CN116942655A - Application of puerarin in preparation of medicine for treating hashimoto thyroiditis - Google Patents

Abstract

The invention discloses application of puerarin in preparing medicines for preventing and/or treating hashimoto thyroiditis, and the puerarin can effectively improve hashimoto phenotype.

Description

Application of puerarin in preparation of medicine for treating hashimoto thyroiditis

Technical Field

The invention relates to a new application of a medicine, in particular to an application of puerarin in preparing a medicine for treating hashimoto thyroiditis.

Background

There is no effective treatment means at present for hashimoto thyroiditis, and most patients suffer from hypothyroidism later, and clinical main suggestions are that the L-thyroxine tablet (L-T4) is orally taken for life to relieve symptoms. However, the use of such drugs is controversial, mainly because of their side effects in clinical applications. Thyroidectomy is the preferred treatment when the bridge patient has severe thyromegaly and affects functions such as swallowing, however, traumatic surgery can bring complications and the thyroxine needs to be taken for life after the surgery, affecting the quality of life of the patient. However, the greatest disadvantage of the above-mentioned treatments is the lack of treatment for pathogenic autoimmune abnormalities. Thus, in addition to symptomatic treatment, improving current symptoms, new therapeutic strategy studies need to be developed.

Puerarin is an isoflavone active ingredient extracted from dry root of Pueraria lobata Ohwi and Pueraria thomsonii of Leguminosae, and is mainly used for treating cardiovascular and cerebrovascular diseases clinically, and has new research on action and mechanism in type 2 diabetes and its complications in recent years.

Disclosure of Invention

The invention aims to: the invention aims to provide a puerarin monomer which can improve the occurrence and development of the bridge disease.

The technical scheme is as follows: the application of puerarin in preparing medicine for preventing and/or treating hashimoto thyroiditis is provided.

The application of the medicine is that the medicine is added with pharmaceutically acceptable auxiliary materials to prepare a preparation.

The application is that the preparation is injection preparation.

The application comprises that the pharmaceutically acceptable auxiliary materials comprise solubilizer, cosolvent, PH regulator, surfactant, antioxidant, isotonic regulator and bacteriostat.

The application is that the preparation is an oral preparation.

The pharmaceutically acceptable auxiliary materials comprise diluents, adhesives, solubilizers, disintegrants, glidants, lubricants, flavoring agents, inclusion materials and adsorption materials.

The application is that the preparation is granule, powder, tablet, capsule, pill and oral liquid.

The use, the prevention and/or treatment of hashimoto thyroiditis is characterized by a reduced degree of lymphocyte infiltration and autoantibody levels in the thyroid tissue of hashimoto thyroiditis.

The use of said prevention and/or treatment of hashimoto's thyroiditis is shown to inhibit the expression of chemokines and adhesion molecules in thyroid tissue.

The use, the prevention and/or treatment of hashimoto thyroiditis is manifested by inhibiting serum inflammatory factor levels of hashimoto thyroiditis.

The beneficial effects are that: compared with the prior art, the invention has the following technical advantages: our findings expand the current new approach to HT treatment, puerarin can combat the expression of chemokines and adhesion molecules in HT thyroid tissues, effectively inhibit the effects of inflammatory lesions of thyroid tissues, and have significant differences from the control group.

Drawings

FIG. 1 is a diagram of a bridge-like mouse model construction (7 week old female mice, day 0 for the first antigen immunostimulatory mice, day 14 for the second antigen immunostimulatory mice, day 21 for the treatment mice);

FIG. 2 shows that puerarin treatment improves HT mouse phenotype. A) Mouse thyroid pathological tissue HE staining with 400 amplification factor; b) Mouse thyroid tissue immunohistochemical CD45 staining, magnification 400; C. d) mouse serum levels of anti-thyroglobulin antibody (TG-ab) and anti-thyroperoxidase antibody (TPO-ab) levels; e) Grade index of mononuclear cell infiltration degree in thyroid gland tissues of mice; (vs NC, <0.05; vs HT, #p < 0.05);

FIG. 3 shows puerarin improvement of thyroid tissue and systemic inflammatory levels in HT mice. A) The qpcr method detects the transcription level of chemokines CCL21, CXCL10 and adhesion molecule ICAM1 in the thyroid tissue of the mice; b) Detecting the transcription level of a regulatory T cell important transcription factor Foxp3 in a mouse thyroid tissue by a qpcr method; c) The qpcr method detects the transcription level of IL-10 in thyroid tissues of mice; d) The qpcr method detects the transcriptional level of IL-17A in thyroid tissues of mice; e) Detecting the transcription level of MCP-1 in thyroid tissues of mice by a qpcr method; F-H) ELISA method for detecting the expression level of IL-10, IL-17A and MCP-1 in serum level of mice. * P <0.05; vs HT, #P < 0.05);

FIG. 4 shows that puerarin effectively inhibits IFNγ -induced macrophage inflammatory response in mice; ifnγ induced mouse macrophages (3 hours); puerarin intervention (3 hours).

Detailed Description

Example 1

Establishing a mouse model

A bridge mouse model was constructed by two swine thyroglobulin (pig TG) immune stresses using classical mouse experimental thyroiditis modeling methods (figure 1).

1. EAT model establishment and puerarin intervention

A classical model method for experimental thyroiditis (Experimental autoimmune thyroiditis, EAT) of mice is adopted, and B6 female 7-week-old mice are selected. On day 0, mice were subcutaneously injected (100. Mu.g/ml) after emulsification of 100. Mu.g of porcine thyroglobulin (dissolved in PBS at a concentration of 1 mg/ml) and 100. Mu.l of Freund's complete adjuvant. On day 14, the mice of the above group were given a second subcutaneous injection near the back of the tail, again emulsified with the same dose of porcine thyroglobulin and Freund's incomplete adjuvant.

Different interventions were given during the molding process, animals were treated and harvested on day 21. Puerarin treatment group (HP group) was intraperitoneally injected with 150mg/kg puerarin injection (Baiyunshan pharmaceutical Co., ltd.) according to body weight, NC group (control group), HT group (bridge not administered group) mice were intraperitoneally injected with equal volume of propylene glycol (puerarin solvent). The model was started with the drug at a fixed time daily, once a day, for three consecutive weeks until the model ended.

Example 2

Pathological morphological analysis:

1) Mouse thyroid tissues were fixed with 4% paraformaldehyde, paraffin embedded and sectioned. Dewaxing in xylene for 5-10 min. Fresh xylene solution was replaced and dewaxed for another 5-10 minutes. Soaking in 100% absolute ethanol for 5min. Soaking in 90% ethanol for 2 min, soaking in 80% ethanol for 2 min, soaking in 70% ethanol for 2 min, and soaking in distilled water for 2 min.

2) Hematoxylin Eosin (HE) staining for 5min was self-adjusting according to the status of the slide. Excess staining solution was removed by double distilled water washing for about 5 minutes. Distilled water is washed once again (controlled time). At this time, if direct observation is required, washing with 70% ethanol may be performed 2 times. If the dehydration and transparent sealing sheet are needed, the dehydration, transparent sealing sheet treatment and sealing sheet treatment can be carried out according to the subsequent steps after 70% ethanol washing.

3) Dehydrating, transparentizing and sealing: 70% ethanol for 10 seconds, 80% ethanol for 10 seconds, 90% ethanol for 10 seconds, and absolute ethanol for 10 seconds. The xylene was transparent for 5 minutes. Fresh xylene was used instead and the mixture was further transparent for 5 minutes. The tablet is encapsulated with a neutral gum or other encapsulating agent. When observed under a microscope, the nuclei appear blue, while the cytoplasm appears pink or red.

4) Pathology scores (infiltration Index, i.i.) for mononuclear cell infiltration were performed on each set of sections, as follows: no mononuclear cell infiltration is found in 0 minutes;

1 minute 2-3 thyroid follicular gaps have aggregation of mononuclear cells;

2 mononucleosis of at least follicular size;

3-division mononuclear cells are widely infiltrated, and the total area of the mononuclear cells is 10% -40%:

4, single nuclear cells are widely infiltrated, and the total area is 40% -80%;

5 minutes of mononuclear cells are widely infiltrated, and the total area is more than 80 percent.

The pathology scoring results of the different groups were statistically analyzed. The degree of lymphocyte infiltration by each thyroid pathology was observed. And immunohistochemical staining for CD45 expression.

Results: the thyroid pathology results are shown in FIG. 2, where Control group, NC group, HP group was improved compared to HT group. Puerarin reduces the infiltration degree of lymphocytes in thyroid tissues of HT-like mice, and obviously reduces the grading index of the infiltration degree, thereby prompting effective improvement of thyroid pathology. Puerarin can be used for treating HT effectively.

Example 3

Cytokine detection

ELISA was used to detect the levels of IFNγ, IL-6, TNF- α, IL-1β, etc. in the relevant sera.

According to Bioscience IL-6, IL-1 beta, IL-10, IFN-gamma instructions

1) The serum of the mice to be tested is diluted 5 times with the diluent.

2) Diluted samples and diluted standard curve standards were added to 96-well plates and incubated at 37℃for 1 hour at 100. Mu.l per well.

3) The 96-well plates were washed 3 times with wash solution for 5min each.

4) Enzyme-labeled antibody was added and incubated at 37℃for 1 hour in 100. Mu.l per well.

5) The 96-well plates were washed 3 times with wash solution for 5min each.

6) TMB substrate was added and 100. Mu.l per well incubated at room temperature for 15min and the liquid chromogenic reaction in the wells was observed.

7) After a significant blue gradient of substrate in the wells, 50. Mu.l of 2N H2SO4 was added to each well to terminate the chromogenic reaction.

8) The degrees are measured on an enzyme-labeled instrument, the two wavelengths are used for calibrating 450nm and 630nm, 450nmOD minus 630nmOD is taken as the degrees, a standard curve is drawn, and the concentration of the cytokines in the serum sample is calculated through the standard curve.

Enzyme-linked immunosorbent assay (ELISA) for detecting autoantibody level and thyroid function

A. Mouse Anti-thyroglobulin antibody (Mouse Anti-tyroid-globulin antibody, TG-Ab)

1) After equilibration for at least 20 minutes from room temperature, the desired strip was removed and the remainder returned to the 4 ° refrigerator.

2) Standard wells and sample wells were set, and standard wells were added with 50 μl of standard of different concentration gradients.

3) Sample Kong Xianjia the sample to be tested is 10 microliters, and 40 microliters of sample diluent is added, i.e., the sample is diluted 5 times. Blank holes are not added.

4) In addition to the blank wells, 100 microliters of horseradish peroxidase (HRP) -labeled detection antigen was added to each of the standard wells and sample wells, and the whole plate was sealed with a sealing plate membrane and incubated for 60 minutes in a 37 ℃ water bath or incubator.

5) The liquid was discarded. The plates were dried by beating on absorbent paper, 250. Mu.l of washing liquid was filled in each well, and the plates were left to stand for 1 minute, the washing liquid was thrown off, dried by beating on absorbent paper, and washed 3 times repeatedly.

6) Substrate A, B was added 50 μl each per well and incubated at 37℃for 15 minutes in the absence of light.

7) The OD of each well was measured at a wavelength of 450nm within 15 minutes by adding 50. Mu.l of stop solution to each well.

8) And calculating the concentration value of each sample according to a curve equation according to the concentration of the standard substance.

B. Mouse Anti-thyroid peroxidase antibody (Mouse Anti-tyroid-peroxidase antibody, TPO-Ab)

The steps are the same as those of TG-Ab, specifically referred to above

C. Mouse thyroxine (T4)

1) The kit is equilibrated for at least 30 minutes.

2) Both standard wells and samples were 50 microliters, and a blank well was placed without any liquid.

3) 50. Mu.l of biotin tag was added to each well, and the mixture was thoroughly mixed except for the blank wells and incubated at 37℃for 1 hour.

4) And (5) manually discharging a gun, adding 250 microliters of washing plates, rapidly discarding the liquid in the holes, filling the holes with the washing liquid, standing and spin-drying, and repeating for three times.

5) 50 microliters of horseradish peroxidase-labeled avidin was added to each well, and the mixture was thoroughly mixed except for blank wells and incubated at 37℃for 15 minutes in the absence of light.

6) The plate was manually washed three times and 50. Mu.l of developer A and 50. Mu.l of developer B were added to each well, mixed by shaking, and incubated at 37℃for 15 minutes in the absence of light. 50 microliters of stop solution was added to each well.

7) The detection was carried out within 10 minutes after the termination of the reaction, and the optical density (OD value) of each well was measured at a wavelength of 450nm by a microplate reader.

Results: as shown in FIG. 2, puerarin has obvious reduction effect on TG-Ab and TPO-Ab. Fig. 3 shows that puerarin significantly inhibits expression of chemokines and adhesion molecules in thyroid tissues of HT mice, suggesting an effective inhibition of inflammatory lesions of thyroid tissues. Puerarin inhibited serum proinflammatory factor levels in mice with bridge samples (FIGS. 3F-H), indicating improved peripheral inflammatory status.

Example 4

Extraction of tissue Total RNA

1) Total RNA was extracted using the Qiagen kit. And (3) cells: less than 5X 106 (disk <6 cm) requires 350ul of RLT buffer, 5X 10X 6 to 1X 10X 7 (disk: 6-10 cm) requires 600ul of RLT buffer. Animal tissue: less than 20mg of RLT buffer 350ul and 20-30mg of RLT buffer 600ul.

2) The tissue or cell homogenates were centrifuged at maximum speed for 3 min and the supernatant was transferred to 2ml gDNA Eliminator spin column.

3) After centrifugation at maximum speed for 30 seconds, the inner column was discarded, the outer collection tube liquid was collected, and one volume of 70% absolute ethanol was added.

4) After mixing, 700. Mu.l of the sample was placed in a 2ml RNeasy spin column collection tube (provided). The lid was gently capped and then centrifuged for 15s at > 8000 Xg (. Gtoreq.10,000 rpm). The liquid was discarded.

5) 700 μl Buffer RW1 was loaded into RNeasy spin columns. The spin column membrane was washed by gently centrifuging the solution with the lid closed for 15s at > 8000 Xg (. Gtoreq.10,000 rpm). The liquid was discarded.

6) 500 μl Buffer RPE was added to the RNeasy spin column. The spin column membrane was washed by gently centrifuging the solution with the lid closed for 15s at > 8000 Xg (. Gtoreq.10,000 rpm). The liquid was discarded.

7) 500 μl Buffer RPE was added to the RNeasy spin column. The spin column membrane was gently washed by centrifugation at > 8000 Xg (. Gtoreq.10,000 rpm) for 2 minutes with the lid closed.

8) The RNeasy spin column was placed in a new 1.5ml collection tube (provided). 30-50. Mu.l of RNase-free water was directly added to the spin-column membrane. The lid was gently closed and centrifuged at ≡8000 Xg (. Gtoreq.10,000 rpm) for 1 minute to elute RNA. If the expected RNA yield is > 30. Mu.g, step 9 is repeated with an additional 30-50. Mu.l of RNase-free water or with the eluate from step 9 (if high RNA concentrations are required).

9) The RNA concentration was measured by a Nanodrop instrument and stored in a-20℃refrigerator.

Reverse transcription and Q-PCR

1) Extracting RNA of corresponding tissues according to RNA extraction instruction, measuring the concentration and purity of the RNA by using a Nanodrop instrument, and storing in a low-temperature refrigerator to prevent repeated freezing and thawing.

2) The extracted RNA was reverse transcribed into cDNA, and the amount of RNA added to a 10. Mu.l system (500 ng RNA) was calculated for each sample at the corresponding RNA concentration using Takara reverse transcription kit. And (3) after the system in the specification is uniformly mixed and completed, carrying out a light-throwing centrifugation, carrying out an RT-PCR reaction, measuring the concentration and purity of cDNA after the completion, and diluting to the concentration required by the experiment for the next step.

5×PrimeScript RT Master Mix	2μl
		Total RNA	500ng
Double distilled water	Added to 10 μl

The reaction procedure:

15min at 37℃C (reverse transcription reaction)

85 ℃ for 5sec (reverse transcriptase inactivation reaction)

4℃

3) Primers of target genes and reference genes required for experiments are designed in advance, and the primers are diluted to proper experimental concentrations. After the samples are determined, the plate layout of the sample adding plate is designed, the sample is carefully added, 3 complex holes are formed in each sample, the PCR reaction is carried out on the machine, and a proper program is selected to meet the experimental requirements.

Q-PCR reaction system:

the reaction procedure:

reagent(s)	Volume of
		SYBR Green PCR Master Mix(2X)	10μl
Forward primer	1μl
		Reverse primer	1μl
Template	1μl
		Water and its preparation method	7μl
Total volume of	20μl

4) Finally, determining the specificity and effectiveness of the reaction by using a dissolution curve, and processing and analyzing the result. The corresponding ratios were calculated and statistically analyzed using the ΔΔct method, as shown in fig. 3A-E.

Example 5

Macrophage culture and intervention

The mouse macrophage line RAW264.7 was purchased from the national academy of sciences of china (Shanghai, china). Cells were grown in DMEM supplemented with 10% (v/v) Fetal Bovine Serum (FBS) and 1% penicillin streptomycin (Gibco, USA) (Thermo, USA) in a 5% co2 incubator at 37 ℃. Macrophage activation (RAW 264.7 cells) model was induced with INF-gamma (0-20 ng/m 1) and on this basis was interfered with puerarin (10 uM).

Immunoblotting (western blot) experiment

Total protein extraction

(1) Pre-cooling the 4-degree centrifugal machine in advance, and preparing an ice box.

(2) And (3) cells: after the number of cells and the time to drug intervention reached the experimental requirements, the cells in the 6-well plate were washed 2 times with PBS after discarding the medium, 200 μl of RIPA lysate containing protease and phosphatase inhibitor (one tablet was dissolved in 10 ml of strong lysate) was added, and the ice-bath was performed for 10min. Centrifuge 13300rpm/min at low temperature of 4 ℃ for 20min, taking supernatant, sub-packaging in an EP tube, and storing at-80 ℃ for standby.

(3) Tissue: 20mg of tissue was minced and placed in a 1.5ml EP tube, homogenized with a homogenizer, and then incubated on ice for 15 minutes. Centrifuge 13 at 4deg.C, centrifugation at 300rpm for 20min, collecting supernatant, packaging in EP tube, and storing at-80deg.C.

BCA method for measuring protein concentration and protein denaturation

(1) Taking 5ul of protein supernatant, adding the solution into 45 microliter (ten times diluted) PBS solution, measuring the protein concentration by using a kit, and preparing working solution (solution A and solution B are mixed according to the volume ratio of 50:1), so as to ensure 200 microliter of the solution A and solution B are mixed in each hole; BSA protein standard (2000, 1000, 500, 250, 125, 62.5,0 ng/ul) was diluted in a gradient.

(2) 10 microliters of standard protein sample or protein sample to be tested is added to each well of the 96-well plate in sequence, and blank control is left. After 200ul BCA working solution is added into each hole, the mixture is vibrated and mixed uniformly.

(3) After incubation at 37 ℃ for 25 minutes, absorbance values are read at 540nm wavelength of the microplate reader, protein concentration of the samples is calculated according to a standard curve, and loading amount of each sample is calculated according to 50 micrograms of protein.

(4) The metal bath machine was opened to 95℃in advance, and the remaining supernatant was denatured by adding protein SDS denaturation buffer (5X) in the metal bath at 95℃for five minutes.

(5) Cooling to room temperature, collecting, and storing in a low temperature refrigerator.

Preparing a solution:

(1) 1 XSDS running buffer:

taking 3.02g of Tris,18.8g of glycine, 1.0g of SDS dry powder, using ddH ₂ O is fixed to 1000mL.

(2) 1 x transfer buffer:

3g Tris,14.4g glycine was taken and subjected to ddH ₂ O constant volume to 100mL, adding 200mL methanol, 700mL ddH ₂ O to 1000mL.

(3) 1 XTBST buffer:

2.42g Tris-base and 8g NaCl were taken and subjected to ddH ₂ O was diluted to about 900mL, the pH was adjusted to 7.6 with hydrochloric acid, 1mL Tween-20 was added, and the mixture was stored at room temperature after constant volume up to 1000mL.

(4) Blocking solution (5% bsa):

5.0g of BSA was dissolved in 100mL of 1 XTBE buffer and prepared for use. Also used as an anti-dilution.

(5) Secondary antibody diluent:

1% BSA was formulated, i.e., 1g BSA was dissolved in 100mL 1 XTBST buffer.

(6) 10% ammonium persulfate (ammonium persulfate, AP) (bi yun day):

0.1. 0.1gAP was dissolved in 1mL of ultrapure water, and the mixture was dispensed into 1.5mL of EP tube and stored in a refrigerator at-20℃in the absence of light.

(7) Preparing 10% concentration glue plate (2 glue blocks):

and after the gel plate is solidified, placing the gel plate in electrophoresis buffer solution for 4 DEG for preservation.

Electrophoresis

(1) And (3) aligning the special glass plate for electrophoresis, inserting the special glass plate into the fixing frame, clamping the special glass plate, and injecting the prepared separating glue into the gap of the glass plate.

(2) A liquid seal was made by adding 1ml of isopropanol to the gel. The mixture was left at room temperature for 30min. When there is a clear line of demarcation between water and release gum, the top isopropyl alcohol on the gum is removed and the water is blotted with blotter paper.

(3) After the comb is cleaned, the concentrated glue is prepared. The remaining space was filled with concentrated glue and then vertically inserted into a comb.

(4) Left at room temperature for 45 minutes.

(5) And (3) taking out the prepared glass plate adhesive after the concentrated adhesive is solidified, clamping the glass plate adhesive into an electrophoresis tank, adding a proper amount of electrophoresis buffer solution, slightly pulling out the comb, and filling the electrophoresis buffer solution.

(6) Adding 50ug protein and Marker, constant voltage running at 80V to the limit of concentrated gel and separation gel, and changing to 120V voltage running to gel bottom until bromophenol blue just running out to stop electrophoresis.

Rotating film (Wet rotating)

(1) In the prepared transfer buffer, a foam-rubber cushion, a filter paper, a 0.45 μm PVDF membrane (immersed in methanol for 1 minute), a gel, a filter paper, and a foam-rubber cushion were placed in this order of positive and negative electrodes, taking care to remove air bubbles between the gel and the PVDF membrane.

(2) The clips were closed and fastened and placed in the transfer tank with the black face of the clip facing the black face of the tank and the white face of the clip facing the red face of the tank. And (5) placing an ice bag at one side of the transfer tank for cooling, and placing the ice bag in an ice bath.

(3) Constant current 350mA for 70 minutes.

Antibody blocking and antibody incubation

(1) PVDF membranes were transferred to a blocking cassette containing 5% bsa blocking solution (1×tbst dissolved) and shake-blocked for 2 hours at room temperature.

(2) After the primary antibody (blocking solution can also be used as primary anti-dilution solution) is diluted to a proper concentration, a PVDF membrane is cut into specific strips according to the known strip sizes of protein markers, and the specific strips are placed in different antibody dilutions.

(3) Shaking table mixing overnight at 4deg.C, then rinsing with 1 XTBE for 3 times each for 5 minutes, adding appropriate secondary antibody to the diluent, shaking table mixing for 1.5h (shaking table speed reduced), and rinsing with 1 XTBE for 5min×3 times.

Chemiluminescent, developing and fixing

(1) Equal volumes of the A solution and the B solution of the ECL luminous solution are mixed for standby, and note that the B solution is generally required to be preserved in a dark place.

(2) The luminous liquid is uniformly coated on the protein surface of the film, each strip is 200 microliters or so, proper exposure time is selected, the film is exposed in a darkroom, the gray value of a target strip is analyzed by Image processing software Image J, and the gray value of the target strip and the gray value of a reference strip in the same lane are used as comparison results, so that the semi-quantitative analysis of the protein can be performed.

(3) If the strip needs to be recycled, the strip is rinsed in distilled water for 5 minutes after the completion of the Western chemiluminescent detection. After distilled water is discarded, a proper amount of Western primary and secondary antibody removing solution (proper PH is selected) is added to completely cover the membrane. Rinsing on a shaker for 5-10 minutes. The Western primary antibody secondary antibody removing liquid is discarded, and the liquid is sucked and removed cleanly. TBS, TBST or PBS was added and rinsed 3 times, each for 5 minutes on a shaker. Then, a subsequent Western operation such as blocking is performed, so that a new primary antibody can be incubated.

The results of the cell experiments show that IFN-gamma induces inflammatory response of mouse macrophages in a dose-dependent manner (3 hours), activates p-stat3 and increases the expression of NF- κB, and the puerarin can down regulate the expression of p-stat3 and NF- κB after intervention, thus indicating that puerarin has anti-inflammatory effect (figure 4).

Claims (10)

1. The application of puerarin in preparing medicine for preventing and/or treating hashimoto thyroiditis is provided.

2. The use according to claim 1, wherein the medicament is formulated with pharmaceutically acceptable excipients.

3. The use according to claim 2, wherein the formulation is an injectable formulation.

4. The use according to claim 2, wherein the pharmaceutically acceptable excipients comprise solubilizers, co-solvents, PH modifiers, surfactants, antioxidants, isotonicity modifiers, bacteriostats.

5. The use according to claim 2, wherein the formulation is an oral formulation.

6. The use according to claim 5, wherein the pharmaceutically acceptable excipients comprise diluents, binders, solubilizers, disintegrants, glidants, lubricants, flavoring agents, inclusion materials, adsorption materials.

7. The use according to claim 5, wherein the formulation is a granule, powder, tablet, capsule, pill, oral liquid.

8. The use according to claim 1, wherein the prevention and/or treatment of hashimoto thyroiditis is manifested by a reduction of lymphocyte infiltration and autoantibody levels in thyroid tissue of hashimoto thyroiditis.

9. The use according to claim 1, wherein the prevention and/or treatment of hashimoto thyroiditis is manifested by inhibition of expression of chemokines and adhesion molecules in thyroid tissue.

10. The use according to claim 1, wherein the prevention and/or treatment of hashimoto thyroiditis is manifested by inhibiting serum inflammatory factor levels of hashimoto thyroiditis.