CN114288281A - Application of triolein in preparation of medicine for preventing and/or treating atopic dermatitis - Google Patents

Abstract

The invention relates to the technical field of medicines, in particular to an application of triolein in preparing a medicine for preventing and/or treating atopic dermatitis. The invention has the advantages that: the trilinolein of the invention can effectively inhibit the expression of TSLP from the prior clinical curative effect or the early cell experiment, thereby regulating the immune balance abnormality of the skin of atopic dermatitis patients with Th2 cells as the main factor and repairing the skin barrier dysfunction.

Description

Application of triolein in preparation of medicine for preventing and/or treating atopic dermatitis

Technical Field

The invention relates to the technical field of medicines, in particular to an application of triolein in preparing a medicine for preventing and/or treating atopic dermatitis.

Background

Atopic Dermatitis (AD) is a disease characterized by a predominance of Th2 cells with eosinophil infiltration and IgE elevation, and dysregulation of Th1/Th2 cell balance is an important pathogenesis. Thymic Stromal Lymphopoietin (TSLP) is a novel interleukin 7(IL-7) -like cytokine, primarily expressed by epithelial cells. Recent studies have shown that TSLP plays an important role in regulating the balance between Th1 and Th2, and is thought to play a role as a master switch and master regulator in the initiation of Th 2-driven inflammatory diseases (CORREN J, ZIEGLER S F. TSLP: from alloergy to cancer [ J ]. Nature Immunology,2019,20(12):1603 1609.). Among them, TSLP variant rs1898671 has been shown to be significantly associated With acute AD in children (CHANG J, MINRA N, HOFFSTAD O, et al.Association of Filaggrin Loss of Function and third Linear lymphopoetin Variation With Treatment Use in Pediatric antigenic Dermatitis [ J ]. JAMA Dendrology, 2017,153(3): 275.).

Recent studies have shown that TSLP is involved in the pathogenesis of allergic diseases through the following pathways: TSLP functions by activating dendritic cells (TSLP-DCs) (ADHIKARY P P, TAN Z, PAGE BD G, et al. TSLP as soluble target-a silver-linking for atomic diseases? [ J ]. Pharmacology & Therapeutics,2021,217: 107648.). (ii) TSLP acts directly on mast cells, promoting their secretion of Th2 cytokines and proinflammatory factors (HAN H, HEADLEY M B, XU W, et al, Thymic Stromal lymphopeptides amplifiers The Differentiation of alternative Activated macromolecules [ J ] The Journal of Immunology,2013,190(3): 904-. The Th2 cytokine and proinflammatory factor can induce the skin to produce more TSLP (Zelixia, Zengbo, Royal, etc.. Thymus stromal lymphopoietin's role in atopic dermatitis mouse model [ J ]. China J. dermatosis, 2013,27(10): 996-999.). One study indicated that IL-33 is a prerequisite for promoting the elevation of TSLP in AD-like dermatitis (HAN H, ROAN F, JOHNSTON L K, et al, IL-33 proteins structural integrity in TSLP-independent manner [ J ]. Mucosal Immunology, 201394 8,11(2): 403.), and that TSLP can then indirectly cause pruritus by activating immune cells, which secrete inflammatory mediators and cytokines that stimulate sensory neurons, such as IL-4, IL-13, and IL-31(MACK M R, KIM B S.the Itch-Scatch Cycle: A neurosimulant perfect in neurons, 201980, 39(12): 991.). Taken together, TSLP is involved in allergic disease development primarily through the priming of both dendritic cell and mast cell pathways, and may act as a master switch in both pathways.

However, the application of triolein in the preparation of a medicament for treating atopic dermatitis has not been reported at present.

Disclosure of Invention

The invention aims to provide a novel medical application of triolein.

The skin lesion and the serum of the Atopic Dermatitis (AD) in the acute stage can show increased expression of cytokines such as IL-4 and the like and reduced expression of IFN-gamma, and TSLP and Th2 cells are dominant, so that the pathological development of AD can be quickly and effectively controlled if the TSLP target can be intervened in the acute stage.

In a first aspect of the present invention, there is provided use of triolein for the preparation of a medicament for the prevention and/or treatment of atopic dermatitis.

Further, the preventive and/or atopic dermatitis remedy is a trilinolein as the only active ingredient or a pharmaceutical composition containing trilinolein.

Further, the content of triolein in the medicament for preventing and/or treating atopic dermatitis is 5 to 10 percent.

Further, the agent for preventing and/or atopic dermatitis is an emulsion, an ointment or a transdermal absorbent.

Further, the preventive and/or atopic dermatitis remedy is triolein cream.

Further, in the application, the trilinolein improves the thickness of skin lesions with skin inflammation.

Further, in the application, the trilinolein reduces the expression of TSLP protein at the skin inflammation and skin damage tissue site.

Furthermore, in the application, the trilinolein regulates the Th2 cell-dominated abnormal immune balance of the skin of the atopic dermatitis patient, and repairs the skin barrier dysfunction.

Further, in the application, the triglycerol improves the moisture content of the skin, but does not increase the oil content of the skin.

In a second aspect of the invention, there is provided the use of triolein in the manufacture of a medicament for inhibiting expression of Thymic Stromal Lymphopoietin (TSLP).

In a third aspect of the present invention, there is provided a pharmaceutical preparation for external use for preventing and/or treating atopic dermatitis, which comprises triolein and pharmaceutically acceptable excipients.

Further, the pharmaceutical preparation for preventing and/or treating atopic dermatitis is triolein cream.

In a fourth aspect of the present invention, there is provided use of an agent for inhibiting Thymic Stromal Lymphopoietin (TSLP) expression in the manufacture of a medicament for the prevention and/or treatment of atopic dermatitis.

Further, the agent for inhibiting the expression of Thymic Stromal Lymphopoietin (TSLP) is triolein.

The invention has the advantages that:

1. the invention provides a new application of the triolein and a new idea for preventing and treating the atopic dermatitis. With the future intensive research on the biological characteristics, the signal transduction pathway and the action mechanism of the TSLP in the immune system of the body, the TSLP can become a new intervention target for clinically treating autoimmune diseases.

2. The trilinolein of the invention can effectively inhibit the expression of TSLP from the prior clinical curative effect or the early cell experiment, thereby regulating the immune balance abnormality of the skin of atopic dermatitis patients with Th2 cells as the main factor and repairing the skin barrier dysfunction.

Drawings

FIG. 1. warp¹HNMR and¹³CNMR identified the structure of the obtained glycerol trilinoleate.

FIG. 2 TRIOLE glyceride reduces TSLP mRNA expression in vitro.

FIG. 3 in vitro reduction of TSLP protein expression by triolein.

Figure 4 triolein improves skin inflammation model lesion thickness.

FIG. 5 TRI-GLYCOL reduces TSLP protein expression at the skin lesion tissue site in mice model for skin inflammation.

FIG. 6. moisture content changes were measured at various time points after application of the triolein cream.

FIG. 7. the change in oil content was measured at different time points after the trilinolein cream was applied.

FIG. 8. patients with scrotal atopic dermatitis who improved after treatment with TRI-OLE glyceride cream (a: before treatment, b: after treatment).

Figure 9 atopic dermatitis in children was maintained after treatment with trilinolein for moisturization with no recurrence at follow-up for 10 weeks.

Detailed Description

The following examples are provided to illustrate specific embodiments of the present invention.

Example 1: extraction and separation of triolein

1.1 preparation of three grass oils

Weighing 60g of lithospermum, 30g of madder and 30g of liquorice which are 120g in total, heating, drying, crushing and sieving by a 40-mesh sieve for later use; adding 6 times of soybean oil (720mL), soaking at room temperature for 24 hours, and shaking during the soaking process to fully soak the soybean oil and completely extract the soybean oil; extracting in 80 deg.C water bath for 2 hr while infrared-assisted heating; filter paper filtration was performed overnight. (the yield of three-herb oil extract is shown in Table 1, and the content of three-herb oil extract is shown in Table 2)

TABLE 1 three herb oil extraction yield

Note: except the three-herb combined oil extract of the batch No. 20170413, the yield is lower, the yield of other batches is similar, the obtained oil extract is purple, and the content detection result is shown in Table 2.

TABLE 2 determination of three herb oil extracts

Note: compared with the Sage oil extract of lot 20160906, lot 20170413 has higher total pigment contents of rubiadin, laevorotatory alkannin and hydroxynaphthoquinone due to high pulverization degree of the medicinal materials and infrared auxiliary heating.

1.2 extraction and separation of Tricenoleracein from Trifolium Pratense L oil

6kg of 200-mesh 300-mesh silica gel was loaded into a 23cm diameter glass column, which was then wetted with petroleum ether (boiling range 60-90 ℃ C.) and vented. Sancao oil (750g) was poured into the column, and 500g of quartz sand was poured into the column when the liquid surface flowed to the silica gel surface. Eluting with ethyl acetate/petroleum ether (0/100-5/95), performing dot-on-plate tracing, collecting dots with Rf being about 0.45, and concentrating to obtain colorless oil (122g), wherein the developing agent is ethyl acetate/petroleum ether (1/10).

¹H NMR(400MHz,Chloroform-d)δ5.49–5.15(m,13H),4.29(dd,J＝11.9,4.3Hz,2H),4.14(dd,J＝11.9,5.9Hz,2H),2.77(t,J＝6.5Hz,4H),2.31(td,J＝7.5,2.6Hz,8H),2.07-2.00(m,12H),1.62-1.53(m,8H),1.37-1.29(m,16H),1.28-1.25(m,24H),0.91-0.86(m,9H).

Example 2: triinolein reduces Thymic Stromal Lymphopoietin (TSLP) expression in vitro

2.1 in vitro cell model

2.1.1 cell recovery

1) The cell house clean bench was opened, and DMEM complete medium, cell culture dish, etc. were prepared.

2) Taking out HaCaT cells from-80 deg.C refrigerator, and adding CO₂Incubator, wait for cells to melt.

3) Add 8ml of complete medium to the dish and aspirate the thawed cells into the dish with a 1ml pipette and then blow well.

4) Carefully place the petri dish into the incubator and return the medium to the 4 ℃ refrigerator, wipe the clean bench top with 75% ethanol and close.

2.1.2 cell exchange fluids

1) A clean bench, PBS, DMEM complete medium was prepared.

2) The growth state and density of the cells were observed under the mirror.

3) The original medium was discarded, an appropriate amount of PBS was added, the cells were washed by gentle shaking, and the PBS was aspirated off.

4) 8ml of complete medium was added and the cells were again placed in the incubator for incubation.

2.1.3 cell plates (24-well plate)

1) Cells were observed under the mirror. And (5) when the cell density is more than 80 percent and the state is good, namely plating.

2) A clean bench, PBS, trypsin, medium, 24-well plate was prepared.

3) The culture dish is aspirated to remove the original medium, and an appropriate amount of PBS is added to wash the cells, followed by addition of 800. mu.l to 1000. mu.l of trypsin, depending on the number of cells.

4) Placing the cells in an incubator for digestion, lightly tapping the cells to accelerate the cell detachment, and slightly tilting the culture dish to ensure that the cells are fully infiltrated by trypsin.

5) When the cells are observed under a mirror to become round and come off the wall, the cells are observed to be in a quicksand shape by naked eyes, and the digestion is finished.

6) Add the appropriate amount of complete medium and pipette with 10ml pipette.

7) By 2X 10⁵Taking cell suspension plate from each living cell/hole, putting the 24-hole plate into culture after markingAnd (4) incubating in a culture box, and fully and adherently growing the cells after 14-17 hours at night.

2.1.4 construction of models of cellular inflammation

1) After the cells are completely attached to the wall, exogenous recombinant human TNF-a protein is added for stimulation treatment so as to induce a cell inflammation model; tritrilinoleate treatment group Tritrilinoleate intervention treatment was performed based on stimulation by recombinant human TNF-a protein.

2) After the cells are treated for 12 hours, discarding the supernatant, gently cleaning the cells for 2 times by using precooled PBS, adding TRIzol solution into a cell culture plate, and collecting cell RNA samples for subsequent RT-qPCR detection; after 24 hours of cell treatment, the supernatant was discarded, the cells were gently washed 2 times with pre-cooled PBS, and a protein lysate was added to the cell culture plate and a protein sample was collected.

2.2 Experimental procedures

2.2.1 TSLP protein detection (Western blot method)

(1) Preparation of protein samples

1) Preparation of lysate (1 mL): 800. mu.l RIPA, 100. mu.l lPI, 100. mu.l phosphatase inhibitor, 10. mu.l sodium glycerophosphate, 10. mu.l sodium fluoride. 5 XLoading buffer DTT 50. mu.l, bromophenol blue 950. mu.l.

2) Harvesting the protein sample

First, crushed ice is prepared and the corresponding ep tube is marked.

Absorbing cell culture solution, adding lysis solution, blowing lysis solution repeatedly with a gun, scraping the culture dish with the gun tip to make the cell wall-removed, sucking in the ep tube after the lysis solution becomes viscous, adding sample buffer solution and storing on crushed ice.

Thirdly, heating the protein sample on a metal bath at 100 ℃ for about 7min until the protein sample is not sticky and has strong fluidity. After boiling, the sample is centrifuged for a short time and can be stored at-20 ℃.

(2) Electrophoresis

1) Reagent preparation

10 × electrophoresis buffer: 144.000g of Glycine, 30.000g of Tris base and 10.000g of SDS were diluted to 1L and left at room temperature.

10 times membrane transferring buffer solution: 28.8g of Glycine, 6g of Tris base and 400mL of methanol are added to the solution until the volume is 2L, and the solution is placed at the normal temperature.

③ 10 × TBS: 24g of Tris base, 88g of NaCl and 13mL of HCl are metered to 1L and placed at normal temperature.

10% APS: weighing 1.000g of ammonium persulfate to a constant volume of 10mL, subpackaging and storing at-20 ℃.

1 × TBST: 100mL of 10 XTSS, 900mL of pure water, and 1mL of Tween were left at room temperature.

2) Preparation of SDS-PAGE gel

First, purified water, 4 × SDS-PAGE gel separation buffer (pH 8.8), 4 × SDS-PAGE gel concentration buffer (pH 6.8), 30% gel preparation solution, 10% APS, and TEMED were prepared.

Secondly, fixing the clean rubber plate on a rubber frame, and preparing the separation rubber. Adding different amounts of pure water, 4 xSDS-PAGE gel separation buffer (pH 8.8), 30% gel preparation solution, 10% APS and TEMED, mixing, injecting into gel preparation plate, adding isopropanol to fill the gel preparation plate, pouring off the isopropanol after the gel is solidified for about 25min, and washing residual isopropanol with pure water. Then adding the concentrated gum (the preparation method is the same as that of the separation gum), inserting a comb, and pulling out the comb after about 25 min. According to the experimental requirements, the glue with different concentrations can be prepared.

3) Spotting is carried out

The 10 Xelectrophoresis buffer solution is diluted to 1 Xand then added into an electrophoresis tank, and the prepared gel is put in.

② taking out the protein sample from-20 ℃, heating on a metal bath at 100 ℃ until the protein sample has no large block adhesion and strong fluidity, the time is about 5min, and centrifuging for a short time after heating.

Thirdly, sequentially dropping the protein sample into the comb teeth, wherein each piece of glue contains at least one marker, and then performing 136V constant voltage electrophoresis for about 1 h.

(3) Rotary film

1) A1 Xtransmembrane buffer (previously placed in a refrigerator at 4 ℃), methanol, a 9X 6cm filter paper, a 8.5X 5.5cm PVDF membrane, and a sponge pad were prepared.

2) Firstly putting a PVDF membrane into methanol for activation, then taking out the glue and cutting off the concentrated glue, soaking filter paper, the glue, the PVDF membrane and a spongy cushion in a membrane conversion buffer solution, opening a clamping plate for membrane conversion, putting a layer of spongy cushion under a negative electrode, then putting three layers of filter paper, sequentially putting the glue, the membrane, the three layers of filter paper and the spongy cushion, and finally fastening the clamping plate, wherein no bubble is contained between every two layers, and the membrane conversion effect is influenced otherwise. Then the whole body is placed in a film-rotating groove for constant flow of 200mA film, crushed ice for cooling is placed outside the groove during film rotation, and film rotation is carried out in a refrigerator at 4 ℃.

(4) Sealing of

1) Preparing 5% of skim milk: 20ml of LTBST was added to 1.000g of skim milk powder.

2) And (3) taking the membrane out of the membrane rotating device, placing the membrane in 5% skimmed milk, and sealing the membrane on a shaking table at normal temperature for 1 h.

(5) Incubation of Primary anti-TSLP

1) 5mL of 5% BSA solution was prepared, and 5. mu.l of TSLP antibody (Waana, ER1918-22, rabbit, 1: 1000) after mixing, 20. mu.l of sodium azide was added and the mixture was stored at 4 ℃.

2) The blocked membranes were washed free of excess milk with TBST solution and then incubated overnight on a shaker at 4 ℃ in primary antibody.

3) Primary antibody was recovered and the membrane was soaked in TBST solution and washed on a shaker for 5X 5 min.

(6) Incubation secondary antibody

1)0.500g blocking mill was added with 10ml of LTBST to make a 5% solution, and 1. mu.l of secondary antibody (HRB Goat Anti-mouse IgG) was added to make a 1: 10000 ratio.

2) The washed membrane was incubated in a secondary antibody on a shaker at room temperature for 1 h.

3) After the secondary antibody incubation was completed, the cells were washed with TBST solution for 5X 5 min.

(7) Development

And (3) uniformly dripping ECL luminous liquid on the membrane, and then developing on a western blot developing instrument.

2.2.2 RNA extraction

(1) After aspirating the cell culture medium, trizol was added to the dish, gently whipped until the liquid became viscous and then aspirated into ep tubes (which can be stored at-80 ℃ for two weeks).

(2) The added volume of the lysate 1/5 of chloroform was added, followed by vortexing for 15-20s, followed by centrifugation at 13000rpm for 10-15min at 4 ℃.

(3) After centrifugation, 3 layers can be seen, the uppermost layer is colorless RNA, the middle white layer is flocculent protein, and the lower layer is pink DNA.

(4) RNA was transferred to an enzyme-free ep tube and mixed by vortexing with equal volume of isopropanol and then precipitated in a freezer at-20 ℃ for 2 h.

(5) After completion of the precipitation, the mixture was centrifuged at 13000rpm at 4 ℃ for 10min, and after completion of the centrifugation, a white RNA precipitate was observed.

(6) After the isopropanol was aspirated, the mixture was washed twice with 70% ethanol (kept at a low temperature), and then centrifuged at 12000rpm for 10 min.

(7) After washing, the RNA was dried on a clean bench for about 3 hours, and the dried RNA became colorless and transparent and was stored at-80 ℃.

2.2.3 reverse transcription

(1) The air-dried RNA was dissolved in 20. mu.l of enzyme-free ultrapure water, and the concentration thereof was measured using a microplate reader (to prevent RNA degradation, the ep tube should be placed on ice).

(2) The measured RNA concentration was first diluted to 100 ng/. mu.l with enzyme-free ultrapure water.

(3) Reverse transcription was performed using Thermo (AB-1453/A) reverse transcription kit. According to the quantity of samples needing reverse transcription, firstly mixing a mix system needed by the reverse transcription, then adding RNA, uniformly mixing, centrifuging for a short time, then carrying out reverse transcription on a PCR instrument, and storing the obtained cDNA for a long time at the temperature of minus 20 ℃.

2.2.4 RT-PCR

(1) The obtained cDNA was diluted with enzyme-free ultrapure water, and 10. mu.l of the cDNA was diluted with 90. mu.l of enzyme-free ultrapure water.

(2) Mu.l of sybergreen mix (Takara, RR820), 0.3. mu.l of TSLP-target primer F5 '-CATGGGTGAATAAGGGCTTC-3' (SEQ ID NO.1), 0.3. mu.l of TSLP-target primer R5'-CACACTAAACTCTTCCCACC-3' (SEQ ID NO.2), and 5. mu.l of 100ng cDNA were mixed.

(3) And (3) putting the mixed system into a real time pcr instrument for amplification. The process is repeated 40 times according to the program of 95 ℃ for 15min to 95 ℃ for 15s to 60 ℃ for 20s to 72 ℃ for 45 s.

2.3 results

As shown in FIG. 2, after the Hacat cells are subjected to TNF-a modeling and drying, the TSLP mRNA expression is remarkably increased by more than 3 times, and after the co-drying and drying of the triolein, the TSLP mRNA expression is remarkably reduced to about 50% of that of a normal group. It was suggested that triolein could reduce TSLP expression at the transcriptional level.

Example 3: tricenoyl glycerol ester for reducing skin injury reaction of skin inflammation animal model

3.1 construction of animal models

5mg of MC903(S3739, seleckchen) dry powder (13000 rpm) were centrifuged for 3min, 242.363. mu.l of absolute ethanol were added to prepare a 50mM stock solution, which was stored in aliquots at-20 ℃. Each time, 1. mu.l of 50mM MC903 was added to 999. mu.l of absolute ethanol to make a working concentration of 50 μm.

A6-week-old SPF-grade C57BL/6 male mouse (purchased from Chengdu medicine Kangbio, Co., Ltd.) was selected, bred under a constant temperature and humidity environment of 25 ℃ and 55% humidity for 12h in a light/dark cycle for three days under standard conditions to adapt to the environment, and then randomly divided into three mice per cage.

After numbering the mice, 40 μ l of 50 μm MC903 was pipetted onto the ears of C57BL/6 mice and spread evenly over the ears in two portions, 20 μ l each inside and outside the ears, and the mice were stimulated for three weeks at the same time interval on Monday, Wednesday and Friday. The ear condition of the mouse is observed every day, and the record is well made.

3.2 results

As shown in fig. 4, after intervention, the epidermis of the model group mice was hyperplastic, and more inflammatory cells were found in the dermis, compared with the normal group, while the epidermis of the treatment group was significantly thinned, and inflammatory cells in the dermis were decreased, indicating that the inflammatory reaction was suppressed to some extent.

Furthermore, we examined the effect of different concentrations (5% and 10%) of the cream on the mouse model and found that there was no significant difference in pathological improvement between the 5% low dose and the 10% high dose (fig. 5), suggesting that both concentrations were effective concentrations and that the subsequent clinical patients were developed with 5% concentration.

Example 4: preparation of triolein cream

Preparation process of 10% triolein cream:

oil phase: 30g of stearic acid, 10g of glyceryl monostearate, 45g of liquid paraffin, 15g of vaseline and 29.61g of glyceryl trilinoleate

Water phase: 0.6g of triethanolamine, 0.6g of sodium dodecyl sulfate, 30g of glycerol, 135g of distilled water and 0.3g of ethylparaben

The method comprises the following steps: melting all components of the oil phase in a water bath, and cooling to 70 ℃; heating the water phase to boiling with direct fire, cooling to 70 deg.C, slowly adding the oil phase into the water phase in stirring water bath, stirring, mixing, transferring the beaker to room temperature, and stirring to cool. (small amounts can be made by reducing the respective components in equal proportions) Tricenolenic acid glyceride is present in the ointment in a proportion of 10%.

Preparation process of 5% triolein cream:

oil phase: 30g of stearic acid, 10g of glyceryl monostearate, 45g of liquid paraffin, 15g of vaseline and 14.03g of glyceryl trilinoleate

Water phase: 0.6g of triethanolamine, 0.6g of sodium dodecyl sulfate, 30g of glycerol, 135g of distilled water and 0.3g of ethylparaben

The method comprises the following steps: melting all components of the oil phase in a water bath, and cooling to 70 ℃; heating the water phase to boiling with direct fire, cooling to 70 deg.C, slowly adding the oil phase into the water phase in stirring water bath, stirring, mixing, transferring the beaker to room temperature, and stirring to cool. (small amounts can be made by reducing the respective components in equal proportions) Triinolein is present in the ointment at a level of 5%.

Example 5: tricenol cream for improving skin moisture content

According to the invention, healthy volunteers are adopted to respectively apply the triolein on the skin fixing part, the water content and the oil content of the part are detected again at the moment of application and 15min, 30min, 1h and 2h after application, and the water content is found to be increased from 19.42 +/-1.55 to 29.14 +/-2.06 and increased by about 50.05% (P is 0.0008) and maintained for 2h (27.21 +/-0.97), and is still increased by 40.11% (P is 0.0009) compared with that before application. Similarly, the content of the oil is detected, and although the oil content is increased from 11.39 +/-0.80 to 14.47 +/-1.36 and is increased by 27.04% immediately after the application (P is 0.047), the oil content is recovered to 11.34 +/-0.58 after 15min, and the oil content is not statistically different from that before the application (P is 0.9549). The tests indicate that the trilinolein cream for external use has good moisturizing effect, but does not increase the oil content of the skin.

Example 6: tricenol improves clinical symptoms in patients with clinical atopic dermatitis

Case 1, patient male, ge x, 22 years old, atopic dermatitis manifested by scrotal itching, history of more than 10 years, serum IgE 376.80ng/ml (0-240ng/ml), VAS score > 6. After 8 weeks of external treatment by the trilinolein cream, the pruritus is obviously improved, and the VAS is divided into 0; the clinic is shown in figure 8.

Case 2, infant, young, 5 years old, dry skin of the whole body with pruritus for more than 4 years, no cure for long term treatment. The results are shown in figure 9 for 4 and 10 weeks of treatment with trilinolein cream.

The present invention has been made to treat atopic dermatitis patients and also found to have good moisturizing effect (intractable scrotum lesions, case 1, fig. 8; neck lesions, case 2, fig. 9).

The trilinolein can effectively inhibit the expression of TSLP from the prior clinical curative effect or the early cell experiment, thereby regulating the immunologic balance abnormality of the skin of AD patients with Th2 cells as the main factor and repairing the skin barrier dysfunction.

The invention provides a new application of the triolein and a new idea for preventing and treating the atopic dermatitis.

While the preferred embodiments of the present invention have been described in detail, it will be understood by those skilled in the art that the invention is not limited thereto, and that various changes and modifications may be made without departing from the spirit of the invention, and the scope of the appended claims is to be accorded the full range of equivalents.

Sequence listing

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Application of TRI (120) triolein in preparation of medicine for preventing and/or treating atopic dermatitis

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Claims (10)

1. Use of triolein in the preparation of a medicament for the prevention and/or treatment of atopic dermatitis.

2. The use of triolein according to claim 1 in the preparation of a medicament for the prevention and/or treatment of atopic dermatitis, wherein said medicament for the prevention and/or treatment of atopic dermatitis comprises triolein as the only active ingredient or is a pharmaceutical composition comprising triolein.

3. Use of triolein according to claim 1 for the preparation of a medicament for the prevention and/or treatment of atopic dermatitis, characterized in that triolein improves the thickness of skin lesions of skin inflammation.

4. Use of triolein according to claim 1 for the preparation of a medicament for the prevention and/or treatment of atopic dermatitis, characterized in that triolein reduces TSLP protein expression at skin inflammatory lesion tissue sites.

5. The use of triolein according to claim 1 in the preparation of a medicament for the prevention and/or treatment of atopic dermatitis, wherein triolein regulates the Th2 cell-dominated dysimmune balance in the skin of atopic dermatitis patients and repairs skin barrier dysfunction.

6. Use of triolein according to claim 1 for the preparation of a medicament for the prevention and/or treatment of atopic dermatitis, characterized in that triolein increases the water content of skin without increasing the oil content of skin.

7. The use of triolein in the preparation of a medicament for inhibiting the expression of thymic stromal lymphopoietin.

8. An external pharmaceutical preparation for preventing and/or treating atopic dermatitis, which is characterized by comprising triolein and pharmaceutically acceptable auxiliary materials.

9. Application of a reagent for inhibiting expression of thymic stromal lymphopoietin in preparing a medicament for preventing and/or treating atopic dermatitis.

10. The use of an agent for inhibiting the expression of thymic stromal lymphopoietin according to claim 9 in the manufacture of a medicament for the prevention and/or treatment of atopic dermatitis, wherein said agent for inhibiting the expression of thymic stromal lymphopoietin is glycerol trilinoleate.

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