Lycium barbarum polysaccharide and betaine self-assembled nanoparticles as well as preparation method and application thereof

Abstract

The invention discloses lycium barbarum polysaccharide and betaine self-assembled nanoparticles and a preparation method and application thereof, and belongs to the technical field of medicines.The preparation method of the lycium barbarum polysaccharide and betaine self-assembled nanoparticles comprises the following steps that betaine is dissolved in absolute ethyl alcohol to obtain an organic phase, lycium barbarum polysaccharide is dissolved in deionized water to obtain a water phase; slowly dropwise adding the organic phase into the water phase under an ultrasonic condition, and carrying out rotary evaporation to remove the organic solvent, so as to obtain the lycium barbarum polysaccharide and betaine self-assembled nanoparticles. The lycium barbarum polysaccharide and betaine self-assembled nanoparticles have good in-vivo and in-vitro anti-inflammatory activity, and the mode of preparing the combined self-assembled nanoparticles can achieve the effect-enhancing and toxicity-reducing effects, so that a foundation is laid for research on the anti-inflammatory activity of medicine and food homologous medicines, especially polysaccharide extracted from lycium barbarum; the invention provides a new strategy for further administration treatment of colitis, and has a great application prospect.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, in particular to self-assembled nanoparticles of Lycium barbarum polysaccharide and betaine, a preparation method and application thereof, and more particularly to self-assembled nanoparticles of Lycium barbarum polysaccharide and betaine, a preparation method thereof and application thereof in a drug for treating ulcerative colitis. Background Art

[0002] Ulcerative colitis (UC) is a chronic, recurrent inflammatory gastrointestinal disease characterized by immune system disorders and gastrointestinal inflammation. It can cause inflammatory damage to the colon mucosa, destroy the gastrointestinal mucosal barrier, and increase the risk of colorectal cancer. Its clinical manifestations mainly include abdominal pain, diarrhea, blood in the stool, and weight loss. At present, UC is still difficult to cure, and the treatment strategy is mainly to target the colon mucosa and reduce systemic inflammation to regulate the aggravated host immune response. Generally, for UC patients, long-term uninterrupted drug treatment is required, and even surgical intervention is required in severe cases. The main clinical treatment for this disease is mesalazine (5-ASA), which can effectively inhibit the inflammatory response of the intestinal wall, but there is a high recurrence rate after discontinuation of this drug, and the patient's acceptance is low. After the patient's systemic distribution, it will cause certain adverse reactions, and the long-term intake of these drugs will lead to impaired immunity, making patients more susceptible to infection and disease.

[0003] Lycium barbarum polysaccharides (LBP) are derived from the traditional Chinese medicinal and edible plant Lycium barbarum L., and are one of the main bioactive components of Lycium barbarum. They have a wide range of biological properties, including anti-inflammatory, immunomodulatory, attenuation, and regulation of intestinal flora. In addition, combined administration is considered an effective strategy for the treatment of UC, which can reduce adverse drug reactions while improving the body's immunity and increasing anti-inflammatory effects. Betaine is a zwitterionic quaternary ammonium compound and a methyl derivative of glycine. It has certain transmethylation, osmotic pressure regulation, anti-inflammatory, antioxidant and immune functions. Lycium barbarum polysaccharide is an acidic polysaccharide. BET-LBP-1NPs are formed by self-assembly of acidic polysaccharides and alkaloids. The combination of the two drugs can have a certain promoting effect on the treatment of inflammation.

[0004] Therefore, how to develop a self-assembled nanoparticle of Lycium barbarum polysaccharide and betaine and a preparation method thereof and use thereof in a drug for treating ulcerative colitis are technical problems that need to be solved urgently by those skilled in the art. Summary of the invention

[0005] In view of this, the present invention provides a self-assembled nanoparticle of Lycium barbarum polysaccharide and betaine, a preparation method thereof, and an application thereof in a drug for treating ulcerative colitis.

[0006] In order to achieve the above object, the present invention adopts the following technical solution:

[0007] A method for preparing self-assembled nanoparticles of wolfberry polysaccharide and betaine comprises the following steps:

[0008] (1) dissolving betaine in anhydrous ethanol to obtain an organic phase, and dissolving wolfberry polysaccharide in deionized water to obtain an aqueous phase;

[0009] (2) The organic phase is slowly dripped into the aqueous phase under ultrasonic conditions, and the organic solvent is removed by rotary evaporation to obtain the above-mentioned Lycium barbarum polysaccharide and betaine self-assembled nanoparticles.

[0010] The beneficial effects of the present invention are as follows: Lycium barbarum polysaccharide is an acidic polysaccharide, and BET-LBP-1NPs are formed by self-assembly of acidic polysaccharide and alkaloids, and the combination of the two drugs can have a certain promoting effect on the treatment of inflammation. The present invention constructs a Lycium barbarum polysaccharide betaine self-assembled nanoparticle by self-assembling betaine (BET) and Lycium barbarum polysaccharide (LBP), an effective ingredient extracted from Lycium barbarum, into nanoparticles by ultrasonic instillation, and uses ulcerative colitis mice as a model to study the in vivo anti-inflammatory activity of the self-assembled nanoparticles by oral administration through gavage, and investigates the drug efficacy comparison with mesalazine (5-ASA); experimental studies have shown that BET / LBP-1NPs have good in vitro and in vivo anti-inflammatory activity, and this method of preparing combined self-assembled nanoparticles can achieve the effect of enhancing synergy and reducing toxicity, which lays a foundation for the study of the anti-inflammatory activity of medicines and edibles, especially polysaccharides extracted from Lycium barbarum, and provides a new strategy for further drug administration and treatment of colitis, and has great application prospects.

[0011] Furthermore, in step (1), the mass volume ratio of betaine, wolfberry polysaccharide, anhydrous ethanol and deionized water is (20-60) mg: (10-30) mg: (1-2) mL: 5 mL.

[0012] The beneficial effects of adopting the above further technical solution are as follows: when the ratio of betaine to wolfberry polysaccharide is 2:1, the preparation effect is best, the particle size of the prepared nanoparticles is uniform and small, and the oral absorption utilization can be better increased.

[0013] Furthermore, in step (2), the ultrasonic power is 25w.

[0014] Furthermore, in step (2), the organic phase is slowly dripped into the aqueous phase drop by drop at a dripping speed of 2 drops / s.

[0015] Furthermore, in step (2), the rotary evaporation temperature is 45° C., the rotary evaporation pressure is 90 bar, and the rotary evaporation time is 3-5 minutes.

[0016] The present invention also provides self-assembled nanoparticles of Lycium barbarum polysaccharide and betaine prepared by the method.

[0017] The present invention also provides an application of the self-assembled nanoparticles of Lycium barbarum polysaccharide and betaine in a drug for treating ulcerative colitis. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 Daily body weight changes of mice in each group;

[0019] Figure 2 Disease activity index (DAI) of mice in each group;

[0020] Figure 3 Evaluation of colon pathological changes in each group (compared with the Model group, *P < 0.05; compared with the normal group, #P < 0.05);

[0021] Figure 4 Colon anatomical images and colon length of each group (compared with the Model group, *P < 0.05; compared with the Blank group, #P < 0.05);

[0022] Figure 5 HE staining of colon pathological sections of each group;

[0023] Figure 6 In the figure, a is the cytokine IL-6 Elisa detection result in the tissues of each group (n=6); b is the cytokine IL-6 Elisa detection result in the serum of each group (n=6); c is the cytokine IL-10 Elisa detection result in the tissues of each group (n=6); d is the cytokine IL-10 Elisa detection result in the serum of each group (n=6); e is the cytokine IFN-γ Elisa detection result in the tissues of each group (n=6); f is the cytokine IFN-γ Elisa detection result in the serum of each group (n=6); (compared with the Model group, *P<0.05). DETAILED DESCRIPTION

[0024] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0025] The method for extracting Lycium barbarum polysaccharides in the embodiments and comparative examples of the present invention (see Chinese patent application number: 202410863239.5) comprises the following steps:

[0026] (1) 500 g of wolfberry was crushed and defatted with 95% ethanol, dried and weighed; the mixture was extracted with 100° C. hot water in a decoction pot for 2 h at a solid-liquid ratio of 1:10, and the extraction was repeated 3 times, and the filtrate was combined and concentrated under reduced pressure, and precipitated with 3 times the volume of 95% ethanol, dried and weighed to obtain crude LBP polysaccharide;

[0027] (2) The crude LBP polysaccharide solution prepared in step (1) is repeatedly subjected to the Sevag reagent (V chloroform: V n-butanol = 4:1) to remove the protein until the white middle layer disappears; the supernatant is collected, concentrated, and freeze-dried to obtain the deproteinized LBP; the deproteinized LBP solution is depigmented using D101 macroporous adsorption resin, and the eluate is then concentrated by rotary evaporation and freeze-dried to obtain the decolorized LBP;

[0028] (3) Weigh 100 mg of the decolorized LBP prepared in step (2) and dissolve it in 10 mL of distilled water. After it is fully dissolved, centrifuge at 5000 r / min for 10 min to obtain the supernatant; chromatograph on a DEAE-52 cellulose column (Φ2.6×50 cm), and elute with distilled water, 0.1 mol / L NaCl, 0.2 mol / L NaCl, and 0.5 mol / L NaCl in sequence to obtain LBP-1, LBP-2, LBP-3, and LBP-4, respectively, at an elution flow rate of 2 mL / min, and collect 10 mL in each tube; collect the eluate, and detect it by phenol-sulfuric acid method in separate tubes, and draw an elution curve with the tube number as the horizontal axis and the absorbance value as the vertical axis; based on the elution curve, combine the eluates of the same elution peak, concentrate under reduced pressure, and freeze-dry to obtain four refined LBPs with different molecular weights, and take LBP-1 with a relative molecular mass of 2726 for subsequent preparation.

[0029] Example 1

[0030] The preparation method of Lycium barbarum polysaccharide and betaine self-assembled nanoparticles comprises the following steps:

[0031] (1) dissolving 20 mg of betaine in 1 mL of anhydrous ethanol to obtain an organic phase, and dissolving 10 mg of Lycium barbarum polysaccharide in 5 mL of deionized water to obtain an aqueous phase;

[0032] (2) Under ultrasonic conditions with an ultrasonic power of 25 W, the organic phase was slowly added dropwise into the aqueous phase at a dropping speed of 2 drops / s, and the organic solvent was removed by rotary evaporation at a rotary evaporation temperature of 45 ° C, a rotary evaporation pressure of 90 bar, and a rotary evaporation time of 3 minutes to obtain a solution of self-assembled nanoparticles of Lycium barbarum polysaccharide and betaine, recorded as BET / LBP-1NPs.

[0033] Example 2

[0034] The preparation method of Lycium barbarum polysaccharide and betaine self-assembled nanoparticles comprises the following steps:

[0035] (1) Dissolve 40 mg of betaine in 1.5 mL of anhydrous ethanol to obtain an organic phase, and dissolve 20 mg of Lycium barbarum polysaccharide in 5 mL of deionized water to obtain an aqueous phase;

[0036] (2) Under ultrasonic conditions with an ultrasonic power of 25 W, the organic phase was slowly added dropwise into the aqueous phase at a dropping speed of 2 drops / s, and the organic solvent was removed by rotary evaporation at a rotary evaporation temperature of 45°C, a rotary evaporation pressure of 90 bar, and a rotary evaporation time of 4 minutes to obtain a solution of Lycium barbarum polysaccharides and betaine self-assembled nanoparticles.

[0037] Example 3

[0038] The preparation method of Lycium barbarum polysaccharide and betaine self-assembled nanoparticles comprises the following steps:

[0039] (1) dissolving 60 mg of betaine in 2 mL of anhydrous ethanol to obtain an organic phase, and dissolving 30 mg of Lycium barbarum polysaccharide in 5 mL of deionized water to obtain an aqueous phase;

[0040] (2) Under ultrasonic conditions with an ultrasonic power of 25 W, the organic phase was slowly added dropwise into the aqueous phase at a dropping speed of 2 drops / s, and the organic solvent was removed by rotary evaporation at a rotary evaporation temperature of 45°C, a rotary evaporation pressure of 90 bar, and a rotary evaporation time of 5 minutes to obtain a solution of self-assembled nanoparticles of Lycium barbarum polysaccharides and betaine.

[0041] Comparative Example 1

[0042] The preparation method of Lycium barbarum polysaccharide and betaine self-assembled nanoparticles comprises the following steps:

[0043] (1) dissolving 10 mg of betaine in 1 mL of anhydrous ethanol to obtain an organic phase, and dissolving 10 mg of Lycium barbarum polysaccharide in 5 mL of deionized water to obtain an aqueous phase;

[0044] (2) Under ultrasonic conditions with an ultrasonic power of 25 W, the organic phase was slowly added dropwise into the aqueous phase at a dropping speed of 2 drops / s, and the organic solvent was removed by rotary evaporation at a rotary evaporation temperature of 45°C, a rotary evaporation pressure of 90 bar, and a rotary evaporation time of 5 minutes to obtain a solution of self-assembled nanoparticles of Lycium barbarum polysaccharides and betaine.

[0045] Comparative Example 2

[0046] The preparation method of Lycium barbarum polysaccharide and betaine self-assembled nanoparticles comprises the following steps:

[0047] (1) dissolving 40 mg of betaine in 1.5 mL of anhydrous ethanol to obtain an organic phase, and dissolving 10 mg of Lycium barbarum polysaccharide in 5 mL of deionized water to obtain an aqueous phase;

[0048] (2) Under ultrasonic conditions with an ultrasonic power of 25 W, the organic phase was slowly added dropwise into the aqueous phase at a dropping speed of 2 drops / s, and the organic solvent was removed by rotary evaporation at a rotary evaporation temperature of 45°C, a rotary evaporation pressure of 90 bar, and a rotary evaporation time of 5 minutes to obtain a solution of self-assembled nanoparticles of Lycium barbarum polysaccharides and betaine.

[0049] Comparative Example 1

[0050] The preparation method of the physical mixture group (BET+LBP-1) comprises the following steps:

[0051] (1) 20 mg of betaine and 10 mg of Lycium barbarum polysaccharide (LBP-1) were mixed to obtain a physical mixture group, which was recorded as BET+LBP-1.

[0052] Effect Experiment

[0053] 1. Basic characterization

[0054] The Lycium barbarum polysaccharide and betaine self-assembled nanoparticles in Example 1, Comparative Example 1 and Comparative Example 2 were taken, and the particle size, polydispersity index (PDI) and Zeta potential were measured using a ZetasizerNano ZS Malvern particle size potential analyzer, and each sample was measured three times.

[0055] Table 1 Changes in the particle size of nanoparticles at different drug ratios

[0056] LBP-1:BET LBP-1:BET Particle size (nm) PDI Zeta(mV) Comparative Example 1 1:1 310.9±9.496 0.319±0.036 -16.0±3.51 Example 1 1:2 171.2±10.78 0.239±0.036 -14.5±1.91 Comparative Example 2 1:4 400.7±33.42 0.530±0.050 -11.8±2.33

[0057] As can be seen from Table 1, when LBP-1:BET=1:2, the particle size and PDI are the smallest, indicating that the nanoparticles formed at this dosage are stable and uniform, and have a small particle size. They can accumulate more at the disease site in the body and have better bioavailability to exert anti-inflammatory effects.

[0058] 2. Determination of drug loading of BET / LBP-1 NPs

[0059] Take 1 mL of the nanoparticle solution of Example 1 and freeze-dry (pre-freezing temperature -20°C, main drying temperature 20°C, vacuum 0.370 mbar, final drying temperature 30°C, vacuum 0.05 bar, drying time 48 h) and accurately weigh the weight of the freeze-dried powder as W. Dissolve the freeze-dried powder with 10 volumes of chromatographic methanol, vortex, centrifuge at 13000 rpm for 30 min, take the supernatant for HPLC analysis, and substitute the measured standard curve y=0.062x+0.5742(R 2 =0.9994) to obtain the content C of betaine in 1 mL of nanoparticle solution. The drug loading capacity (DLC) was calculated according to the following formula.

[0060]

[0061] The drug loading of BET / LBP-1NPs was measured by HPLC to be 22.53%.

[0062] From the above, it can be seen that the drug loading of BET / LBP-1NPs is 22.53%, which improves the solubility of the monomer, indicating that the drug is successfully encapsulated, which is convenient for subsequent drug administration.

[0063] 3. Animal Experiment Design

[0064] 70 mice were acclimated for one week and randomly divided into 7 groups, with 10 mice in each group: normal control group (Blank), model control group (Model), positive drug mesalazine group (150 mg / kg) (5-ASA), Lycium barbarum polysaccharide group (LBP-1), betaine group (BET), physical mixture group of comparative example 1 (BET+LBP-1), and self-assembled nanoparticles group of Lycium barbarum polysaccharide and betaine of Example 1 (BET / LBP-1NPs);

[0065] One day before the experiment, mice in each group were fasted but not watered for 12 h. During the experiment, all mice were given standard feed with a daily gavage volume of 1 mL / 100 g for 13 consecutive days. On the 14th day, samples were collected from each group and relevant indicators were tested.

[0066] Among them, mice in the Blank group were given sterile water and drank freely during the experiment;

[0067] The model group was given sterile water containing 4% dextran sulfate sodium (DSS) for 7 consecutive days to induce the establishment of an ulcerative colitis model. After the model was successfully established, the mice were first given normal water for 3 days and then 3.5% DSS for continued free drinking.

[0068] The LBP-1 group, BET group, BET+LBP-1 group, and BET / LBP-1NPs group were given sterile water containing 4% dextran sulfate sodium (DSS) for 7 consecutive days to induce the establishment of ulcerative colitis models. After the model was successfully established, the mice were first given normal water for free drinking for 3 days, and then changed to 3.5% DSS for continued free drinking. At the same time, they were gavaged every day at a dose of 300 mg / kg.

[0069] The 5-ASA group was given water containing 4wt% dextran sulfate sodium (DSS) for 7 consecutive days to induce the establishment of an ulcerative colitis model. After the model was successfully established, the mice were first given normal water for 3 days, and then 3.5% DSS for continued drinking. The mice were gavaged daily at a dose of 150 mg / kg.

[0070] 3.1 Disease Activity Index Score (DAI Score)

[0071] The body weight of mice in each group was recorded regularly every day, and the spirit, physical signs and defecation of mice were observed twice a day. The mental state, stool characteristics, hair smoothness, neatness and glossiness, perianal lesions and fecal occult blood of mice were observed and recorded. The disease activity index (DAI) scoring standard is shown in Table 1, DAI = (body weight loss score + stool characteristics score + blood in stool score) / 3.

[0072] Table 2 DAI scoring criteria

[0073]

[0074]

[0075] like Figure 1 As shown, the weight of mice in the Model group showed the most obvious downward trend. As time went on, the weight of the Model group and the 5-ASA group continued to decrease, while the weight of the drug-treated group showed an upward trend. On the 14th day, the weight of mice in the Model group was significantly reduced compared with the Blank group (P<0.01); compared with the Model group, there were statistical differences between the 5-ASA group and each experimental group (P<0.05); the body weight of the BET / LBP-1NPs group was more uniform, which was greatly improved compared with the Model. There was no obvious change in the general physical signs such as body weight and hair of mice in the Blank group, and no deaths occurred; there were deaths in mice in both the Model group and the BET group. Figure 2 As shown, the DAI score of mice in the Model group was higher than that in the Blank group (P<0.05); compared with the Model group, the DAI score of mice in the 5-ASA group, LBP-1 group, LBP-1+BET group, and BET / LBP-1NPs group was lower (P<0.05), among which the stool shape and degree of blood in the stool of the BET / LBP-1NPs group were significantly improved compared with the Model group. In summary, LBP-1 may be the main component that exerts anti-inflammatory effects and improves the weight loss of colitis mice. The BET / LBP-1NPs group is superior to the 5-ASA group to a certain extent, and there is no significant statistical difference between it and the BET+LBP-1 group, but it can inhibit the aggravation of inflammation and weight loss earlier, because the smaller particle size of the nanoparticles can promote the passive transport and absorption of drugs.

[0076] 3.2 Macroscopic observation of colon tissue and evaluation of pathological changes

[0077] On the 14th day of the experiment, the mice in each group were killed by cervical dislocation, and the entire colon was quickly dissected out. The length of the colon of each group of mice was measured with a ruler; the colon was opened longitudinally along the mesentery, rinsed several times with pre-cooled saline, and the water was absorbed with filter paper. The colon was visually observed and scored for colon mucosal inflammation and ulceration according to the standards listed in Table 3.

[0078] Table 3 Macroscopic observation and pathological scoring criteria of colon tissue

[0079]

[0080]

[0081] like Figure 3 The colon lesion scores of mice in the Model group were higher than those in the Blank group (P<0.01). Compared with the Model group, the lesion scores of all experimental groups were lower, and the colon lesion scores of mice in the BET / LBP-1NPs group were significantly lower (P<0.05). The results showed that the combination of LBP-1 and BET had a certain effect on alleviating colon intestinal damage in mice.

[0082] 3.3 Colon length measurement and tissue preservation

[0083] After the experiment, blood was collected from the mouse orbital capillaries and the colon was dissected to measure the colon length. The blood was centrifuged to obtain the supernatant to obtain serum samples. Part of the colon tissue specimens were fixed in 4% paraformaldehyde solution for H&E staining. The remaining colon tissue was ground with tissue grinding beads, centrifuged (12000r / min, 10min), and the supernatant and collected serum were used together for subsequent ELISA detection.

[0084] like Figure 4 As shown, compared with the Blank group, the colon length of mice in the Model group was significantly shortened (P<0.05). Compared with the Model group, the colon length of mice in the 5-ASA group and the LBP-1 group increased slightly, the increase in the BET group was not obvious, and the colon length of the BET+LBP-1 group and the BET / LBP-1NPs group increased (P<0.05). The colon length of the BET / LBP-1NPs group was closest to that of the Blank group and was significantly better than that of the 5-ASA group.

[0085] 3.4 H&E staining and histopathological examination of colon tissue

[0086] The fixed colon tissue was dehydrated and embedded in paraffin, and 4 μm paraffin sections were prepared. After dewaxing and hydration, HE staining was performed, and the sections were observed and photographed with an optical biological microscope for colon tissue pathology analysis.

[0087] Depend on Figure 5The analysis showed that the mucosal epithelial structure of the Blank group was neat and orderly, with no degeneration or shedding. The crypt structure was obvious, the glands were orderly arranged, the number did not decrease, and the glandular epithelial cells did not change. There were scattered inflammatory cells in the stroma, and no congestion, hemorrhage, or edema. In the Model group, large pieces of mucosal epithelial cells fell off, the ulcers enlarged and fused; the crypts were severely destroyed and the structure was unclear; the glands degenerated and necrotic, and the number decreased significantly; large pieces of inflammatory cells infiltrated the entire layer of the intestinal wall, with obvious congestion and edema, and local abscess formation, indicating that the colitis model was successfully established. Compared with the Model group, the 5-ASA group, the LBP-1 group, the BET+LBP-1 group, and the BET / LBP-1NPs group had less mucosal epithelial shedding and smaller ulcers; the crypts were slightly to moderately destroyed, and the structure was visible; the glands degenerated, necrotized, and the number decreased slightly; the entire layer of the intestinal wall was infiltrated with inflammatory cells of varying amounts, with mild congestion and edema, and focal abscess formation. Among them, compared with other groups, the BET / LBP-1NPs group showed smaller mucosal ulcers, mild destruction of crypts, reduced glandular degeneration, and no significant reduction in number; a large number of inflammatory cells infiltrated under the mucosa, and submucosal edema was still obvious. This shows that BET / LBP-1 self-assembled nanoparticles have a good therapeutic effect on ulcerative colitis.

[0088] 3.5ELISA analysis of the expression of inflammatory factors in colon tissue and serum

[0089] After the colon tissue was homogenized in PBS containing protease inhibitors, IL-6, IL-10, IFN-γ, NO, and reactive oxygen species were detected according to the steps recommended by the ELISA kit, and the OD value was read at a wavelength of 450 nm.

[0090] like Figure 6 As shown in the figure, compared with the Blank group, the expression levels of IL-6 and IFN-γ in the colon tissue of the mice in the Model group were significantly increased (P<0.05), and the expression level of IL-10 was significantly decreased (P<0.05); compared with the Model group, the expression levels of IL-6 and IFN-γ in the colon tissue of the mice in the 5-ASA group and each experimental group were decreased (P<0.05), and the expression level of IL-10 was increased (P<0.05). Among the cytokines in the serum, only the expression level of IL-6 in the LBP-1 group was significantly reduced (P<0.05). The results showed that BET and LBP-1 mainly reduce the expression levels of pro-inflammatory factors IL-6 and IFN-γ through LBP-1, and increase the expression level of anti-inflammatory factor IL-10, thereby achieving the effect of treating colitis. The anti-inflammatory effect of BET itself is not obvious, but the combination with LBP-1 can improve the anti-inflammatory effect to a certain extent.

[0091] The above description of the disclosed embodiments enables one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A method for preparing self-assembled nanoparticles of Lycium barbarum polysaccharide and betaine, characterized in that: The steps include: (1) dissolving betaine in anhydrous ethanol to obtain an organic phase, and dissolving wolfberry polysaccharide in deionized water to obtain an aqueous phase; (2) slowly dripping the organic phase into the aqueous phase drop by drop under ultrasonic conditions, and removing the organic solvent by rotary evaporation to obtain the Lycium barbarum polysaccharide and betaine self-assembled nanoparticles.

2. The method for preparing self-assembled nanoparticles of Lycium barbarum polysaccharide and betaine according to claim 1, characterized in that: In step (1), the mass volume ratio of betaine, wolfberry polysaccharide, anhydrous ethanol and deionized water is (20-60) mg: (10-30) mg: (1-2) mL: 5 mL.

3. The method for preparing self-assembled nanoparticles of Lycium barbarum polysaccharide and betaine according to claim 1, characterized in that: In step (2), the ultrasonic power is 25w.

4. The method for preparing self-assembled nanoparticles of Lycium barbarum polysaccharide and betaine according to claim 1, characterized in that: In step (2), the organic phase is slowly dripped into the aqueous phase drop by drop at a dripping speed of 2 drops / s.

5. The method for preparing self-assembled nanoparticles of Lycium barbarum polysaccharide and betaine according to claim 1, characterized in that: In step (2), the rotary evaporation temperature is 45° C., the rotary evaporation pressure is 90 bar, and the rotary evaporation time is 3-5 minutes.

6. Lycium barbarum polysaccharide and betaine self-assembled nanoparticles prepared by the method according to any one of claims 1 to 5.

7. Use of the Lycium barbarum polysaccharide and betaine self-assembled nanoparticles according to claim 6 in a drug for treating ulcerative colitis.

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