阅读说明：本技术 一种茯苓多糖水凝胶及其制备方法和应用 (Pachyman hydrogel and preparation method and application thereof ) 是由 李菀 叶晓川 袁浩 罗心遥 方可欣 于 2021-09-16 设计创作，主要内容包括：本发明公开了一种茯苓多糖水凝胶及其制备方法和应用,其制备方法包括如下步骤：将碱溶性茯苓多糖溶于碱液中,加酸调至pH＜11,制备浓度≥4mg/mL的碱溶性茯苓多糖分散液；将所得碱溶性茯苓多糖分散液于不高于50℃的条件下静置,得茯苓多糖水凝胶。本发明的茯苓多糖水凝胶的制备方法简单、生产成本低,所制备的茯苓多糖水凝胶具有良好的力学性能、保水性和抗炎性,该茯苓多糖水凝胶负载的药物可在50小时左右释放出70％～80％,因此,该茯苓多糖水凝胶作为药物控缓释放载体具有良好的应用前景。(The invention discloses pachyman hydrogel and a preparation method and application thereof, wherein the preparation method comprises the following steps: dissolving alkali soluble pachyman in alkali solution, adding acid to adjust pH to less than 11, and preparing alkali soluble pachyman dispersion solution with concentration of more than or equal to 4 mg/mL; standing the obtained alkali soluble pachyman dispersion liquid at a temperature of not higher than 50 ℃ to obtain pachyman hydrogel. The pachyman hydrogel disclosed by the invention is simple in preparation method and low in production cost, the prepared pachyman hydrogel has good mechanical property, water retention property and anti-inflammation property, and the medicine loaded on the pachyman hydrogel can be released for about 50 hours by 70% -80%, so that the pachyman hydrogel has good application prospect as a medicine controlled slow release carrier.)

1. The preparation method of pachyman hydrogel is characterized by comprising the following steps:

dissolving alkali soluble pachyman in alkali solution, adding acid to adjust pH to less than 11, and preparing alkali soluble pachyman dispersion solution with concentration of more than or equal to 4 mg/mL; standing the obtained alkali soluble pachyman dispersion liquid at a temperature of not higher than 50 ℃ to obtain pachyman hydrogel.

2. The method of preparing pachyman hydrogel according to claim 1, wherein: the alkali soluble pachyman dispersion has a concentration of C₁When the pH value is more than 0 and less than or equal to 1; the alkali soluble pachyman dispersion has a concentration of C₂When the pH value is more than 0 and less than or equal to 7.8; the alkali soluble pachyman dispersion has a concentration of C₃When the pH value is more than 0 and less than or equal to 10.5; wherein, C is less than or equal to 4mg/mL₁＜10mg/mL，10mg/mL≤C₂＜25mg/mL，C₃≥25mg/mL。

3. The method of preparing pachyman hydrogel according to claim 2, wherein: the pH value of the alkali-soluble pachyman dispersion liquid is more than or equal to 4 and less than or equal to 6.

4. The method for preparing pachyman hydrogel according to claim 1 or 2, wherein: the concentration of the alkali-soluble pachyman dispersion liquid is 4-30 mg/mL.

5. The method of preparing pachyman hydrogel according to claim 1, wherein: the temperature of the alkali soluble pachyman dispersion liquid is less than or equal to 30 ℃ when the alkali soluble pachyman dispersion liquid is stood.

6. The method of preparing pachyman hydrogel according to claim 1, wherein: the alkali liquor is sodium hydroxide solution or potassium hydroxide solution, and the acid is at least one of hydrochloric acid, phosphoric acid, sulfuric acid, citric acid and glacial acetic acid.

7. The method of preparing pachyman hydrogel according to claim 1, wherein: the alkali soluble pachyman is prepared by the following method:

extracting Poria dry powder with ethanol, oven drying the residue to obtain primary Poria residue;

decocting the primary Poria residue in water, oven drying the residue to obtain secondary Poria residue;

extracting the residue with alkali solution, filtering to remove residue, adjusting pH with acid, and standing to obtain precipitate;

desalting the precipitate with pure water, and drying to obtain alkali soluble pachyman.

8. The method of preparing pachyman hydrogel according to claim 1, wherein: the alkali soluble pachyman dispersion has a concentration of 1 wt% + -0.1 wt% and a pH of 7 + -0.2.

9. A pachyman hydrogel is characterized in that: the pachyman hydrogel according to any one of claims 1 to 8, which is produced by the method.

10. Use of the pachyman hydrogel of claim 9 for the preparation of a sustained release medicament.

Technical Field

The invention relates to the field of biological polymer materials, and particularly relates to pachyman hydrogel and a preparation method and application thereof.

Background

The gel is a soft solid with a three-dimensional network structure formed by covalent or non-covalent crosslinking. In recent years, gels have been widely used in drug delivery systems due to their good water retention, biocompatibility, degradability and environmental responsiveness, especially for some chronic diseases requiring sustained treatment. However, most of the gel matrixes are synthetic materials at present, and more importantly, the gel matrixes have little synergistic effect with the loaded drugs.

Polysaccharides are natural macromolecular compounds formed by connecting monosaccharides through glycosidic bonds, and widely exist in animal cell membranes, plants and microbial cell walls. It not only has the function of regulating immunity, but also has wide pharmacological actions in the aspects of resisting tumor, resisting oxidation, reducing blood fat and the like. The polysaccharide molecules have rich binding sites and good biocompatibility, so the polysaccharide is an excellent material for preparing natural hydrogel. When the compound is used as a drug release system, the use of ineffective carriers can be reduced, and the effect of drug-assisted synergy can be expected to be realized.

Poria cocos (Schw.) Wolf is oven-dried sclerotia of Poria cocos (a fungus in Polyporaceae), is a traditional Chinese medicine in China, and is also a bulk Chinese medicinal material used as both medicine and food. The pachyman is the main component of the tuckahoe medicinal material, and the content of the pachyman accounts for 70 to 90 percent of the sclerotium of tuckahoe. Modern pharmacological studies show that pachyman is the main active component of Poria, and has effects of resisting tumor, improving immunity, etc. At present, researches on pachyman mainly focus on the aspects of pharmacological activity and action mechanism, and related researches on preparation of gel by using pachyman are not seen.

Disclosure of Invention

The inventor finds that the pachyman hydrogel can be obtained by standing an alkali-soluble pachyman dispersion liquid with a certain concentration under the conditions that the pH is less than 11 and the temperature is less than 50 ℃. The pachymaran hydrogel is thermally stable at a temperature of less than 50 ℃, a leaching solution of the pachymaran hydrogel has biological activity, and 70-80% of loaded drugs can be released in about 50 hours at 37 ℃ under the condition that the pH value is 5.8-7.4, so that the pachymaran hydrogel can be used for preparing sustained-release drugs.

The technical scheme provided by the invention is as follows:

in a first aspect, a method for preparing pachyman hydrogel comprises the following steps:

dissolving alkali soluble pachyman in alkali solution, adding acid to adjust pH to less than 11, and preparing alkali soluble pachyman dispersion solution with concentration of more than or equal to 4 mg/mL; standing the obtained alkali soluble pachyman dispersion liquid at a temperature of less than 50 ℃ to obtain pachyman hydrogel.

The temperature of the alkali liquor is preferably less than or equal to 35 ℃, more preferably less than or equal to 30 ℃, and even more preferably less than or equal to 25 ℃; the temperature at which the alkali-soluble pachyman dispersion is allowed to stand is preferably 35 ℃ or less, more preferably 30 ℃ or less, still more preferably 25 ℃ or less, and most preferably 4 ℃.

Preferably, the alkali-soluble pachyman dispersion has a concentration of 4 to 30 mg/mL.

Preferably, the alkali-soluble pachyman dispersion has a concentration of C₁When the pH value is more than 0 and less than or equal to 1; the alkali soluble pachyman dispersion has a concentration of C₂When the pH value is more than 0 and less than or equal to 7.8; the alkali soluble pachyman dispersion has a concentration of C₃When the pH value is more than 0 and less than or equal to 10.5; wherein, C is less than or equal to 4mg/mL₁＜10mg/mL，10mg/mL≤C₂＜25mg/mL，C₃≥25mg/mL。

Preferably, the alkali-soluble pachyman dispersion has a pH of 4-6, and the time for the alkali-soluble pachyman to form pachyman hydrogel is less than 30 s.

Preferably, the alkali-soluble pachyman dispersion has a concentration of 1 wt% + -0.1 wt% and a pH of 7 + -0.2, the release time of the pachyman hydrogel prepared under the conditions is as long as 50 hours, the pH of the prepared pachyman hydrogel is relatively mild, and the complete gelation time is less than 45 min.

Preferably, the alkali solution is sodium hydroxide solution or potassium hydroxide solution, and the acid is at least one of hydrochloric acid, phosphoric acid, sulfuric acid, citric acid and glacial acetic acid.

Preferably, the alkali-soluble pachyman is prepared by the following method:

extracting Poria dry powder with ethanol, oven drying the residue to obtain primary Poria residue;

decocting the primary Poria residue in water, oven drying the residue to obtain secondary Poria residue;

extracting the residue with alkali solution, filtering to remove residue, adjusting pH with acid, and standing to obtain precipitate;

desalting the precipitate with pure water, and drying to obtain alkali soluble pachyman.

In a second aspect, the present invention provides a pachyman hydrogel prepared by the above preparation method, which has a high elastic modulus and a high strength and is capable of releasing bioactive pachyman.

In a third aspect, the invention provides the use of the pachymaran hydrogel in the preparation of a sustained-release drug, specifically, the drug is dispersed in an alkaline solution, and the dispersion state and activity of the drug are not changed when the pH of the alkaline solution is changed; the slow release medicine prepared from the pachyman hydrogel can release 70-80% of medicine within about 50 hours under the condition of pH5.8-7.4, and the pachyman hydrogel releases pachyman with biological activity.

Compared with the prior art, the invention has the following advantages and beneficial effects:

(1) the invention firstly provides pachyman hydrogel with a three-dimensional network structure prepared by taking alkali-soluble pachyman as a raw material, and the pachyman hydrogel has good mechanical property, water retention property and anti-inflammation property.

(2) The slow release medicine prepared by utilizing the pachyman hydrogel can release 70-80% of medicine within about 50 hours, and the pachyman hydrogel releases pachyman with biological activity.

(3) The preparation method of the pachymaran hydrogel provided by the invention is environment-friendly, simple in process and low in cost, is expected to be applied to the field of controlled and sustained release of medicines, and expands the application range of pachymaran.

Drawings

In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings needed to be used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without creative efforts.

FIG. 1 is a photomicrograph of an alkali-soluble pachyman dispersion and a pachyman hydrogel according to example 1 of the present invention; FIG. 1(a) is a photomicrograph of an alkali-soluble pachyman dispersion and FIG. 1(b) is a photomicrograph of a pachyman hydrogel.

FIG. 2 is a scanning electron micrograph of pachyman hydrogel of example 1 according to the present invention.

FIG. 3 shows the minimum gelation concentration of alkali soluble pachyman dispersions of varying pH.

FIG. 4 shows the phase transition temperatures of pachyman hydrogels prepared from different concentrations of alkali soluble pachyman dispersions.

FIG. 5 shows the gelation time of alkali soluble pachyman dispersions at concentrations of 2.5 wt% at different pH.

FIG. 6 is a graph of storage modulus and loss modulus of a pachyman hydrogel.

FIG. 7 shows the cell viability of RAW264.7 cells cultured for 24h after diluted by 1:1, 1:2, 1:10, 1:20 times.

FIG. 8 is a graph showing the effect of pachyman hydrogel on the level of inflammatory factors tumor necrosis factor (TNF-. alpha.) production by LPS-induced RAW264.7 cells.

FIG. 9 is a graph showing the effect of pachyman hydrogel on LPS-induced production of interleukin 1 β (IL-1 β) inflammatory factor levels by RAW264.7 cells.

FIG. 10 is the release curve of pachyman drug-loaded gel to baicalin at 37 deg.C and different pH.

Detailed Description

To make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions of the present invention will be clearly and completely described below with reference to specific embodiments, and it is obvious that the described embodiments are some, but not all embodiments of the present invention. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention.

The invention extracts alkali soluble pachyman from tuckahoe medicinal materials, and dissolves a certain amount of alkali soluble pachyman into alkali liquor to prepare alkali soluble pachyman dispersion liquid; adding acid into the alkali soluble pachyman dispersion liquid to adjust pH to less than 11, and standing at room temperature or 4 deg.C to obtain pachyman hydrogel.

The alkali solution in the above technical scheme is strong alkaline solution such as sodium hydroxide solution or potassium hydroxide solution, and the carbonate solution can not dissolve the alkali-soluble pachyman.

In the above technical scheme, phosphoric acid, hydrochloric acid, sulfuric acid, citric acid or acetic acid is used as the acid, and in order to avoid excessive dilution of the alkali-soluble pachyman dispersion after adding the acid, concentrated phosphoric acid, concentrated hydrochloric acid, concentrated sulfuric acid or glacial acetic acid is suggested to be used, and concentrated phosphoric acid is used in the following examples.

In the technical scheme, after the pH is adjusted by adding acid, the alkali-soluble pachyman dispersion liquid needs to keep the concentration within a certain range, and when the concentration is too low, the alkali-soluble pachyman dispersion liquid keeps a sol state all the time, and hydrogel cannot be generated; at too high a concentration, the alkali soluble pachyman dispersion is difficult to dissolve completely; and the alkali soluble pachyman dispersion liquid needs to be kept still, otherwise, the network structure is easy to be damaged, and the cross-linked hydrogel can not be formed.

Specifically, the alkali-soluble pachyman is re-dissolved in 0.3-2 mol/L sodium hydroxide solution, concentrated acid is added to adjust the pH value to 1-9, the concentration of the alkali-soluble pachyman after the pH value is adjusted is ensured to be 2-3 wt%, and the alkali-soluble pachyman is kept stand for 5 s-3 h at room temperature or 4 ℃ to obtain the pachyman hydrogel.

The invention extracts alkali soluble pachyman from tuckahoe medicinal material by the following steps:

(1) pulverizing Poria, sieving with 40 mesh sieve, soaking in 100% ethanol for 12 hr, and oven drying to obtain primary Poria residue; decocting the primary Poria residue with 20 times of water for 2 hr, filtering to remove filtrate, repeating for 3 times, and oven drying the residue to obtain secondary Poria residue;

(2) dispersing the secondary poria cocos residues in 0.5-2 mol/L sodium hydroxide solution according to the weight ratio of the material liquid of 1:30, stirring at room temperature, filtering to remove filter residues, adding 1mol/L hydrochloric acid into filtrate to adjust the pH value to 7, standing for 1h, desalting the precipitate with pure water, and drying to obtain the alkali-soluble pachyman.

Example 1: preparation of pachyman hydrogel

(1) Pulverizing Poria, and sieving with 40 mesh sieve; firstly, ethanol leaching: soaking in 100% ethanol for 12 hr, filtering, and oven drying the residue to obtain primary Poria residue; and (3) reheating water leaching: decocting Poria residue with 20 times of water for 2 hr, filtering, and removing filtrate; repeating the hot water extraction step for 3 times, and drying the filter residue to obtain secondary poria residue;

(2) dispersing the secondary poria cocos residues in 0.5mol/L sodium hydroxide solution according to the weight ratio of 1:30 of the material liquid, stirring at room temperature, and filtering to remove filter residues; adding 1mol/L hydrochloric acid into the filtrate to adjust the pH value to 7, standing for 1h, and filtering out a precipitate; desalting the precipitate with pure water, and oven drying to obtain alkali soluble pachyman;

(3) redissolving the alkali soluble pachyman in 0.5mol/L sodium hydroxide solution, then adding concentrated phosphoric acid to adjust the pH to 1 to obtain alkali soluble pachyman dispersion liquid with the concentration of 2 wt%, and standing the alkali soluble pachyman dispersion liquid for 0.5h at room temperature to obtain the pachyman hydrogel.

The appearances of the alkali-soluble pachyman dispersion and the pachyman hydrogel were recorded by a digital camera, and as a result, as shown in fig. 1, fig. 1(a) is a macroscopic photograph of the alkali-soluble pachyman dispersion and fig. 1(b) is a macroscopic photograph of the pachyman hydrogel. After being freeze-dried, the pachyman hydrogel is adhered to a conductive gel, and subjected to electron microscope scanning after gold spraying, and fig. 2 is an electron microscope photograph of the pachyman hydrogel in the embodiment, and as shown in fig. 2, the pachyman hydrogel has a typical three-dimensional network structure.

Example 2: selection of conditions for gelation

Alkali-soluble pachyman dispersion solutions of different concentrations were prepared by redissolving the alkali-soluble pachyman obtained in example 1 in 0.5mol/L sodium hydroxide solution and adjusting the pH by adding concentrated phosphoric acid. Discussion the minimum gelation concentration of alkali soluble pachyman dispersions at different gelation pH.

The results of the experiments show that the gelation behavior of the alkali soluble pachyman dispersion is pH dependent: when the pH of the alkali soluble pachyman dispersion liquid is less than or equal to 0, the alkali soluble pachyman dispersion liquid is difficult to gel; when the pH value of the alkali soluble pachyman dispersion liquid is more than 0 and less than or equal to 1, the lowest gelation concentration is 4mg/mL, and the pH value is closer to 0, and the lower the gelation concentration is higher and is closer to 16 mg/mL; when the pH value of the alkali soluble pachyman dispersion liquid is more than 1.5 and less than 7.8, the lowest gelation concentration is 9 mg/mL; when the pH of the alkali soluble pachyman dispersion liquid is 7.8, the lowest gelation concentration is 10 mg/mL; when the pH of the alkali soluble pachyman dispersion liquid is 10.50, the lowest gelation concentration is 25 mg/mL; when the alkali-soluble pachyman dispersion was adjusted to a pH of about 11.3, the alkali-soluble pachyman dispersion was in a semi-gel state and could not be completely inverted without flowing. FIG. 3 shows the minimum gelation concentration of alkali soluble pachyman dispersions at different pH.

Example 3

The alkali-soluble pachyman obtained in example 1 was redissolved in 0.5mol/L sodium hydroxide solution, concentrated phosphoric acid was added to adjust pH to 1, alkali-soluble pachyman dispersions of different concentrations (4mg/mL, 8mg/mL, 12mg/mL, 16mg/mL, 20mg/mL) were prepared, and the alkali-soluble pachyman dispersion was allowed to stand at room temperature for 0.5h to obtain a pachyman hydrogel. Heating pachyman hydrogel, and investigating the disintegration temperature (phase transition temperature T) of pachyman hydrogel prepared from alkali soluble pachyman dispersion liquid with different concentrations_GS)。

The results of the phase transition temperature experiments shown in fig. 4 show that the phase transition temperature of the pachyman hydrogel is positively correlated with the concentration of the alkali-soluble pachyman dispersion. Rhizoma Smilacis GlabraeWhen the concentration of the sugar dispersion liquid is more than 8mg/mL, the phase transition temperature T of the obtained pachyman hydrogel_GSIs 52 ℃; when the concentration of the alkali soluble pachyman dispersion is more than 12mg/mL, the phase transition temperature T of the obtained pachyman hydrogel_GSAt 58 ℃; when the concentration of the alkali soluble pachyman dispersion is more than 16mg/mL, the phase transition temperature T of the obtained pachyman hydrogel_GSIs 62 ℃; when the concentration of the alkali soluble pachyman dispersion is more than 20mg/mL, the phase transition temperature T of the obtained pachyman hydrogel_GSIt was 64 ℃.

Example 4

Redissolving the alkali-soluble pachymaran obtained in example 1 in 0.5mol/L sodium hydroxide solution, adding concentrated phosphoric acid to adjust the pH to 0-10, preparing alkali-soluble pachymaran dispersion liquid with the concentration of 25mg/mL, standing the alkali-soluble pachymaran dispersion liquid at room temperature to obtain pachymaran hydrogel, and recording the time from standing to inversion of the alkali-soluble pachymaran dispersion liquid with different pH values, wherein the time is the gelation time of the alkali-soluble pachymaran dispersion liquid.

Table 1 shows the time required for 25mg/mL alkali soluble pachyman dispersions of different pH to form pachyman hydrogels, and the results are shown in FIG. 5.

TABLE 1

pH(25mg/mL)	Required time of
		0.62	30min
0.95	12min
		2.14	20-30s
3.19	20s
		4.2	4s
5.25	5s
		6.06	5s
7	15～20min
		7.79	30min
10.17	40min
		10.5	3h

Example 5: rheological property experiment of pachyman hydrogel

Redissolving the alkali-soluble pachyman obtained in example 1 in 0.5mol/L sodium hydroxide solution, adding concentrated phosphoric acid to adjust pH to 1, and preparing alkali-soluble pachyman dispersion solutions with different concentrations (1 wt%, 1.5 wt%, 2.0 wt%, 2.5 wt%); and standing the alkali-soluble pachyman dispersion liquid at room temperature for 0.5h to obtain the pachyman hydrogel. The measurement is carried out at 25 ℃ by adopting a rheometer, a clamp with the thickness of 25mm is selected, the amplitude is set to be 1%, and the distance between two plates is set to be 1 mm. Under the measurement mode of frequency scanning, the storage modulus G 'and the loss modulus G' of the pachyman hydrogel are measured with the angular frequency of 0.1-10 rad/s.

FIG. 6 shows the rheological properties of the pachyman hydrogel of the present example, and as shown in FIG. 6, the storage modulus G 'of the pachyman hydrogel prepared is greater than the loss modulus G' at an angular frequency of 0.01 to 10rad/s, thus demonstrating that the pachyman hydrogel is formed under the conditions and has good elastic properties. As the concentration of the alkali soluble pachyman dispersion increased, both G' and G "increased, indicating that the mechanical properties of the pachyman hydrogel were better.

Example 6: anti-inflammatory test of pachyman hydrogel

Redissolving the alkali-soluble pachyman obtained in example 1 in 0.5mol/L sodium hydroxide solution, adding concentrated phosphoric acid to adjust pH to 7, and preparing an alkali-soluble pachyman dispersion liquid with a concentration of 2 wt%; and standing the alkali-soluble pachyman dispersion liquid at room temperature for 2h to obtain the pachyman hydrogel.

Macrophage RAW264.7 was used as the subject, and Lipopolysaccharide (LPS) was used for stimulation to induce inflammation to create a cellular inflammatory response model. RAW264.7 cells were cultured at 37 ℃ with 5% CO₂Culturing in DMEM medium containing fetal calf serum under saturated humidity environment. Adding DMEM culture medium of fetal calf serum above 0.25g pachyman hydrogel, and soaking for 24 hr to obtain leaching solution of pachyman hydrogel; culturing cells in the leaching solution of pachyman hydrogel, stimulating with 1 μ g/mL LPS for 24h, and observing the reaction of the leaching solution of pachyman hydrogel on cell inflammation.

TABLE 2 anti-inflammatory Experimental groups

Extracting miRNA in cells, adding Trizol for full lysis, adding chloroform into 200 mu L of chloroform/mL of Trizol for full mixing, centrifuging at 12000rpm for 15min, adding 80% isopropanol into filtrate, mixing uniformly, and centrifuging at 12000rpm for 10 min. The pellet was dissolved in 75% ethanol without miRNA enzyme. After the RNA concentration is determined, 1.0 mu g of RNA is taken for reverse transcription according to the instruction of the kit, and the miRNA expression of TNF-alpha and 1L-1 beta is detected by a fluorescent real-time quantitative PCR instrument. The upstream and downstream primer sequences of each gene are respectively TNF-alpha: 5'-GGCGGTGCCTATGTCTCA-3' and 5'-CCTCCACTTGGTGGTTTGT-3'; 1L-1. beta.: 5'-GTTCCCATTAGACAACTGC-3' and 5'-GATTCTTTCCTTTGAGGC-3'; GAPDH: 5'-TGTTTCCTCGTCCCGTAGA-3', and 5'-GATGGCAACAATCTCCACTTTG-3'. And calculating the expression difference of the target gene by taking the GAPDH as an internal reference gene and taking the difference between the saturation state cycle value of the target gene and the saturation state cycle value of the GAPDH gene as a basis.

FIG. 7 shows the cell viability of RAW264.7 cells cultured in 24h medium diluted 1:1, 1:2, 1:10, 1:20 times respectively.

FIG. 8 is a graph showing the effect of pachyman hydrogel on the level of tumor necrosis factor-alpha (TNF-alpha) production by LPS-induced RAW264.7 cells, as can be seen in FIG. 8: compared with the blank group, the TNF-alpha level of the LPS group is obviously increased (p is less than 0.001), which indicates that the TNF-alpha level generated by RAW264.7 cells stimulated by LPS is obviously increased; the pachyman hydrogel group had a decreased TNF- α level (p <0.05) compared to the LPS group, indicating that the pachyman hydrogel group was able to inhibit the expression level of TNF- α.

FIG. 9 is a graph showing the effect of pachyman hydrogel on the level of interleukin 1 β (1L-1 β) production by LPS-induced RAW264.7 cells, as can be seen in FIG. 9: compared with the blank group, the LPS group has the advantages that the 1L-1 beta level is obviously increased (p is less than 0.001), which shows that the LPS stimulates the RAW264.7 cells to generate the 1L-1 beta level which is obviously increased; compared with LPS group, the pachyman hydrogel group has obviously reduced level of 1L-1 beta (p is less than 0.001), which indicates that the pachyman hydrogel group can obviously inhibit the expression level of 1L-1 beta.

The analysis shows that a certain amount of pachyman hydrosol is wrapped in the pachyman hydrogel, and the pachyman hydrosol slowly seeps out under the soaking of a DMEM culture medium of fetal calf serum.

Example 7: application of pachyman hydrogel in-vitro drug release

Redissolving the alkali-soluble pachyman obtained in example 1 in 0.5mol/L sodium hydroxide solution containing 1.5mg/mL baicalin, adding concentrated phosphoric acid to adjust pH to 7, and preparing an alkali-soluble pachyman dispersion solution with a concentration of 1 wt%; and standing the alkali soluble pachyman dispersion liquid at room temperature for 2h to obtain pachyman medicine carrying gel.

Fig. 10 shows the drug release behavior of pachyman drug loaded gels at 37 ℃ in different release medium environments. As can be seen from fig. 10, the pachyman drug-loaded gel can effectively realize the long-term release and controlled release of baicalin, which indicates that the pachyman hydrogel prepared by the invention can be used as a carrier of drugs.

The foregoing are merely exemplary embodiments of the present invention, which enable those skilled in the art to understand or practice the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.