Coagulator for treating enteritis as well as preparation method and application thereof

Abstract

The invention provides a condensate for treating enteritis as well as a preparation method and application thereof, and belongs to the technical field of biological composite materials. According to the preparation method, the aggregate is prepared by dissolving the lipoic acid powder in the spermidine solution and stirring, so that the problem of low solubility of the lipoic acid in water is solved, and the problem of intestinal intolerance caused by too strong alkalinity when the spermidine is singly used is also solved. According to the present invention, the obtained coacervate has good spreadability on the intestinal tract surface, the coacervate can be fused to form the hydrogel with strong adhesion so as to form the intestinal tract barrier, and the enteritis treatment effect can be jointly performed through different treatment mechanisms; the process for preparing the condensate is green and safe, does not need to adopt complex chemical substances and synthesis processes, does not introduce toxic and harmful substances, and is simple to operate, controllable and efficient.

Description

Technical Field

[0001] The invention belongs to the technical field of biocomposite materials, and in particular relates to a coagulant for treating enteritis, and a preparation method and application thereof. Background Art

[0002] Lipoic acid (ALA), also known as 1,2-dithiolane-3-pentanoic acid, contains 1,2-dithiolane and carboxylic acid structures. It is a natural sulfur-containing fatty acid that was isolated from liver tissue in 1951 and was originally considered to be a cofactor of dihydrofatty acid acyltransferase in the mitochondrial tricarboxylic acid cycle. Various biological activities of lipoic acid have been confirmed. It is well known that it has a strong antioxidant function and is manufactured as a drug or supplement to prevent aging. In ulcerative colitis, it can scavenge reactive oxygen species (ROS) and maintain a healthy cellular redox state to achieve the purpose of treatment. Due to the hydrophobic structure of 1,2-dithiolane, the solubility of lipoic acid in water is low, only 1g / L, which limits its application in colitis.

[0003] The polyamine spermidine is a small positively charged molecule that exists naturally in all organisms. It is an important component of the human diet and plays a beneficial role in human diseases. Studies have shown that spermidine may play a therapeutic role in colitis through autophagy, regulating macrophage polarization, preventing intestinal flora imbalance, and protecting the intestinal mucosal barrier. However, due to the strong alkalinity of spermidine, it can cause intestinal intolerance.

[0004] Hydrogel is composed of hydrophilic polymers, and its three-dimensional network structure can absorb a large amount of water and can also be used to carry drugs. It has been widely used in drug delivery, wound dressings, tissue engineering, and sanitary products. Hydrogel as a bioadhesive has the following bottlenecks in biomedical applications. First, the adhesion ability is weak in a wet environment, especially in the intestinal mucosal environment, which is extremely difficult to adhere to. There are a lot of mucous contents in patients with enteritis, and the water layer on the mucosal surface usually weakens the interaction between existing adhesive materials and interface molecules, and the adhesion ability is insufficient in a wet environment. Second, the mechanical strength is low and it is easy to fall off on the peristaltic intestinal surface. The adhesion strength of the hydrogel on the substrate depends on the interfacial adhesion force and the cohesion of the hydrogel. The hydrogel has a high water content and low mechanical strength. Intestinal peristalsis causes the intestinal wall to produce continuous contraction and relaxation forces, which can easily cause the adhered hydrogel network to rupture, and even the intestinal mucosal surface to fall off prematurely. Third, the flow spread is poor, and it is difficult to cover irregular wounds. The colitis ulcer mucosa is mostly scattered in dots, the ulcer wound is irregular in shape, and the molded hydrogel spreads poorly on the ulcer mucosa after implantation, making it difficult to establish a tight hydrogel-tissue fit, and the coverage efficiency is low. In response to these problems, studies have constructed thermosensitive hydrogels to improve the coverage efficiency of irregular wounds, but these in situ hydrogels usually require a higher polymer concentration, are not resistant to dilution in the intestinal cavity, and have the risk of increasing barrier permeability. Fourth, the constituent materials of the hydrogel lack biological activity, and the mucosal repair effect is limited. Therefore, constructing deformable, wet environment-adherent bioactive materials is the key to improving the therapeutic effect of physical barriers. Summary of the invention

[0005] In view of this, the object of the present invention is to provide a coagulant for treating enteritis, and a preparation method and application thereof. The present invention prepares a coagulant by dissolving lipoic acid powder in a spermidine solution and stirring, which solves the problem of low solubility of lipoic acid in water, and also solves the problem of intestinal intolerance caused by the excessive alkalinity of spermidine used alone. The obtained coagulant has good spreadability on the intestinal surface, and the coagulant will fuse to form a hydrogel with stronger adhesion, forming an intestinal barrier, and jointly play the role of treating enteritis through different therapeutic mechanisms.

[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0007] The invention provides a coagulant for treating enteritis. The coagulant comprises the following components in mass percentage: 1% to 35% lipoic acid, 0.5% to 10% spermidine, and the balance is water.

[0008] Preferably, the coagulant comprises the following components in mass percentage: 7% to 16% lipoic acid, 2% to 5% spermidine, and the balance is water.

[0009] Preferably, the coagulant comprises the following components in percentage by mass: 10% lipoic acid, 3% spermidine, and the balance is water.

[0010] Preferably, the coagulant is in a gel state.

[0011] The present invention provides a method for preparing the coagulant, comprising the following steps:

[0012] 1) preparing a spermidine solution with a concentration of 0.5% to 10%;

[0013] 2) Add lipoic acid to the spermidine solution, and wait until the lipoic acid is completely dissolved to obtain a gel-like coagulant.

[0014] Preferably, the lipoic acid is in powder form.

[0015] Preferably, the dissolving is carried out by shaking or stirring.

[0016] The present invention also provides the use of the coagulant or the coagulant obtained according to the preparation method in preparing a drug for treating enteritis.

[0017] The present invention also provides the use of the coagulant or the coagulant obtained according to the preparation method in preparing hemostatic and / or digestive tract mucosal repairing materials.

[0018] Compared with the prior art, the present invention has the following beneficial effects:

[0019] (1) The present invention prepares a coagulant by dissolving lipoic acid powder in a spermidine solution and stirring, thereby solving the problem of low solubility of lipoic acid in water and the problem of intestinal intolerance caused by the excessive alkalinity of spermidine used alone.

[0020] (2) The coacervate obtained by the present invention has good spreadability on the surface of the intestine, and the coacervate will fuse to form a hydrogel with stronger adhesion to form an intestinal barrier. The present invention utilizes lipoic acid to remove excess free radicals in the intestine and regulate oxidative stress, and spermidine can regulate macrophage polarization and prevent intestinal flora imbalance through autophagy, so that the coacervate obtained can jointly play a role in treating enteritis through different therapeutic mechanisms. Provide a material with both antioxidant, anti-inflammatory and autophagy regulation for the treatment of clinical enteritis.

[0021] (3) The process for preparing the coagulant of the present invention is green and safe, does not require the use of complex chemical substances and synthesis processes, does not introduce toxic and harmful substances, is simple to operate, and is controllable and efficient. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 It is the process of preparing the coagulant;

[0023] Figure 2 is the condensate obtained in Example 1, wherein A is the recovery of the fluorescence of the entire condensate within 200 s after irreversible quenching, and B is the recovery of the fluorescence of a part of the condensate within 200 s after irreversible quenching;

[0024] Figure 3 The contact angles of the coagulant and water on various materials and the statistical results obtained in Example 1;

[0025] Figure 4 This is a graph showing the oscillation frequency scanning results of the condensate obtained in Example 1 before and after the phase transition;

[0026] Figure 5 Inverted microscope images and scanning electron microscope images of the coagulants obtained in Example 1 and Comparative Example 5, respectively, wherein A is an inverted fluorescence microscope image of the sample obtained in Example 1, B is an inverted fluorescence microscope image of the sample obtained in Comparative Example 5, C is a scanning electron microscope image of the sample obtained in Example 1, and D is a scanning electron microscope image of the sample obtained in Comparative Example 5;

[0027] Figure 6 The results of the condensate obtained in Example 1 scavenging endogenous free radicals in macrophages, wherein the scale of the combined image is 50 μm, and the scale of the white light image is 50 μm;

[0028] Figure 7 The results of using different treatment groups to treat dextran sodium sulfate enteritis mice in the biological performance evaluation experiment, where the upper scale in the H&E figure is 800μm, and the lower scale is 150μm;

[0029] Figure 8 The effects of different treatment groups on the immunofluorescence staining of LC3 and P62 in mouse colon. DETAILED DESCRIPTION

[0030] The present invention provides a coagulant for treating enteritis, wherein the coagulant comprises the following components in percentage by mass:

[0031] Lipoic acid 1% to 35%, preferably 7% to 16%, more preferably 10%. Lipoic acid has the effects of scavenging free radicals, anti-oxidation, anti-inflammatory, and regulating oxidative stress;

[0032] Spermidine 0.5% to 10%, preferably 2% to 5%, more preferably 3%. Spermidine is an anti-aging molecule that can improve the self-protection of cells under stress conditions through autophagy, regulate macrophage polarization, and prevent intestinal flora imbalance;

[0033] The balance is water.

[0034] In the present invention, the carboxyl group of lipoic acid and the amino group of spermidine form a physical coacervate through electrostatic interaction and hydrophobic interaction between lipoic acid molecules. The coacervate is in the form of a spherical droplet, has a low surface tension, has good fluidity, spreadability and cohesive gel-forming properties at various tissue interfaces, has good adhesion to various moist wound surfaces, can be used for hemostasis and repair of digestive tract mucosa, and has the ability to scavenge free radicals on a broad spectrum and induce cell autophagy. The spontaneous aggregation of hydrophobic 1,2-dithiolane in the coacervate causes the coacervate to transform into a hydrogel, so that the obtained coacervate is in a gel state.

[0035] The present invention provides a method for preparing the coagulant, comprising the following steps:

[0036] 1) preparing a spermidine solution with a concentration of 0.5% to 10%;

[0037] 2) Add lipoic acid to the spermidine solution, and wait until the lipoic acid is completely dissolved to obtain a gel-like coagulant.

[0038] In the present invention, a spermidine solution with a concentration of 0.5% to 10% is prepared, and spermidine is dissolved in pure water to obtain a spermidine solution with a concentration of 0.5% to 10%.

[0039] In the present invention, lipoic acid is added to a spermidine solution, and a gelatinous coagulant is obtained after the lipoic acid is completely dissolved; lipoic acid powder is added to the prepared spermidine solution, and the added lipoic acid powder is completely dissolved by shaking or stirring to obtain a turbid liquid, thereby obtaining a gelatinous coagulant.

[0040] The present invention also provides the use of the coagulant or the coagulant obtained according to the preparation method in preparing a drug for treating enteritis.

[0041] In the present invention, after the coagulant is administered by enema, it can be spread on the surface of the intestine, and the droplets fuse to form a hydrogel with stronger adhesion to form an intestinal barrier. The lipoic acid and spermidine in the coagulant work together to treat enteritis through different therapeutic mechanisms.

[0042] The present invention also provides the use of the coagulant or the coagulant obtained according to the preparation method in preparing hemostatic and / or digestive tract mucosal repairing materials.

[0043] In the present invention, after the coacervate is covered on the surface of the intestine as a mucosal material, it removes excessive toxic free radicals, regulates oxidative stress and autophagy pathways, and restores intestinal redox and immune homeostasis.

[0044] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0045] Example 1

[0046] Prepare 3% spermidine solution: weigh 3 g of spermidine, dissolve it in distilled water, and dilute to 100 mL to obtain a 3% spermidine solution.

[0047] Preparation of coacervate ( Figure 1 ): Weigh 10 g of lipoic acid powder and add it to 100 mL of the prepared 3% spermidine solution. Stir until the lipoic acid powder disappears completely and a turbid liquid is obtained, which is the coagulant.

[0048] Example 2

[0049] Prepare 0.5% spermidine solution: weigh 0.5 g spermidine, dissolve it in distilled water, and dilute to 100 mL to obtain 0.5% spermidine solution.

[0050] Preparation of coagulants: Weigh 1 g of lipoic acid powder and add it to 100 mL of the above-prepared 0.5% spermidine solution, stir until the lipoic acid powder disappears completely, and obtain a turbid liquid, which is the coagulant.

[0051] Example 3

[0052] Prepare 10% spermidine solution: weigh 10 g of spermidine, dissolve it in distilled water, and dilute to 100 mL to obtain 10% spermidine solution.

[0053] Preparation of coagulants: Weigh 35 g of lipoic acid powder, add it to 100 mL of the above-prepared 10% spermidine solution, stir until the lipoic acid powder disappears completely, and obtain a turbid liquid, which is the coagulant.

[0054] Example 4

[0055] Prepare 2% spermidine solution: weigh 2 g of spermidine, dissolve it in distilled water, and dilute to 100 mL to obtain a 2% spermidine solution.

[0056] Preparation of coagulants: Weigh 7 g of lipoic acid powder, add it to 100 mL of the above-prepared 2% spermidine solution, shake it until the lipoic acid powder disappears completely, and obtain a turbid liquid, which is the coagulant.

[0057] Example 5

[0058] Prepare 5% spermidine solution: weigh 5 g of spermidine, dissolve it in distilled water, and dilute to 100 mL to obtain a 5% spermidine solution.

[0059] Preparation of coacervate: Weigh 16 g of lipoic acid powder, add it to 100 mL of the above-prepared 5% spermidine solution, shake it until the lipoic acid powder disappears completely, and obtain a turbid liquid, which is the coacervate.

[0060] Example 6

[0061] Prepare 0.5% spermidine solution: weigh 0.5 g spermidine, dissolve it in distilled water, and dilute to 100 mL to obtain 0.5% spermidine solution.

[0062] Preparation of coacervate: Weigh 2.5 g of lipoic acid powder, add it to 100 mL of the above-prepared 0.5% spermidine solution, shake it until the lipoic acid powder disappears completely, and obtain a turbid liquid, which is the coacervate.

[0063] Example 7

[0064] Prepare 10% spermidine solution: weigh 10 g of spermidine, dissolve it in distilled water, and dilute to 100 mL to obtain 10% spermidine solution.

[0065] Preparation of coagulants: Weigh 31 g of lipoic acid powder, add it to 100 mL of the 10% spermidine solution prepared above, shake it until the lipoic acid powder disappears completely, and obtain a turbid liquid, which is the coagulant.

[0066] Comparative Example 1

[0067] Prepare 0.5% spermidine solution: weigh 0.5 g spermidine, dissolve it in distilled water, and dilute to 100 mL to obtain 0.5% spermidine solution.

[0068] Preparation of coacervate: Weigh 0.5 g of lipoic acid powder, add it into 100 mL of the above-prepared 0.5% spermidine solution, stir until the lipoic acid powder disappears completely, and continue stirring to obtain no gel-like coacervate.

[0069] Comparative Example 2

[0070] Prepare 11% spermidine solution: weigh 11 g of spermidine, dissolve it in distilled water, and dilute to 100 mL to obtain 11% spermidine solution.

[0071] Preparation of coacervate: Weigh 30 g of lipoic acid powder, add it into 100 mL of the above-prepared 11% spermidine solution, stir until the lipoic acid powder disappears completely, and continue stirring to obtain no gel-like coacervate.

[0072] Comparative Example 3

[0073] Weigh 3 g of spermidine, dissolve it in distilled water, and make up to 100 mL to obtain a 3% spermidine solution.

[0074] Comparative Example 4

[0075] Weigh 10 g of lipoic acid powder and add it to distilled water, make the volume to 100 mL, grind and suspend to obtain a lipoic acid suspension with a concentration of 10%.

[0076] Comparative Example 5

[0077] Prepare 3% polylysine solution: weigh 3 g of polylysine, dissolve it in distilled water, and dilute to 100 mL to obtain a 3% polylysine solution.

[0078] Preparation of a mixed solution of lipoic acid and polylysine: Weigh 10 g of lipoic acid powder, add it to 100 mL of the prepared 3% polylysine solution, and stir until the lipoic acid powder disappears completely to obtain a mixed solution of lipoic acid and polylysine.

[0079] Comparative Example 6

[0080] Prepare 3% chitosan solution: weigh 3 g chitosan, dissolve it in 1% acetic acid aqueous solution, and make up to 100 mL to obtain 3% chitosan solution.

[0081] Preparation of a mixed solution of lipoic acid and chitosan: Weigh 10 g of lipoic acid powder, add it to 100 mL of the prepared 3% chitosan solution, and stir until the lipoic acid powder disappears completely to obtain a mixed solution of lipoic acid and chitosan solution.

[0082] Experimental Example 1 Determination of fluidity of agglomerates

[0083] Fluorescence photobleaching recovery technology is a technology used to study the material exchange capacity on the surface of condensates and inside and outside the condensates. The principle of fluorescence photobleaching recovery technology is to irradiate a specific area of ​​the condensate with a high-energy laser to irreversibly quench the fluorescent molecules labeled in the area. This area is called the fluorescence bleaching (photobleaching) area. Subsequently, due to the movement of the condensate molecules, the fluorescent molecules in the surrounding non-bleaching areas continue to migrate to the fluorescence bleaching area. As a result, the fluorescence intensity in the fluorescence bleaching area gradually recovers to its original level. The present invention uses FITC-labeled condensate molecules obtained in Example 1, first irradiates the whole or part of the condensate with a high-energy laser confocal (C2si, Nikon, Japan) to quench the fluorescence, then stops the laser irradiation, and observes the recovery of the fluorescence intensity of the condensate for 200 seconds. The results are shown in FIG. Figure 2 .

[0084] Depend on Figure 2 From A in the figure, it can be seen that after the fluorescence of the condensate as a whole is irreversibly quenched, it can be restored in 200s, indicating that the molecules inside the condensate and the molecules on the surface of the condensate can flow and exchange; Figure 2From the result B in the figure, we can see that after the local fluorescence of the condensate is irreversibly quenched, it can be restored in 200s, indicating that the molecules on the surface of the condensate can flow and exchange.

[0085] Experimental Example 2 Determination of Spreadability of Agglomerates

[0086] The contact angle test is a surface measurement technology for studying the surface wettability and interface properties of materials. The principle of the contact angle test is to drop a liquid on the surface to be tested, and then measure the angle of contact between the liquid and the surface to be tested. The larger the contact angle, the worse the wettability of the droplet on the surface to be tested; the smaller the contact angle, the better the wettability of the droplet on the surface to be tested. In the present invention, the condensate and water obtained in Example 1 are dropped on a hydrophilic plate (glass plate), a hydrophobic plate (chlorinated polyethylene plate, polypropylene plate, polystyrene plate) and a tissue (rat skin), and a contact angle meter (Theta, Sweden) is used to record and measure the results. Figure 3 .

[0087] Depend on Figure 3 The results show that the contact angles of the coagulants on all the above test plates are smaller than water and smaller than 90°, indicating that the wettability (spreadability) of the coagulants on these materials is good and better than that of aqueous solutions.

[0088] Experimental Example 3 Determination of the gelling properties of the coagulant

[0089] Preparation of non-gelled coagulants: The only difference from Example 1 is that sampling is immediately performed after the added lipoic acid powder disappears completely, and the sample is used as a sample.

[0090] The rheological characterization of the sample and the coagulant obtained in Example 1 was performed using a rotational rheometer (DHR-2, TA, USA) within 30 min. The cone-plate mold used in the present invention had a diameter of 40 mm and an inclination angle of 2°. The parameters set for the oscillation frequency sweep were: temperature 25°C, strain 0.1%, angular frequency 0.1 to 10 rad / s. The storage modulus (G′) and loss modulus (G″) values ​​were detected by frequency sweeping. The results are shown in Figure 4 .

[0091] Depend on Figure 4 The results show that the storage modulus (G′) of the ungelled coagulant (sample) is lower than the loss modulus (G″) during the entire test process, indicating that the ungelled coagulant (sample) exhibits fluid properties. In Example 1, the storage modulus (G′) of the colloid state converted from the ungelled coagulant is higher than the loss modulus (G″) during the entire test process, indicating that the ungelled coagulant can be converted into a colloid state and exhibits solid-like properties and the gel structure is relatively stable.

[0092] Experimental Example 4: Results of inverted fluorescence microscopy and scanning electron microscopy of condensates

[0093] The state of the samples obtained in Example 1 and Comparative Example 5 was observed using an inverted fluorescence microscope (TI-S, Nikon, Japan). Figure 5 A and B in the figure. Using a scanning electron microscope (Hitachi SU8600, Japan) with a voltage of 8000V, the state of the samples obtained in Example 1 and Comparative Example 5 was observed. The results are shown in Figure 5 C and D in.

[0094] Figure 5 A and B in the figure are inverted fluorescence microscope images of Example 1 and Comparative Example 5. Figure 5 As shown in A, after 10 g of lipoic acid was dissolved in 3% spermidine solution, droplets were observed under the microscope, indicating that condensates were formed by the electrostatic interaction between the carboxyl group of lipoic acid and the amino group of spermidine and the hydrophobic interaction between lipoic acid molecules. Figure 5 As can be seen from B, after 10 g of lipoic acid was dissolved in 3% polylysine solution, no droplets were observed under the microscope, indicating that 10% lipoic acid and 3% polylysine did not form coagulation products.

[0095] Figure 5 C and D in the figure are scanning electron microscope images of Example 1 and Comparative Example 5. Figure 5 As shown in C, the product obtained in Example 1 is a smooth film under electron microscope, which is the surface of the gel formed by the fusion of the coacervate droplets, indicating that 10% lipoic acid can form coacervates with 3% spermidine. Figure 5 As can be seen from D in FIG. 1 , the product obtained in Comparative Example 5 has a rough surface under an electron microscope, indicating that 10% lipoic acid and 3% polylysine cannot form coagulation products.

[0096] Experimental Example 5: Aggregates scavenging endogenous free radicals in macrophages

[0097] RAW264.7 cells (purchased from Shanghai Fuheng Biotechnology Co., Ltd., FH0328) were cultured at 1×10 5The density of cells was inoculated into a 24-well plate, and 500 μL of 10% FBS (Gibco, USA) and 1% double antibody (penicillin sodium salt: 10 kU / mL; streptomycin: 10 mg / mL) DMEM medium was added, and cultured in an incubator at 37°C, 5% CO2 and 70% relative humidity. After the cells were cultured for 24 hours, the cell culture medium was discarded, and fresh culture medium containing 0.7 mM H2O2 but not containing serum and double antibodies was added to each well for incubation for 12 hours. The culture medium was then removed, the cells were washed twice with PBS, and then 500 μL of serum-free and double antibody DMEM culture medium containing Example 1 (100 μg / mL lipoic acid concentration), Comparative Example 3 (100 μg / mL spermidine concentration) and Comparative Example 4 (50 μg / mL lipoic acid concentration) was added to each well and continued to incubate for 12 hours. The culture medium was discarded and the cells in each well were washed with PBS. Then, 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescent probe (final concentration was 10 μM) was added to the serum-free and double-antibody DMEM culture medium and the cells were incubated in a constant temperature incubator for 30 minutes. After discarding the culture medium and washing the cells three times with fresh culture medium, the intracellular ROS level was quantified using an inverted fluorescence microscope. The results are shown in Figure 6 .

[0098] Depend on Figure 6 It can be seen that normal cells without H2O2 stimulation have no DCF fluorescence, while strong DCF fluorescence appears after H2O2 stimulation, indicating the generation of endogenous free radicals. Example 1 and Comparative Example 4 significantly reduce the intensity of DCF fluorescence, while the DCF fluorescence of Comparative Example 3 does not decrease significantly, indicating that lipoic acid is the basis for the prepared coagulant to exert its antioxidant effect.

[0099] Experimental Example 6 Evaluation of the coagulant in the treatment of dextran sodium sulfate enteritis in mice

[0100] The dextran sulfate sodium (DSS) enteritis model is one of the most widely used enteritis models. By continuously feeding male C57BL / 6 mice (purchased from Zhejiang Muke Biotechnology Co., Ltd., 6-8 weeks old) with 3% to 5% DSS solution, enteritis symptoms similar to human pathological characteristics, such as diarrhea, bloody stools, and weight loss, can be quickly formed. The mice were fed 3.5% DSS for 12 consecutive days, during which time they were given 0.2 mL PBS (control group), Example 1, Comparative Example 3, Comparative Example 4, Comparative Example 5, and Comparative Example 6 by enema every other day. The mice in the normal group were fed pure water. On the 12th day, the mice were humanely euthanized, and the colons were taken for photographing and sectioning and staining to observe the microscopic morphology and barrier integrity of the intestinal epithelium. The results are shown in Table 1. Figure 7 .

[0101] The results are as follows Figure 7As shown, comparative examples 3 to 6 cannot stay in the intestine for a long time, cannot effectively restore the intestinal shortening caused by enteritis, and epithelial cell apoptosis, barrier defect, crypt necrosis, and inflammatory cell infiltration are found in HE staining. In contrast, the coacervate prepared in Example 1 can treat enteritis by anti-oxidation and regulating autophagy, and the colon length is significantly restored. The epithelial cells and barrier in HE staining are close to normal, the crypts are intact, and there is no obvious inflammation.

[0102] Experimental Example 7: Condensates can activate the autophagy pathway

[0103] After dewaxing, paraffin sections of animal tissues were immersed in citric acid buffer (concentration of 10mM, pH=6), and then heated in a pressure cooker at 100°C and 170kPa for 3min. After the temperature dropped to room temperature, 20μL of 5% BSA solution was added to each tissue for blocking for 30min. 10μL of primary antibody (LC3 and p62) was added to each tissue and incubated in a humidified box at 4°C overnight. After removing the primary antibody the next day, 10μL of fluorescent secondary antibody was added to each tissue. After sealing, the slides were photographed using a laser confocal microscope (C2si, Nikon, Japan).

[0104] Autophagy is a process in which eukaryotic cells use lysosomes to degrade their own cytoplasmic proteins and damaged organelles under the regulation of autophagy-related genes. LC3, also known as MAP1LC3, runs through the entire autophagy process and is currently a recognized autophagy marker. p62 is a widely studied autophagy substrate. The autophagy indicators LC3 and p62 were co-stained, and the results are as follows: Figure 8 shown.

[0105] Depend on Figure 8 It can be seen that compared with the control group, the two indicators of the condensate group obtained in Example 1 increased, indicating that the condensate prepared by the present invention can regulate autophagy to treat enteritis.

[0106] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A coagulant for treating enteritis, characterized in that: The condensate comprises the following components in mass percentage: 1% to 35% lipoic acid, 0.5% to 10% spermidine, and the balance is water.

2. The condensate according to claim 1, characterized in that The coagulant comprises the following components in mass percentage: 7% to 16% lipoic acid, 2% to 5% spermidine, and the balance is water.

3. The condensate according to claim 2, characterized in that The coagulant comprises the following components in mass percentage: 10% lipoic acid, 3% spermidine, and the balance is water.

4. The agglomerate according to any one of claims 1 to 3, characterized in that: The coacervate is in a gel state.

5. The method for preparing a coagulant according to any one of claims 1 to 4, characterized in that: The following steps are involved: 1) preparing a spermidine solution with a concentration of 0.5% to 10%; 2) Add lipoic acid to the spermidine solution, and wait until the lipoic acid is completely dissolved to obtain a gel-like coagulant.

6. The preparation method according to claim 5, characterized in that: The lipoic acid is in powder form.

7. The preparation method according to claim 5, characterized in that: The dissolving is carried out by shaking or stirring.

8. Use of the coagulant according to any one of claims 1 to 4 or the coagulant obtained according to the preparation method according to any one of claims 5 to 7 in the preparation of drugs for treating enteritis.

9. Use of the coagulant according to any one of claims 1 to 4 or the coagulant obtained according to the preparation method according to any one of claims 5 to 7 in the preparation of hemostatic and / or digestive tract mucosal repair materials.

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