] Specifically, in the medicine, the Morinda officinalis polysaccharide-inulin composite gel described in any one of the above items or the Morinda officinalis polysaccharide-inulin composite gel obtained by any one of the above preparation methods is the sole active ingredient or the main active ingredient.

[0061] Preferably, the medicine may further include other active ingredients, including any one or more of alkaloid active ingredients, glycoside active ingredients, volatile oil active ingredients, flavonoid active ingredients, terpenoid active ingredients, steroid active ingredients, polysaccharide active ingredients or organic acid active ingredients.

[0062] Further preferably, the other active ingredients include but are not limited to: any one or more of alkaloid active ingredients, glycoside active ingredients, volatile oil active ingredients, flavonoid active ingredients, terpenoid active ingredients, steroid active ingredients, polysaccharide active ingredients or organic acid active ingredients.

[0063] Still further preferably, the alkaloid active ingredients include but are not limited to: any one or more of morphine, codeine, atropine, ephedrine, berberine, matrine, colchicine, caffeine, arecoline, aconitine, tetrahydropalmatine, tetrandrine, magnopoic acid, rhynchophylline, evodiamine, strychnine, strychnine, veratrine, fritillary pine, vinblastine, vincristine, camptothecin, cinchona alkaloids or ergometrine.

[0064] Still further preferably, the glycoside active ingredients include but are not limited to any one or more of cardiac glycosides, flavonoid glycosides, saponins, cyanogenic glycosides, phenolic glycosides, indole glycosides, anthraquinone glycosides, coumarin glycosides, lignan glycosides, iridoid glycosides, steroidal saponins, triterpenoid saponins or polysaccharide glycosides.

[0065] Still further preferably, the volatile oil active ingredients include but are not limited to: any one or more of terpenoids, aromatic compounds, aromatic compounds, sulfur-containing compounds, and nitrogen-containing compounds.

[0066] Still further preferably, the flavonoid active ingredients include but are not limited to: any one or more of flavonoid active substances, flavonol active substances, dihydroflavonoid active substances, dihydroflavonol active substances, isoflavone active substances, anthocyanidin active substances, chalcones active substances, diflavonol active substances or aurone active substances.

[0067] More preferably, the flavonoid active ingredients include but are not limited to any one or more of apigenin, luteolin, quercetin, kaempferol, hesperidin, naringin, dihydroquercetin, soy isoflavones, puerarin, cyanidin, delphinidin, safflower yellow, isoliquiritigenin, liquiritigenin, ginkgo biloba flavonoids, aconitum flavonoids, catechin, epicatechin, sulphurein or antirrhinol.

[0068] Still more preferably, the terpene active ingredients include but are not limited to: any one or more of monoterpene active ingredients, sesquiterpene active ingredients, diterpene active ingredients, triterpene active ingredients or tetraterpene active ingredients.

[0069] More preferably, the terpene active ingredients include but are not limited to any one or more of geraniol, nerol, terpineol, farnesol, farnesol, santosan, andrographolide, paclitaxel, stevioside, oleanolic acid, ursolic acid, ginsenosides, carotene, and lycopene.

[0070] Still further preferably, the steroid active ingredients include but are not limited to any one or more of cardiac glycoside active ingredients, steroid saponin active ingredients, steroid alkaloid active ingredients, bile acid active ingredients or steroid hormone active ingredients.

[0071] More preferably, the steroid active ingredients include but are not limited to: any one or more of digoxin, cedilol, dioscin, anemarrhena saponin, solanine, solanine, bile acid, ursodeoxycholic acid, corticosteroid active ingredients or sex hormone active ingredients.

[0072] Still more preferably, the polysaccharide active ingredients include but are not limited to: any one or more of fungal polysaccharides, animal polysaccharides, plant polysaccharides or algae polysaccharides.

[0073] More preferably, the fungal polysaccharide includes but is not limited to any one or more of: Poria cocos polysaccharide, Tremella fuciformis polysaccharide, Flammulina velutipes polysaccharide, Auricularia auricularia polysaccharide, Cordyceps sinensis polysaccharide, Phellinus igniarius polysaccharide or Coriolus versicolor polysaccharide.

[0074] More preferably, the animal polysaccharide includes but is not limited to any one or more of glycogen, chondroitin sulfate, hyaluronic acid, heparin, chitosan, chitin or glycosaminoglycan.

[0075] More preferably, the plant polysaccharides include but are not limited to any one or more of starch, cellulose, hemicellulose, pectin, inulin, konjac polysaccharide, astragalus polysaccharide, wolfberry polysaccharide, tea polysaccharide, kelp polysaccharide or jujube polysaccharide.

[0076] More preferably, the algae polysaccharide includes but is not limited to any one or more of green algae polysaccharide, red algae polysaccharide or brown algae polysaccharide.

[0077] Still more preferably, the organic acid active ingredients include, but are not limited to, any one or more of aliphatic organic acids, aromatic organic acids, terpenoid organic acids or nucleic acid organic acids.

[0078] In another aspect, the present invention provides a method for using the Morinda officinalis polysaccharide-inulin composite gel described in any one of the above items, the Morinda officinalis polysaccharide-inulin composite gel obtained by any one of the above preparation methods, or the medicine described in any one of the above items.

[0079] Specifically, the method of use includes administering the Morinda officinalis polysaccharide-inulin composite gel or drug to a subject.

[0080] Preferably, the subject includes humans or animals.

[0081] Preferably, the dosage of the drug is adjusted according to the severity of the subject's condition, age, administration method and treatment course.

[0082] In another aspect, the present invention provides a method for preventing or treating renal fibrosis, which comprises administering the Morinda officinalis polysaccharide-inulin composite gel or drug to a subject.

[0083] The beneficial effects of the present invention are:

[0084] (1) The Morinda officinalis polysaccharide-inulin composite gel provided by the present invention combines Morinda officinalis polysaccharide with inulin, giving full play to the synergistic effect of the two polysaccharides and improving the physicochemical properties and biological activity of the hydrogel.

[0085] (2) The Morinda officinalis polysaccharide-inulin composite gel material provided by the present invention has good biocompatibility and multifunctionality, and can simultaneously intervene in the intestinal microenvironment and renal fibrosis, filling a gap in existing treatment methods. Its potential therapeutic effects in intestinal barrier repair, inflammation regulation, microbial regulation, and renal fibrosis inhibition provide new clinical treatment options.

[0086] (3) The Morinda officinalis polysaccharide-inulin composite gel material provided by the present invention is based on natural polysaccharides and has good safety and degradability, meeting the high safety requirements of biomaterials in the medical field. Compared with existing single-component hydrogels, its composite design can achieve multi-target effects, significantly improving therapeutic effects and possessing a significant market competitive advantage. BRIEF DESCRIPTION OF THE DRAWINGS

[0087] Figure 1 is the glucose standard curve.

[0088] Figure 2 This is a characterization of the Morinda officinalis polysaccharide-inulin composite gel; A in the figure shows the phenomenon of the gel being injected into water through a syringe; B shows the phenomenon of water at a 45° angle; and C shows the phenomenon of the gel at a 45° angle.

[0089] Figure 3 3 FT-IR spectra.

[0090] Figure 4 This is the SEM scanning result.

[0091] Figure 5 Rheological test results.

[0092] Figure 6 Figure 2 is the weight growth trend and kidney index; A in the figure is the weight change trend from the beginning of modeling to sampling; B is the weight change of mice at the time of sampling compared with the beginning of modeling; C is the kidney index at the time of sampling; in the figure, ns means no significant difference between the groups; * means there is a significant difference compared with the RIF model group, p < 0.05; ** means there is a significant difference compared with the RIF model group, p < 0.01; *** means there is a significant difference compared with the RIF model group, p < 0.001; **** means there is a significant difference compared with the RIF model group, p < 0.0001; # means there is a significant difference compared with the Morinda officinalis polysaccharide group, p < 0.05; ### means there is a significant difference compared with the Morinda officinalis polysaccharide group, p < 0.001.

[0093] Figure 7 are serum biochemical indicators; ns in the figure means no significant difference between the groups; * means significant difference compared with the RIF model group, p < 0.05; # means significant difference compared with the Morinda officinalis polysaccharide group, p < 0.05; ### means significant difference compared with the Morinda officinalis polysaccharide group, p < 0.001.

[0094] Figure 8 are urine biochemical indicators; ns in the figure means no significant difference between the groups; * means there is a significant difference compared with the RIF model group, p < 0.05; ** means there is a significant difference compared with the RIF model group, p < 0.01; *** means there is a significant difference compared with the RIF model group, p < 0.001; # means there is a significant difference compared with the Morinda officinalis polysaccharide group, p < 0.05.

[0095] Figure 9 The results are histomorphology and pathology. DETAILED DESCRIPTION

[0096] The present invention will be further clarified and fully described below by way of examples. The following examples are only a portion of the present invention and are not intended to limit the present invention, but are merely for illustration. The experimental methods used in the following examples are all routine experiments unless otherwise specified, and the materials and reagents used in the following examples are all commercially available unless otherwise specified.

[0097] Example 1 Preparation of Morinda officinalis polysaccharide and determination of polysaccharide content

[0098] 1. Preparation of Morinda officinalis polysaccharide

[0099] Ultrapure water (100 g) and Morinda officinalis (sliced) (100 g) were mixed and extracted under reflux at 90°C for 2 hours. The extracted solution was centrifuged at 4000 rpm for 20 minutes, and the supernatant was harvested and filtered to obtain the filtrate. This process was repeated for a second extraction, centrifugation, and filtration, and the two filtrates were combined. The mixture was treated with 80% ethanol by volume and allowed to stand at 4°C for 24 hours to precipitate polysaccharides, and the precipitate was collected.

[0100] The precipitate was washed twice with 95% ethanol and then dialyzed against distilled water (using a molecular weight cutoff of 3.5 kDa). The product was decolorized using activated carbon. Protein was removed using the Savage method. The purified polysaccharide was lyophilized to produce an off-white powder, which is Morinda officinalis polysaccharide.

[0101] 2. Draw the glucose standard curve

[0102] Take 10 mg of glucose reference substance, accurately weigh it, dissolve it in distilled water, and then transfer it to a 100 mL volumetric flask to make up to the volume to prepare a glucose reference substance stock solution with a concentration of 0.100 mg / mL.

[0103] The prepared glucose reference stock solution was diluted to a series of concentrations of 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.08, and 0.10 mg / mL, and the color was developed and the absorbance (A) at 490 nm was measured. A standard curve was drawn with glucose concentration (C) as the horizontal axis and absorbance (A) as the vertical axis. The standard curve is shown in the figure below. Figure 1 shown.

[0104] 3. Determination of polysaccharide content

[0105] Take Morinda officinalis polysaccharide and quantitatively redissolve it in ultrapure water. Determine the polysaccharide content in Morinda officinalis polysaccharide. The polysaccharide content is qualified if it is greater than 85%. The specific steps are as follows:

[0106] Accurately weigh approximately 0.02 g of Morinda officinalis polysaccharide into a 100 mL volumetric flask. Dissolve it in distilled water and dilute to the mark. Remove 10 mL and transfer it to the 100 mL volumetric flask. Dilute to the mark again with distilled water. Then, pipette 1.0 mL into a test tube. Add 1.0 mL of 5% phenol reagent and 5.0 mL of concentrated sulfuric acid, shake well, and boil for 30 minutes. Measure the absorbance (A) at 490 nm using a UV spectrophotometer. Substitute the A value into the linear regression equation of the standard curve to calculate the polysaccharide concentration (C).

[0107] The polysaccharide absorbance values ​​were measured three times and were 1.462, 1.548, and 1.473, respectively, which were converted to a polysaccharide concentration of 94.15%. Therefore, the polysaccharide content of the Morinda officinalis polysaccharide prepared by the present invention was >85%, which was qualified.

[0108] Example 2 Preparation of Morinda officinalis polysaccharide-inulin composite gel

[0109] Mix 10g of inulin and 0.8g of Morinda officinalis polysaccharide to obtain 10.8g of the mixture. Heat 10g of water to 72°C and slowly add the mixture. Mix with a magnetic stirrer until a homogeneous, viscous consistency forms. Allow the sample to cool to 26°C and allow to stand at 4°C overnight. This will yield the Morinda officinalis polysaccharide-inulin composite gel. Figure 2 The gel morphology, injectability, and tilt angle stability of the Morinda officinalis polysaccharide-inulin composite gel were demonstrated. Visually, it exhibited good injectability, demonstrating the fluidity characteristics of the Morinda officinalis polysaccharide-inulin composite gel. Comparison with water at a 45° tilt angle and attempts at shaping demonstrated the excellent adhesion of the Morinda officinalis polysaccharide-inulin composite gel.

[0110] Example 3 FT-IR spectrum measurement

[0111] The KBr-disk method was performed using a Nicolet iZ-10 spectrometer (Thermo Nicolet, USA). Potassium bromide (KBr) powder was pre-dried at 104°C for 12 hours. Then, freeze-dried Morinda officinalis polysaccharide-inulin composite gel (FTZP) sample was mixed with pre-dried KBr powder (100:1) in an agate mortar and pressed into 1 mm pellets. The results were obtained at 400 to 4000 cm -1 Collect spectra within the range.

[0112] The FT-IR spectrum of FTZP sample is as follows Figure 3 As shown, the test shows:

[0113] 3368cm -1 The corresponding point is the OH stretching vibration absorption peak, and a broad absorption band is formed;

[0114] 2931cm -1 The corresponding peak is the stretching vibration peak of CH3, CH2 or CH;

[0115] 1419cm -1 The corresponding peak is the variable angle vibration peak of CH3, CH2 or CH;

[0116] 1031cm -1 The corresponding peak is the stretching vibration peak of C-OH;

[0117] 936cm -1 The corresponding peak is the symmetrical stretching vibration peak of COC;

[0118] 1636cm -1 The corresponding peak is the OH angle vibration peak, indicating that the sample contains a small amount of water;

[0119] The above characteristic peaks can be used to determine the presence of sugar compounds in the FTZP sample.

[0120] Example 4 SEM scanning

[0121] Freeze-dried Morinda officinalis polysaccharide-inulin composite gel samples were coated with gold under vacuum and imaged using a scanning electron microscope at magnifications ranging from 100 to 4000 times using an accelerating voltage of 5.0 kV.

[0122] Scanning electron microscopy (SEM) results are as follows Figure 4 As shown, the SEM image shows an interconnected and uniform porous network structure, which is the microstructural characteristic of the Morinda officinalis polysaccharide-inulin composite gel.

[0123] Example 5 Rheological test

[0124] The rheometer was used to test the amplitude of Morinda officinalis polysaccharide-inulin composite gel at a fixed frequency; the shear rate was linearly changed; and the temperature was swept at a fixed strain and frequency.

[0125] The results are as follows Figure 5 As shown, Figure 5 In Figure A, G' > G'' is a typical sign of a gel state. G' reaches approximately 35,000 Pa, indicating a highly strong, densely structured gel. At 30-40°C, the material's elastic behavior dominates, exhibiting solid-like properties (maintaining shape and resisting deformation). Upon heating, the gel's network structure is destroyed, suggesting that the Morinda officinalis polysaccharide-inulin composite gel is a typical thermosensitive gel. At approximately 65-70°C, G'' exceeds G' (G'' > G'). is the melting temperature (Tm) of the gel or the gel-sol transition temperature;

[0126] Figure 5 The B stress scan results in Figure 2 show that the Morinda officinalis polysaccharide-inulin composite gel exhibits stable linear viscoelastic behavior in the low strain region (γ<1%), and its storage modulus (G' ≈ 10 5 The yield modulus (γ_yield ≈ 1%) is significantly higher than the loss modulus (G''), confirming a robust three-dimensional network structure. When the strain exceeds the yield point (γ_yield ≈ 1%), G' drops sharply, indicating the onset of irreversible network failure. A crossover between G' and G'' is observed at the flow point (γ_flow ≈ 30%), marking the material's transition from a gel to a sol state.

[0127] Figure 5 C in low shear rate ( <1 s -1 ): At high viscosity platform (gel network is complete, resistance is large). Medium shear rate (1 - 100 s -1): Viscosity drops sharply (network destruction, structural alignment). High shear rate ( >100 s -1 ): Low viscosity plateau (fully fluid state). This indicates disentanglement / destruction of the gel network or macromolecular chains under shear stress, a behavior consistent with biomedical gels such as injectable hydrogels.

[0128] Example 6 Therapeutic Effect of Morinda officinalis Polysaccharide-Inulin Composite Gel on Renal Fibrosis (RIF)

[0129] 1. Experimental Animals

[0130] Sixty 8-week-old male C57BL / 6J mice were purchased from Guangdong Ruige Biotechnology Co., Ltd. Mice were housed in an SPF environment at the Animal Experimental Center of Guangdong Pharmaceutical University with a 12-h light / dark cycle, a temperature of 20–25°C, and a humidity of 60 ± 5%. Animals had free access to food and water. All mice were acclimated to a diet for 1 week before the experiment. The experimental protocol and procedures were approved by the Animal Experimentation Ethics Committee of Guangdong Pharmaceutical University on October 16, 2023, and were in compliance with the Guide for the Care and Use of Laboratory Animals.

[0131] 2. Induced RIF Model, Grouping, and Dosage

[0132] The mice were divided into healthy control group, RIF model group, positive control group, Morinda officinalis polysaccharide-inulin composite gel group, inulin group, and Morinda officinalis polysaccharide group.

[0133] Except for the healthy control group of mice, which were fed a normal diet, the mice in the remaining groups were pre-fed a diet containing 0.25% adenine for 5 weeks, followed by a diet containing 0.1% adenine until week 11, to induce the RIF model. Drug administration began at week 6 for a 6-week period. The dosing schedule for each group was as follows:

[0134] Healthy control group: gavage with distilled water (100 μL / d);

[0135] RIF model group: intragastric administration of distilled water (100 μL / d);

[0136] Positive control group: oral administration of losartan (100 μL / d, i.e., losartan 30 mg / kg / d);

[0137] Morinda officinalis polysaccharide-inulin composite gel group: Oral administration of Morinda officinalis polysaccharide-inulin composite gel (100 μL / d);

[0138] Morinda officinalis polysaccharide group: Oral administration of Morinda officinalis polysaccharide aqueous solution (100 μL / d, i.e. Morinda officinalis polysaccharide 200 mg / kg / d).

[0139] Inulin group: oral administration of inulin aqueous solution (100 μL / d, i.e., inulin 3125 mg / kg / d);

[0140] 3. Growth curve

[0141] The weight growth trend of the experimental animals from the beginning of modeling to sampling and the renal index at the time of sampling were recorded. Figure 6 As shown in the figure, A is the weight growth trend from the beginning of modeling to sampling; B is the weight change of mice at the time of sampling compared with the beginning of modeling; C is the kidney index.

[0142] Except for the mice in the healthy control group, the body weight of the mice in the RIF model group, positive control group, Morinda officinalis polysaccharide-inulin composite gel group, Morinda officinalis polysaccharide group, and inulin group showed a downward trend in the first 5 weeks. After medication, the body weight of the mice in the Morinda officinalis polysaccharide-inulin composite gel group, Morinda officinalis polysaccharide group, and inulin group continued to recover, indicating that the Morinda officinalis polysaccharide-inulin composite gel, Morinda officinalis polysaccharide, and inulin can all improve the weight loss caused by RIF, among which the Morinda officinalis polysaccharide-inulin composite gel had the most obvious improvement effect.

[0143] The renal index of mice in the RIF model group was significantly lower than that of mice in the healthy control group (p < 0.0001), indicating that the renal fibrosis model was successfully established. The renal index of mice in the Morinda officinalis polysaccharide-inulin composite gel group, Morinda officinalis polysaccharide group, and inulin group all increased to varying degrees, among which the renal index of mice in the Morinda officinalis polysaccharide-inulin composite gel group had the best improvement effect.

[0144] 4. Biochemical indicators

[0145] One day after the last dose, blood samples were collected from mice and centrifuged at 3000 rpm for 15 minutes to obtain serum. Serum urea nitrogen (BUN) and creatinine (Scr) were measured using biochemical marker kits. Urine was collected from mice for 24 hours in metabolic cages. Urine protein, BUN, creatinine, and other biochemical markers were measured using biochemical marker kits. The BUN kit (purchased from Nanjing Jiancheng, Cat. No. C013-2-1); the creatinine kit (purchased from Nanjing Jiancheng, Cat. No. C011-2-1); and the urine protein kit (purchased from Nanjing Jiancheng, Cat. No. C035-2-1) were used.

[0146] Serum biochemical indicators such as Figure 7 As shown in the figure, urine biochemical indicators are as follows Figure 8 The test results showed that the Morinda officinalis polysaccharide-inulin composite gel significantly reduced the levels of serum creatinine and urea nitrogen; inhibited the increase of 24-hour urine protein, and significantly enhanced the kidney's metabolic rate of urea nitrogen and creatinine.

[0147] 5. Histomorphology and pathology

[0148] Representative photographs of collected mouse kidney and colon tissues were taken, fixed in 4% paraformaldehyde overnight at 4°C, embedded in paraffin, and cut into 4μm sections. Glomerular structure was visualized using hematoxylin and eosin (HE) staining. Masson staining was used to assess the extent of renal fibrosis.

[0149] like Figure 9 As shown, histomorphologically, the kidneys of mice in the RIF model group showed significant atrophy and fibrosis compared to the healthy control group, while the Morinda officinalis polysaccharide-inulin composite gel group showed an improvement trend. H&E staining revealed glomerular hypertrophy in RIF mice, which was alleviated by Morinda officinalis polysaccharide-inulin composite gel treatment. Masson staining showed that the Morinda officinalis polysaccharide-inulin composite gel group significantly reduced the positive area of ​​fibrosis.

[0150] The above detailed description is a specific description of one feasible embodiment of the present invention and is not intended to limit the scope of the present invention. It should be noted that any equivalent implementation or modification that does not depart from the present invention should be included within the scope of the technical solution of the present invention. Therefore, the scope of protection of the patent of this invention should be based on the attached requirements.

Claims

1. A Morinda officinalis polysaccharide-inulin composite gel, characterized in that: The Morinda officinalis polysaccharide-inulin composite gel is composed of 0.5-1.5 parts of Morinda officinalis polysaccharide, 5-15 parts of inulin and 5-15 parts of water in parts by weight; The preparation method of the Morinda officinalis polysaccharide comprises: (1) Slice Morinda officinalis and mix it with water in a certain proportion to obtain a mixed solution; (2) After the mixed solution is refluxed for extraction, the extract is collected, centrifuged, the supernatant is collected, and filtered to obtain filtrate 1; (3) Repeat step (2) to obtain filtrate 2; (4) Combine filtrate 1 and filtrate 2, add ethanol for precipitation, and collect the precipitate; (5) The precipitate was washed twice with ethanol, and after dialysis, decolorization, and protein removal, polysaccharide was obtained, and then freeze-dried to obtain Morinda officinalis polysaccharide.

2. The Morinda officinalis polysaccharide-inulin composite gel according to claim 1, characterized in that The Morinda officinalis polysaccharide-inulin composite gel comprises, by weight, 0.8 parts of Morinda officinalis polysaccharide, 10 parts of inulin and 10 parts of water.

3. The method for preparing the Morinda officinalis polysaccharide-inulin composite gel according to any one of claims 1 to 2, characterized in that: The preparation method comprises the following steps: S1. Mixing Morinda officinalis polysaccharide and inulin according to parts by weight to obtain a mixture; S2. Heat water and add the mixture obtained in step S1 to the water according to parts by weight; S3. Stir until the mixture becomes homogeneous and viscous, cool down, and let it stand to obtain a Morinda officinalis polysaccharide-inulin composite gel.

4. The preparation method according to claim 3, characterized in that The heating temperature in step S2 is 65-85°C.

5. The preparation method according to claim 3, characterized in that The temperature of the cooling step S3 is 20-30°C.

6. The preparation method according to claim 3, characterized in that The standing temperature in step S3 is 0-4° C. and the standing time is ≥24 h.

7. Use of the Morinda officinalis polysaccharide-inulin composite gel according to any one of claims 1 to 2 or the Morinda officinalis polysaccharide-inulin composite gel obtained by the preparation method according to any one of claims 3 to 6 in the preparation of a drug for preventing or treating renal fibrosis.

8. A drug for preventing or treating renal fibrosis, characterized in that: The medicine comprises the Morinda officinalis polysaccharide-inulin composite gel according to any one of claims 1-2 or the Morinda officinalis polysaccharide-inulin composite gel obtained by the preparation method according to any one of claims 3-6.

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