CN106692037B - Rhein supramolecular hydrogel and preparation method and application thereof - Google Patents
Rhein supramolecular hydrogel and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses rhein supramolecular hydrogel and a preparation method and application thereof, wherein the rhein supramolecular hydrogel comprises rhein and NaHCO3A solution of said rhein dissolved in NaHCO3In solution. The preparation method comprises the following steps: dissolving rhein in NaHCO3In the solution, rhein supermolecule hydrogel is obtained by ultrasonic dispersion. The rhein supermolecule hydrogel has the characteristics of anti-inflammation, antibiosis, anti-tumor and the like, can solve the problems of poor solubility and short half-life period of rhein, retains the biological activity of rhein, and has obvious treatment effect when being applied to the medicines for treating microglial cell mediated inflammatory encephalopathy such as Alzheimer disease, Parkinson disease, cerebral trauma, cerebral edema and the like.
Description
Technical Field
The invention relates to the technical field of traditional Chinese medicines, in particular to a preparation method and application of rhein supermolecule hydrogel.
Background
Rhein is a free anthraquinone derived from radix et rhizoma Rhei15H8O6. Rhein is a natural antioxidant and anti-inflammatory substance, has little toxic and side effects, and has good antioxidant and anti-inflammatory effects, so that the rhein has high activity in the aspects of tumor resistance, anti-inflammation, purgation, neuroprotection, lipid regulation and the like.
In the inflammatory diseases mediated by microglia such as Alzheimer's disease, Parkinson's disease, brain trauma and cerebral edema, the microglia in a resting state is activated and migrates to the damaged brain cell region after the central nervous system is damaged, and the activated microglia can secrete various cell inflammatory factors such as Inducible Nitric Oxide Synthase (iNOS), tumor necrosis factor (TNF- α), interleukin 1 β (IL-1 β), interleukin 6 (IL-6) and interleukin 12 (IL-12), which are involved in the immune response of the inflammatory reaction.
Rhein has good anti-inflammatory pharmacological action on microglia-mediated central nervous system diseases, but rhein has poor solubility due to a rigid structure, so that the bioavailability is reduced. On the other hand, the traditional Chinese medicine preparation has a certain medicine half-life period in a human body, and the acting time at a pathological part is short, so that the due curative effect is difficult to achieve. Therefore, a new formulation is needed to improve the sustained release effect of rhein.
Disclosure of Invention
The invention aims to overcome the defects of the prior art, provides the rhein supermolecule hydrogel with the characteristics of anti-inflammation, antibiosis, anti-tumor and the like, can solve the problems of poor solubility and short half-life period of rhein, and retains the biological activity of rhein. The invention also provides a preparation method of the rhein supermolecule hydrogel, and the preparation process is simple. The invention also provides application of the rhein supermolecule hydrogel in preparing a medicament for treating microglial cell-mediated inflammatory encephalopathy such as Alzheimer disease, Parkinson disease, cerebral trauma, cerebral edema and the like, and the treatment effect is obvious.
In order to solve the technical problems, the invention adopts the following technical scheme:
a rhein supermolecule hydrogel comprises rhein and NaHCO3A solution of said rhein dissolved in NaHCO3In solution.
Preferably, the rhein supramolecular hydrogel is prepared by adding rhein to a solution with a concentration of more than 4mg/mL and NaHCO3The concentration of the solution is 0.2-0.5 mol/L.
Preferably, the concentration of the rhein supermolecule hydrogel is 5-8 mg/mL.
As a general technical concept, the invention also provides a preparation method of the rhein supermolecule hydrogel, which is to dissolve rhein in NaHCO3In the solution, rhein supermolecule hydrogel is obtained by ultrasonic dispersion.
In the above preparation method, preferably, the concentration of the rhein is more than 4mg/mL, and the NaHCO is3The concentration of the solution is 0.2-0.5 mol/L.
In the preparation method, the concentration of the rhein is preferably 5-8 mg/mL.
In the preparation method, preferably, the power of the ultrasonic dispersion is 70-120W; the time of ultrasonic dispersion is 30-60 min, and the temperature of ultrasonic dispersion is not more than 37 ℃.
As a general technical concept, the invention also provides an application of the rhein supramolecular hydrogel or the rhein supramolecular hydrogel prepared by the preparation method in preparation of a medicine for treating related diseases mediated by microglia or a medicine for treating inflammatory encephalopathy mediated by the microglia.
As a general technical concept, the invention also provides an application of the rhein supramolecular hydrogel or the rhein supramolecular hydrogel prepared by the preparation method in preparation of a medicine for relieving or treating microglial inflammatory diseases.
As a general technical concept, the invention also provides application of the rhein supramolecular hydrogel or the rhein supramolecular hydrogel prepared by the preparation method in preparation of medicines for treating Alzheimer disease, Parkinson disease, cerebral trauma and cerebral edema or medicines for protecting nerve cells.
Compared with the prior art, the invention has the advantages that:
(1) the invention provides rheinic acid supermolecule hydrogel, which is in a nanofiber shape and has self slow release, so that the problem of poor solubility of rheinic acid is solved, the bioactivity of rheinic acid is retained, the action time of rheinic acid on a pathological change position is prolonged, and the treatment effect of a medicament is enhanced. The rhein supermolecule hydrogel prepared by the invention firstly provides that rhein serving as a traditional Chinese medicine monomer is taken as a gel factor and is added into NaHCO3The rhein supermolecule hydrogel with a three-dimensional network structure is self-assembled by utilizing non-covalent bond actions such as pi-pi bond, hydrogen bond, ionic bond, hydrophobic action and the like in the solution,
(2) the invention provides a preparation method of rhein supermolecule hydrogel, in the hydrogel forming process, the ultrasonic action promotes the system to rapidly form tiny crystal nucleus, then the tiny crystal nucleus grows into a fibrous skeleton structure, fibers are mutually lapped to form a network structure, and then the network structure is combined with water to form gel based on the surface tension effect, the preparation process is simple, the cost is low, the method can be commercialized, and the method is suitable for large-scale production and is expected to be applied to the biomedical fields of tissue engineering, drug slow release, wound healing, inflammation inhibition, neuroprotection and the like.
Drawings
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention.
Fig. 1 is a Scanning Electron Microscope (SEM) image of rhein solution and rhein supramolecular hydrogel in example 1 of the present invention.
Fig. 2 is a digital photograph of rhein solution and rhein supramolecular hydrogel in example 2 of the invention.
Fig. 3 is a fourier infrared spectrum of the rhein supramolecular hydrogel and rhein powder in example 2 of the present invention.
Fig. 4 is an ultraviolet spectrum of the rhein supramolecular hydrogel in example 3 of the invention.
FIG. 5 is a graph showing the effect of rhein supramolecular hydrogel and rhein on LPS-induced levels of Inducible Nitric Oxide Synthase (iNOS) in BV2 cells in example 4 of the present invention.
FIG. 6 is a graph showing the effect of rhein supramolecular hydrogel and rhein on LPS-induced generation of tumor necrosis factor α (TNF- α) inflammatory factor by BV2 cells in example 4 of the present invention.
FIG. 7 is a graph showing the effect of rhein supramolecular hydrogel and rhein LPS in inducing BV2 cells to generate interleukin 6 (IL-6) inflammatory factor in example 4 of the present invention.
FIG. 8 is a graph showing the effect of rhein supramolecular hydrogel and rhein LPS in inducing BV2 cells to generate interleukin 12 (IL-12) inflammatory factor levels in example 4 of the present invention.
Detailed Description
The invention is further described below with reference to the drawings and specific preferred embodiments of the description, without thereby limiting the scope of protection of the invention.
The materials and equipment used in the following examples are commercially available.
Example 1
The rhein supermolecule hydrogel comprises rhein and NaHCO3Solution, rhein dissolved in NaHCO3The solution is prepared by the following preparation method:
(1) 1.68 g NaHCO are weighed3Adding ultrapure water into a beaker, stirring and dissolving, transferring into a 100 mL volumetric flask, adding water to a constant volume, and preparing NaHCO with the concentration of 0.2 mol/L3And (3) solution.
（2）Weighing 1 mg, 2 mg and 6mg rhein in corresponding screw bottles respectively, and adding 1 mL of NaHCO with the concentration of 0.2 mol/L into each screw bottle while shaking3And (3) carrying out ultrasonic dispersion on the solution for 30min at the ultrasonic power of 100W, wherein the temperature is not more than 37 ℃, and preparing 1 mg/mL rhein solution and 2 mg/mL rhein supermolecule hydrogel respectively.
Scanning the rhein solution of 1 mg/mL and 2 mg/mL and the rhein supermolecule hydrogel of 6mg/mL respectively by an electron microscope.
Experimental procedures of electron microscope scanning:
1. cleaning a silicon wafer: firstly, ultrasonically cleaning the silicon wafer by using goby (concentrated sulfuric acid (v): hydrogen peroxide (v) = 7: 3) for 15 min, then ultrasonically cleaning the silicon wafer by using ethanol for 15 min, finally ultrasonically cleaning the silicon wafer by using distilled water for 15 min (2 times), and drying the silicon wafer by using nitrogen.
2. Respectively taking 10 mu L of rheinic acid solution of 1 mg/mL or 2 mg/mL and rheinic acid supramolecular hydrogel of 6mg/mL on a blown-clean silicon chip, freezing for 12 h, putting the silicon chip into a freeze dryer for drying for 12 h, and then scanning by an electron microscope. Because the biological sample has poor conductivity, the biological sample is sprayed with gold before scanning.
FIG. 1 shows the scanning results of electron microscopy on 1 mg/mL rhein solution, 2 mg/mL rhein solution and 6mg/mL rhein supramolecular hydrogel in this example. a. b and c are 1 mg/mL, 2 mg/mL and 6mg/mL rhein supramolecular hydrogel respectively, as shown in the figure, when the concentration of rhein is 1 mg/mL (solution state), the obtained SEM appearance is a needle-shaped nanorod structure. When the concentration is increased to 2 mg/mL, the nano rod becomes thicker gradually, and when the concentration is increased to 6mg/mL continuously, rhein supramolecular hydrogel with a three-dimensional network space structure is finally obtained. Along with the increase of the concentration, rhein molecules form nanofibers through non-covalent self-assembly from the acicular nanorods.
Example 2
The rhein supermolecule hydrogel comprises rhein and NaHCO3Solution, rhein dissolved in NaHCO3The solution is prepared by the following preparation method:
(1) 1.68 g NaHCO are weighed3In a beakerAdding ultrapure water, stirring for dissolving, transferring into a 100 mL volumetric flask, adding water for constant volume, and preparing NaHCO with concentration of 0.2 mol/L3And (3) solution.
(2) Weighing 4mg and 5 mg rhein respectively in a screw bottle, and adding 1 mL NaHCO obtained in the step (1) into the screw bottle while vibrating3And carrying out ultrasonic dispersion on the solution for 30min at the ultrasonic power of 70W at the temperature of not more than 37 ℃ to prepare 4mg/mL rhein solution and 5 mg/mL rhein supermolecule hydrogel.
The appearance of rhein solution and rhein supramolecular hydrogel was recorded with a digital camera. In fig. 3, (a) is a digital photo of rhein solution, and (b) is a digital photo of rhein supramolecular hydrogel. Adding 0.2 mol/L NaHCO into rhein powder3The solution is prepared into emulsion, when the concentration reaches 5 mg/mL, rhein supermolecule hydrogel is formed by self-assembly of rhein molecular monomer through non-covalent bond actions such as pi-pi bond, hydrogen bond, ionic bond, hydrophobic action and the like after ultrasonic treatment for 30min, when the concentration is 4mg/mL, rhein supermolecule hydrogel cannot be formed after ultrasonic treatment, and rhein solution is obtained, as shown in figure 2 (a), and therefore 5 mg/mL is the critical gel concentration of the rhein supermolecule hydrogel.
Fourier infrared test experiment steps: freeze-drying 6mg/mL rhein supermolecule hydrogel to obtain xerogel, then weighing the freeze-dried hydrogel with equal mass and tabletting with certain mass of potassium bromide to obtain a flaky sample, and carrying out infrared test, wherein rhein powder solid is used as a reference.
Fig. 3 is a fourier infrared spectrum of rhein supramolecular hydrogel and rhein powder. Curve a in fig. 3 is rhein powder; curve b is rhein supramolecular hydrogel. 3448 cm in curve a-1Belongs to the stretching vibration absorption peak of O-H, 1962 cm-1And 1630 cm-1Asymmetric and symmetric stretching vibration absorption peaks respectively belonging to C = O and appearing at 1692 cm-1And 1452 cm-1The absorption peak of (2) belongs to the stretching vibration peak of the aromatic ring framework and is positioned at 1268 cm-1The peak belongs to the C-O stretching vibration peak and appears at 808 cm-1The peak of (A) belongs to the bending vibration absorption of O-HA peak; curve b is Rhein in basic NaHCO3And (3) self-assembling to form a spectrum of the rhein supermolecule hydrogel. At 3446 cm-1The peak is the stretching vibration absorption peak of O-H and appears at 1635cm-1The peak belongs to a C = O name telescopic vibration absorption peak, which indicates that the rhein supermolecule hydrogel is formed through hydrogen bond interaction and is at 1692 cm-1The peak at 1378 cm disappeared-1Relatively wide peaks appear, which indicates that pi-pi accumulation and hydrogen bond interaction occur in the process of self-assembly forming rhein supermolecule hydrogel.
Example 3
The rhein supermolecule hydrogel comprises rhein and NaHCO3Solution, rhein dissolved in NaHCO3The solution is prepared by the following method:
(1) 1.68 g NaHCO are weighed3Adding ultrapure water into a beaker, stirring and dissolving, transferring into a 100 mL volumetric flask, adding water to a constant volume, and preparing NaHCO with the concentration of 0.2 mol/L3And (3) solution.
(2) Weighing 5 mg, 6mg, 7 mg and 8 mg rhein respectively into corresponding screw bottles, and adding 1 mL NaHCO of example 1 into the screw bottles while vibrating3And ultrasonically dispersing the solution for 30min at the ultrasonic power of 100W at the temperature of not more than 37 ℃ to respectively prepare 5 mg/mL, 6mg/mL, 7 mg/mL and 8 mg/mL rhein supramolecular hydrogel.
The rhein supramolecular hydrogel of 5 mg/mL, 6mg/mL, 7 mg/mL and 8 mg/mL in this example was subjected to UV testing.
The test result is shown in fig. 4, the ultraviolet spectrum of the rhein supramolecular hydrogel under different concentrations shows that the ultraviolet absorption peak of the rhein supramolecular hydrogel is red-shifted with the increase of the concentration, and rhein molecules may be subjected to pi-pi stacking. The more significant the pi-pi stacking, the more pronounced the red shift phenomenon.
Example 4
The invention relates to an application of rhein supermolecule hydrogel in treating microglial cell mediated inflammatory diseases, which specifically comprises the following steps:
1. preparing the medicine:
preparation of a medicated medium 1: adding rhein supramolecular hydrogel mother liquor with the concentration of 6mg/mL into a culture medium for dilution to obtain a culture medium 1 containing the medicine, wherein the rhein supramolecular hydrogel concentration in the culture medium 1 containing the medicine is 4.5 mu g/mL.
Preparation of culture medium 2: adding rhein monomer mother liquor with concentration of 6mg/mL into culture medium to prepare culture medium 2 containing medicine, wherein the rhein monomer concentration in the culture medium 2 containing medicine is 4.5 mug/mL.
2. Preparation of rat microglia (BV 2 cell line) inflammatory model:
rhein group: digesting BV2 cells in logarithmic phase with pancreatin digestive juice to prepare cell suspension; and (4) paving to ensure 70% coverage rate of each hole. Overnight, cells adhere to the wall; removing culture medium of BV2 cells, adding culture medium 2 containing medicine, and labeling; after the treatment of the culture medium containing 2 for 30min, 1 mu g/mL Lipopolysaccharide (LPS) is added into each hole, and after 48 h of treatment, cells and supernatant are collected for subsequent detection.
Rhein supramolecular hydrogel group: digesting BV2 cells in logarithmic phase with pancreatin digestive juice to prepare cell suspension; and (4) paving to ensure 70% coverage rate of each hole. Overnight, cells adhere to the wall; removing culture medium of BV2 cells, adding culture medium 1 containing medicine, and labeling; and after the drug-containing culture medium 1 is treated for 30min, 1 mu g/mL LPS is added into each hole, and after the treatment is carried out for 48 h, cells and supernate are collected for subsequent detection.
LPS group: digesting BV2 cells in logarithmic phase with pancreatin digestive juice to prepare cell suspension; and (4) paving to ensure 70% coverage rate of each hole. Overnight, cells adhere to the wall; removing culture medium of BV2 cells, adding fresh culture medium without drug, labeling, adding 1 μ g/mL LPS 30min later, treating for 48 h, collecting cells and supernatant, and performing subsequent detection.
Blank group: digesting BV2 cells in logarithmic phase with pancreatin digestive juice to prepare cell suspension; and (4) paving to ensure 70% coverage rate of each hole. Overnight, cells adhere to the wall; the BV2 cell medium was removed and fresh drug-free medium was added and labeled.
3. ELISA method for determining the expression of microglial-Inducible Nitric Oxide Synthase (iNOS), tumor necrosis factor (TNF- α), interleukin 6 (IL-6) and interleukin 12 (IL-12):
(1) an experimental instrument: a desk-top high-speed refrigerated centrifuge Hunan apparatus (H1650R); collecting and loosening a full-automatic enzyme labeling plate washing machine (PW-812); gathering pine by a multifunctional enzyme-labeled analyzer (MB-530); constant temperature incubator glowing (DHP-500); an automatic balancing centrifuge Hunan instrument (L530); cleaning a test tube and a centrifuge tube; a volumetric flask; a series of adjustable pipettors and pipette tips.
(2) Collecting a sample; respectively collecting cell supernatant in each pore plate, centrifuging at 10000 rpm for 10min, carefully sucking the supernatant, preserving in a refrigerator at-20 ℃ for reserving samples for experiments, taking out from the refrigerator before the experiments, and naturally thawing.
(3) Preparing an experimental reagent:
in this embodiment, the kit is an ELISA kit produced by wuhan huamei biotechnology limited, wherein the batch number of each kit is as follows: a16039818, and the kit of the tumor necrosis factor is Lot: z06939817, the interleukin 6 kit is Lot: Z01039820, and the interleukin 12 kit is Lot: Z01039822.
And (3) standard substance: one standard was removed from the kit and centrifuged at 10000 rpm for 30 s. Dissolving the sample with 1 mL of a sample diluent (10 mmol/L PBS (disodium hydrogen phosphate-potassium dihydrogen phosphate), pH 7.3, 0.05% Tween-20 (by volume), and 0.5% Bovine Serum Albumin (BSA), repeatedly sucking the sample at the bottom of the vial with a pipette tip for 5 times to aid dissolution, and mixing to obtain standard S7Standing for later use; 7 centrifuge tubes of 1.5 mL are taken and numbered as S6～S0. The tubes were aligned in order and 250. mu.L of each sample dilution was added. Sucking 250 microliter of standard substance S7Into the first centrifuge tube (S)6) Lightly blow, beat and mix evenly. From S6Draw 250. mu.L into a second EP tube (S)5) And lightly blowing, beating and uniformly mixing, and performing multiple dilution on the standard product by analogy. S0Is a sample dilution.
Washing the working solution: the concentrated wash (0.02 mol/L PBS (pH =7.4) plus 0.05% Tween-20 in this example) was diluted 1: 25 times with deionized water, specifically: 240 mL of deionized water is measured by a measuring cylinder, poured into a beaker or other clean containers, 10 mL of concentrated washing solution is measured, added uniformly, stirred and mixed uniformly, and prepared before use. The concentrated washing solution can be salted out when stored at low temperature, and can be heated in a water bath to assist dissolution when diluted.
Biotin-labeled antibody working solution: the biotin-labeled antibody solution (in this example, 10 mg/mL biotin N-hydroxysuccinimide ester solution) was diluted 1: 100 times with a biotin-labeled antibody diluent, specifically: add 990. mu.L of biotin-labeled antibody dilution to 10. mu.L of biotin-labeled antibody solution, mix gently, and prepare within 10min before use.
Horse radish peroxidase-labeled avidin working solution: the horseradish peroxidase-labeled avidin (horseradish peroxidase-labeled avidin produced by Wuhan Huamei Biotech Co., Ltd. in the example) was diluted by horseradish peroxidase-labeled avidin diluent at a ratio of 1: 100, specifically: mu.L of horseradish peroxidase-labeled avidin and 990. mu.L of horseradish peroxidase-labeled avidin diluent are added, and the mixture is gently mixed and mixed for 10min before use.
(4) The method comprises the following operation steps:
4.1, moving the various reagents to room temperature (18-25 ℃) for balancing for at least 30min, and preparing the reagents according to the method for later use.
4.2, sample adding: respectively arranging a standard hole and a sample hole to be detected, adding 100 mu L of standard substance or sample to be detected into each hole, slightly shaking and uniformly mixing, covering a plate and sticking, and incubating for 2 h at 37 ℃.
4.3, discarding the liquid, and drying by spin without washing.
4.4 adding 100 μ L of biotin-labeled antibody working solution to each well, covering with a new plate, and incubating at 37 ℃ for 1 h.
4.5, discarding liquid in the holes, spin-drying, and washing the plate for 3 times by adopting the prepared washing working solution. Washing the plate each time, soaking the washing working solution for 2 min, and spin-drying at 200 μ L per hole.
4.6, 100 mu L of horseradish peroxidase labeled avidin working solution is added into each hole, a new plate is covered, and the mixture is incubated for 1 h at 37 ℃.
4.7, discarding liquid in the holes, spin-drying, and washing the plate for 5 times. Soaking for 2 min each time, soaking in 200 μ L per hole, and drying.
4.8, sequentially adding 90 μ L of substrate solution to each well, and developing in the dark at 37 ℃ for 15 min.
4.9 adding stop solution (2 mol/L H in this example) to each well in sequence2SO4Solution) 50. mu.L, the reaction was terminated.
4.10, measuring the optical density (OD value) of each hole in turn by using a microplate reader within 5 min after the reaction is ended and at the wavelength of 450 nm, and calculating the content of each inflammatory factor. The experimental results are shown in fig. 5, 6, 7 and 8, respectively.
Fig. 5 is a graph of the effect of rhein supramolecular hydrogel and rhein on LPS-induced BV2 cell-Induced Nitric Oxide Synthase (iNOS) levels, as can be seen in fig. 5: to blank set ratio (#p is less than 0.01), the iNOS level of LPS group is obviously increased, which shows that the iNOS level is obviously increased when the LPS stimulates BV2 cells to generate iNOS; (significantly reduced levels of iNOS expression in rhein group compared with LPS group△p is less than 0.05), the expression level of the rhein supramolecular hydrogel group iNOS is also obviously reduced (p is less than 0.01), which indicates that rhein and rhein supramolecular hydrogel can inhibit the expression level of iNOS; compared with rhein group, rhein supermolecular hydrogel group iNOS expression level is obviously reduced (□p is less than 0.01), which shows that the rhein supermolecular hydrogel is obviously superior to rhein in inhibiting the expression of iNOS.
FIG. 6 shows the effect of rhein supramolecular hydrogel and rhein on LPS-induced BV2 cell tumor necrosis factor α (TNF- α) levels, as seen in FIG. 6, compared to blank group (R: (R) (R))#p is less than 0.01), the TNF- α level of LPS group is obviously increased, which shows that the LPS stimulates BV2 cells to generate TNF- α level to be obviously increased, and the TNF- α expression level of rhein group is obviously reduced compared with LPS group (△p is less than 0.05), the expression level of the rhein supramolecular hydrogel group TNF- α is also obviously reduced (*p is less than 0.01), which shows that the rhein and the rhein supermolecular hydrogel can inhibit the expression level of TNF- α, and compared with rhein, the rhein supermolecular hydrogel group TNF- αIs obviously reduced (□p is less than 0.01), which shows that the rhein supermolecular hydrogel is obviously superior to rhein in inhibiting the expression of TNF- α.
FIG. 7 is a graph of the effect of rhein supramolecular hydrogel and rhein on LPS-induced BV2 cell interleukin 6 (IL-6) levels, as can be seen in FIG. 7: to blank set ratio (#p is less than 0.01), the IL-6 level of the LPS group is obviously increased, which shows that the IL-6 level of BV2 cells stimulated by LPS is obviously increased; the rhein group showed a significant reduction in IL-6 expression levels compared to the LPS group (△p is less than 0.05), the expression level of rhein supramolecular hydrogel group IL-6 is also obviously reduced (*p is less than 0.01), which indicates that rhein and rhein supermolecular hydrogel can inhibit the expression level of IL-6; compared with rhein group, rhein supermolecular hydrogel group IL-6 expression level is obviously reduced (□p is less than 0.01), which shows that the rhein supermolecular hydrogel is obviously superior to rhein in inhibiting the expression of IL-6.
FIG. 8 is a graph of the effect of rhein supramolecular hydrogel and rhein on LPS-induced BV2 cell interleukin 12 (IL-12) levels, as can be seen in FIG. 8: to blank set ratio (#p is less than 0.01), the IL-12 level of the LPS group is obviously increased, which shows that the IL-12 level of BV2 cells stimulated by LPS is obviously increased; the rhein group showed a significant reduction in IL-12 expression levels compared to the LPS group (△p is less than 0.05), the expression level of rhein supramolecular hydrogel group IL-12 is also obviously reduced (*p is less than 0.01), which indicates that rhein and rhein supermolecular hydrogel can inhibit the expression level of IL-12; compared with rhein group, rhein supramolecular hydrogel group IL-12 expression level is obviously reduced (□p is less than 0.01), which shows that the rhein supermolecular hydrogel is obviously superior to rhein in inhibiting the expression of IL-12.
In conclusion, the rhein supramolecular hydrogel can reduce the expression of cell inflammation factors such as nitric oxide synthase, tumor necrosis factor, interleukin 6, interleukin 12 and the like, and shows that the rhein supramolecular hydrogel can be used for treating or relieving microglia-mediated central nervous system diseases such as Alzheimer disease, Parkinson disease, brain trauma, cerebral edema and the like, and plays an important role in preparing medicaments for treating microglia-mediated related diseases, microglia inflammatory diseases, Alzheimer disease, Parkinson disease, brain trauma, cerebral edema, microglia-mediated inflammatory encephalopathy and medicaments for protecting nerve cells.
The foregoing is merely a preferred embodiment of the invention and is not intended to limit the invention in any manner. Although the present invention has been described with reference to the preferred embodiments, it is not intended to be limited thereto. Those skilled in the art can make many possible variations and modifications to the disclosed embodiments, or equivalent modifications, without departing from the spirit and scope of the invention, using the methods and techniques disclosed above. Therefore, any simple modification, equivalent replacement, equivalent change and modification made to the above embodiments according to the technical essence of the present invention are still within the scope of the protection of the technical solution of the present invention.
Claims (6)
1. The rhein supramolecular hydrogel is characterized in that raw materials of the rhein supramolecular hydrogel comprise rhein and NaHCO3The preparation method of the rhein supermolecule hydrogel comprises the step of dissolving rhein serving as a gel factor in NaHCO3In the solution, rhein supermolecule hydrogel is obtained by ultrasonic dispersion; the concentration of the rhein is 5-8 mg/mL, and the NaHCO is3The concentration of the solution is 0.2-0.5 mol/L.
2. A preparation method of rhein supermolecule hydrogel is characterized in that rhein is used as a gel factor to be dissolved in NaHCO3In the solution, rhein supermolecule hydrogel is obtained by ultrasonic dispersion; the concentration of the rhein is 5-8 mg/mL, and the NaHCO is3The concentration of the solution is 0.2-0.5 mol/L.
3. The preparation method according to claim 2, wherein the power of the ultrasonic dispersion is 70-120W; the time of ultrasonic dispersion is 30-60 min, and the temperature of ultrasonic dispersion is not more than 37 ℃.
4. An application of the rhein supramolecular hydrogel according to claim 1 or the rhein supramolecular hydrogel prepared by the preparation method according to claim 2 or 3 in preparation of a medicament for treating diseases related to microglia-mediated diseases or in preparation of a medicament for treating the inflammatory encephalopathy mediated by the microglia.
5. Use of the supramolecular rhein hydrogel according to claim 1 or the supramolecular rhein hydrogel prepared by the preparation method according to claim 2 or 3 in preparation of a medicament for alleviating or treating microglial inflammatory diseases.
6. Use of the supramolecular hydrogel of rhein according to claim 1 or the supramolecular hydrogel of rhein prepared by the preparation method according to claim 2 or 3 in preparation of medicines for treating alzheimer disease, parkinson disease, brain trauma and cerebral edema or medicines for protecting nerve cells.
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