A sodium alginate-laminarin-ganoderic acid D nanoparticle for targeted treatment of ulcerative colitis, and a preparation method and application thereof

Abstract

The application discloses sodium alginate-kelp polysaccharide-ganoderic acid D nanoparticles for targeted treatment of ulcerative colitis and a preparation method and application thereof. Raw materials of the nanoparticles include sodium alginate, kelp polysaccharide and ganoderic acid D with a mass ratio of 20-30:2-5:8-12. The sodium alginate-kelp polysaccharide-ganoderic acid D nanoparticles are prepared by taking sodium alginate gel as a carrier and loading kelp polysaccharide and ganoderic acid D in the carrier, and the nanoparticles are used for preventing and / or relieving ulcerative colitis, can have antioxidant effect, target regulation of intestinal barrier function, reduction of inflammatory reaction and improvement of intestinal health, and provide a new method for ulcerative colitis.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to a sodium alginate-laminarin-ganoderic acid D nanoparticle for targeted treatment of ulcerative colitis, its preparation method, and its application. Background Technology

[0002] Inflammatory bowel disease (IBD) is a group of idiopathic, chronic, relapsing inflammatory bowel diseases, mainly including ulcerative colitis and Crohn's disease, which can occur in people of all ages. Its pathogenesis involves complex interactions between genetic susceptibility, environmental factors, and the gut microbiome, typically leading to impaired intestinal barrier function and subsequent abnormal intestinal immune activation. Current clinical treatment primarily relies on anti-inflammatory drugs, immunomodulators, and antibiotics; however, these methods generally suffer from problems such as high relapse rates, significant side effects, and unstable efficacy due to large individual variability in treatment outcomes. Therefore, the treatment of IBD still faces many challenges.

[0003] Nanoparticles, ultrafine particles with sizes ranging from 1 to 100 nm, have demonstrated significant application value in drug delivery due to their unique size advantages and properties. Drug delivery in nanoparticle form can effectively enhance the delivery efficiency of therapeutic drugs to the target treatment site, maximizing therapeutic efficacy and reducing the incidence of adverse reactions. Currently commonly used nanoparticle carriers include liposomes, mesoporous silica nanoparticles, polymer micelles, and hydrogels. Among these, sodium alginate, as a biocompatible and biodegradable natural polymer, is one of the ideal materials for constructing targeted delivery systems. Sodium alginate gel microspheres formed through ionic cross-linking can effectively prevent drug burst release and possess characteristics such as pH-responsive drug release, controllable particle size, and high oral safety.

[0004] Laminaria polysaccharides are macromolecular polysaccharides extracted from brown algae. Their molecular chains contain sulfate groups at the ends, exhibiting good biocompatibility. Studies have shown that laminarin polysaccharides not only exert anti-inflammatory effects by regulating immune responses, but also directly promote the repair and regeneration of intestinal mucosal epithelial cells, enhancing the integrity of the intestinal barrier.

[0005] Ganoderic acid D is an active ingredient in Ganoderma lucidum (reishi mushroom) and possesses various medicinal functions, including antioxidant, anti-inflammatory, hypoglycemic, anti-tumor, and immunomodulatory effects. Studies have shown that ganoderic acid D can regulate cellular energy metabolism and alleviate oxidative stress.

[0006] Currently, there are no reports on the synergistic treatment of ulcerative colitis with sodium alginate, laminarin, and ganoderic acid D. Summary of the Invention

[0007] In order to reduce the side effects of anti-inflammatory drugs, immunomodulators and antibiotics, expand the product range for ulcerative colitis, and treat ulcerative colitis more safely and effectively, the present invention provides the following technical solutions.

[0008] In a first aspect, the present invention provides sodium alginate-laminarin-ganoderic acid D nanoparticles, wherein the raw materials of the nanoparticles include sodium alginate, laminarin and ganoderic acid D in a mass ratio of 20~30:2~5:8~12.

[0009] Preferably, the preparation process of the nanoparticles includes the following steps: S1, Weigh out sodium alginate, laminarin and polyethyleneimine, and disperse them in water to obtain sodium alginate solution, laminarin solution and polyethyleneimine solution; S2, the kelp polysaccharide solution and the polyethyleneimine solution are added to the sodium alginate solution, and after the reaction is allowed to stand, dialyzed, and the dialysate is dried to obtain a solid sample; S3, dissolve the solid sample in dimethyl sulfoxide to obtain a reconstituted solution; S4, weigh out ganoderic acid D, dissolve it in dimethyl sulfoxide, add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, let stand, and obtain ganoderic acid D solution; S5, the ganoderic acid D solution is added to the reconstituted solution in S3, and the mixture is subjected to ultrasonication, shaking, centrifugation, resuspension of the precipitate with water, and drying in sequence to obtain the sodium alginate-laminarin-ganoderic acid D nanoparticles.

[0010] Preferably, the nanoparticles have a particle size of 80~100 nm.

[0011] In a second aspect, the present invention provides a composition comprising the sodium alginate-laminarin-ganoderic acid D nanoparticles described in the first aspect.

[0012] Thirdly, the present invention provides a method for preparing sodium alginate-laminarin-ganoderic acid D nanoparticles, the preparation method comprising the following steps: S1, Weigh out sodium alginate, laminarin and polyethyleneimine, and disperse them in water to obtain sodium alginate solution, laminarin solution and polyethyleneimine solution; S2, the kelp polysaccharide solution and the polyethyleneimine solution are added to the sodium alginate solution, and after the reaction is allowed to stand, dialyzed, and the dialysate is dried to obtain a solid sample; S3, dissolve the solid sample in dimethyl sulfoxide to obtain a reconstituted solution; S4, weigh out ganoderic acid D, dissolve it in dimethyl sulfoxide, add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, let stand, and obtain ganoderic acid D solution; S5, the ganoderic acid D solution is added to the reconstituted solution in S3, and the mixture is subjected to ultrasonication, shaking, centrifugation, resuspension of the precipitate with water, and drying in sequence to obtain the sodium alginate-laminarin-ganoderic acid D nanoparticles.

[0013] Preferably, the mass ratio of sodium alginate, laminarin and ganoderic acid D in step S1 is 20~30:2~5:8~12, for example: 20:3:8, 24:3:10, 25:2:10, 27:4:9, 30:5:12.

[0014] Preferably, the kelp polysaccharide solution and the polyethyleneimine solution in step S2 are added to the sodium alginate solution at the same rate.

[0015] Furthermore, the method of addition is dropwise addition.

[0016] Preferably, the molecular weight of the dialysis bag used in step S2 is ≤100 kDa.

[0017] Preferably, the drying method in step S2 is air drying, low-temperature drying, or freeze drying, and more preferably freeze drying.

[0018] Preferably, the concentration of ganoderic acid D in the ganoderic acid D solution in step S4 is 15~50 g / L, for example: 15 g / L, 20 g / L, 30 g / L, 40 g / L, 50 g / L.

[0019] Fourthly, the present invention provides the use of the sodium alginate-laminarin-ganoderic acid D nanoparticles described in the first aspect or the composition described in the second aspect in the preparation of products for treating and / or alleviating ulcerative colitis.

[0020] Preferably, the product is a medicine or feed.

[0021] Furthermore, the drug is either a human or veterinary drug.

[0022] Furthermore, the product also includes excipients that are permitted to be added to pharmaceuticals or feed.

[0023] Preferably, the application includes at least one of the following: (1) Application in the preparation of products that alleviate weight loss; (2) Application in the preparation of products that alleviate colonic shortening; (3) Application in the preparation of products that reduce colonic mucosal stratification and crypt atrophy.

[0024] (4) Application in the preparation of products that reduce the content of myeloperoxidase in colonic tissue; (5) In the preparation of colon-lowering... and Applications in horizontal products; (6) Application in the preparation of products that enhance the expression levels of Occluding-1 and ZO-1; (7) Application in the preparation of products that reduce intestinal permeability.

[0025] The beneficial effects of this invention are: This invention utilizes sodium alginate gel as a carrier to load laminarin and ganoderic acid D, preparing sodium alginate-laminarin-ganoderic acid D nanoparticles. These nanoparticles are used to prevent and / or alleviate ulcerative colitis, exhibiting antioxidant effects, targeted regulation of intestinal barrier function, reduction of inflammatory responses, and improvement of intestinal health. This provides a novel approach for treating ulcerative colitis. Attached Figure Description

[0026] Figure 1 The image shows transmission electron microscopy (TEM) images of sodium alginate-laminarin-ganoderic acid D nanoparticles with different ganoderic acid loadings; A represents 100 mg of ganoderic acid D, B represents 20 mg of ganoderic acid D, C represents 30 mg of ganoderic acid D, and D represents 40 mg of ganoderic acid D. Figure 2 The DLS diagram of sodium alginate@laminarin is shown below; Figure 3 The image shows the DLS diagram of sodium alginate@laminarin loaded with ganoderic acid D; Figure 4 The standard curve for testing the coupling amount of ganoderic acid is shown. Figure 5 The diagram shows the experimental flowchart of sodium alginate-laminarin-ganoderic acid D nanoparticles regulating intestinal barrier function in DSS-induced colitis mice. Figure 6 The figure shows the changes in body weight of colitis mice after intervention with sodium alginate-laminarin-ganoderic acid D nanoparticles; Figure 7 The figure shows the colon length of mice with colitis after intervention with sodium alginate-laminarin-ganoderic acid D nanoparticles; Figure 8 The image shows HE staining of the colon of mice with colitis after intervention with sodium alginate-laminarin-ganoderic acid D nanoparticles. Figure 9 The figure shows the MPO content in the colonic tissue of mice with colitis after intervention with sodium alginate-laminarin-ganoderic acid D nanoparticles. Figure 10 The figure shows the cytokine content in the colon tissue of colitis mice after intervention with sodium alginate-laminarin-ganoderic acid D nanoparticles; Figure 11 The image shows the immunofluorescence staining results of colonic proteins in colitis mice after intervention with sodium alginate-laminarin-ganoderic acid D nanoparticles. Figure 12 The image shows the expression of colonic barrier function genes in colitis mice after intervention with sodium alginate-laminarin-ganoderic acid D nanoparticles. Figure 13 The figure shows the serum FITC-dextran content in colitis mice after intervention with sodium alginate-lamb polysaccharide-ganoderic acid D nanoparticles. Figure 14 The following is the safety experimental procedure for sodium alginate-laminarin-ganoderic acid D nanoparticles; Figure 15 The figure shows the changes in mouse body weight during a safety experiment of sodium alginate-laminarin-ganoderic acid D nanoparticles. Figure 16 The image shows the changes in organ indices in mice during a safety experiment of sodium alginate-laminarin-ganoderic acid D nanoparticles. In the figure, * indicates a significant difference from the model group (P < 0.05), and ** indicates a highly significant difference from the model group (P < 0.01). Detailed Implementation

[0027] The technical solution of the present invention will be clearly and completely described below with reference to the embodiments and accompanying drawings. The described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0028] Example 1: Preparation of sodium alginate-laminarin-ganoderic acid D nanoparticles (1) Take 240 mg of sodium alginate (purchased from MCE), disperse it in 80 mL of water, stir overnight at room temperature, and record it as solution A.

[0029] (2) Take 30 mg of laminarin (purchased from MCE), disperse it in 40 mL of water, stir overnight at room temperature, and record it as solution B.

[0030] (3) Take 20 mg of polyethyleneimine (PEI, purchased from Aladdin), disperse it in 40 mL of water, and denote it as solution C. At room temperature, add solutions B and C at the same dropping rate to solution A simultaneously, react at room temperature for 45 minutes, and then let it stand at room temperature for 8 hours. The resulting mixed solution is placed in a dialysis bag with a molecular weight cutoff of 100 kDa and dialyzed in 100 times its volume of deionized water for 4 days, changing the dialysate every 12 hours. After dialysis, add sucrose to the dialysate as a lyophilization protectant to a final concentration of 10% (w / v), pre-freeze, and then lyophilize for 72 hours to obtain a solid sample.

[0031] (4) The lyophilized solid sample was reconstituted in 15 mL of dimethyl sulfoxide (DMSO) to obtain a reconstituted solution; (5) Take 20 mg, 30 mg, 40 mg and 100 mg of ganoderic acid D (purchased from MCE), dissolve them in 4 mL of DMSO, add 1 mL of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC, purchased from Aladdin), and let stand at room temperature for 30 minutes to obtain ganoderic acid D solutions of different concentrations.

[0032] (6) Add the different concentrations of ganoderic acid D solution from (5) to the reconstituted solution from (4), sonicate for 10 min under ice bath conditions, power 100 W, frequency 30 kHz; react overnight on a shaker at room temperature, speed 100~120 rpm, time 12 h; centrifuge at 4℃, 15,000 g, 15 min, and wash. Then redisperse in 50 mL of water, add sucrose as a freeze-drying protectant to make the final concentration 10% (w / v), freeze-dry for 72 h after pre-freezing to obtain sodium alginate-laminarin-ganoderic acid D nanoparticle solid powder.

[0033] (7) The size and morphology of sodium alginate-laminarin-ganoderic acid D nanoparticles were detected by transmission electron lens. (8) Take the solid sample from step (3) and the sodium alginate-laminarin-ganoderic acid D nanoparticle sample from step (6) respectively, and use dynamic light scattering (DLS) to detect the particle size distribution. After acid hydrolysis and destruction of the nanoparticles, use HPLC to detect the content of ganoderic acid and perform ganoderic acid D coupling amount test to detect unbound ganoderic acid D, so as to prove that ganoderic acid D has entered the nano system.

[0034] like Figure 1As shown, based on the analysis of morphological uniformity, dispersibility, particle size distribution, and structural integrity, nanoparticle A is mainly composed of regular spherical / quasi-spherical shapes, with almost no rod-shaped or irregular shapes, meeting the morphological requirements of an ideal nanocarrier. There is no obvious aggregation between particles, and the boundaries are clear, avoiding the problems of increased particle size, decreased drug loading efficiency, and uneven in vivo distribution caused by aggregation. The relatively consistent particle size is beneficial to ensuring batch stability and predictable in vivo pharmacokinetic behavior. The clear edges and complete outlines of the particles indicate that the structure was stable during preparation and no obvious breakage or deformation occurred. Nanoparticle B is mainly composed of rod-shaped particles with only a small number of spherical particles interspersed, exhibiting poor overall morphological uniformity and significant large-scale aggregation. Particle C is mainly composed of spherical particles, but significant aggregation is visible. The overall particle size is larger than the aforementioned ideal particles, the boundary clarity is reduced, and some particles show breakage or incomplete morphology. Particle D has an excessively high overall particle density, with particles distributed too closely together, making it difficult to distinguish the boundaries clearly, suggesting significant particle accumulation, adhesion, or local fusion. Therefore, nanoparticle A will be used for further experiments.

[0035] like Figure 2 and 3 As shown in the data, the DLS of sodium alginate@lamb polysaccharide was 112.7 nm, while the DLS of ganoderic acid-loaded was 129.6 nm. The increase in DLS indicates that the ganoderic acid-D loading was successful.

[0036] like Figure 4 As shown in the standard curve, the OD value of the supernatant is 0.259. Substituting into the formula, the calculated concentration is 2.6 mg / mL. The total volume is 20 mL, the ganoderic acid in the supernatant is 52 mg, and the total added amount is 100 mg. Therefore, the modification amount is 48 mg.

[0037] Example 2: Experiment on the regulation of intestinal barrier function in mice with DSS-induced colitis by sodium alginate-laminarin-ganoderic acid D nanoparticles. according to Figure 5 The following experiments will be conducted according to the experimental procedure shown: Thirty-six 6-week-old female C57BL / 6 mice (purchased from Beijing Huafukang Biotechnology Co., Ltd.) were divided into 6 groups of 6 mice each. The groups were: normal control group (PBS + normal drinking water, hereinafter referred to as PBS group), ulcerative colitis model group (PBS + DSS + drinking water, hereinafter referred to as DSS group), sodium alginate group (SA + DSS + drinking water, hereinafter referred to as SA group), laminarin group (KP + DSS + drinking water, hereinafter referred to as KP group), ganoderic acid D group (GAD + DSS + drinking water, hereinafter referred to as GAD group), and sodium alginate-laminarin-ganoderic acid D group (SA-KP-GAD + DSS + drinking water, hereinafter referred to as SA-KP-GAD group). The mice were allowed to acclimatize for three days before the formal experiment began. The normal control group received normal drinking water, while the other five groups received 3% sodium dextran sulfate (DSS) in their drinking water. After 6 days, all groups returned to normal drinking water. During the experiment, mice were gavaged every two days. The normal group and the ulcerative colitis model group were administered PBS by gavage. The SA group was administered 1 mg / mL sodium alginate by gavage, the KP group was administered 1 mg / mL laminarin by gavage, the GAD group was administered 1 mg / mL ganoderic acid D by gavage, and the SA-KP-GAD group was administered 1 mg / mL sodium alginate-laminarin-ganoderic acid D by gavage. PBS was used as the solvent for all reagents. All mice were weighed daily during the formal experiment. On day 9, FITC-dextran was administered by gavage. Four hours later, blood was collected, the mice were anesthetized, euthanized, and dissected to measure colon length. Colonic proteins were extracted and detected using an enzyme-linked immunosorbent assay (ELISA) kit (all kits were provided by Shanghai Hengyuan Biotechnology Co., Ltd.). (HB1931-Mu) (HB049-Mu), IL-6 (HY1061-Mu), IL-10 (HY1087-D), The levels of (HY027-Mu) and myeloperoxidase (MPO) (HB1200-Mu) were measured. Paraffin sections of mouse colon tissue were prepared and immunofluorescence staining was performed to detect Occludin-1 (Proteintech, 27260-1-AP) and ZO-1 (Proteintech, 21773-1-AP). (Abcam, ab6302) and The protein expression level of (Abcam, ab14425) was measured; colonic RNA was extracted using the RNA Easy Fast Animal Tissue RNA Extraction Kit (DP451) from Tiangen Biotech (Beijing) Co., Ltd. to detect the mRNA expression of ZO-1 and Occluding-1; intestinal permeability was detected based on the brightness of FITC-dextran (purchased from MCE, HY-128868) under a fluorescence microscope.

[0038] like Figure 6 As shown, the normal group mice showed a steady increase in body weight without significant fluctuations. However, the model group mice given 3% DSS showed stagnant weight gain from day 3 onwards, and began to decline significantly after day 4, with their body weight significantly lower than the initial level by the end of the experiment, indicating a successful establishment of the ulcerative colitis model. In the first 6 days, there were no significant differences in body weight changes among the intervention groups compared to the model group; on day 7, the SA-KP-GAD gavage group mice had significantly higher body weight than the model group (P<0.05); on day 8, the SA-KP-GAD gavage group mice had extremely significantly higher body weight than the model group (P<0.01), and also significantly higher than the SA, KP, and GAD gavage groups (P<0.05). This indicates that the use of sodium alginate-laminarin-ganoderic acid D nanoparticles can effectively alleviate DSS-induced weight loss in mice.

[0039] Compare the efficacy of SA-KP-GAD with conventional IBD treatments, such as sulfasalazine (SASP) and berberine (BBR), see the literature (Zhang et al. Journal of Nanobiotechnology (2024) 22:607, doi.org / 10.1186 / s12951-024-02856-z; Yang et al. Journal of Nanobiotechnology (2025)23:444, doi.org / 10.1186 / s12951-025-03466-z; J. Agric. Food Chem.2025,73, 12690-12701, doi.org / 10.1021 / acs.jafc.4c11976). SA-KP-GAD treatment significantly enhanced the efficacy against DSS-induced acute colitis, achieving complete weight recovery compared to clinical doses of SASP and BBR (P>0.05). These results indicate that sodium alginate-laminarin-ganoderic acid D nanoparticles (SA-KP-GAD) are a promising candidate drug with high therapeutic potential, effectively addressing the symptoms of ulcerative colitis and showing promising clinical application prospects.

[0040] like Figure 7As shown, compared with the normal group, the colon of mice in the model group was significantly shortened (P<0.01), which is a typical pathological feature of DSS-induced acute colitis, further confirming the successful establishment of the ulcerative colitis model. Compared with the model group, each treatment group showed varying degrees of relief from colon shortening. Among them, the colon length of mice in the SA-KP-GAD gavage group was significantly higher than that in the model group (P<0.01), while there was no statistically significant difference compared with the normal group (P>0.05). Compared with commonly used clinical treatment drugs sulfasalazine (SASP) and berberine (BBR), sodium alginate-laminarin-ganoderic acid D nanoparticles have a better intervention effect on DSS-induced colon shortening, and can significantly restore colon length to near normal levels.

[0041] like Figure 8 As shown, HE staining of the colon revealed that in the PBS gavage group, the intestinal mucosa was intact, epithelial cells were neatly arranged without post-sloughing necrosis, and the muscularis propria and serosa showed no structural changes. In the DSS model group, the submucosa was separated, intestinal crypts atrophied, and inflammatory cells infiltrated the mucosa. In the SA group, intestinal crypts partially atrophied. In the KP group, intestinal mucosal epithelial cells showed diffuse and severe degeneration and necrosis, intestinal crypt atrophy, submucosa edema, and extensive inflammatory cell infiltration. In the GAD group, intestinal mucosal epithelial cells showed localized mild degeneration and necrosis, and intestinal villi detached. In the SA-KP-GAD group, intestinal crypts were intact, and intestinal mucosal epithelial cells showed localized mild degeneration and necrosis. Compared with commonly used clinical treatment drugs sulfasalazine (SASP) and berberine (BBR), sodium alginate-laminarin-ganoderic acid D nanoparticles showed similar effects in the recovery of the colonic mucosa and muscularis propria induced by DSS. This indicates that sodium alginate-laminarin-ganoderic acid D nanoparticles can improve the pathological changes in colonic tissue after DSS treatment, reduce mucosal stratification and crypt atrophy, and promote intestinal health.

[0042] like Figure 9 As shown, the MPO content in the colonic tissue of mice in the model group was significantly higher than that in the normal group (P<0.01), indicating that there was a large amount of neutrophil infiltration in the colonic tissue after DSS induction. In contrast, the MPO content in the colonic tissue of mice in the SA-KP-GAD gavage group was significantly lower than that in the model group (P<0.05). This indicates that sodium alginate-laminarin-ganoderic acid D nanoparticles can alleviate the regression of neutrophil infiltration.

[0043] like Figure 10 As shown, compared with the normal group, the colon of the model group had... IL-6 and The levels were significantly elevated (P<0.05), indicating that DSS induction successfully triggered a strong pro-inflammatory response. The anti-inflammatory factor IL-10 was significantly elevated in the model group, while... There were no significant differences in the level groups among the treatment groups. Each intervention group showed different regulatory effects on pro-inflammatory factors. Compared with the model group, the colon of the KP gavage group... The level decreased significantly (P<0.05) in the SA-KP-GAD gavage group. The levels decreased significantly (P<0.01). This indicates that the body's inflammatory response has been alleviated. Both KP and SA-KP-GAD can effectively inhibit the overexpression of key pro-inflammatory factors, thereby alleviating the local inflammatory response, and the intervention effect of SA-KP-GAD is more significant.

[0044] like Figure 11 As shown, immunofluorescence staining was performed on mouse colon tissue to detect Occludin-1, ZO-1, and... and The protein expression levels of Occludin-1 and ZO-1 were assessed to evaluate changes in intestinal barrier function. Results showed that, compared to the control group, the DSS model group exhibited significantly higher levels of these proteins. and The fluorescence signals of the proteins were all weakened; however, in the SA and GAD groups, the fluorescence signals recovered to varying degrees; the recovery of fluorescence signals was most significant in the SA-KP-GAD group. This indicates that sodium alginate-laminarin-ganoderic acid D nanoparticles can effectively repair intestinal barrier function damage caused by DSS.

[0045] like Figure 12 As shown, compared with the normal group, the expression of ZO-1 gene in the model group was significantly reduced (P<0.01), indicating that DSS-induced colitis severely disrupts the tight junction structure of the intestinal epithelium. In contrast, the SA, KP, GAD, and SA-KP-GAD gavage groups all significantly increased ZO-1 expression (P<0.05). Meanwhile, the Occluding-1 gene expression in the SA-KP-GAD gavage group was significantly higher than that in the model group (P<0.05). This indicates that all treatment groups effectively improved intestinal barrier function and alleviated local inflammatory responses, with the SA-KP-GAD intervention showing the most significant effect.

[0046] like Figure 13 As shown, the serum FITC-dextran signal intensity in the model group was significantly higher than that in the normal group (P<0.05), indicating that DSS-induced colitis leads to increased intestinal permeability and damage to the intestinal barrier function. The serum FITC-dextran signal intensity in the SA-KP-GAD gavage group was significantly lower than that in the model group (P<0.05), which is consistent with the upregulation of mRNA expression of tight junction protein genes (ZO-1, Occludin-1). This further confirms from a functional perspective that SA-KP-GAD intervention can effectively reduce intestinal permeability and repair the damaged intestinal barrier function.

[0047] Example 3: Safety Experiment of Sodium Alginate-Laminaria Polysaccharide-Ganoderic Acid D Nanoparticles according to Figure 14 The following experiments will be conducted according to the experimental procedure shown: Sixteen 6-week-old female C57BL / 6 mice were selected and divided into two groups of eight each: a control group (PBS) and a sodium alginate-laminarin-ganoderic acid D group (SA-KP-GAD). After three days of acclimatization, the formal gavage experiment began. The control group was administered PBS by gavage, while the treatment group was administered 1 mg / mL sodium alginate-laminarin-ganoderic acid D by gavage. Gavage was performed every two days. All mice were weighed daily. On day 9, they were administered FITC-dextran by gavage. Four hours later, the mice were sacrificed. The heart, liver, spleen, lungs, and left and right kidneys were weighed, and blood was collected to perform complete blood count and blood biochemistry tests.

[0048] like Figure 15 As shown, the body weight of mice in the control group fluctuated within a certain range, while the body weight of mice in the SA-KP-GAD group showed a continuous and stable growth trend. There was no statistically significant difference in body weight between the two groups during the experimental period (P>0.05). This indicates that sodium alginate-laminarin-ganoderic acid D nanoparticles did not have a negative impact on the body weight gain of mice.

[0049] like Figure 16 As shown, the mice were dissected and weighed, and the differences in organ indices between the groups were calculated. The results showed no statistically significant difference in the weight of organs (heart, liver, spleen, lungs, and left and right kidneys) between the control group and the SA-KP-GAD group (P>0.05). This indicates that the sodium alginate-laminarin-ganoderic acid D nanoparticles did not have a significant negative impact on the major organs of the mice.

[0050] The results of the biochemical indicators in Table 1 and the blood routine indicators in Table 2 show that, compared with the control group, the glucose content in the blood biochemical indicators of the SA-KP-GAD group mice was significantly increased (P<0.01), while there were no statistically significant differences in the other blood biochemical and routine indicators (P>0.05). This may be because the mice were not fasted before blood collection, suggesting that sodium alginate-laminarin-ganoderic acid D nanoparticles may have a certain impact on the glucose metabolism process in mice, but will not cause significant adverse effects on other physiological functions.

[0051] Table 1 Blood biochemical indicators of mice Table 2. Blood routine indicators of mice The above description is merely an embodiment of the present invention and does not limit the patent scope of the present invention. Any equivalent structural transformations made based on the content of the present invention specification, or direct or indirect applications in other related technical fields, are similarly included within the patent protection scope of the present invention.

Claims

1. A sodium alginate-laminarin-ganoderic acid D nanoparticle, characterized in that, The raw materials for the nanoparticles include sodium alginate, laminarin, and ganoderic acid D in a mass ratio of 20-30:2-5:8-12; the preparation process of the nanoparticles... Includes the following steps: S1, Weigh out sodium alginate, laminarin and polyethyleneimine, and disperse them in water to obtain sodium alginate solution, laminarin solution and polyethyleneimine solution; S2, the kelp polysaccharide solution and the polyethyleneimine solution are added to the sodium alginate solution, and after the reaction is allowed to stand, dialyzed, and the dialysate is dried to obtain a solid sample; S3, dissolve the solid sample in dimethyl sulfoxide to obtain a reconstituted solution; S4, weigh out ganoderic acid D, dissolve it in dimethyl sulfoxide, add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, let stand, and obtain ganoderic acid D solution; S5, the Ganoderma lucidum acid D solution is added to the reconstituted solution in S3, and the mixture is subjected to ultrasonication, shaking, centrifugation, resuspending the precipitate in water, and drying to obtain the sodium alginate-laminarin-Ganoderma lucidum acid D nanoparticles.

2. The sodium alginate-laminarin-ganoderic acid D nanoparticles according to claim 1, characterized in that, The nanoparticles have a particle size of 80~100 nm.

3. A composition, characterized in that, The composition comprises sodium alginate-laminarin-ganoderic acid D nanoparticles as described in any one of claims 1-2.

4. A method for preparing sodium alginate-laminarin-ganoderic acid D nanoparticles, characterized in that, The preparation method includes the following steps: S1, Weigh out sodium alginate, laminarin and polyethyleneimine, and disperse them in water to obtain sodium alginate solution, laminarin solution and polyethyleneimine solution; S2, the kelp polysaccharide solution and the polyethyleneimine solution are added to the sodium alginate solution, and after the reaction is allowed to stand, dialyzed, and the dialysate is dried to obtain a solid sample; S3, dissolve the solid sample in dimethyl sulfoxide to obtain a reconstituted solution; S4, weigh out ganoderic acid D, dissolve it in dimethyl sulfoxide, add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, let stand, and obtain ganoderic acid D solution; S5, the Ganoderma lucidum acid D solution is added to the reconstituted solution in S3, and the mixture is subjected to ultrasonication, shaking, centrifugation, resuspending the precipitate in water, and drying to obtain the sodium alginate-laminarin-Ganoderma lucidum acid D nanoparticles.

5. The preparation method according to claim 4, characterized in that, The mass ratio of sodium alginate, laminarin, and ganoderic acid D is 20-30:2-5:8-12.

6. The preparation method according to claim 4, characterized in that, In step S2, the kelp polysaccharide solution and the polyethyleneimine solution are added to the sodium alginate solution at the same rate.

7. The preparation method according to claim 6, characterized in that, The method of addition is dropwise addition.

8. The preparation method according to claim 4, characterized in that, The dialysis bag used in step S2 has a molecular weight ≤100 kDa.

9. The preparation method according to claim 4, characterized in that, The concentration of ganoderic acid D in the ganoderic acid D solution in step S4 is 10~50 g / L.

10. The use of the sodium alginate-laminarin-ganoderic acid D nanoparticles of claim 1 or the composition of claim 3 in the preparation of a medicament for treating and / or alleviating ulcerative colitis.

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